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  • 1990-1994  (4,897)
  • 1970-1974  (3,868)
  • 1860-1869  (4,025)
  • Organic Chemistry  (11,970)
  • Ultrastructure  (820)
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  • 1
    ISSN: 1432-2307
    Keywords: Mast cell ; Compound 48/80 ; Ultrastructure ; Quick-freezing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The ultrastructure of mast cells stimulated with compound 48/80 was examined by quick-freezing and deep-etching (QF-DE) or freeze-substitution (QF-FS) methods. Peritoneal cells including mast cells of adult male rats were stimulated in vitro with compound 48/80 at 17° C for 0, 10, 30, 60 or 180 s. The QF-DE replicas revealed that the mast cells stimulated with compound 48/80 for 30 s decreased filamentous actin around secretory granules. In the QF-FS specimens, perigranular membranes in mast cells stimulated for 60 s formed pentalaminar structures between adjacent granules in their cytoplasm prior to degranulation. These findings suggest that preparatory states for degranulation occur in the whole cytoplasm of stimulated mast cells at early stages. Moreover, both QF-FS specimens and QF-DE replicas revealed a compact morphological appearance of discharged granules in the extra-cellular space, indicating the existence of considerable content within the granules. Skeletal structures in the granules were also demonstrated on QF-DE replicas prepared after extracting soluble elements from the cytoplasm. It is suggested that the granular contents associated with the skeletal structures are gradually detached from the discharged granules to ensure local concentration in the tissues.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Virchows Archiv 425 (1994), S. 93-99 
    ISSN: 1432-2307
    Keywords: Pituitary adenoma ; Sellar gangliocytoma ; Immunohistology ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Three cases of a composite sellar tumour composed of a gangliocytoma and an adenoma are presented. Two patients who showed acromegaly and hyperprolactinaemia had a gangliocytoma and a growth hormone (GH)-prolactin cell adenoma in close proximity. The gangliocytoma contained growth hormone-releasing hormone (GHRH) by immunohistochemistry. At the electron microscopical level, the gangliocytoma was characterized by numerous synaptic vesicles. The third patient, a child with Cushing's disease, presented a corticotropin-releasing hormone (CRH)-positive gangliocytoma in close contact with an adrenocorticotropic hormone (ACTH) secreting adenoma, the latter a typical densely granulated ACTH cell adenoma. Ultrastructurally, the gangliocytoma revealed synaptic vesicles and sparse secretory granules. The results suggest that gangliocytomas may promote the development of pituitary adenomas by hypersecretion of releasing hormones. Whereas 20 cases of sellar GHRH producing gangliocytomas in acromegaly are reported in the literature, the combination of a CRH-positive gangliocytoma and an ACTH cell adenoma in Cushing's disease is apparently the first case.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Virchows Archiv 425 (1994), S. 305-313 
    ISSN: 1432-2307
    Keywords: Rat ; Pancreatic beta cells ; Immunocytochemistry ; Ultrastructure ; Insulin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract When studied morphologically in semi-thin sections in the rat in vivo, pancreatic beta cells displayed heterogeneous immunoreactivities for insulin and amylin, depending on the islet size and the intra-islet position of the beta cells. In larger islets, cortical beta cells (beta cells with contacts with all islet cell types and with the exocrine parenchyma) which are located in the periphery were more densely immunostained for insulin and amylin than medullary beta cells (beta cells with contacts only with other beta cells) which are located in the centre of the islet. Ultrastructurally, these findings were accompanied by differences in the number of secretory granules and mitochondria. Beta cells in small islets and at extra-islet sites exhibited a dense immunoreactivity. After administration of glibenclamide, immunoreactivities for insulin and amylin were diminished in a time-dependent manner, decreasing first in medullary and thereafter in cortical beta cells of larger islets. Ultrastructurally, the beta cells exhibited the typical signs of stimulation. A minority of beta cells in small islets and all beta cells in extra-islet locations remained unchanged. Thus pancreatic beta cells under basal and stimulatory conditions in vivo exhibit heterogeneity in hormone content and in ultrastructural features. These differences may represent the basis for a functional heterogeneity of the insulin secretory response of the individual beta cell both in vivo and in vitro in states of normal and impaired insulin secretion. As heterogeneity was observed only among beta cells in islets, while single beta cells surrounded by acinar cells exhibited no changes in insulin immunoreactivity, interactions between beta cells as well as between beta cells and other endocrine cells may be critical for expression of heterogeneity within the beta cell population.
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  • 4
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Invertiertes Harnblasenpapillom ; Wachstumstypen ; Ultrastruktur ; Maligne Transformation ; Klinischer Verlauf ; Key words Inverted urinary bladder papilloma ; Growth types ; Ultrastructure ; Malignant transformation ; Clinical course
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary The histopathology, ultrastructure, pathological status and clinical course are described in a total of 29 cases of inverted papilloma of the urinary bladder (IP). The IP was isolated in 16 cases and occurred in combination with a papillary urothelial carcinoma in 13 cases. Histologically, a trabecular, a glandular and a mixed type can be differentiated. The trabecular from predominates in a ratio of 4 : 1. The glandular form is further subdivided into a cystic form and an adenomatous form containing cylindrical cells. Contrary to earlier assumptions, dysplasia and malignant transformation also occur in IP. Amongst the 16 isolated IP observed, four showed slight and four showed moderate dysplasia. One isolated IP was malignant and invasive. In IP in combination with papillary urothelial carcinomas, malignant transformation is somewhat more frequent. Four malignant IP and up to 85 % dysplasias were found among the 13 cases. The ultrastructure of IP reveals two cell types: a light cell form which corresponds to a slightly dysplastic urothelium and a darker cell form with or without microvilli which is observed in the glandular type. A frequent characteristic is a thickening of the basement membrane besides abundant “tight junctions”. The immunohistochemistry is relatively uncharacteristic. This also applies to the blood group isoantigens, which showed irregular and relative uninformative results in the SCRA test. IP is observed in all age groups and the sex ratio (M/F) is 3 : 1. The average age of manifestation is 56 years, about 10 years earlier than bladder carcinoma. Just under two-thirds of IPs are in the region of the trigonum, the ureteral ostia at the neck of the bladder and the prostatic part of the urethra, and IPs rarely exceed 3 cm in size. Malignant transformation and recurrence are only rarely observed. With regard to the formal pathogenetics, there are correlations with Brunn cell clusters and with the basal urothelial cell, which can be characterized as the urothelially determinative cell.
    Notes: Zusammenfassung Auf der Basis von insgesamt 29 Fällen mit einem invertierten Harnblasenpapillom (IP), von denen bei 16 das IP isoliert vorkam und in weiteren 13 eine Kombination mit einem papillären Urothelkarzinom vorlag, wird zur Histopathologie, Ultrastruktur, Dignität und zum klinischen Verlauf Stellung genommen. Histologisch zeigen sich Wachstumstypen, die sich in einem trabekulären, glandulären und einem Mischtyp äußern. In einem Verhältnis von 4 : 1 überwiegt die trabekuläre Form. Entgegen früheren Annahmen kommen beim IP auch Dysplasien sowie eine maligne Transformation vor. Die Ultrastruktur des IP läßt 2 Zellarten erkennen: eine helle Zellform, die einem gering dysplastischen Urothel entspricht und eine dunklere Zellform mit oder ohne Mikrovilli, die beim drüsigen Typ zur Beobachtung kommt. Häufiges Merkmal ist eine Basalmembranverdickung neben reichlich „tight junction“. Die Immunhistochemie zeigt eine Differenzierung in Richtung Basalzelle, ist jedoch wenig charakteristisch, so auch die Blutgruppenisoantigene, die sich irregulär verhalten. Das IP wird in allen Altersgruppen beobachtet und überwiegt mit 3 : 1 beim männlichen Geschlecht. Durchschnittliches Manifestationsalter liegt im 56. Lebensjahr, etwa 10 Jahre früher als beim Harnblasenkarzinom. Knapp zwei Drittel sind im Bereich Trigonum, der Ureterostien des Blasenhalses und der Pars prostatica urethrae lokalisiert und überschreiten nur sehr selten die Größe von 3,0 cm. Ebenso wie die maligne Transformation werden auch Rezidive nur selten beobachtet. Formalpathogenetisch bestehen Beziehungen zu den Brunn-Zellnestern und zur basalständigen Urothelzelle, die als urotheliale Determinativzelle charakterisiert werden kann.
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  • 5
    ISSN: 1432-1351
    Keywords: Electric fish ; Pacemaker ; GABA ; Glutamate ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The medullary pacemaker nucleus of Hypopomus triggers each electric organ discharge (EOD) by a single command pulse. It consists of electrotonically coupled ‘pacemaker’ cells, which generate the rhythm, and ‘relay’ cells, which follow the pacemaker cells and excite the spinal motoneurons of the electric organ. The pacemaker cells receive two inputs from the complex of the diencephalic prepacemaker nucleus (PPn), a GABA-ergic inhibition and a glutamatergic excitation. Relay cells, on the other hand, receive two glutamatergic inputs, one from a subnucleus of the PPn, the PPn-C, and a second from the sublemniscal prepacemaker nucleus (SPPn). We have labelled afferents to the pacemaker nucleus by injecting HRP to specific sites of the prepacemaker complex. By using immunogold-labelled antibodies and en-grid staining techniques, we demonstrated GABA and glutamate immunoreactivity in labelled synaptic profiles of ultra-thin sections of the pacemaker nucleus. The two types of synapses were interspersed on the surfaces of pacemaker cells, with GABA-immunoreactive synapses apparently representing the GABA-mediated input of the ‘PPn-I’, an inhibitory subdivision of the PPn, and glutamate-immunoreactive synapses representing the input of the ‘PPn-G’, an excitatory subdivision of the PPn. Only glutamate-immunoreactive synapses were found on relay cells.
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  • 6
    ISSN: 1432-0827
    Keywords: In vitro ; Bioactive glass ceramic ; Mineralization ; Bone bonding mechanisms ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Rat bone cells were cultured in the presence of bioactive glass-ceramic containing crystalline apatite and wollaston te. Scanning electron microscopy observations of the surface of the seeded ceramic disks revealed that cells attached, spread, and proliferated on the material surface. Soaking in cell-free culture medium showed that no change occurred in the surface structure. However, when cultured with bone cells and observed under a transmission electron microscope, an electron-dense layer was noted initially at the surface of the material, before bone formation occurred. In addition, energy-dispersive X-ray microanalysis demonstrated the presence of calcium and phosphorus in this layer. Progressively, during the following days of culture, active osteoblasts synthetized and laid down an osteoid matrix composed of numerous collagen fibrils arranged either parallel or perpendicularly to the first-formed electron-dense layer. Mineralization initiated on the ceramic surface dispersed then along the collagenous fibrils, leading to a mineralized matrix which surrounded the ceramic particles. These results demonstrate the capacity of apatite-wollastonite glass ceramic to initiate biomineralization in osteoblast cultures and to achieve a direct bond between the surface apatite layer of the bioactive glass-ceramic and the mineralized bone matrix.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 55 (1994), S. 180-189 
    ISSN: 1432-0827
    Keywords: Collagen ; Crystal habit ; Ultrastructure ; Turkey leg tendon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Transmission electron micrographs of fully mineralized turkey leg tendon in cross-section show the ultrastructure to be more complex than has been previously described. The mineral is divided into two regions. Needlelike-appearing crystallites fill the extrafibrillar volume whereas only platelike crystallites are found within the fibrils. When the speciment is tilted through a large angle, some of the needlelike-appearing crystallites are replaced by platelets, suggesting that the needlelike crystallites are platelets viewed on edge. If so, these platelets have their broad face roughly parallel to the fibril surface and thereby the fibril axis, where the intrafibrillar platelets are steeply inclined to the fibril axis. The projection of the intrafibrillar platelets is perpendicular to the fibril axis. The extrafibrillar volume is at least 60% of the total, the fibrils occupying 40%. More of the mineral appears to be extrafibrillar than within the fibrils. Micrographs of the mineralized tendon in thickness show both needlelike-appearing and platelet crystallites. Stereoscopic views show that the needlelike-appearing crystallites do not have a preferred orientation. From the two-dimensional Fourier transform of a selected area of the cross-sectional image, the platelike crystallites have an average dimension of 58 nm. The needlelike-appearing crystallites have an average thickness of 7 nm. The maximum length is at least 90 nm. Atomic force microscopy (AFM) of unstained, unmineralized turkey leg tendon shows collagen fibrils very much like shadow replicas of collagen in electron micrographs. AFM images of the mineralized tendon show only an occasional fibril. Mineral crystallites are not visible. Because the collagen is within the fibrils, the extrafibrillar mineral must be embedded in noncollagenous organic matter. When the tissue is demineralized, the collagen fibrils are exposed. The structure as revealed by the two modalities is a composite material in which each component is itself a composite. Determination of the properties of the mineralized tendon from the properties of its elements is more difficult than considering the tendon to be just mineral-filled collagen.
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  • 8
    ISSN: 1432-0533
    Keywords: Vacuolization ; Neurotoxicity ; Neuropathology ; Electron microscopy ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Cytoplasmic vacuoles appear in neurons of the posterior cingulate/retrosplenial cortex (PC/RS) of rats after treatment with N-methyl-d-aspartate (NMDA) receptor antagonists. Prominent dilatation of mitochondria and endoplasmic reticulum has been described within 2 h; however, the ultrastructural features of vacuole formation are unknown. To investigate this, the present study examined the PC/RS cortex of male rats (age 60–70 days) at 15, 30, 45, 60, 90, and 120 min after subcutaneous treatment with 1 mg/kg of the noncompetitive NMDA antagonist MK-801 (dizocilpine maleate, 5-methyl-10, 11-dihydro-5H-dibenzo [a,d] cyclohepten-5,10-imine). Subtle mitochondrial dilatation was identified in a few neurons as early as 15 min postdose (MPD). By 30 MPD, dilatation was more pronounced in mitochondria and also involved the endoplasmic reticulum and perinuclear space. Ribosomal disaggregation and degranulation were also evident by 30 MPD. At all subsequent time points, dilatation of mitochondria and endoplasmic reticulum progressed in severity. Although the relative involvement of mitochondria and endoplasmic reticulum varied, glia were not involved. These ultrastructural data suggest that after treatment with MK-801, mitochondrial dilatation precedes involvement of endoplasmic reticulum in vacuolization of susceptible PC/RS cortical neurons. The early mitochondrial effects identified in this study suggest an initial metabolic insult that rapidly progresses to affect endoplasmic reticulum and ribosomes. This strengthens the relationship between the ability of certain NMDA antagonists to induce energy perturbations and neuronal vacuoles in the same region of the rat cerebral cortex.
