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  • 1
    ISSN: 1434-0879
    Keywords: Bladder cancer ; Urothelium ; CD44V2 ; Alternative splicing ; Immunohistochemistry ; Diagnostic marker
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract CD44 is the principal cell surface receptor for hyaluronate. Variant forms of the receptor, produced by alternative splicing, have been found to be associated with tumor progression in a variety of cancers. Based on investigations at the RNA level, it has recently been proposed that expression of CD44 variant V2 was present in urothelial cancer but not in normal urothelium. Since a distinctive marker for urothelial cancer would be extremely useful, frozen sections of normal urothelium and urothelial cancer were examined for expression of standard CD44 and CD44V2. Frozen sections of specimens of 35 patients with transitional cell carcinoma of the bladder, 16 specimens of normal bladder and 5 ureters were examined. Immunohistochemical staining was performed using a polyclonal antibody to CD44V2 (PAB CD44V2), a monoclonal antibody to CD44V2 (MAB CD44V2) and a monoclonal antibody to CD44S (MAB CD44S). CD44V2 and CD44S were also measured in lysates of urine sediments from 21 patients by enzyme-linked immunoabsorbent assay (ELISA). All investigated transitional cell carcinomas expressed CD44V2. There was no differentiation between invasive and noninvasive carcinoma. CD44V2 was also expressed in normal urothelium. Standard CD44 was expressed by the transitional cell carcinoma, normal urothelium, musculature and interstitial tissue. The amount of CD44V2 and CD44S in lysates of urine sediments is not correlated to diagnosis. In contrast to investigations at the RNA level, CD44V2 on the protein level seems not to be a distinctive marker for urothelial cancer. Therefore, CD44V2 will not be a useful diagnostic marker for detection of transitional cell carcinoma.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0533
    Keywords: Brain ; Cerebral ischemia ; Gerbil ; Immunohistochemistry ; Hippocampus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Anesthetized Mongolian gerbils were subjected to 5-min ischemia and 8 h of recirculation. Vibratiom sections were taken for studying changes in ornithine decarboxylase (ODC) immunoreactivity using an antiserum to ODC, and tissue samples were taken for measuring ODC activity. After 5-min ischemia and 8-h recirculation ODC activity increased 11.5-, 5.9-, and 7.9-fold in the cerebral cortex, striatum and hippocampus, respectively (P≤0.05 to 0.01). In the cortex, striatum and hippocampus of control animals immunoreactivity was low but clearly above the detection limit. The reaction was confined to neurons. After 5-min ischemia and 8-h recirculation a sharp increase in immunoreactivity was observed confined to neurons, indicating that the postischemic activation of polyamine metabolism is a neuronal response to ischemia. The immunoreactivity was markedly increased in the perinuclear cytoplasm and the dendrites. In the striatum the density of neurons exhibiting a sharp increase in immunoreactivity was more pronounced in the lateral than in the ventral part. In the hippocampus a strong reaction was present in all subfields but the CA1 subfield was particularly affected. The present study demonstrates for the first time that biosynthesis of a protein is markedly activated during the first 24 h of recirculation after 5-min cerebral ischemia of gerbils even in the vulnerable CA1 subfield, in which the overall protein synthesis is sharply reduced at the same time. Studying polyamine metabolism after ischemia may, thus, provide new information about the basic molecular mechanisms responsible for the altered gene expression after metabolic stress.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Intensive care medicine 22 (1996), S. 1075-1081 
    ISSN: 1432-1238
