Library

Notes: Summary Samples from renal and thyroid oncocytomas were studied with antibodies against intermediate filament proteins, nephron site specific antigens and nephron segment specific lectins to gather information on the immunohistological and carbohydrate histochemical features of these tumours. The results show a surprising failure of most antibodies and lectins used to react with the oncocytomas, although readily staining the surrounding normal tissue areas. No immunohistological evidence for derivation of oncocytomas from proximal tubular epithelial cells could thus be found. Instead,Triticum vulgaris (wheat germ agglutinin; WGA) andConcanavalin A (ConA) lectins were seen to stain the oncocytes specifically, suggesting that these lectins are useful to further characterize oncocytomas.

Notes: Summary The purpose of the work was to develop an in vitro model for the study of human kidney development. Human metanephric explants from foetuses 10–18 weeks of gestational age were cultured in serum-free Leibovitz L-15 medium without hormones. Under the current minimal conditions for growth, the system permitted to maintain the renal tissues in culture for periods up to 9 days, although no evident sign of morphological differentiation was observed. However, during the studied period the overall architecture of the explants was preserved as well as the ultrastructural features of cytoplasmic organelles. The incorporation of 3H-thymidine and 3H-leucine indicated that DNA and protein synthesis was maintained or increased. Glycoprotein synthesis evaluated by 3H-glucosamine incorporation and radioautography continued in mesangium as well as in glomerular and tubular basement membranes. Alkaline phosphatase, γ-glutamyltranspeptidase (brush border) and catalase (peroxisomes) activities remained histochemically active. The proposed organ culture system appears as a reliable and promising model that will provide basic data on the morphology and functional characteristics of the developing kidney. Since it is achieved in a completely controlled environment, it will permit to study the role of growth factors and hormones in proliferation and differentiation of the cell populations during development of the human foetal kidney.

Notes: Abstract This study presents the result of 12–21 years' follow-up in a group of children with neonatal urinary tract infection (onset within 1 month after birth) in whom early renal growth retardation was noted without concomitant classical renal scarring. In all cases the neonatal infection was diagnosed and treated within a few days of onset and the patients were closely supervised thereafter. Renal length, parenchymal thickness and area were measured at urography. At first follow-up (22 children, mean age 4.1 years) a significant reduction of renal parenchymal thickness was noted. Long-term follow-up (18 patients, mean age 17 years) demonstrated a normalization of renal size in the entire group, although less complete in the subgroup with reflux. There were two major findings in the present study. Firstly, renal growth retardation was seen after neonatal infection, both with and without reflux. Secondly, normalization of renal size in previously small kidneys was demonstrated, suggesting that growth retardation can be a reversible phenomenon. The tendency for such normalization was slightly more marked in children without reflux. Reduction of parenchymal thickness without calyceal deformity, therefore, does not necessarily mean irreversible damage, and differentiation between permanent scarring and temporary growth retardation can thus only be made at later follow-up, possibly not until after puberty. The demonstration of renal growth retardation in spite of early diagnosis and treatment emphasizes the great vulnerability of the kidney in the newborn.

Notes: Abstract We briefly review what appear to be the most important elements responsible for renal cell injury during and after oxygen deprivation. Recent studies in numerous laboratories have vastly improved our understanding of the changes in cell function that occur during ischemia and yet, the underlying mechanisms by which tubule damage and cell death occur remain elusive. We attempt to separate the effects that occur during ischemia or anoxia from those occurring during reperfusion (reoxygenation). These are not always separable, especially because it appears that ischemia initiates a series of complex events that may only become manifested during reperfusion. Ischemia-induced renal dysfunctions are clearly multifactorial events that will require major efforts to unravel.