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  • 9
    ISSN: 1432-0533
    Keywords: Extraskeletal osteosarcoma ; Brain neoplasms ; Ultrastructure ; Multinucleated giant cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A 57-year-old woman with primary intracerebral osteosarcoma is reported. The tumor was identified by computed tomography as a mass with hemorrhage in the right parietal lobe. The surgical and pathological examinations confirmed an osteosarcoma of intracerebral origin. She suffered from repeated local recurrence of the tumor and died about 1 year after the onset. The pathological findings showed features of osteoblastic osteosarcoma with numerous osteoclastlike multinucleated giant cells. Immunohistochemically, tumor cells were positive for vimentin, and partially for actin. Multinucleated giant cells were reactive with vimentin and CD68 antibodies. Ultrastructurally, tumor cells were rich with rough endoplasmic reticulum. These findings are consistent with the histological features of skeletal or extraskeletal osteosarcoma. This is the third case of primary intracerebral osteosarcoma reported in the literature and the first one analyzed ultrastructurally.
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  • 10
    ISSN: 1432-0533
    Keywords: Key words Vacuolization ; Neurotoxicity ; Neuropathology ; Electron microscopy ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Cytoplasmic vacuoles appear in neurons of the posterior cingulate/retrosplenial cortex (PC/RS) of rats after treatment with N-methyl-D-aspartate (NMDA) receptor antagonists. Prominent dilatation of mitochondria and endoplasmic reticulum has been described within 2 h; however, the ultrastructural features of vacuole formation are unknown. To investigate this, the present study examined the PC/RS cortex of male rats (age 60 – 70 days) at 15, 30, 45, 60, 90, and 120 min after subcutaneous treatment with 1 mg/kg of the noncompetitive NMDA antagonist MK-801 (dizocilpine maleate, 5-methyl-10, 11-dihydro-5H-dibenzo [a,d] cyclohepten-5,10-imine). Subtle mitochondrial dilatation was identified in a few neurons as early as 15 min postdose (MPD). By 30 MPD, dilatation was more pronounced in mitochondria and also involved the endoplasmic reticulum and perinuclear space. Ribosomal disaggregation and degranulation were also evident by 30 MPD. At all subsequent time points, dilatation of mitochondria and endoplasmic reticulum progressed in severity. Although the relative involvement of mitochondria and endoplasmic reticulum varied, glia were not involved. These ultrastructural data suggest that after treatment with MK-801, mitochondrial dilatation precedes involvement of endoplasmic reticulum in vacuolization of susceptible PC/RS cortical neurons. The early mitochondrial effects identified in this study suggest an initial metabolic insult that rapidly progresses to affect endoplasmic reticulum and ribosomes. This strengthens the relationship between the ability of certain NMDA antagonists to induce energy perturbations and neuronal vacuoles in the same region of the rat cerebral cortex.
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  • 11
    Electronic Resource
    Electronic Resource
    Springer
    Acta neuropathologica 88 (1994), S. 222-227 
    ISSN: 1432-0533
    Keywords: Amyotrophic lateral sclerosis ; Anterior horn neuron ; Synapse ; Active zone ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract This report deals with an ultrastructural investigation of the synapses of anterior horn neurons in the lumbar spinal cords of five patients with amyotrophic lateral sclerosis (ALS) who had mild neuronal depletion. Specimens from five age-matched, neurologically normal individuals served as controls. In each instance, the autopsy was performed within 3 h after death. A statistically significant decrease in cell body area, number of synapses and total synaptic length was found in the normal-appearing neurons of the ALS patients. The alterations were more pronounced in neurons with central chromatolysis. However, despite an approximately 20% reduction in the number of synapses, the length of the active synaptic zone of the normal-appearing neurons in the ALS patients was not diminished. This observation may be accounted for by a plasticity to the loss of synapses which maintained the active zone of the remaining synapses to increase synaptic efficiency. It is suggested that when the plasticity of the active zone reaches its limit, the continuing loss of synapses may lead to functional impairment. The capacity of the active synaptic zone to respond to progressive denervation of the anterior horn neurons may preserve motor function or slow the development of motor deficits in the early stage of degeneration of the lower motor neurons.
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  • 12
    Electronic Resource
    Electronic Resource
    Springer
    Acta neuropathologica 88 (1994), S. 222-227 
    ISSN: 1432-0533
    Keywords: Key words Amyotrophic lateral sclerosis ; Anterior horn neuron ; Synapse ; Active zone ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract This report deals with an ultrastructural investigation of the synapses of anterior horn neurons in the lumbar spinal cords of five patients with amyotrophic lateral sclerosis (ALS) who had mild neuronal depletion. Specimens from five age-matched, neurologically normal individuals served as controls. In each instance, the autopsy was performed within 3 h after death. A statistically significant decrease in cell body area, number of synapses and total synaptic length was found in the normal-appearing neurons of the ALS patients. The alterations were more pronounced in neurons with central chromatolysis. However, despite an approximately 20  % reduction in the number of synapses, the length of the active synaptic zone of the normal-appearing neurons in the ALS patients was not diminished. This observation may be accounted for by a plasticity to the loss of synapses which maintained the active zone of the remaining synapses to increase synaptic efficiency. It is suggested that when the plasticity of the active zone reaches its limit, the continuing loss of synapses may lead to functional impairment. The capacity of the active synaptic zone to respond to progressive denervation of the anterior horn neurons may preserve motor function or slow the development of motor deficits in the early stage of degeneration of the lower motor neurons.
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  • 13
    ISSN: 1432-0533
    Keywords: Human pineal gland ; Pineal parenchymal tumors ; Ultrastructure ; Chromogranin A
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We have studied 20 pineal parenchymal tumors (PPT) and 4 normal or cystic pineal glands both by light and electron microscopy and immunohistochemistry with antibodies against glial markers [glial fibrillary acidic protein (GFAP) and protein S-100] or neural/neuroendocrine markers [neurofilaments (NF), synaptophysin and chromogranin A]. Light microscopy revealed the cellular organization of pinealocytes in the normal gland and in different morphological types of pineal tumors (typical pineocytomas, PPT with intermediate differentiation, mixed PPT exhibiting elements of both pineocytoma and pineoblastoma and pineoblastomas). Immunohistochemistry showed the presence of GFAP and protein S-100 in interstitial cells in nonneoplastic pineal gland. Cell processes were labeled with anti-synaptophysin and anti-NF antibodies. No immunoreactivity was found for chromogranin A in non-neoplastic pineal gland. In pineocytomas, GFAP and protein S-100 were observed in interstitial cells. Synaptophysin and NF were present in the large rosettes of pineocytomas. Synaptophysin, NF and chromogranin A were present in pineocytomas with a lobular arrangement of cells. Anti-chromogranin A immuno-reactivity was also seen in lobular areas of some PPT with intermediate differentiation. Analysis of normal human pineal gland by electron microscopy showed the presence of vesicle-crowned rodlets (VCR or synaptic ribbons), fibrous filaments (F), paired twisted filaments but few dense-core vesicles (DCV) in normal pinealocytes. Tumoral pineal cells appeared to differentiate either towards a neurosensory pathway characterized by the presence of sensory cells elements (VCR and F), or towards a neuroendocrine pathway, with the occurrence of many DCV. Immunogold labeling demonstrated the presence of chromogranin A in neurosecretory granules.
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  • 14
    ISSN: 1432-0533
    Keywords: Corticobasal degeneration ; Ultrastructure ; Tau ; Glial inclusions ; Progressive supranuclear palsy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We have studied brain tissues from three patients with corticobasal degeneration (CBD) histologically, ultrastructurally and immunohistochemically. Ballooned neurons in the cerebral cortex and severe degeneration of the substantia nigra were observed in them all and weakly basophilic neurofibrillary tangles (NFTs) were distributed widely in the basal ganglia and brain stem. Ultrastructural examination demonstrated that the NFTs comprised characteristic 15-nm-wide straight tubules, which showed positive immunohistochemical staining with an antibody against tau, but not ubiquitin. Tau-immunoreactive neuronal cell bodies without NFTs also were found in the cerebral cortex and subcortical nuclei, predominantly in the brain stem, and the greatest number of tau-positive glial inclusions occurred in the cerebral gray and white matter of the pre- and post-central gyri. These inclusions comprised tubular structures with diameters of about 15 nm and were localized in the oligodendroglial cellular cytoplasm and processes. These findings indicate that there is a close cytoskeletal pathological relationship between CBD and progressive supranuclear palsy.
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  • 15
    ISSN: 1432-0533
    Keywords: Key words Human pineal gland ; Pineal parenchymal tumors ; Ultrastructure ; Chromogranin A
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We have studied 20 pineal parenchymal tumors (PPT) and 4 normal or cystic pineal glands both by light and electron microscopy and immunohistochemistry with antibodies against glial markers [glial fibrillary acidic protein (GFAP) and protein S-100] or neural/neuroendocrine markers [neurofilaments (NF), synaptophysin and chromogranin A]. Light microscopy revealed the cellular organization of pinealocytes in the normal gland and in different morphological types of pineal tumors (typical pineocytomas, PPT with intermediate differentiation, mixed PPT exhibiting elements of both pineocytoma and pineoblastoma and pineoblastomas). Immunohistochemistry showed the presence of GFAP and protein S-100 in interstitial cells in non-neoplastic pineal gland. Cell processes were labeled with anti-synaptophysin and anti-NF antibodies. No immunoreactivity was found for chromogranin A in non-neoplastic pineal gland. In pineocytomas, GFAP and protein S-100 were observed in interstitial cells. Synaptophysin and NF were present in the large rosettes of pineocytomas. Synaptophysin, NF and chromogranin A were present in pineocytomas with a lobular arrangement of cells. Anti-chromogranin A immunoreactivity was also seen in lobular areas of some PPT with intermediate differentiation. Analysis of normal human pineal gland by electron microscopy showed the presence of vesicle-crowned rodlets (VCR or synaptic ribbons), fibrous filaments (F), paired twisted filaments but few dense-core vesicles (DCV) in normal pinealocytes. Tumoral pineal cells appeared to differentiate either towards a neurosensory pathway characterized by the presence of sensory cells elements (VCR and F), or towards a neuroendocrine pathway, with the occurrence of many DCV. Immunogold labeling demonstrated the presence of chromogranin A in neurosecretory granules.
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  • 16
    ISSN: 1432-0533
    Keywords: Key words Extraskeletal osteosarcoma ; Brain neoplasms ; Ultrastructure ; Multinucleated giant cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A 57-year-old woman with primary intracerebral osteosarcoma is reported. The tumor was identified by computed tomography as a mass with hemorrhage in the right parietal lobe. The surgical and pathological examinations confirmed an osteosarcoma of intracerebral origin. She suffered from repeated local recurrence of the tumor and died about 1 year after the onset. The pathological findings showed features of osteoblastic osteosarcoma with numerous osteoclast-like multinucleated giant cells. Immunohistochemically, tumor cells were positive for vimentin, and partially for actin. Multinucleated giant cells were reactive with vimentin and CD68 antibodies. Ultrastructurally, tumor cells were rich with rough endoplasmic reticulum. These findings are consistent with the histological features of skeletal or extraskeletal osteosarcoma. This is the third case of primary intracerebral osteosarcoma reported in the literature and the first one analyzed ultrastructurally.
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  • 17
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 190 (1994), S. 461-468 
    ISSN: 1432-0568
    Keywords: Oocyte ; Ultrastructure ; Zebu ; Nucleolus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The objective of the study was to investigate the light and ultrastructural morphology of dominant and subordinate oocytes from zebu (Bos indicus) cattle. Healthy cycling animals, which had a well-developed corpus luteum as judged by rectal palpation, were administered cloprostenol to induce luteolysis and therefore ovulation. The animals were slaughtered at days 3–11 post-ovulation, but those slaughtered at days 8–11 received a second injection of cloprostenol at day 7. Cumulus-oocyte complexes were aspirated from the largest (dominant) and the second largest (subordinate) follicles, and processed for transmission electron microscopy. Up to day 7, the dominant oocyte presented a peripherally located spherical oocyte nucleus with a compact dense fibrillar nucleolus. After day 7, the nuclear envelope became undulated and the nucleolus vacuolated. The nuclei contained an average of four nucleoli. In addition to vesicles, mitochondria, smooth endoplasmic reticulum, Golgi complexes and cortical granules, the ooplasm contained annulate lamellae and microtubules. Moreover, mitochondrial granules and pleomorphic forms of mitochondria were commonly observed. Some subordinate oocytes exhibited advanced stages of meiotic maturation. It is concluded that (1) the dominant oocyte undergoes certain prematurational changes, including nucleolus vacuolation, in the period from luteolysis up to the presumptive occurrence of the LH peak and (2) subordinate oocytes may undergo meiotic maturation.
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  • 18
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 190 (1994), S. 439-444 
    ISSN: 1432-0568
    Keywords: Chick embryo ; Bursectomy ; Female gonads ; Steroidogenic cells ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We examined the ultrastructural characteristics of the medullary steroidogenic cells in left and right female gonads of surgically bursectomized chick embryos killed on the 17th day of incubation. The steroidogenic cells of the bursectomized embryos have a more developed system of cisternae in the smooth endoplasmic reticulum than controls, and their mitochondria show some alterations in the density of the matrix and in the shape of the cristae. On the basis of these results, an enhancement of the steroidogenic activity in both gonads is suggested.
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  • 19
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 190 (1994), S. 583-590 
    ISSN: 1432-0568
    Keywords: Cattle ; Epithelium ; Oviduct ; Secretion ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The non-ciliated (NC) cells of the bovine oviduct epithelium, have been shown to release embryotrophic substances to the oviduct lumen. The aim of the present study was to investigate the ultrastructure, focusing on aspects of the secretory machinery, of NC cells in different segments of the oviduct during and after transoviduct migration of zygotes and embryos. Dairy heifers (n=8) were superovulated with an ECG/cloprostenol regimen, and the time of ovulation was estimated by ultrasound scanning. Samples from the infundibulum, ampulla, isthmus and uterotubal-junction of the oviduct were surgically collected from animals at 19–96 h and 7 1/2; –8 1/2 days after ovulation and processed for transmission electron microscopy, following standard procedures. The NC cells contained characteristic membrane-bound secretory granules composed of a lamellar cortex encaging an amorphous medulla. The two components could still be recognized during extrusion of the granule content into the oviduct lumen by exocytosis. During granulogenesis, small maturing granules without the lamellar structure were observed, but distinct condensing vacuoles were absent. An abundance of granules was found in the early versus the late group. In both groups the uterotubual junction was almost free of granules. This segment, on the contrary, was characterized by the presence of primary and secondary lysosome-like bodies. In the early group the intracellular location of the granules varied between oviduct segments. In the infundibulum they were placed in the supranuclear cytoplasm, in the isthmus they were found in the most apical part of the cells, while in the ampulla an intermediate granule position was noticed. In both groups the uterotubal junction was almost free of granules. This segment, on the contrary, was characterized by the presence of primary and secondary lysosome-like bodies. Cytoplasmic protrusions, often containing nuclei, were more frequent in the late than in the early group. This phenomenon may represent epithelial renewal. In conclusion, the NC cell of the bovine oviduct epithelium possesses an extensive capacity for protein synthesis and secretion. The numbers and location of secretory granules show cyclic and segmental variations. Most granules are present in the infundibulum and ampulla during the period of transoviduct migration of zygote and embryo.