    Keywords: Key words Critically ill ; Sepsis ; Trauma ; Volume therapy ; Platelet function ; Aggregometry ; Hydroxyethyl starch solution ; Albumin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Objective: Both albumin and synthetic colloids such as hydroxyethyl starch (HES) solution are used to optimize hemodynamics in the critically ill. The influence of different long-term infusion regimes on platelet function was studied. Design: Prospective, randomized study. Setting: Clinical investigation on a university hospital surgical intensive care unit. Patients: Twenty-eight consecutive trauma patients (injury severity score〉15 points) and 28 consecutive nontraumatized surgical patients with sepsis. Interventions: The patients received either 20% human albumin (HA trauma, n=14; HA sepsis, n=14) or 10% low-molecular-weight HES solution HES 200/0.5 (HES trauma, n=14; HES sepsis; n=14) for 5 days to maintain central venous pressure and/or pulmonary capillary wedge pressure between 12 and 16 mmHg. Measurements and results: Platelet function was assessed by aggregometry (=turbidimetric technique) using adenosine diphosphate 2.0 μmol/l, collagen 4 μl/ml, and epinephrine 25 μmol/l as inductors. Arterial blood was sampled on the day of admission or the day of diagnosis of sepsis (=baseline value) and over the next 5 days. Standard coagulation parameters (antithrombin III, fibrinogen, partial thromboplastin time) were also measured. Total use of HES by the 5th day totalled 4870±990 ml in the trauma and 3260±790 ml in the sepsis patients (HA trauma: 1850±380 ml; HA sepsis: 1790±400 ml). Maximum platelet aggregation decreased significantly during the first 2–3 days after baseline in all groups. At the end of the investigation period, platelet aggregation variables had recovered and reached (or even exceeded) baseline values. Within the entire investigation period, the course of platelet aggregation variables did not differ significantly between HA and HES-treated patients irrespective of whether they were trauma or sepsis patients. Conclusions: Alterations in hemostasis may occur for several reasons in the critically ill. Human albumin is the preferred first-line volume therapy in patients at risk for coagulation disorders. With respect to platelet function, volume replacement with (lower-priced) low-molecular-weight HES solutions can be recommended in this situation without any risk.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Intensive care medicine 22 (1996), S. 1075-1081 
    ISSN: 1432-1238
    Keywords: Critically ill ; Sepsis ; Trauma ; Volume therapy ; Platelet function ; Aggregometry ; Hydroxyethyl starch solution ; Albumin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Objective Both albumin and synthetic colloids such as hydroxyethyl starch (HES) solution are used to optimize hemodynamics in the critically ill. The influence of different long-term infusion regimes on platelet function was studied. Design Prospective, randomized study. Setting Clinical investigation on a university hospital surgical intensive care unit. Patients Twenty-eight consecutive trauma patients (injury severity score〉15 points) and 28 consecutive nontraumatized surgical patients with sepsis. Interventions The patients received either 20% human albumin (HA trauma,n=14; HA sepsis,n=14) or 10% low-molecular-weight HES solution HES 200/0.5 (HES trauma,n=14; HES sepsis;n=14) for 5 days to maintain central venous pressure and/or pulmonary capillary wedge pressure between 12 and 16 mmHg. Measurements and results Platelet function was assessed by aggregometry (=turbidimetric technique) using adenosine diphosphate 2.0 μmol/l, collagen 4 μl/ml, and epinephrine 25 μmol/l as inductors. Arterial blood was sampled on the day of admission or the day of diagnosis of sepsis (=baseline value) and over the next 5 days. Standard coagulation parameters (antithrombin III, fibrinogen, partial thromboplastin time) were also measured. Total use of HES by the 5th day totalled 4870±990 ml in the trauma and 3260±790 ml in the sepsis patients (HA trauma: 1850±380 ml; HA sepsis: 1790±400 ml). Maximum platelet aggregation decreased significantly during the first 2–3 days after baseline in all groups. At the end of the investigation period, platelet aggregation variables had recovered and reached (or even exceeded) baseline values. Within the entire investigation period, the course of platelet aggregation variables did not differ significantly between HA and HES-treated patients irrespective of whether they were trauma or sepsis patients. Conclusions Alterations in hemostasis may occur for several reasons in the critically ill. Human albumin is the preferred first-line volume therapy in patients at risk for coagulation disorders. With respect to platelet function, volume replacement with (lower-priced) low-molecular-weight HES solutions can be recommended in this situation without any risk.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-1238