Notes: Abstract This study investigated pharmacological manipulations of the cholinergic (ACh) and dopaminergic (DA) transmitter systems in monkeys with a long-term lead-induced cognitive deficit on delayed spatial alternation (DSA). Both ACh and DA have been found to be affected by developmental lead exposure and to be involved with performance on spatial learning and memory tasks. The lead-induced deficit in performance accuracy on DSA persisted throughout the 2 years of this experiment, which ended more than 8 years after the end of the postnatal lead exposure. Acute administration of agonists and antagonists of the ACh and DA systems did not elicit differential effects from the lead-exposed and control groups in terms of DSA per cent correct performance. The ACh antagonist, scopolamine, caused a dose-related decline in performance in both groups. Significant amelioration of the lead-induced DSA deficit was achieved by chronic treatment with the DA agonist, L-dopa. After withdrawal from L-dopa, the lead-related deficit reappeared. Improvement in performance of the lead-treated group was also seen after chronic amphetamine administration, but this effect was not significant. These data implicate DA mechanisms in the long-lasting cognitive effects of developmental lead exposure. The alleviation of the deficit with chronic administration of a DA precursor points to a possible line of treatment for the cognitive effects of developmental lead exposure.

Notes: Abstract In 1946 von Euler identified the major transmitter of sympathetic nerve fibers, norepinephrine (NE), and about a decade later Vogt (1954) provided the first evidence that NE may also serve as a neurotransmitter in the central nervous system (CNS). Since that time, a literal explosion in CNS neurotransmitter research has taken place involving histological, biochemical, physiological, pharmacological and clinical investigations, Yet, it is only now that we are beginning to understand the biological function of NE in brain, in particular because of recent advances regarding the physiology and regulation of NE neurons in locus coeruleus (LC), a bilateral pontine structure with a uniquely wide-spread terminal network reaching throughout the neuroaxis and in primates accounting for about 70% of all brain NE. Recently, the neurobiology of the LC noradrenergic network was extensively reviewed by Foote et al. (1983), and its implication in vigilance as well as global orientation of behavior towards imperative, environmental sensory stimuli was outlined. Yet, more recent information regarding the peripheral, autonomic regulation of LC neurons in brain provides fundamentally new biological aspects on behavior and mental function which seem to allow a more integrated view of the rôle of brain NE in the overall function of the individual than previously understood. The purpose of this review is to summarize these findings and, furthermore, to outline some putative implications for psychiatry and neuropsychopharmacology. In particular, the new data seem to allow a better understanding of how autonomic vulnerability or visceral dysfunction may precipitate or aggravate mental symptoms and disorder.

Notes: Abstract An attempt has been made to identify the subcellular localization of renal corticosteroid metabolism. Subcellular fractions were prepared by differential centrifugation, identified by marker enzymes and incubated under different conditions with corticosterone (B). The NADP+/NADPH dependent metabolism of B could be localized in the nuclear and microsomal fraction. The most prominent metabolite was 11-dehydro-B, which is formed by 11β-hydroxysteroid dehydrogenase (EC 1.1.1.146). Enzyme kinetic studies of this enzyme with B as substrate revealed apparentK m-values in the range of 10−7 M for both the nuclear and microsomal fraction.

Notes: Abstract Previous studies in rat kidneys have demonstrated that oscillations in the proximal intratubular pressure are a prominent feature of the tubulo-glomerular feedback mechanism (TGF) operating during free flow conditions. The purpose of the present study was to investigate whether a subpopulation of synchronized, interacting nephrons could be detected. In group A nephrons, i.e., nephrons with a high probability of having their afferent arterioles arising from the same interlobular artery, 29 out of 33 pairs of nephrons were found to show synchronized pressure oscillations. In group B nephrons, not expected to have this common origin of their afferent arterioles, only 1 out of 23 pairs was found to be synchronized. The standard deviation of the frequency differences was 0.063 cycles per minute in group A nephron pairs and 0.202 cycles per minute in group B pairs (p〈0.05), showing the greater homogeneity in frequency in group A. Furthermore, nephrons having synchronized pressure oscillations were found to interact with each other. Thus, perturbation of the proximal tubular pressure oscillations in one nephron by loop microperfusion affected the amplitude of the pressure oscillations in the non-perfused nephron; and reactivation of pressure oscillations in one nephron was followed by reactivation of oscillations in the non-perfused nephron. Thus, the present results show that there exists a well defined subpopulation of nephrons, in which the TGF-mediated proximal pressure oscillations are synchronized. This synchronization is a result of interaction between the different nephrons. It is suggested that the interaction is a consequence of “cross-talk” between the TGF signals from the different nephrons concerned, transmitted along the afferent arterioles, and probably also along a part of the interlobular artery.