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  • 20
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    Archives of microbiology 162 (1994), S. 267-271 
    ISSN: 1432-072X
    Keywords: Extremely thermophilic eubacterium ; Calderobacterium hydrogenophilium ; Ultrastructure ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Calderobacterium hydrogenophilum is an extreme thermophilic, obligately chemoautotrophic, hydrogen-oxidizing bacterium. The cells were shown to be nonmotile straight rods of average size 0.4x2.5 μm. After negative-staining of the whole cells, no flagella were observed. The multilayered cell wall was of type 1 and possessed a crystalline proteinaceous surface layer exhibiting p4 symmetry. The square unit cells had a lattice constant of approximately 11 nm. Cell division occurred by a constriction mechanism. C. hydrogenophilum differred from a similar hydrogen-oxidizing eubacterium, Hydrogenobacter thermophilus, by the absence of intracytoplasmic membrane structures in chemically fixed cells. However, an electron-dense intracytoplasmic hemispherical structure adhering to the inner membrane was frequently observed.
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  • 21
    ISSN: 1432-1106
    Keywords: Dopaminergic grafts ; Neuropeptide Y ; Ultrastructure ; Striatum ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In a previous study we demonstrated that grafted dopamine (DA) neurons are able to induce an early and widespread normalization of DA-neuropeptide Y (NPY) interactions in the host striatum previously deprived of its DA input. Since similar recoveries were found to occur in striatal areas densely or poorly reinnervated by the graft, the question was raised as to what mechanisms (synaptic or volumic release) were involved in these functional effects. Ultrastructural analysis of graft-to-host relationships was performed using single — and double — immunolabelling techniques to detect neurons containing tyrosine hydroxylase (TH) and NPY, with a view to analysing the early establishment of synaptic connectivity in various areas of the host striatum. Within 1 month of the grafting, TH-immunoreactive (TH-IR) neurons showed most of the normal intrinsic morphological features characteristic of adult rat neurons and were found to have established direct relationships with various striatal neuronal populations. TH-NPY relationships were observed only in the area most densely reinnervated by the graft, and their relative frequency was found to be roughly the same as that determined in the intact striatum. Three months after the grafting, this percentage decreased, probably owing to the further elongation in TH-IR axons resulting in a wider distribution of the TH-NPY associations over the host striatum. In the zones distal from the graft, the reinnervation was far from complete and the few TH-IR fibres projected only to some unlabelled elements, mainly of the spiny type, which have been shown to interact normally with both DA afferents and NPY cells and therefore may relay the DA action over the whole striatum on the NPY population. It can be concluded from these data that the rapid and extensive functional normalization of the TH-NPY interactions previously found to occur in the entire striatum may depend on the restoration of direct and indirect synaptic relationships. A diffuse action of DA through non-synaptic mechanisms may also account for the fact that the amine has access to broader striatal populations than to those presumably reached by DA fibres arising from the graft.
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  • 22
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    Archives of microbiology 162 (1994), S. 267-271 
    ISSN: 1432-072X
    Keywords: Key words     Extremely thermophilic eubacterium ; Calderobacterium hydrogenophilium ; Ultrastructure ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract       Calderobacterium hydrogenophilum is an extreme thermophilic, obligately chemoautotrophic, hydrogen-oxidizing bacterium. The cells were shown to be non-motile straight rods of average size 0.4 × 2.5 μm. After negative-staining of the whole cells, no flagella were observed. The multilayered cell wall was of type 1 and possessed a crystalline proteinaceous surface layer exhibiting p4 symmetry. The square unit cells had a lattice constant of approximately 11 nm. Cell division occurred by a constriction mechanism. C. hydrogenophilum differred from a similar hydrogen-oxidizing eubacterium, Hydrogenobacter thermophilus, by the absence of intracytoplasmic membrane structures in chemically fixed cells. However, an electron-dense intracytoplasmic hemispherical structure adhering to the inner membrane was frequently observed.
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  • 23
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    Mycopathologia 125 (1994), S. 33-39 
    ISSN: 1573-0832
    Keywords: Aflatoxin ; Lymphocytes ; Mice ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract This investigation sought to determine whether splenic lymphocytes obtained from Balb/C mice exposed to aflatoxin B1 (AFB1) showed any ultrastructural changes which could account for the immunodysfunction attributable to aflatoxins. Lymphocytes obtained from Balb/C mice administered aflatoxin B1 in olive oil daily for three weeks were studied using both transmission and scanning electron microscopy. The lymphocytes demonstrated ultrastructural changes primarily in the mitochondria where marked internal dissociation of the cristae was revealed by transmission electron microscopy. All other cellular organelles were unaffected. No significant alterations in external structure were observed under scanning electron microscopy. The findings of this study indicate that AFB1 administration does not affect the surface topography of lymphocytes, but AFB1, by causing extensive mitochondrial damage, may affect the way in which these cells function. This could be a possible explanation for the immunodysfunction associated with AFB1.
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  • 24
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    European archives of oto-rhino-laryngology and head & neck 251 (1994), S. 267-270 
    ISSN: 1434-4726
    Keywords: Cochlea ; Ultrastructure ; Stria vascularis ; Development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The mammalian stria vascularis undergoes certain developmental changes in the postnatal rat. The present study was designed to examine the ultrastructure of the stria vascularis in rat pups from immediately after birth to 20 days postpartum. The cochlea were removed with the animals under xylazine (Rompun) anesthesia and were prepared for transmission electron microscopy. Each of the three cell types in the stria were found to contain kinocilia up until 12–17 days of age. The presence of kinocilia in the intermediate and basal cells has not been previously described. Findings suggest that these organelles may serve a motile and/or sensory function to assist in the maturation of cell functions, particularly ion transport, during early stages of development.
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  • 25
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    Mycopathologia 128 (1994), S. 181-192 
    ISSN: 1573-0832
    Keywords: Aflatoxin B1 ; Embryo ; Mature ; Ultrastructure ; Zea mays L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Mature maize (Zea mays L.) embryos were exposed to aflatoxin B1 (AFB1) concentrations ranging from 0.1 to 25 µg/ml for 9 days. With increasing toxin concentration above 2 µg/ml, primary root elongation of germinated embryos was progressively inhibited, to reach a maximum value of 81% at 25 µ/ml toxin. An ultrastructural investigation of the subcellular alterations induced following toxin exposure provided evidence of deteriorative changes in several compartments of the plant cell. Alteration in membrane integrity (e.g., the tonoplast, plasmalemma and inner mitochondrial membrane) was a frequent feature of many cells. Apparent fusion of vacuoles, incorporation of cytoplasmic components into vacuoles and intravacuolar membrane whorls might be interpreted as deteriorative alterations. The results are discussed in the light of ultrastructural findings for other plant systems exposed to similar AFB1 concentrations, as well as findings for animal systems.
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  • 26
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    Methods in cell science 16 (1994), S. 109-115 
    ISSN: 1573-0603
    Keywords: Animal model ; Neurofibroma ; Schwann cell ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Peripheral nerve sheath (PNS) neoplasms, primarily neurofibromas, schwannomas and maliganant schwannomas, are among the most common tumors in fishes. Model systems involving PNS tumors in fishes are also valuable because mammalian models of PNS tumors are rare. Schwann cells, the primary cell type suspected of neoplastic transformation in these tumors, have been difficult to culture. We describe techniques for culturing normal and neoplastic Schwann cells from fish. We also present methods for preparing cells on culture dishes for electron microscopy which are especially useful when specific cells in a culture must be located for ultrastructural examination.
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  • 27
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    Hydrobiologia 292-293 (1994), S. 137-142 
    ISSN: 1573-5117
    Keywords: Ultrastructure ; morphology ; integument ; copepoda ; crustacea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The integument of Parathalestris harpactoides (Claus, 1863) is studied by scanning and transmission electron microscopy. The general structure of the integument conforms to the common pattern known from Copepoda. Emphasis is given to the structural variation of the cuticle in different regions of the body. The cuticle measures about 6 µm in most parts of the body, and shows a laminate appearance. The epicuticle is about 60 nm thick. Numerous pore canals containing muscular tonofilaments penetrate the procuticular layer of the integument. A peculiar feature is the presence of a ‘honeycombed’ layer in the outermost zone of the cuticle of some parts of the body. The epidermal layer, muscle insertions and integumental pores are of common type. The cuticle of some specimens, both males and females, is covered with microorganisms.
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  • 28
    ISSN: 1434-0879
    Keywords: GnRH-DT vaccine ; Testosterone ; Ultrastructure ; Rat ; Prostate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To evaluate the effects of active immunization against gonadotropin-releasing hormone (GnRH) on the ultrastructure of the rat ventral prostate, male Sprague-Dawley rats received three consecutive intramuscular injections of 10 μg/100g body weight (D-Lys6)-GnRH-diphtheria toxoid conjugate (GnRH-DT vaccine). Following immunization, test animals developed sufficiently high antibody titres to block the pituitary gonadal axis. Consequently testosterone values dropped to the levels in castrates. This therapy leads to atrophy of the prostate. Following immunization a strong immunological response, indicating the presence of considerable amounts of a GnRH-like peptide, was observed in the ventral prostates as early as 14 days after the first injection of GnRH-DT. Immunoneutralisation of GnRH-like activity may contribute to the effects observed.
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  • 29
    ISSN: 1615-6102
    Keywords: α-Amylase ; (1-3, l-4)-β-Glucanase ; Hormones ; Monensin ; Transfection ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A procedure has been developed to isolate protoplasts from mature aleurone layers of the malting variety Alexis and four other barley genotypes. It combines induction of endogenous cell wall degrading enzymes together with use of Onuzuka cellulase R 10 and driselase and results in better yields for two varieties than can be obtained with the huskless variety Himalaya. The viability of the freshly isolated protoplasts is greater than 90% and in spite of the presence of gibberellic acid during isolation procedures, most of the protoplasts are at an early developmental stage, as judged by ultrastructure. Gibberellic acid-induced changes in protoplast structure resemble those reported for Himalaya protoplasts. The protoplasts secrete both α-amylase (EC 3.2.1.1) and (1-3, 1-4)-β-glucanase (EC 3.2.1.73) into the surrounding medium. Transfection studies using a low pI α-amylase promoter to direct chloramphenicol acetyltransferase expression in aleurone protoplasts from Alexis and Himalaya revealed significant differences in their hormone responsiveness. In the absence of hormones, low levels of expression of the reporter enzyme were obtained in Alexis protoplasts, while high levels were characteristic for Himalaya protoplasts. An 8-fold increase in the expression of the reporter gene was induced by supplying the transfected Alexis protoplasts with gibberellin A3, whereas expression in Himalaya protoplasts remained unchanged. When Himalaya protoplasts were isolated from aleurone layers that had not been incubated with GA3 during the initial stages of protoplasting (the classical procedure), the hormone response of the promoter was 2.5-fold. It is thus possible to optimize the aleurone protoplast isolation procedure for different barley genotypes and mutants of interest in studies of transgenic gene expression and hormone induced secretion of proteins from this unique secretory plant tissue.
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  • 30
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    Medical molecular morphology 27 (1994), S. 1-7 
    ISSN: 1860-1499
    Keywords: Medulloblastoma ; Primitive glioma ; Differentiation ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To increase our understanding of the differentiation potential and cytogenesis of embryonal tumors in the brain, 12 medulloblastomas and 10 primitive gliomas were studied by electron microscopy. Of the medulloblastomas, 6 cases were undifferentiated tumors that demonstrated fine processes similar to microspikes. The remaining 6 cases were poorly-differentiated tumors. Axon-like processes were seen in 4 of these cases, including one with dense core vesicles. Astroglial processes were seen in 4 cases. Of the primitive gliomas, there were 2 undifferentiated tumors and 8 poorly-differentiated tumors. Some of the undifferentiated tumor cells had broad cytoplasmic processes. Of the 8 poorly-differentiated gliomas, there were groupings of junctional complexes in 7, microvilli in 4 and bundles of intermediate filaments in 5. It is concluded that medulloblastomas have both neuronal and glial differentiation potentials and that primitive gliomas have only glial differentiation potential. The origin of medulloblastomas may be from the remnants of cell nests with a multipotential differentiation capacity, and the origin for primitive gliomas may be from remnants of glial precursor cells.
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  • 31
    ISSN: 1860-1499
    Keywords: Ultrastructure ; Transferrin receptor ; Oral malignant melanoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A rare variety of malignant melanoma in the maxilla was studied by electron microscopic observation and an immunohistochemical investigation of the transferrin receptor (TfR). A number of low-stage melanosomes were seen, and the TfR was found in both the cell membrane and the cytoplasm.
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  • 32
    ISSN: 1860-1499
    Keywords: Vasoactive intestinal polypeptide (VIP) ; Intestinal ischemia ; Immunohistochemistry ; Ganglionic cells ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Alterations in the distribution of vasoactive intestinal polypeptide (VIP) in normal and ischemic small intestines of dogs were studied by using conventional transmission electron microscope, and immunohistochemistry for light and electron microscopy. At the light microscopic level, immunoreactivity was evident in the intestinal ganglionic cells of control segments. At the electron microscopic level using a pre-embedding method, the entire cytoplasm of the ganglionic cells in the control segments was filled with VIP immunoreactive products, while the post-embedding experiment showed positive reactions only within the VIP granules and Golgi vesicles. After 30 min of ischemia, immunoreactivity was greatly decreased in the ganglionic cells and a large amount of VIP immunoreactive product appeared in the striated border of epithelial cells and in nerve fibers of the subepithelial layer. These results suggest that intestinal ischemia might lead to the release of VIP, which seems to bind to the microvillus membrane of epithelial cells. The relationship between the changes in VIP distribution and its protecting mechanisms of ischemic damage is discussed.