    Keywords: Key words Pentoxifylline ; Critically ill ; Sepsis ; Trauma ; Inflammation ; Coagulation ; Platelet function ; Aggregometry ; Collagen ; Epinephrine ; Adenosine diphosphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Objective: The methylxanthine derivative pentoxifylline (PTX) is one of those promising substances which are under current investigation to modify or limit inflammatory response. Antiaggregation activity has also been described that may contribute to the beneficial effects of this substance. Long-term effects on platelet function have not been elucidated yet. Design: Prospective, randomized study. Setting: Clinical investigation on a surgical intensive care unit of a university hospital. Patients: 26 trauma patients and 26 patients suffering from sepsis secondary to major operations were consecutively studied. Interventions: The patients prospectively received either 1.5 mg/kg per h pentoxifylline continuously for 5 days (after a loading dose of 600 mg) (trauma-PTX, n=13; sepsis-PTX, n=13) or saline solution as placebo (trauma-control; n=13; sepsis-control, n=13). Measurements: On the day of admission (trauma patients) or day of the diagnosis of sepsis and at 12:00 p.m. during the next 5 days, platelet aggregation induced by adenosine diphosphate (ADP 2.0 μmol/l), collagen (4 μl/ml), and epinephrine (25 μmol/l) was determined by a turbidimetric method from arterial blood samples. Standard coagulation screen was also monitored. Main results: In untreated trauma and sepsis patients, maximum platelet aggregation induced by all three agonists decreased during the first few days after inclusion in the study [trauma: ADP–17.1±8.0 rel% (% change from baseline); sepsis: ADP –26.1±5.6 rel%]. In due course, maximum platelet aggregation recovered, reaching the baseline value or even exceeding it (trauma patients). In the PTX-treated patients, platelet aggregation was significantly less impaired (sepsis group: ADP –4.4±3.3 rel%) or even increased beyond baseline values in the first few days of the study (trauma group: ADP 16.1±8.0 rel%). Fibrinogen plasma levels were lower in the non-treated control groups (p〈0.05) than in the PTX groups. Conclusions: Continuous infusion of PTX for 5 days did not impair platelet function in critically ill patients. In both trauma and sepsis patients, the usual deterioration in platelet function was even attenuated, which may be due to the effects of PTX on cytokine release (e.g., reduction in tumor necrosis factor and interleukin-1), improvement in microcirculation, or additional fibrinolytic effects.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-1238
    Keywords: Pentoxifylline ; Critically ill ; Sepsis ; Trauma ; Inflammation ; Coagulation ; Platelet function ; Aggregometry ; Collagen ; Epinephrine ; Adenosine diphosphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Objective The methylxanthine derivative pentoxifylline (PTX) is one of those promising substances which are under current investigation to modify or limit inflammatory response. Antiaggregation activity has also been described that may contribute to the beneficial effects of this substance. Long-term effects on platelet function have not been elucidated yet. Design Prospective, randomized study. Setting Clinical investigation on a surgical intensive care unit of a university hospital. Patients 26 trauma patients and 26 patients suffering from sepsis secondary to major operations were consecutively studied. Interventions The patients prospectively received either 1.5 mg/kg per h pentoxifylline continuously for 5 days (after a loading dose of 600 mg) (trauma-PTX,n=13; sepsis-PTX,n=13) or saline solution as placebo (trauma-control;n=13; sepsis-control,n=13). Measurements On the day of admission (trauma patients) or day of the diagnosis of sepsis and at 12:00 p.m. during the next 5 days, platelet aggregation induced by adenosine diphosphate (ADP 2.0 μmol/l), collagen (4 μl/ml), and epinephrine (25 μmol/l) was determined by a turbidimetric method from arterial blood samples. Standard coagulation screen was also monitored. Main results In untreated trauma and sepsis patients, maximum platelet aggregation induced by all three agonists decreased during the first few days after inclusion in the study [trauma: ADP −17.1±8.0 rel% (% change from baseline); sepsis: ADP −26.1±5.6 rel%]. In due course, maximum platelet aggregation recovered, reaching the baseline value or even exceeding it (trauma patients). In the PTX-treated patients, platelet