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  • 33
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    Medical molecular morphology 27 (1994), S. 113-122 
    ISSN: 1860-1499
    Keywords: Breast cancer ; Apocrine carcinoma ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Three cases of apocrine carcinoma of the breast were investigated by light and electron microscopy. Histologically, the tumor cells were shown to have eosinophilic granular cytoplasm and occasional cytoplasmic snouts protruding into the lumen. Ultrastructurally, light and dark tumor cells were identified, and there were many lysosome-like dense bodies of various sizes and small dense granules grouped in the subapical region. Many mitochondria with incomplete cristae and numerous small vacuoles were distributed in the cytoplasm. Occasionally, intracytoplasmic lumen were visible in the tumor cells. These ultrastructural findings are quite similar to those found in normal apocrine sweat glands. However, the presence of an intraductal component of carcinoma suggests the origin to be mammary epithelial cells.
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  • 34
    ISSN: 1860-1499
    Keywords: Human T-lymphotropic virus type I (HTLV-I) ; Ultrastructure ; Basedow's disease ; Lymphocyte ; Thyroid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Thyroid tissue from a patient with Basedow's disease and immunopositive for HTLV-I was examined. Histopathological and immunohistochemical studies revealed that follicular-epithelial cells (FEC) had papillary projections reaching into the follicular lumina which were associated with diffuse T, but not B, lymphocyte infiltration and occasionally formed aggregations intermingled with T and B lymphocytes. Ultrastructurally, infiltrated lymphocytes possessing round or irregularly indented nuclei were detected in the capillary lumen, stroma and, also, FEC layers. The lymphocytes extended irregular processes, attached to long thin processes of FEC and infiltrated into FEC layers. Cocultivation of minced thyroid tissue with normal lymphocytes resulted in the establishment of a type-C virus-producing T-cell line.
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  • 35
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    Medical molecular morphology 27 (1994), S. 254-256 
    ISSN: 1860-1499
    Keywords: Gonad ; Mesonephros ; Histogenesis ; Primordial sex cord ; Folliculogenous sex cord ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The histogenesis and organogenesis of the human gonad in twelve embryos and six fetuses of ovulational ages 5 to 18 weeks was investigated by histological and ultrastructural examination, including observation of almost complete serial Epon-embedded sections of the entire gonads of 10 embryos. This investigation revealed that the main constituent cells of the gonads are derived from the mesonephros, and that the coelomic epithelium is not involved in the formation of the main component at any stage.
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  • 36
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    Medical molecular morphology 27 (1994), S. 260-263 
    ISSN: 1860-1499
    Keywords: Cellular interaction ; Follicular wall ; Granulosa cell ; Theca cell ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Bovine granulosa and theca cells were cultured on the opposite sides of a collagen membrane, and the morphological and structural effects of the interaction between theca and granulosa cells were studied. The apical surface of both granulosa and theca cells in the cocultured group appeared convex, while control cells were thin and flat. Numerous filopodia spread over the cellular surface connecting the cells. Constituents of rough endoplasmic reticulum in the cocultured cells were extended and swollen when compared with those observed in the cells cultured alone. Cocultured granulosa cells formed multilayered sheets, while granulosa cells cultured alone formed monolayered sheets. Theca cells in the cocultured group were more densely packed than cells cultured alone. These results indicate that the levels of cellular activity and intercellular communication among each type of cell are increased by the granulosa and theca cell interactions.
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  • 37
    ISSN: 1860-1499
    Keywords: X-ray microanalysis ; Freeze-drying ; Calcium ; Ultrastructure ; Isehemia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract There is considerable interest of the changes in intracellular electrolytes in various pathophysiological states in cardiology. X-ray microanalysis is a useful method to measure the concentration of intracellular elements, and freeze-drying is a required technique for X-ray microanalysis to preserve elements. In this study, we utilized a new technique by which ultrathin sections were evaporated by carbon in an external freeze-dryer after freeze-drying and we were able to obtain clear electron micrographs of papillary muscles and myocytes. Calcium concentrations of mitochondria and cytosol were measured during 120 min ischemia. We showed that mitochondria played a role in intracellular calcium regulation during 30 min ischemia.
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  • 38
    ISSN: 0899-0042
    Keywords: amylose ; cellulose ; phenylcarbamate ; 3,5-dimethylphenylcarbamate ; 3,5-dimethoxyphenylcarbamate ; trans-stilbene oxide ; chiral sulfoxides ; resolution ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The enantioselective resolution of trans-stilbene oxide and of 23 chiral sulfoxides was investigated on cellulose and amylose tris(arylcarbamate) stationary phases coated on aminopropylated 7 μm spherical silica with 500 Å diameter pores. Cellulose tris-(3,5 dimethylphenylcarbamate) showed good resolving power for many of the sulfoxides and amylose tris-(3,5 dimethoxyphenylcarbamate) showed advantages for the resolution of certain sulfoxides which were not separated on other phases. © 1994 Wiley-Liss, Inc.
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  • 39
    ISSN: 0899-0042
    Keywords: chemoselective reduction ; disiamylborane ; (-)-2′-deoxy-3′-thiacytidine ; (1′R,2′S,5′R)-menthyl-(5R)-acetoxy-1,3-oxathiolan-(2R)-carboxylate ; Lamivudine ; 3TCTM ; chiral HPLC ; chiral stationary phase ; Pirkle β-GEM 1 column ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Chemoselective reduction of one isomer of the 1-menthylester of 1,3-oxathiolan-5-one-2-carboxylic acid produces a mixture of four lactol diastereomers from which the title compound was isolated after acylation. The isomeric purity and absolute stereochemistry were determined by spectroscopic methods, chiral HPLC techniques, and conversion to (-)-2′-deoxy-3′-thiacytidine (Lamivudine, 3TCTM). © 1994 Wiley-Liss, Inc.
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  • 40
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    Chirality 6 (1994), S. 165-168 
    ISSN: 0899-0042
    Keywords: chirality ; chemical evolution ; phase transitions ; optical activity ; spontaneous symmetry breakdown ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: We discuss the origin of the chirality of protein amino acids from the point of view of a phase transition from a racemic mixture into an optically pure state. We assume that Bose-Einstein condensation may act as an amplification mechanism. The original theory is due to Salam. We suggest a new role for the phase transition. Following Quack we distinguish parity violation of two kinds (de facto and de lege symmetry breaking). While the Salam phase transition corresponds to parity violation of the second kind (de lege), the phase transition we discuss in this work corresponds to parity violation of what we may call a third kind. This is suggested by recent experimental phenomena which correlate chiral symmetry breaking and pattern formation (spontaneous symmetry breaking that separates an initial racemic mixture into right- and left-handed space domains by means of a substrate). Tentative comments are given on the eventual design of possible experiments that may test this new hypothesis. © 1994 Wiley-Liss, Inc.
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  • 41
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    Chirality 6 (1994), S. 185-195 
    ISSN: 0899-0042
    Keywords: stereoselective pharmacokinetics ; stereoselective bioavailability ; bioequivalence of chiral drugs ; nonlinear pharmacokinetics ; Michaelis-Menten kinetics ; computer simulation ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Computer simulation was used to test the effects of pulsatile oral input on the stereoselectivity in the area under the blood concentration-time curves (AUCs) of the enantiomers of racemic drugs. The effects of input rate determinants, namely, dose, dosage interval, and formulation on the stereoselectivity were investigated under both steady-state and nonsteady-state conditions. Simulations were carried out for drugs undergoing Michaelis-Menten hepatic metabolism with different enantiomeric maximum velocity (Vmax) or constant (Km) values. With pulsatile input, the enantiomeric AUC ratios of both types of drugs were dependent on all the determinants of input rate. However, in most cases, the direction of input rate-dependent changes in the enantiomeric AUC ratios for drugs with different enantiomeric Vmax was opposite of that for drugs with different enantiomeric Km. The direction and magnitude of changes in the enantiomeric AUC ratios were also dependent on the selected dose, dosage interval, and formulation. Further, different conclusions could be reached based on the nonsteady-state and steady-state data. Additional simulations were then performed to test the effects of input rate-dependent stereoselective pharmacokinetics on the bioequivalence of chiral drugs with nonlinear metabolism. These simulations suggested that bioequivalence studies based on the racemic drug measurement may result in erroneous conclusions for the individual enantiomers. The results of this study may be used as a tool for the design of experiments to test the input rate dependence of stereoselective pharmacokinetics and bioequivalence of racemic drugs in animals and humans. © 1994 Wiley-Liss, Inc.
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  • 42
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    Chirality 6 (1994), S. 213-215 
    ISSN: 0899-0042
    Keywords: ICRF-187 ; ICRF-186 ; ICRF-159 ; dexrazoxane ; doxorubicin ; dihydropyrimidine amidohydrolase ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The enzymatic ring-opening hydrolyses of the doxorubicin cardioprotective agents (+)-(S)-ICRF-187 (dexrazoxane), (-)-(R)-ICRF-186, and rac-ICRF-159 by the enzyme dihydropyrimidine amidohydrolase (DHPase) have been studied. ICRF-187 underwent enzymatic ring-opening hydrolysis by DHPase 4.5 times faster than did ICRF-186. It was also shown that DHPase opens only one ring of ICRF-186 and does not act on this one-ring open hydrolysis product, as has been observed for ICRF-187. Differences in the rates at which the two optical isomers are acted upon by DHPase suggest that they could have differing protective effects. © 1994 Wiley-Liss, Inc.
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  • 43
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    Chirality 6 (1994) 
    ISSN: 0899-0042
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
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  • 44
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    Chirality 6 (1994), S. 239-244 
    ISSN: 0899-0042
    Keywords: HPLC ; reverse-phase additive ; β-cyclodextrin ; methylphenobarbitone ; molecular mechanics ; complex stability ; chiral separation ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Molecular modelling of β-cyclodextrin and optimisation of its potential energy suggests that a favoured conformation is that distorted from a symmetrical torus. The inclusion of water molecules into the torus cavity simulates the increased stability in an aqueous solvent. Complexes of β-cyclodextrin with (R)- and (S)-enantiomers of methylphenobarbitone have been modelled and energetically optimised by the application of molecular mechanics. The simulations suggests that the guest molecules adopt an orientation in which the phenyl ring is projected into the torus cavity, with in each case the plane of the ring parallel to a longer axis of the distorted torus and slightly displaced from the axis through the torus cavity. It is suggested that the asymmetry in the macrocyclic ring contributes to chiral recognition as a result of additional discriminatory binding to the barbiturate ring residue of each enantiomer, which occupy different 3D geometries. The enantiomers form complexes of different minimum potential energies. The resulting difference in complex stability can be related to the behaviour of β-cyclodextrin, as a mobile phase additive in reverse-phase HPLC to effect chiral separation of rac-medthylphenobarbitone during chromatography. © 1994 Wiley-Liss, Inc.
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  • 45
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    Chirality 6 (1994), S. 245-250 
    ISSN: 0899-0042
    Keywords: intermolecular association ; hydrogen-bonding ; π-facial ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: 2,2,2-Trifluoro-(9-anthryl)-ethanol (TFAE) has been extensively used, in its pure enantiomeric forms, as a chiral solvating agent in nuclear magnetic resonance spectroscopy (NMR). It has also played an important role in the development of chiral stationary phases in liquid chromatography (LC). X-ray crystallography of the enantiomeric and racemic crystals shows, in both cases, the formation of an intermolecular hydrogen bond between the O—H and the π-face of one of the rings of the anthracene aromatic system.1 Few examples of such hydrogen bonding have been published previously, and those that have are not as clear cut as in this case. An explanation for the hydrogen bonding is sought using molecular modelling via the PM3 analytically derived molecular electrostatic potentials. Using NMR and dynamic lineshape analysis, the barrier to rotation about the aryl-carbon bond is estimated, indicating the C—CF3 bond to be perpendicular to the anthracene axis in nonpolar solution. This conformation is identical to the conformation in the crystal. Evidence is also presented to support the formation of intermolecular π-facial hydrogen bonding in TFAE solutions. It is thought that such hydrogen bonding may be implicated in chiral recognition using this compound. © 1994 Wiley-Liss, Inc.
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  • 46
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    Chirality 6 (1994), S. 277-282 
    ISSN: 0899-0042
    Keywords: D-amino acids ; D-glutamate ; monosodium glutamate ; food analysis ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Monosodium glutamate (MSG) is added to many processed foods at significant levels for flavor enhancement. It is also naturally occurring at high levels in some foods. The enantiomeric composition of free glutamate in foods was examined and all foods analyzed were found to contain D-glutamate. The relative percent of D-glutamate in the food products studied depended on the origin of the glutamate. Foods to which MSG was added by the manufacturer had a high total level of MSG but a lower relative percentage of the D-enantiomer (usually less than 0.8%). In comparison, fermented foods tend to have high relative levels of D-glutamate but a lower total amount of the amino acid. The relative percent of D-glutamate in nonfermented foods containing no added MSG was also found to be low compared to fermented products. In some cases the percent D-glutamate could be related to the relative amounts of other food ingredients such as cheese. © 1994 Wiley-Liss, Inc.
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  • 47
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    Chirality 6 (1994), S. 314-320 
    ISSN: 0899-0042
    Keywords: amphetamine ; distillation method ; clathrate ; optical activation of bases ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The present paper illustrates the development of an advanced technique in optical resolution. Both of the amphetamine enantiomers can be obtained by a two-step distillation in nearly quantitative yield without any loss of the resolving agent. It is proved that the second-order interactions (H-bond) are sufficient for separation of enantiomers by distillation. © 1994 Wiley-Liss, Inc.
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  • 48
    ISSN: 0899-0042
    Keywords: diastereoisomer ; drugs of abuse ; GC-MS ; automation ; sample preparation ; drug testing ; stereoselective analysis ; enantioselectivity ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: An automated sample preparation system has been applied to the chiral analysis of amphetamine and methamphetamine using derivatization with trifluoracetyl-L-prolyl chloride (L-TPC) and subsequent separation on a gas chromatography-mass spectrometry (GC-MS) system. Tasks automated were the dilution of standards and the off-line preparation of the diastereoisomer derivatives. Chromatographic performance, sensitivity, and reproducibility of the automated procedure were compared to the equivalent values obtained with two existing assays methods which employ manual derivatiation, either on-column or off-line. Chromatographic performance was unaffected by the derivatization procedure and sensitivity was better for both automated and manual off-line derivatization. Qualitative reproducibility as based on enantiomeric composition was equivalent for all three approaches, while quantitative reproducibility as based on peak areas was best for the automated procedure. Considering the fact that the diastereoisomer derivatives are unstable over time, automated sample preparation with “just-in-time” derivatization can increase the overall precision of the analytical method. The procedures described here are general enough in nature that they could be applied to other chiral or even achiral analytes. © 1994 Wiley-Liss, Inc.