aggregation was significantly less impaired (sepsis group: ADP −4.4±3.3 rel%) or even increased beyond baseline values in the first few days of the study (trauma group: ADP 16.1±8.0 rel%). Fibrinogen plasma levels were lower in the non-treated control groups (p〈0.05) than in the PTX groups. Conclusions Continuous infusion of PTX for 5 days did not impair platelet function in critically ill patients. In both trauma and sepsis patients, the usual deterioration in platelet function was even attenuated, which may be due to the effects of PTX on cytokine release (e.g., reduction in tumor necrosis factor and interleukin-1), improvement in microcirculation, or additional fibrinolytic effects.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Intensive care medicine 23 (1997), S. 567-573 
    ISSN: 1432-1238
    Keywords: Key words Critically ill ; Sepsis ; Trauma ; Neurosurgery ; Heparin ; Anticoagulation ; Platelet function ; Aggregometry ; Adenosine diphosphate ; Epinephrine ; Collagen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Objective: To study the influence of continuous administration of heparin on platelet function in intensive care patients. Design: Prospective, serial investigation. Setting: Clinical investigation on a surgical and neurosurgical intensive care unit in a university hospital. Patients: The study included 45 patients: 15 postoperative with patients sepsis (Acute Physiology and Chronic Health Evaluation II score between 15 and 25), 15 trauma patients (Injury Severity Score 15 to 25), and 15 neurosurgical patients. Interventions: Management of the patients was carried out according to the guidelines for modern intensive care therapy. Sepsis and trauma patients received standard (unfractionated) heparin continuously [aim: an activated partial thromboplastin time (aPTT) approximately 2.0 times normal value; sepsis-heparin and trauma-heparin patients], whereas neurosurgical patients received no heparin (neurosurgical patients). Measurements and results: From arterial blood samples, platelet aggregation was measured by the turbidimetric method. Platelet aggregation was induced by adenosine diphosphate (ADP; 2.0 μmol/l), collagen (10 μg/ml), and epinephrine (25 μmol/l). Measurements were carried out on the day of diagnosis of sepsis or 12 h after hemodynamic stabilization (trauma and neurosurgery patients) (baseline) and during the next 5 days at 12.00 noon. Standard coagulation parameters [platelet count and fibrinogen and antithrombin III (AT III) plasma concentrations] were also monitored. Heparin 4–10 U/kg per h (mean dose: approximately 500 U/h) was necessary to reach an aPTT of about 2.0 times normal. Platelet count was highest in the neurosurgical patients, but it did not decrease after heparin administration to the trauma and sepsis patients. AT III and fibrinogen plasma levels were similar in the three groups of patients. In the sepsis group, platelet aggregation variables decreased significantly (e. g., epinephrine-induced maximum platelet aggregation: − 45 relative % from baseline value). Platelet function recovered during the study and even exceeded baseline values (e. g., ADP-induced maximum platelet aggregation: + 42.5 relative % from baseline value). Continuous heparinization did not blunt this increase of platelet aggregation variables. In the heparinized trauma patients, platelet aggregation variables remained almost stable and were no different to platelet aggregation data in the untreated neurosurgical patients. Conclusions: Continuous administration of heparin with an average dose of approximately 500 U/h did not negatively influence platelet function in the trauma patients. Recovery from reduced platelet function in the sepsis group was not affected by continuous heparinization. Thus, continuous heparinization with this dose appears to be safe with regard to platelet function in the intensive care patient.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Experimental brain research 82 (1990), S. 25-32 
    ISSN: 1432-1106