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  • 49
    Electronic Resource
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    New York, NY [u.a.] : Wiley-Blackwell
    Chirality 6 (1994) 
    ISSN: 0899-0042
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
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  • 50
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    New York, NY [u.a.] : Wiley-Blackwell
    Chirality 6 (1994), S. 148-155 
    ISSN: 0899-0042
    Keywords: chiral stationary phase ; dinitrobenzylphenylethylamine ; dinitrobenzoylphenylglycine ; enantiomers ; 2,2,2-trifluoro-1-(9-anthryl) ethanol ; competition ; nonlinear ; liquid chromatography ; system peaks ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Competition between the (+)- and (-) enantiomers of 2,2,2-trifluoro-1-(9-anthryl) ethanol as mobile phase additives was indicated by the chromatographic behavior of their system peaks. Two types of chiral stationary phases were used, one based on dinitrobenzoylphenylglycine and the other on dinitrobenzylphenylethylamine plus tartaric acid. The racemic mixture was used as the mobile phase additive and k′ of their system peaks was studied as a function of the mixture concentration in the mobile phase in both cases. A shift in k′ of the two system peaks was observed and considered as an indication that competition occurred. The areas of the two system peaks were also studied as a function of the concentration of the enantiomers in the samples, using two different compositions of the mobile phase. The dependency of system peaks' area on the sample composition indicated whether competition between the enantiomers occurred. One mobile phase contained 0.1 mM of the racemic mixture, where the area of the two retained system peaks behaved independently, i.e., only the peak corresponding to the enantiomer was affected by its presence in the sample. The other mobile phase contained 0.75 mM of the racemic mixture, and both peaks were affected by the injection of any one of the enantiomers. The interdependency of the system peaks' area on both the enantiomers indicated that their distribution in the chiral system was interrelated due to mutual interactions. A quantitative treatment of the interdependency and competition was excluded, due to the irreversible adsorption of the two enantiomers on the chiral stationary phase after using overloading concentrations. This irreversible adsorption was visualized by the appearance of two retained system peaks of the two residual enantiomers. These system peaks were detected only when the sample contained pure enantiomers due to competition between the enantiomer in the sample with the residual enantiomers in the stationary phase. © 1994 Wiley-Liss, Inc.
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  • 51
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    Chirality 6 (1994), S. 175-184 
    ISSN: 0899-0042
    Keywords: 1,4-benzodiazepines ; oxazepam ; 3-O-methyloxazepam ; 3-O-ethyloxazepam ; stereoselective nucleophilic substitution ; kinetics of racemization ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Enantiomers of 3-O-methyloxazepam (MeOX) and 3-O-ethyloxazepam (EtOX) were resolved by chiral stationary phase high-performance liquid chromatography (CSP-HPLC). Reaction kinetics and deuterium isotope effects of acid-catalyzed racemization of enantiomeric MeOX in ethanol and enantiomeric EtOX in methanol were studied by spectropolarimetry. The acid-catalyzed heteronucleophilic substitution reactions of racemic MeOX in ethanol and racemic EtOX in methanol were studied by reversed-phase HPLC. Thermodynamic parameters involved in the reactions were obtained by temperature-dependent reaction rates. The effects of solvent's dielectric constant on the heteronucleophilic substitution reactions were also determined. A nucleophilically solvated and transient C3 carbocation intermediate resulting from an N4-protonated enantiomer, derived from a 1,4-benzodiazcpine either in M (minus) or P (plus) conformation, is proposed to be an intermediate and responsible for the acid-catalyzed stereoselective nucleophilic substitution and the resulting racemization. © 1994 Wiley-Liss, Inc.
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  • 52
    ISSN: 0899-0042
    Keywords: 77Se NMR spectroscopy ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Previously, a diazaphospholidine has been synthesized and evaluated as a chiral derivatizing reagent for the determination of the optical purity of chiral alcohols via 31P NMR spectroscopy (Alexakis et al., J. Org. Chem. 57:1224-1237, 1992). Our laboratory is interested in the advantageous and practical applications of 77Se NMR spectroscopic studies in many facets of chemistry and biochemistry. To this end we have used this diazaphospholidine as a starting point and have investigated chiral alcohols coupled to an optically pure diazaselenophospholidine. The diastereomers formed were then evaluated by 77Se NMR spectroscopy, and these results were compared to the 31P NMR results published by Alexakis and co-workers. It was found that addition of the Se atom produced diastereomers that were air stable and, in many cases, the individual diastereomers could be distinguished by 77Se NMR spectroscopy. Preliminary results indicate that the 77Se nucleus is somewhat more sensitive to remotely disposed chiral centers than is the 31P nucleus. Furthermore, because of their stability, these compounds do not readily decompose and can, therefore, be studied by a variety of chromatographic and spectroscopic techniques. © 1994 Wiley-Liss, Inc.
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  • 53
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    Chirality 6 (1994), S. 479-483 
    ISSN: 0899-0042
    Keywords: terfenadine metabolite ; enantiomer separation ; HPLC ; pharmacokinetics ; humans ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: A stereoselective and sensitive achiral/chiral method for the determination of terfenadine acid metabolite in human plasma was developed. The metabolite was separated and quantitated using an achiral chromatographic procedure with a cyano column. The mobile phase was 1 mM sodium acetate buffer (pH 4.0) and acetonitrile (25:75% v/v) at a flow rate of 2 ml/min, at ambient temperature. The stereospecific resolution was accomplished using a chiral-AGP column and a mobile phase consisting of sodium acetate (0.01 M): methanol (98.7:1.3% v/v), and 20 mM di-n-butylamine at a flow rate of 1.2 ml/min. The column temperature was maintained at 32°C. The eluent was monitored at 230 nm (excitation) and 300 nm (emission) with a cut-off filter at 270 nm. This assay was used for a pharmacokinetic study in five subjects after administration of a single dose of 60 mg of terfenadine. The t½ values of the two enantiomers were similar, but the AUC values of the (+)-enantiomer were 2.05-2.35 times higher than those of (-)-enantiomer. © 1994 Wiley-Liss, Inc.
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  • 54
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    Chirality 6 (1994), S. 496-509 
    ISSN: 0899-0042
    Keywords: enantiomeric separations ; macrocyclic antibiotics capillary electrophoresis ; vancomycin ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Vancomycin is one of a family of related macrocyclic glycopeptide antibiotics that were discovered by scientists at the Eli Lilly Company in the 1950s. It has been used to treat severe staphylococcal infections, particularly when bacterial resistance to other antibiotics has developed. Vancomycin is a naturally occurring chiral compound and has a number of stereogenic centers. Furthermore, it contains a variety of functionalities that are known to be useful for enantioselective interactions (e.g., hydrogen bonding groups, hydrophobic pockets, aromatic groups, amide linkages, etc.). The physicochemical properties of vancomycin, including its stability in solution, are discussed as they pertain to capillary electrophoresis. Over 100 racemates were resolved including many nonsteroidal antiinflammatory drugs, antineoplastic compounds and N-derivatized amino acids. Many of these compounds had very high resolution factors. Optimization and the effect of different experimental parameters on the enantioselective separations are discussed. © 1994 Wiley-Liss, Inc.
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  • 55
    ISSN: 0899-0042
    Keywords: 2,2′-diisopropylbiphenyl ; racemization and enantiomerization ; rotational energy barrier ; computer simulation ; dynamic chiral gas chromatography ; cyclodextrin stationary phase ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: By computer simulation of experimental dynamic gas chromatographic elution profiles, the rotational energy barrier ΔG= of racemic 2,2′-diisopropylbiphenyl has been determined as 114.6-115.0 kJ/mol (75-100°C). These data are in good agreement with a value that was determined previously by measuring the racemization kinetics of an enriched sample. This indicates that there is no measurable catalytic or inhibitory effect of the stationary phase. © 1994 Wiley-Liss, Inc.
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  • 56
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    Chirality 6 (1994), S. 521-530 
    ISSN: 0899-0042
    Keywords: metallothionein ; copper binding ; glutathione ; circular dichroism ; charge transfer ; metal-ligand stoichiometries ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Metallothionein (MT) is a ubiquitous mammalian protein comprising 61 or 62 nonaromatic amino acids of which 20 are cysteine residues. The high sulfhydryl content imparts to this protein a unique and remarkable ability to bind multiple metal ions in structurally significant metal-thiolate clusters. MT can bind seven divalent metal ions per protein molecule in two domains with exclusive tetrahedral metal coordination. The domain stoichiometries for the M7S20 structure are M4(Scys)11 (α domain) and M3(Scys)9 (β domain). Up to 12 Cu(I) ions can displace the 7 Zn2+ ions bound per molecule in Zn7-MT. The incoming Cu(I) ions adopt a trigonal planar geometry with domain stoichiometries for the Cu12S20 structure of Cu6(Scys)11 and Cu6(Scys)9 for the α and β domains, respectively. The circular dichroism (CD) spectra recorded as Cu+ is added to Zn7-MT to form Cu12-MT directly report structural changes that take place in the metal binding region. The spectrum arises under charge transfer transitions between the cysteine S and the Cu(I); because the Cu(I)-thiolate cluster units are located within the chiral binding site, intensities in the CD spectrum are directly related to changes in the binding site. The CD technique clearly indicates stoichiometries of several Cu(I)-MT species. Model Cu(I)-thiolate complexes, using the tripeptide glutathione as the sulfhydryl source, were examined by CD spectroscopy to obtain transition energies and the Cu(I)-thiolate coordination geometries which correspond to these bands. Possible structures for the Cu(I)-thiolate clusters in the α and β domains of Cu12-MT are proposed. © 1994 Wiley-Liss, Inc.
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  • 57
    ISSN: 0899-0042
    Keywords: resolution ; D,L-amino acid esters ; alkaline protease ; simple separation ; high enantiomeric excess and yields ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: We developed a procedure for the resolution of esters of amino acids in 95% t-butanol, followed by saponification of the unreacted esters to afford both enantiomers with high yield and optical purity. The hydrolysis, catalyzed by alkaline protease, was conducted in a mixture of t-butanol (95%) and water (5%) at 25°C, with a pH controlled at pH 8.5 by the addition of NaOH (2 M). The hydrolyzed L-amino acid, which was insoluble under these conditions, precipitated during the course of hydrolysis. After separation of the precipitate, the pH of the filtrate was adjusted to 11.5 to saponify the unreacted ester. The D-antipode precipitated at pH 6.2-6.5. Both optically pure antipodes were obtained with high enantiomeric excesses and yields by simple filtration. © 1994 Wiley-Liss, Inc.
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  • 58
    ISSN: 0899-0042
    Keywords: chiral HPLC ; resolution ; absolute configuration ; X-ray analysis ; GABAB antagonist ; GABAB receptor affinity ; phaclofen ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Phaclofen, which is the phosphonic acid analogue of the GABAB agonist (RS)-3-(4-chlorophenyl)-4-aminobutyric acid (baclofen), is a GABAB antagonist. As part of our studies on the structural requirements for activation and blockade of GABAB receptors, we have resolved phaclofen using chiral chromatographic techniques. The absolute stereochemistry of (-)-(R)-phaclofen was established by X-ray crystallographic analysis. (-)-(R)-Phaclofen was shown to inhibit the binding of [3H]-(R)-baclofen to GABAB receptor sites on rat cerebellar membranes (IC50 = 76 ± 13 μM), whereas (+)-(S)-phaclofen was inactive in this binding assay (IC50 〉 1000 μM). (-)-(R)-Phaclofen (200 μM) was equipotent with (RS)-phaclofen (400 μM) in antagonizing the action of baclofen in rat cerebral cortical slices, while (+)-(S)-phaclofen (200 μM) was inactive. The structural similarity of the agonist (R)-baclofen and the antagonist (-)-(R)-phaclofen suggests that these ligands interact with the GABAB receptor sites in a similar manner. Thus, it may be concluded that the different pharmacological effects of these compounds essentially result from the different spatial and proteolytic properties of their acid groups. © 1994 Wiley-Liss, Inc.
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  • 59
    ISSN: 0899-0042
    Keywords: thalidomide enantiomers ; HPLC ; hydroxylated metabolites ; mass spectrometry ; EM 12 ; in vitro metabolism ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The stereoselective metabolism of the former sedative thalidomide and the metabolism of its analogue EM 12 were studied in vitro with liver homogenates. In our study we focused on hydroxylated nonhydrolyzed metabolites of thalidomide. An analytical HPLC method was developed to determine these metabolites directly. The investigations showed a highly stereoselective biotransformation of thalidomide. 5-Hydroxy thalidomide was preferentially formed by (-)-(S)-thalidomide, whereas (+)-(R)-thalidomide was metabolized to two hitherto unknown compounds (Met A and B). Mass spectrometry of these metabolites Met A and B indicated that oxidation or hydroxylation took place in the glutarimide moiety. Biotransformation studies with the more stable thalidomide analogue EM 12 revealed four new metabolites (Met C—F) whose quantities differed in the selected liver homogenate. © 1994 Wiley-Liss, Inc.
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  • 60
    ISSN: 0899-0042
    Keywords: chiral HPLC ; experimental design ; quantitative substituent effects ; recognition mechanism ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The determination of lipophilicity of the title compounds allowed treatment of the data for chiral separation (capacity factors) on CTA and CTPB according to these parameters. A linear correlation between In k′(+) and log k′w was found on both CTA and CTPB, as far as the substituents are situated in the plane of the aryl ring or the heterocycle. This correlation with a nonchiral descriptor allows treatment of capacity factors for (-)-enantiomers as deviations from the lipophilicity line or derived parallels. It results in a clear description of the molecular area affecting enantioselectivity. Application to larger alkyl derivatives shows that the effect of the substituent should be treated on a basis of attractive effect in the case of CTA and on the basis of attractive and repulsive effects for CTPB. © 1994 Wiley-Liss, Inc.
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  • 61
    ISSN: 0899-0042
    Keywords: indirect and direct enantioseparation ; chromatographic assignment of absolute configuration ; econazole ; miconazole ; imidazolylethanol ; protein type CSPs (OVM) ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: A method is described for the synthesis and optical purity determination of (-)-(R)- and (+)-(S)-econazole via the optically pure intermediates, (R)- and (S)-imidazolylethanol, which are available by chromatographic resolution or by fractional crystallization of diastereomeric O,O′-disubstituted (R*;R*)- or (S*;S*)-tartaric acid monoesters of the parent imidazolylethanol racemate. Furthermore, this method allows the chromatographic assignment of the absolute configuration of the chiral center of the imidazolylethanol enantiomers and consequently of econazole enantiomers. In addition, a direct liquid chromatographic enantioseparation method for the determination of the optical purity of (R)- and (S)-econazole and other chiral imidazoles on a protein type CSP (OVM) is described and applied to confirm chromatographically the absolute configuration evaluations. © 1994 Wiley-Liss, Inc.
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  • 62
    ISSN: 0899-0042
    Keywords: preparative chromatography ; chiral separation ; polarimetric detection ; dual detection technique ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Pure enantiomers of an agrochemical process intermediate, (RS)-1-(4-chlorophenyl)-4,4-dimethyl-2-(1H-1,2,4-triazol-1-yl)-pentan-3-one (1), have been prepared on the milligram scale under overload chromatographic conditions on an analytical chiral column (250 × 4.6 mm i.d.). The effects of variation of temperature and mobile phase composition on retention factor, separation factor, and peak resolution have been investigated. Effects of flow rate, enantiomer ratio, sample concentration, and column load on productivity are also studied. Seven milligrams of the less retained (+)-enantiomer and 5 mg of the (-)-enantiomer were obtained from a single injection of 21 mg of (RS)-1. © 1994 Wiley-Liss, Inc.
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  • 63
    ISSN: 0899-0042
    Keywords: SFC ; brush-type chiral stationary phase ; Whelk-O CSP ; pharmaceutical analysis ; chiral analysis ; chiral preparative chromatography ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Good results have been obtained with use of the new bonded chiral stationary phase Whelk-O 1 in analytical and preparative subcritical fluid chromatography. A wide variety of enantiomeric pairs of compounds with different functional groups that are of pharmaceutical and biological interest have been resolved. This Pirkle-concept CSP appears to be more rugged than cellulosic phases (e.g., Chiralcel) with regards to solvents and pressure. In comparing the usefulness of the column for SFC versus HPLC chiral analysis, we have observed a clear superiority of SFC in terms of higher speed and efficiency of analysis, and faster method development. This is consistent with our experience with Chiralcel CSPs. With the Whelk-O 1 we have shown that the effects of temperature and modifier on SFC separations are similar to what has been reported for most other CSPs. We also observed a unique selectivity advantage of SFC for verapamil. We had good success with using a 1-in. diameter column packed with Whelk-O 1 to perform preparative SFC separations of a number of enantiomeric mixtures. The advantages of preparative SFC over preparative HPLC will be discussed. The feasibility of preparative SFC is dependent on how well we meet the practical challenges such as sample introduction issues, special hardware requirements due to the high pressure, and fraction collection issues. © 1994 Wiley-Liss, Inc.
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  • 64
    ISSN: 0899-0042
    Keywords: pharmacokinetics ; antimalarials ; enantiomers ; pharmacodynamics ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Hydroxychloroquine, a slow acting antirheumatic drug, is administered as the racemic mixture. Blood concentrations of the two enantiomers of hydroxychloroquine were measured in two studies, one study of eight patients, in whom blood and urine concentrations were measured during the first 6 months of therapy with rac-hydroxychloroquine, and one of 43 patients who had received rac-hydroxychloroquine therapy for at least 6 months. In the latter study rheumatoid disease activity was also measured. The pharmacokinetics of hydroxychloroquine were found to be enantioselective. The concentrations of (-)-(R)-hydroxychloroquine were higher than those of the (+)-(S)-antipode in all patients at all time points, although the ratios of the two enantiomers did display a two to three fold variability between patients. Both total and renal clearance were greater for the (+)-(S)-enantiomer. From the observational, cross-sectional study design used, it was not possible to differentiate concentration-effect relationships of the two enantiomers. The 11-fold range of drug concentrations swamped any effect of variability between patients in enantiomer proportions. Blood concentrations of both enantiomers were significantly higher in groups of patients with less active disease. © 1994 Wiley-Liss, Inc.
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  • 65
    ISSN: 0899-0042
    Keywords: hydroxychloroquine enantiomers ; absorption ; pharmacokinetics ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The disposition of hydroxychloroquine enantiomers has been investigated in nine patients with rheumatoid arthritis following administration of a single dose of the racemate. Blood concentrations of (-)-(R)-hydroxychloroquine exceed those of (+)-(S)-hydroxychloroquine following both an oral and intravenous dose of the racemate. Maximum blood concentrations of (-)-(R)-hydroxychloroquine were higher than (+)-(S) -hydroxychloroquine after oral dosing (121 ± 56 and 99 ± 42 ng/ml, respectively, P = 0.009). The time to maximum concentration and the absorption half-life, calculated using deconvolution techniques, were similar for both enantiomers. The fractions of the dose of each enantiomer absorbed were similar, 0.74 and 0.77 for (-)-(R)- and (+)-(S)-hydroxychloroquine, respectively (P = 0.77). The data suggest that absorption of hydroxychloroquine is not enantioselective. The stereoselective disposition of hydroxychloroquine appears to be due to enantioselective metabolism and renal clearance, rather than stereoselectivity in absorption and distribution. © 1994 Wiley-Liss, Inc.
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  • 66
    ISSN: 0899-0042
    Keywords: 2-phenylpropionic acid ; peroxisome proliferation ; rat liver ; acyl CoA ; stereoselectivity ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The significance of disturbances of lipid metabolism caused by xenobiotic acyl-CoAs as possible causes of peroxisomal proliferation has been studied with the enantiomers of 2-phenylpropionic acid (2-PPA), the (R)-enantiomer of which is converted to the acyl-CoA in rats while its (S)-antipode is not. rac-2-PPA (250 mg/kg/day ip × 3) was shown to be an hepatic peroxisomal proliferator in male Sprague-Dawley rats on the basis of increases in microsomal cytochrome P-450 content and lauric acid hydroxylation and hepatic CN--insensitive palmitoyl-CoA oxidation, a peroxisomal marker activity, while electron microscopy revealed a rise in the peroxisome/mitochondria ratio in hepatocytes. Further studies established the dose-response relationships for these biochemical changes. The (R)- and (S)-enantiomers were administered at a dose of 50 mg/kg/day ip × 3 and both were peroxisome proliferators of very similar potency. The effects of 100 mg/kg/day ip × 3 of the racemate, a dose giving ca. 75% of maximal response, were essentially additive of those of 50 mg/kg/day ip × 3 of its two component isomers. The stereoselectivity of acyl-CoA formation from the enantiomers of 2-PPA was confirmed by their differential inhibition of microsomal palmitoyl-CoA synthesis. Taken together, these data indicate that it is very unlikely that the acyl-CoA of 2-PPA plays any role in the peroxisomal proliferation which this compound causes in the rat. © 1994 Wiley-Liss, Inc.
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  • 67
    ISSN: 0899-0042
    Keywords: β-adrenergic blocking agent ; calcium antagonist ; enantiomers ; inflammation ; protein binding ; pharmacokinetics ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The influence of endotoxin-induced inflammation on the enantioselective pharmacokinetics of propranolol, oxprenolol, and verapamil, which bind to α1-acid glycoprotein, was studied in the rat. The racemic mixtures were given orally. In the control animals, for propranolol and oxprenolol, the plasma concentrations of the (R)-enantiomer were higher than those of the (S)-enantiomer, while for verapamil the reverse was true. Protein binding and intrinsic clearance are the main factors responsible for this enantioselectivity. After endotoxin treatment, for the three drugs tested the plasma concentrations and the plasma binding of both enantiomers were significantly increased. This effect was more pronounced for (R)-propranolol, (R)-oxprenolol, and (S)-verapamil than for their respective antipodes. The enantioselective effect of endotoxin on the plasma concentrations of the drugs studied seems mainly due to the enantioselective increase in binding to α1-acid glycoprotein. © 1994 Wiley-Liss, Inc.
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  • 68
    ISSN: 0899-0042
    Keywords: synthesis of enantiopure 2-, 3-, and 4-alkyl-branched acids ; diastereomeric phenylglycinol amides ; chain elongation by Arndt-Eistert synthesis ; malonic ester synthesis or via 2-alkylated alkyl carbonitriles ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: A general approach to the synthesis of 2-, 3-, and 4-alkyl-branched acids of high enantiomeric purity is described. The enantiopure 2-alkyl-branched acids are prepared via liquid chromatographic resolution of diastereomeric phenylglycinol amides and their absolute configuration is deduced from the 1H-NMR data of the separated diastereomers. Chain elongation methods, by Arndt-Eistert synthesis, via 2-alkylated alkyl carbonitrile or by malonic ester synthesis, are used to prepare 3- and 4-alkyl-branched acids of high configurational purity and known absolute configuration starting from the enantiomeric 2-alkyl-branched acids. © 1994 Wiley-Liss, Inc.
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  • 69
    ISSN: 0899-0042
    Keywords: Ibuprofen ; nonsteroidal antiinflammatory drug (NSAID) ; chiral inversion ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Ibuprofen (IB) is a chiral 2-arylpropionic acid derivative used as a nonsteroidal antiinflammatory drug (NSAID). It undergoes substantial R to S chiral inversion in humans and rats. In addition to systemic inversion, presystemic chiral inversion has been suggested for IB in humans but only after administration of formulations with slow absorption rates. In search for a suitable animal model, the absorption rate dependency of the extent of inversion was examined in male Sprague-Dawley rats given 20 mg/kg of racemic IB in aqueous solution (Tmax, 0.6 h), suspension (Tmax, 1 h) or as sustained release granules (Tmax, 2.3 h). In addition, (R)-IB (5 mg/liter) was incubated in the presence of everted rat gut segments in an organ bath at 37°. After sustained release granules, the S:R AUC ratios (7.3 ± 1.5) were significantly higher than suspension (3.6 ± 1.1) and solution (3.5 ± 0.2). Accordingly, AUCS and AUCR, as percent of the total AUC (S + R), significantly increased and decreased, respectively, after administration of the sustained released granules as compared with the solution and suspension. A significant positive linear correlation was found between the S:R AUC ratios and the corresponding Tmax for (R)-IB (r = 0.82). In vitro, (R)-IB was inverted by everted jejunum (12.2 ± 1.6%), ileum (14.2 ± 2.0%), and colon (4.4 ± 0.6%) segments. IB was also glucuronidated in the presence of the intestinal segments. Therefore, similar to earlier observations made in humans, in the rat, the S:R AUC ratio was positively and significantly correlated with the absorption rate from the dosage form. Rat small intestine was capable of inverting and conjugating (R)-IB. Hence, rat is a suitable model for studying the chiral inversion of IB. © 1994 Wiley-Liss, Inc.
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  • 70
    ISSN: 0899-0042
    Keywords: chiral pharmacokinetics ; rac-flurbiprofen ; rat ; bone pharmacodynamics ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The route of administration of the NSAID, flurbiprofen (sq vs. po) resulted in positive and negative results respectively with regard to enhanced cancellous and cortical bone accumulation in the immature rat. This pharmacokinetic study was an effort to understand the pharmacodynamic difference between the two routes of administration observed when the same dose range of drug, given as single daily doses, had been employed in both studies. Conventional chiral pharmacokinetics were evaluated in young rats. A significant difference was observed in the Tmax of the active (S)-enantiomer by both administration routes (sq 4 h and po 1 h). The bioavailability, as evaluated by AUCs favored the sq route as expected. The plasma concentrations over 18 h, at steady state, for one po dose group (0.5 mg/kg/day) fell well within the therapeutic window described by the 0.1 and 0.5 mg/kg sq doses which had demonstrated anabolic bone activity. Oral dosing had exhibited no significant bone activity. We conclude that the pharmacodynamic difference between routes of administration cannot be simply explained on a pahrmacokinetic basis. Consequently, experiments detailing the pharmacodynamics and pharmacokinetics of single and multiple dose administration of aryl-propionic acids in normal and osteopenic states need further pharmacologic study. © 1994 Wiley-Liss, Inc.
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  • 71
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    Chirality 6 (1994), S. 472-478 
    ISSN: 0899-0042
    Keywords: nonproteinogenic α-amino acid ; pronase ; optical resolution ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: We describe a practical method to produce pure enantiomers of non-proteinogenic α-amino acids by pronase-catalyzed hydrolysis of their methyl esters. Each of the two pure enantiomers was obtained in high yield, while the reaction conditions were optimized with a view to large scale production. Part of this work was devoted to conceiving an analytical procedure especially designed to monitor the steric course of enzymatic reactions. © 1994 Wiley-Liss, Inc.
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  • 72
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    Chirality 6 (1994), S. 513-513 
    ISSN: 0899-0042
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
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  • 73
    ISSN: 0899-0042
    Keywords: stereoselective synthesis ; chiral synthon ; Hantzsch esters ; calcium channels ; smooth muscle relaxation ; positive inotropes ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The (+)- and (-)-enantiomers of 3-isopropyl 5-(4-methylphenethyl) 1,4-dihydro-2,6-dimethyl-4-(2-pyridyl)-3,5-pyridinedicarboxylate were synthesized using an efficient highly enantioselective (ee ≥ 96%) variant of the Hantzsch dihydropyridine synthesis. The key step in this procedure involved the asymmetric Michael addition of a metalated chiral aminocrotonate, derived from D-valine or L-valine, respectively, to the Knoevenagel acceptor (Z)-2-isopropoxycarbonyl-1-(2-pyridyl)-but-1-en-3-one. Both enantiomers exhibited a dual cardioselective partial calcium channel agonist (positive inotropic)/smooth muscle selective calcium channel antagonist effect. The relative in vitro smooth muscle calcium channel antagonist activities of the (-):(+) enantiomers was 26:1. In contrast, the (+)-enantiomer exhibited a greater in vitro positive inotropic effect on guinea pig left atrium where the contractile force was maximally increased by 14.8% at a concentration of 1.63 × 10-8 M. © 1994 Wiley-Liss, Inc.