    Keywords: Retina ; Development ; In situ hybridization ; Gene expression ; Immunohistochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The expression of the synapsin I gene was studied during postnatal development of the rat retina at the mRNA and protein levels. In situ hybridization histochemistry showed that synapsin I mRNA was expressed already in nerve cells in the ganglion cell layer of the neonatal retina, while it appeared in neurons of the inner nuclear layer from postnatal day 4 onward. Maximal expression of synapsin I mRNA was observed at P12 in ganglion cells and in neurons of the inner nuclear layer followed by moderate expression in the adult. At the protein level a shift of synapsin I appearance was observed from cytoplasmic to terminal localization during retinal development by immunohistochemistry. In early stages (P4 and P8), synapsin I was seen in neurons of the ganglion cell layer and in neurons of the developing inner nuclear layer as well as in the developing inner plexiform layer. In the developing outer plexiform layer synapsin I was localized only in horizontal cells and in their processes. Its early appearance at P4 indicated the early maturation of this cell type. A shift and strong increase of labelling to the plexiform layers at P12 indicated the localization of synapsin I in synaptic terminals. The inner plexiform layer exhibited a characteristic stratified pattern. Photoreceptor cells never exhibited synapsin I mRNA or synapsin I protein throughout development.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 265 (1991), S. 175-184 
    ISSN: 1432-0878
    Keywords: Nervous system ; Nervous system, peripheral ; Catecholamines ; Immunohistochemistry ; Glyoxylic acid fluorescence ; Ophryotrocha puerilis (Annelida)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The complex catecholaminergic (CA) nervous system of the polychaete Ophryotrocha puerilis is documented using glyoxylic acid induced fluorescence (GIF) and immunohistochemistry. CA-neurons are found both in the central and peripheral nervous system. In the brain, about 50 CA-neurons are present in the perikaryal layer together with numerous CA fibres. Two pairs of CA perikarya are characteristic for each ganglion of the ventral nerve cord. CA-neurites in the ventral nerve cord are mainly arranged in 4 strands paralleling the longitudinal axis of the worm. Fluorescent neurons with receptive ciliary structures are present in body appendages (antennae, palps, urites, parapodial cirri), in the body-wall, and within the oesophageal wall. Furthermore, a subepidermal nerve net of free CA nerve endings has been found. After incubation of specimens with dopamine prior to the development of GIF more fluorescent perikarya could be observed; the fluorescence was also intensified. Pre-incubation with reserpine reduced the intensity of GIF. Results of high pressure liquid chromatography and immunostaining with a polyclonal antibody against a dopamine-glutaraldehyde-complex suggest that dopamine is the major CA transmitter. It is thought that dopaminergic neurons together with ciliary receptive structures act as mechano- and/or chemoreceptors.
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  • 10
    ISSN: 1432-0878
    Keywords: Neurosecretion ; Catecholamines ; HPLC ; Immunohistochemistry ; Glyoxylic acid fluorescence ; Ophryotrocha puerilis (Annelida)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the posterior part of the brain of the protandric polychaete Ophryotrocha puerilis neurosecretory cells form prominent axon terminals. The terminals are arranged in two complexes. The perikarya of these presumably monopolar neurons are scattered in the anterior part of the cerebral perikaryal layer. In females the terminals store large amounts of neurosecretory material. It has been suggested earlier that neurosecretions of the terminals may play a role during sex reversal from females to males. Application of histamine caused the release of neurosecretory material from the respective terminals in females. However, this discharge was not followed by sex reversal. Application of reserpine had no influence on the terminals. Neither by in vivo observation nor by ultrastructural analysis any effect of reserpine on the terminal complexes could be observed. In isolated terminals filled with neurosecretory material from females, catecholamines could not be detected by HPLC. Also, polyclonal antibodies against dopamine did not stain the terminal complexes. Furthermore, the complexes did not develop any fluorescence after glyoxylic acid treatment. Therefore, the present results contradict the hypothesis that the neurosecretory material of the respective axon terminals is catecholaminergic and that it is involved in sex differentiation. The function of the secretory neurons studied here remains unclear.
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