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  • 74
    ISSN: 0899-0042
    Keywords: arylpropionic acid ; ketoprofen ; chiral inversion ; stereoselective pharmacokinetics ; monkey ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Pharmacokinetic studies are reported after single oral administration of 3 mg/kg of stereochemically pure (S)-ketoprofen [(S)-KP] and (R)-ketoprofen [(R)-KP] to three male Cynomolgus monkeys and after repeated administration for 6 months of 3, 15 and 75 mg/kg/day of (S)-KP to both male and female monkeys. A high-performance liquid chromatographic (HPLC) analysis was performed without derivatization of the samples, using a chiral column. The pharmacokinetic parameters for (S)-KP after administration of (S)-KP and for (R)-KP after administration of (R)-KP were, respectively, elimination half-life 2.32 ± 0.36 and 1.64 ± 0.40 h; oral clearance 3.50 ± 0.66 and 7.50 ± 3.20 ml/min/kg; apparent volume of distribution 0.74 ± 0.24 and 1.16 ± 0.76 liter/kg; mean residence time 1.79 ± 0.77 and 1.41 ± 0.65 h; area under the concentration/time curve 14.16 ± 2.93 and 7.31 ± 2.98 μg·h/ml. Forty-nine percent unidirectional bioinversion of (R)-KP to (S)-KP was observed in this species and the pharmacokinetic parameters for the (S)-KP resulting from this inversion were also calculated. In the study of 6-month repeated administration of (S)-KP, linear pharmacokinetic behavior and no evidence of drug accumulation were observed at the three dose levels. © 1994 Wiley-Liss, Inc.
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  • 75
    ISSN: 0899-0042
    Keywords: mianserin ; enantiomer ; metabolism ; human plasma ; urine ; HPLC ; CYP2D6 ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: An HPLC method is presented which allows the measurement in the same run of the enantiomers of mianserin, desmethylmianserin, and 8-hydroxymianserin in plasma and urine of mianserin-treated patients. Limits of quantitation for the (S)- and (R)- enantiomers of mianserin and desmethylmianserin were 4 and 2.5 ng/ml, respectively, in plasma, and for the (S)- and (R)-enantiomers of mianserin, desmethylmianserin, and 8-hydroxymianserin 5, 2.5, and 5 ng/ml, respectively, in urine. The measured ratios of (S)-mianserin/(R)-mianserin and (S)-desmethylmianserin/(R)-desmethylmianserin in the plasmas of 10 mianserin-treated patients, all extensive metabolizers of debrisoquine as determined by CYP2D6 genotyping, varied, respectively, from 1.0 to 4.06 and from 0.19 to 0.64. As the enantiomers of mianserin differ in their pharmacological profile, these results could partially explain why, until now, no consistent relationship has been established between the therapeutic response and total [(S) + (R)] plasma levels of this antidepressant. © 1994 Wiley-Liss, Inc.
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  • 76
    ISSN: 0899-0042
    Keywords: microsomal epoxide hydrolase ; kinetic parameters ; enantiomeric excesses ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The microsomal epoxide hydrolase (mEH)-catalyzed hydrolysis of cis-4,4′-dimethylstilbene oxide (1a), cis-4,4′-diethylstilbene oxide (1b), cis-4,4′-diisopropylstilbene oxide (1c), and cis-4,4′-dichlorostilbene oxide (1d) have been investigated using rabbit liver microsomal preparations. The kinetic parameters, Km and Vmax, and the absolute stereochemistry of the reactions have been determined and compared with those of cis-stilbene oxide (1e). All epoxides 1a-d are hydrolyzed by mEH with high product enantioselectivity to give (R,R)-(+)-diols with ee ≥ 90%. The presence of the substituents on the phenyl rings markedly reduces the rates of mEH catalyzed hydrolysis with respect to cis-stilbene oxide, by increasing Km and reducing Vmax in the cases of 1a, 1b, and 1d, or reducing only the Vmax in the case of 1c. The very low Vmax, together with a persistent ability to fit into the mEH active site, make all these epoxides, and particularly 1c, inhibitors of cis-stilbene oxide hydrolysis. The kinetic and stereochemical results are interpreted on the basis of the proposed topology of the mEH active site. © 1994 Wiley-Liss, Inc.
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  • 77
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    Chirality 6 (1994), S. 605-605 
    ISSN: 0899-0042
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
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  • 78
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    Chirality 6 (1994), S. 615-622 
    ISSN: 0899-0042
    Keywords: absolute configuration ; chiral stationary phase ; enantiomer ; chiral recognition ; NSAID ; HPLC ; NMR ; CD ; X-ray ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Assignment of absolute configuration to a recently developed chiral selector useful in the separation of the underivatized enantiomers of naproxen and other nonsteroidal anti-inflammatory drugs (NSAIDs) is described. Circular dichroism, 1H NMR, and X-ray diffraction have been used to confirm the original assignment which was based solely upon elution orders from HPLC chiral stationary phases. All of these techniques agree in the assignment of the (S,S) absolute configuration to the enantiomer of the chiral selector which associates preferentially with (S)-naproxen. © 1994 Wiley-Liss, Inc.
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  • 79
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    Chirality 6 (1994), S. 654-657 
    ISSN: 0899-0042
    Keywords: successive optical resolution ; replacing crystallization ; threonine ; optically active cosolute ; L-serine ; 4-hydroxy-L-proline ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: DL-Threonine [DL-Thr; (2RS,3SR)-2-amino-3-hydroxybutanoic acid] was optically resolved by replacing crystallization using L-serine (L-Ser) and 4-hydroxy-L-proline (L-Hyp) as optically active cosolutes. D-Thr was allowed to crystallize preferentially from racemic aqueous solutions in the presence of these L-α-amino acids. The optical resolution of DL-Thr was more successfully achieved by using L-Ser, whose structure is more similar to that of DL-Thr than L-Hyp, and successively gave D- and L-Thr of 87 - 92% optical purities. The D- and L-Thr obtained were then recrystallized from water to give optically pure D- and L-Thr. © 1994 Wiley-Liss, Inc.
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  • 80
    ISSN: 0899-0042
    Keywords: methanephosphonic acid dichloride ; N,N′-bis(1-phenylethyl)diamidomethylphosphonate ; chiral discrimination on self-association ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Enantiomeric (S,S)- and (R,R)-bis(1-phenylethyl)diamidomethylphosphonates, the phosphorus atom being prochiral, have been synthesized, starting from the methanephosphonic acid dichloride and corresponding chiral amines. The splitting observed in the 31P-NMR spectra of their nonracemic mixtures may be accounted for the self-discrimination of chiral species. This effect can be very useful for enantiomeric analysis of amines based on coupling reactions with methanephosphonic acid dichloride. © 1994 Wiley-Liss, Inc.
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  • 81
    ISSN: 0899-0042
    Keywords: peroxisomal fatty acid β-oxidation ; peroxisomal lauroyl-CoA oxidase ; catalase ; ω-hydroxylation ; epoxide hydrolases ; induction ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Male C57BL/6 mice were exposed to 1% (w/w) (+)- or (-)-2-ethylhexanoic acid or an equimolar mixture of these enantiomers in their diet for 4 or 10 days. A significant increase in liver weight and a 2- to 3-fold increase in the protein content of the mitochondrial fraction were seen in all cases. Peroxisomal palmitoyl-CoA oxidation was increased 2- to 3.5-fold after 4 days of treatment and 4- to 5-fold after 10 days, while the corresponding increases in peroxisomal lauroyl-CoA oxidase activity were 2- to 3-fold and 9- to 12-fold, respectively. Peroxisomal catalase activity was unchanged, whereas the microsomal and cytosolic activities were increased 2- to 3-fold and 6- to 16-fold, respectively. These treatments also induced microsomal ω-hydroxylation of lauric acid 7-fold and soluble epoxide hydrolase activity in the mitochondrial and cytosolic fractions, as well as microsomal epoxide hydrolase activity about 50-100%. The only significant differences observed between the effects of (+)-2-ethylhexanoic acid and its (-)-enantiomer were on peroxisomal palmitoyl-CoA oxidation and lauroyl-CoA oxidase activity after 4 days of treatment. In both these cases the (+)-enantiomer resulted in increases which were 50-75% greater than those seen with the (-)-form. © 1994 Wiley-Liss, Inc.
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  • 82
    ISSN: 0899-0042
    Keywords: chiral HPLC ; Chiralpak AD ; amylose carbamate stationary phase ; antiestrogen ; breast cancer ; dichlorotriarylcyclopropane ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: (Z)-1,1-Dichloro-2-(4-benzyloxyphenyl)-2,3-bis(4-methoxyphenyl)cyclopropane (5), a potential antitumor agent designed to treat breast cancer, was prepared in three steps. A stereospecific palladium-catalyzed cross coupling reaction which provided the intermediate (Z)-triaryl alkene 4 was a crucial step in the synthesis. Makosza phase transfer reaction on 4 gave the enantiomeric (Z)-dichlorocyclopropane derivatives 5 which were resolved by semipreparative HPLC on a chiral stationary phase consisting of amylose tris-3,5-dimethylphenyl carbamate coated on silica gel. © 1994 Wiley-Liss, Inc.
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  • 83
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    Chirality 6 (1994), S. 63-71 
    ISSN: 0899-0042
    Keywords: agrochemicals ; isolation ; enantiomers ; diode laser polarimeter ; SFC ; preparative isolation ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The development of chiral HPLC methods and isolation techniques within Zeneca Agrochemicals (formerly ICI Agrochemicals) is reviewed. The use of low temperature to improve chiral separations has been successfully applied to production analysis, but although useful for some compounds it is regrettably not a universal panacea for all poor separations. The need to isolate small quantities of individual enantiomers from new compounds for research evaluation has led us to devise a more universal and cheap chiral stationary phase (CSP) for Preparative-LC. Joint academic research produced a CSP based on tartaric acid which was made commercially available and it was gratifying to find it was the only phase able to resolve a novel insecticide. However, as new CSPs emerged almost every month, our attention turned to using a universal chiral detector for analysis, rather than via separation of individual enantiomers. Diode laser-based polarimeters offered the opportunity of cheap, sensitive chiroptical detectors for HPLC and the ability to move away from chiral columns in both research and production analysis. Jointly sponsored research with a university has successfully explored the versatility of chiroptical detectors in agrochemical and food analysis. Comparison of chiral SFC with chiral HPLC and an extensive evaluation of established and research agrochemicals on a wide range of commercial CSPs have led to a revised method development strategy. Current work with high load displacement chiral chromatography will be described as a potential means of isolating pure enantiomers from racemates. © 1994 Wiley-Liss, Inc.
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  • 84
    ISSN: 0899-0042
    Keywords: high-performance liquid chromatography ; chiral stationary-phase ; flavin-containing monooxygenase ; N-ethyl-N-methylaniline N-oxide, Chiralcel OD ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The prochiral tertiary amine N-ethyl-N-methylaniline (EMA) is known to be metabolically N-oxygenated in vitro with microsomal preparations. This biotransformation is thought to be mediated predominantly by the flavin-containing monooxygenase (FMO) enzyme system. Microsomal N-oxygenation of EMA is known to be stereoselective and varies between species. In order to further characterise this metabolic transformation, we have examined the in vitro metabolism of EMA using purified porcine hepatic FMO. Following incubation of EMA with purified FMO, EMA N-oxide, the only metabolite detected, was found to be produced stereoselectively [ratio (-)-(S):(+)-(R), ca. 4:1]. The enantiomeric ratio of the N-oxide product did not change markedly with respect to time, enzyme or substrate concentration. Determination of the kinetics of formation of the N-oxide indicated a single affinity for the prochiral substrate with differential rates of formation of the enantiomers. The extent of EMA N-oxide formation was shown to be affected by activators and inhibitors of FMO and pH, but its stereoselectively was unaltered. © 1994 Wiley-Liss, Inc.
    Additional Material: 4 Ill.
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  • 85
    ISSN: 0899-0042
    Keywords: enantioselective ; chromatography ; validation ; column-switching ; robotic ; pharmacokinetic ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Lifibrol, a new drug for the treatment of hypercholesterolemia, contains a stereogenic center bearing a secondary alcohol group. A normal-phase achiral-chiral HPLC separation of the enantiomers of lifibrol and two of its metabolites was developed and validated for quantitation in dog plasma. A silica and a Chiralcel OD-H column were operated in series and all six enantiomeric components and internal standard were directly separated. An initial solid-phase extraction (phenyl) clean-up step and a column-switching step to eliminate late-eluting compounds were also utilized. The solid-phase extraction step was automated using a robotic system. Assay development, validation, and application of the method to a bioavailability study of the racemate and enantiomers of lifibrol in dogs are described. The lower limit of quantitation was 0.0125 μg/ml for each enantiomer of lifibrol using 200 μl of dog plasma with UV detection (255 nm). In dog plasma following oral or intravenous administration of the racemate, the (R)/(S) ratio of the enantiomers of lifibrol was greater than one and increased with time. Following administration of the individual enantiomers, chiral inversion of the (S)-enantiomer but not the (R)-enantiomer was observed. © 1994 Wiley-Liss, Inc.
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  • 86
    ISSN: 0899-0042
    Keywords: substrate enantioselectivity ; product enantioselectivity ; chirality ; activation enthalpy ; activation entropy ; chiral HPLC ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The steady-state kinetic parameters for pig liver carboxylesterase (PLE)-catalyzed hydrolysis of the prochiral substrate dimethyl phenylmalonate (DMPM) (product enantioselectivity) and the separate enantiomers of three chiral 2-phenylpropionic acid esters (substrate enantioselectivity) were measured at seven temperatures between 288 K and 312 K. Arrhenius plots of turnover numbers against the reciprocal of experimental temperatures yielded enthalpies and entropies of activation at enzyme saturation. (+)-(S)-methyl-2-phenylpropionate, (+)-(S)-4-nitrophenyl 2-phenylpropionate, and both enantiomers of phenyl 2-phenylpropionate showed very similar activation enthalpies and entropies (approximately 50 kJ mol-1 and -50 J mol-1 K-1, respectively), but differences were observed for (-)-(R)-methyl 2-phenylpropionate and (-)-(R)-4-nitrophenyl 2-phenylpropionate. Whereas the entropies of activation of all 2-phenylpropionates were negative, positive entropies of activation were measured in the formation of monomethyl phenylmalonate enantiomers from DMPM. Enthalpy-entropy compensation analysis of the data indicates a common mechanism of PLE substrate and product enantiospecificity in the reactions studied here. © 1994 Wiley-Liss, Inc.
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  • 87
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    New York, NY [u.a.] : Wiley-Blackwell
    Chirality 6 (1994) 
    ISSN: 0899-0042
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
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  • 88
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    Chirality 6 (1994), S. 58-62 
    ISSN: 0899-0042
    Keywords: ion channels ; calcium channels ; sodium channels ; state-dependent interactions ; voltage-dependent interactions ; stereoselectivity ; local anesthetics ; calcium antagonists ; 1,4-dihydropyridines ; verapamil ; antifreeze proteins ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Ion channels are pharmacological receptors with specific drug binding sites. These binding sites define specific structure-function relationships for the actions of drug classes. Interpretation of these structure-function relationships may be complex because of state-dependent drug-channel interactions. These state-dependent interactions determine affinity and access of drug to binding sites and may result in both quantitative and qualitative changes in structure-function relationships including stereoselectivity. A channel-active drug may exhibit antagonist or activator properties according to membrane potential and the stereoselectivity of interaction may also change with channel state. © 1994 Wiley-Liss, Inc.
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  • 89
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    Chirality 6 (1994), S. 51-57 
    ISSN: 0899-0042
    Keywords: arabinitol enantiomers ; disseminated candidiasis ; chiral separation ; gas chromatography-mass spectrometry ; diagnosis/monitoring ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Disseminated candidiasis, a devastating disease with high morbidity and mortality in immunosuppressed patients, is difficult to diagnose because of the protean nature of symptoms and the lack of rapid and reliable laboratory diagnostic procedures. The subject of this review is the status of gas chromatographic-mass spectrometric techniques for the determination of D-arabitinol, a unique metabolite of pathogenic Candida species, in serum and urine. The enantiomers are separated by chiral chromatography followed by specific and sensitive detection using chemical ionization and selected ion monitoring. Using D/L-arabinitol ratios, instead of individual concentrations, eliminates the need for knowing the volume of samples and for calibration curves. A new filter paper technique requires only an unmeasured drop of whole blood (venous or finger/heel puncture) or urine; paper spots are mailable. Parallel determinations of D/L-arabinitol ratios in serum and urine in normal subjects and cancer patients with both normal and increased D/L-arabitinol ratios revealed constant (1.2-1.3 range) ratios of serum D/L-arabitinol/urine D/L-arabinitol for all populations studied. Analyzing two body fluids taken at the same time increases reliability by reducing false positives. © 1994 Wiley-Liss, Inc.
    Additional Material: 4 Ill.
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  • 90
    ISSN: 0899-0042
    Keywords: chirality ; fluoxetine ; norfluoxetine ; desipramine ; iprindole ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The antidepressant fluoxetine (FLU) and its N-demethylated metabolite, norfluoxetine (NFLU), each contains a chiral center. The combination of FLU and desipramine (DMI), another antidepressant, has been reported to be useful in treatment of depression, to dramatically increase plasma levels of DMI and also to produce more rapid β-adrenergic receptor down-regulation in brain than caused by DMI alone. We have now begun studies on the effects of this drug combination on the levels of FLU and NFLU enantiomers in the rat. In addition, the combination of FLU and iprindole (IPR) was also investigated. Male Sprague-Dawley rats were treated intraperitoneally with either normal saline vehicle, DMI (5 mg/kg/day), (R,S)-FLU (10 mg/kg/day) or DMI (5 mg/kg/day) + (R,S)-FLU (10 mg/kg/day) for 4 days. Following the last treatment, 24 h urine samples were collected. Rats were sacrificed and brains were removed. For the IPR study, rats were pretreated with either saline or IPR-HCl (11.2 mg/kg) and then treated 1 h later with (R,S)-FLU. After 5 h, the rats were sacrificed and brains were removed. Brain and urine samples were analyzed by gas chromatography with electron-capture detection for free (R)- and (S)-FLU and (R)- and (S)-NFLU after extraction and reaction with (-)-(S)-N-(trifluoroacetyl)prolyl chloride. The results from the brains of the rats treated with DMI/FLU indicate that levels of the enantiomers of both FLU and NFLU were significantly increased over those seen in the animals receiving (R,S)-FLU alone. In the IPR/FLU treated rats, an increase in the brain levels of both (R)- and (S)-FLU was noted when compared with rats receiving (R,S)-FLU alone; however, there appeared to be no increase in the brain levels of NFLU enantiomers. © 1994 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
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  • 91
    ISSN: 0899-0042
    Keywords: atypical β-adrenoceptors ; Chinese hamster ovary cells ; E. coli ; cAMP accumulation ; heart ; lung ; brown adipocytes ; colon ; esophageal smooth muscle ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Trimetoquinol [1-(3′,4′,5′-trimethoxybenzyl)-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline, TMQ] exists as two enantiomers, and the (-)-(S)-isomer is a potent β-adrenergic receptor (β-AR) agonist. Experiments were conducted to examine the functional and biochemical potencies of the (S)- and (R)-enantiomers of TMQ for interaction with β-AR subtypes in tissues, membrane fractions, and cell systems. The isomeric-activity ratios (IARs) of the TMQ isomers [(S)-isomer ≫ (R)-isomer] for stimulation of β1- and β2-AR of guinea pig right atria and trachea were 224 and 1585, respectively; these IARs were similar to those observed on atypical β-AR systems of rat distal colon (575), rat brown adipocytes (398), but differed from that of rat esophageal smooth muscle (2884) in the presence of pindolol. In the absence of pindolol, the potencies for the TMQ enantiomers were slightly increased; however, the IARs remained unchanged in rat distal colon, rat brown adipocytes, and rat esophageal smooth muscle. Similarly, radioligand binding studies demonstrated that the TMQ isomer β-AR affinities were stereoselective for the (-)-(S)-isomer in membranes of guinea pig left ventricle (β1) and lung (β2) giving IARs of 115 and 389, respectively; and in E. coli expressing human β1- and β2-AR giving IARs of 661 and 724, respectively. Corresponding IARs of receptor affinities and stimulation of cAMP accumulation in Chinese hamster ovary cells expressing human β2-AR and rat β3-AR were 331 and 282, and 118 and 4678, respectively. These results indicate that the (-)-(S)-isomer of TMQ exhibits high affinity, and is a potent and highly stereoselective agonist for each β-AR subtype, that the isomers generally fail to differentiate between the β-AR subtypes, and that, based upon differences in IAR within β3-AR containing systems, subtypes of atypical β (or β3)-AR may exist in adipose tissue and smooth muscle. © 1994 Wiley-Liss, Inc.
    Additional Material: 4 Ill.
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  • 92
    ISSN: 0899-0042
    Keywords: chiral recognition ; diphenylethylendiamine ; benzodiazepines ; amino acids ; phthalide ; sulfoxide ; drug ; carbinol ; tetrahydropyrimidine ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Undecanoyl bound 3,5-dinitrobenzoyl-(S,R)-1,2-diphenylethane-1,2-diamine [(1S,2R)-DNB-DPEDA] as chiral selector (SO) has been synthesized and used as a chiral stationary phase (CSP II) for normal-phase enantioselective HPLC. It is compared with the already published diastereomeric (1S,2S)-DNB-DPEDA-derived CSP I and with the “standard” Pirkle DNB-(R)-phenylglycine-derived CSP III. Chromatographic data for about 100 racemic analytes reveal that CSP II is able to separate especially well enantiomers of derivatized aromatic carboxylic acids and analytes having a benzyl substituent bound at the chiral center. However, CSP I was found to be superior to CSP II and III in its general applicability and its ability to resolve enantiomers of heterocyclic drugs. © 1994 Wiley-Liss, Inc.
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  • 93
    ISSN: 0899-0042
    Keywords: enantiomers ; propranolol ; enantioselective pharmacokinetics ; protein binding ; nicardipine ; drug-drug interaction ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The influence of a single oral dose of 30 mg nicardipine on the pharmacokinetics of (R)- and (S)-propranolol, given orally as rac-propranolol 80 mg, was studied in 12 healthy volunteers. The plasma concentrations were higher for the (S)-enantiomer than for the (R)-enantiomer. The Clo and the Cl′intr of (S)-propranolol were significantly lower than the Clo and Cl′intr of (R)-propranolol. The unbound fraction of (R)-propranolol was significantly higher than that of (S)-propranolol. Coadministration of nicardipine significantly increased the AUC and Cmax and significantly decreased the Clo and Cl′intr for unbound drug of (R)- and (S)-propranolol. These changes were more important for (R)- than for (S)-propranolol. The protein binding was not altered by nicardipine. The enantioselective effect of nicardipine on the metabolic clearance of propranolol appears to be due to an interaction at the level of the metabolizing enzymes. The effect on blood pressure of rac-propranolol was little affected when nicardipine was coadministered with rac-propranolol, and its bradycardic effect was reduced. © 1994 Wiley-Liss, Inc.
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  • 94
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Chirality 6 (1994), S. 25-40 
    ISSN: 0899-0042
    Keywords: Review ; capillary electrophoresis ; enantiomeric resolution ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The preconditions are outlined for enantioselective separations in capillary electrophoresis (CE) with chiral selectors as additives to the background electrolyte. Free solution capillary electrophoresis conditions are characterised by a single solution phase. Chiral separations are reviewed by selector type (chiral ligand exchange, cyclodextrins, crown ethers, glycoproteins) with the extensive studies on cyclodextrins grouped into sections on amino acids, pharmaceuticals, and speciality chemicals, optimisation, biological fluids, and quantitative aspects. In micellar electrokinetic capillary chromatography, enantioselective discrimination occurs by partition in a two-phase system, with a chiral micellar phase as selector. Optimum separation conditions can be readily predicted for a given selector-selectand combination, and absolute values of binding constants determined by CE. Advantages of CE in comparison with HPLC using a chiral stationary phase include robust, rapid assays and the use of small volumes of aqueous solutions; disadvantages include less favourable detection limits. © 1994 Wiley-Liss, Inc.
    Additional Material: 12 Ill.
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  • 95
    ISSN: 0899-0042
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 96
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Chirality 6 (1994), S. 47-50 
    ISSN: 0899-0042
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Additional Material: 5 Ill.
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  • 97
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Chirality 6 (1994), S. 467-471 
    ISSN: 0899-0042
    Keywords: enantiomers ; interaction ; pharmacokinetics ; pharmacodynamics ; pharmacology ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Although the implications of stereochemistry for pharmacokinetics are relatively well appreciated only recently has its influence on pharmacodynamics begun to be examined. The implications of different pharmacological interactions between enantiomers with similar and different kinetic properties are examined through the use of simulation of the pharmacological effect vs. time profile. The influences of assuming that the pharmacological effect is solely the result of the more active enantiomer are also discussed. The simulations demonstrate that the less pharmacologically active enantiomer may have a significant influence on the observed effect vs. time profile and that the assumption that all of the observed pharmacological activity arises from the more active enantiomer may lead to highly inaccurate prediction of the pharmacodynamic parameters. Finally, these observations suggest that the pharmacodynamic profiles of a drug administered as a racemate or as a “pure” formulation of the more active enantiomer may be significantly different. © 1994 Wiley-Liss, Inc.
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  • 98
    ISSN: 0899-0042
    Keywords: cytochrome P-450 ; microsomal epoxide hydrolase ; epoxidation rates ; epoxide hydrolysis rates ; enantiomeric excesses ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The rabbit liver microsomal biotransformation of α-methylstyrene (1a), 2-methyl-1-hexene (1b), 2,4,4-trimethyl-1-pentene (1c), and 2,3,3-trimethyl-1-butene (1d) has been investigated with the aim at establishing the enantioface selection of the cytochrome P-450-promoted epoxidation of the double bond and the enantioselectivity of microsomal epoxide hydrolase (mEH)-catalyzed hydrolysis of the resulting epoxides. GLC on a Chiraldex G-TA (ASTEC) column was used to determine the enantiomeric composition of the products. The epoxides 2 first produced in incubations carried out in the presence of an NADPH regenerating system were not detected, being rapidly hydrolyzed by mEH to diols 3. The enantiomeric composition of the latter showed that no enantioface selection occurred in the epoxidation of 1c and 1d, and a very low (8%) ee of the (R)-epoxide was formed from 1b. Incubation of racemic epoxides 2b-d with the microsomal fraction showed that the mEH-catalyzed hydrolysis of 2c and 2d was practically nonenantioselective, while that of 2b exhibited a selectivity E = 4.9 favoring the hydrolysis of the (S)-enantiomer. A comparison of these results with those previously obtained for linear and branched chain alkyl monosubstituted oxiranes shows that the introduction of the second alkyl substituent suppresses the selectivity of the mEH reaction of the latter and reverses that of the former substrates. © 1994 Wiley-Liss, Inc.
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  • 99
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Chirality 6 (1994), S. 225-229 
    ISSN: 0899-0042
    Keywords: warfarin ; phenprocoumon ; CE ; chiral ; maltooligosaccharides ; enantiomeric excess ; assay ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The separation and quantitation of coumarinic anticoagulant drug enantiomers were achieved by direct chiral capillary electrophoresis using complex maltooligosaccharide mixtures as stereoselective electrolyte modifiers. Chiral separations were characterized by a high selectivity and efficiency, enabling enantiomeric excess determinations. In addition, preliminary results indicate the applicability of the method for the determination of individual enantiomers in biological samples. So the method can be used to perform stereoselective pharmacokinetic studies. © 1994 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
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  • 100
    ISSN: 0899-0042
    Keywords: human serum albumin ; electrokinetic chromatography ; chiral drugs ; ligand-ligand interactions ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: When a protein such as human serum albumin is added to the separation buffer in capillary electrophoresis, the mobility of solutes which bind to that protein may be altered. The change in mobility of the solute is a function both of the strength of the binding interaction, and the difference in mobility between the free solute and protein additive. By adding other ligands which themselves bind to the protein, the strength of the solute-protein binding may be modified, leading to a measurable change in the mobility of the solute. These effects are particularly striking for chiral compounds, where enantioselectivity may be completely lost on addition of a competitive ligand. Capillary electrophoresis with human serum ablumin as a buffer additive was used to separate the enantiomers of benzoin and three phenothiazine derivatives. A comparison of the binding of (S)-benzoin to human serum albumin as determined by capillary electrophoresis and by ultrafiltration was made. A variety of other ligands were then added to the buffer along with the protein, and the effects on mobility and enantioselectivity were studied. The displacers included (R)- and (S)-oxazepam hemisuccinate, (R)- and (S)-warfarin, nitrazepam, phenylbutazone, and octanoic acid. From the results obtained, it seems that capillary electrophoresis may be a useful, rapid method to screen for drug-drug interactions. There are some advantages of using this technique to study protein-ligand interactions: Only very small amounts of ligand are needed (useful when dealing with metabolites); for chiral compounds, if protein binding is stereoselective, then the method is also stereoselective, so single enantiomers are not needed; finally, measurements are obtained in solution, without the need for immobilization of the protein. A disadvantage is that the ligand and protein must have significantly different electrophoretic mobilities. © 1994 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
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