Notes: Abstract A method for automated image capture and analysis of shrinking drop sequences is described. The procedure can be managed by a single operator and allows estimates of proximal tubule volume flux to be calculated during the experiment. Speed of analysis is increased considerably (mean time 1.5 min per sequence) compared with previous methods. The potential exists for measurement of droplets in tubules that follow a tortuous path in the horizontal plane and further increases in capture and analysis rate would allow estimation of volume flux under non-steady state conditions. Film processing is eliminated and the removal of operator errors, bias and fatigue associated with manual measurement, coupled with the greatly reduced time required for analysis of sequences makes the shrinking drop method much more reliable and attractive.

Notes: Abstract Drug transport in intestine, liver and kidney is similar, because in each case transport occurs across a barrier of epithelial cells. However, the physiological conditions differ in each organ: intestinal drug absorption is largely influenced by physicochemical conditions in the intestinal lumen; actual transport across the epithelial barrier occurs mainly by diffusion; carrier-mediated transport plays a subordinate role. In contrast, hepatic uptake is mediated by specific carriers, which transport a wide variety of drugs into the liver cell and then release them either into bile, or back into the portal blood. It is unclear how many carrier systems are involved, how they are organized in the liver cell membrane, and to what extent their substrate specificities overlap. Renal secretion and reabsorption of drugs is mediated by highly active carrier systems for cations and anions. Their cooperative action results in either active reabsorption or active secretion of drugs.

Notes: Abstract A single dose of clofibrate (400 mg/kg), given to rats, increased the incorporation of (3H)thymidine into liver DNA, in a period of 20–30 h after administration. However, (3H)thymidine incorporation into hepatic DNA of rats treated repeatedly was identical to that of control animals. After the administration of a single dose of clofibrate a small increase in (3H)thymidine incorporation also occurred in kidney DNA; repeated doses, however, resulted in a marked suppression of labeling.

Notes: Summary Attempts by early workers to induce liver tumours by the local implantation of carcinogens had by and large not been successful, so that the liver came to be viewed as being “resistant” to tumourigenesis by this means. A review of these early studies showed not only that fibrosarcomas could be easily induced by the local application of 3-methylcholanthrene (3-M.C.), but that there were also reasons why the apparently low susceptibility of the liver to the localised induction of hepatocellular tumours should not be accepted as established dogma. In an attempt to re-investigate this problem pellets made of chlolesterol (CHOL), anthracene (ANT), α-naphthylisothiocyanate (ANIT), 3-M.C. or 4-dimethylaminoazobenzene (DAB) were implanted into the livers of male litter-mate weanling rats. The evolution of the response was studied by histological examination of the implantation site at varying intervals. In each instance the liver responded with the formation of a firm, complete connective tissue capsule which, however, did not prevent the gradual degradation of the implants. No tumours or other significant changes were observed with the control implants of CHOL or ANT. ANIT, known to damage biliary ducts, elicited what appeared to be an intense serous exudation which was separated from the adjacent parenchyma by a shell-like deposition of calcium in the connective tissue capsule. No significant biliary changes were observed, however, and no tumours were produced. Attention should be drawn to this reproducible, regularly occurring, in vivo model of extra-osseous calcification. The 3-M.C. induced a high incidence of large solitary bosselated tumours associated with the carcinogenic pellet which was found embedded in the tumour mass. The architectural arrangement and bizzare cytological appearance of the tumours led to the currently widely used diagnosis of malignant fibrous histiocytoma (M. F. H.) rather than the fibrosarcoma or rhabdomyosarcoma of the early workers. Some tumours produced large numbers of implantation metastases in the peritoneal cavity, but no distant metastases were observed in this series. Of particular interest is the fact that it was not possible to determine the site of origin of these tumours despite histological sampling at intervals of the site of implantation of the pellets. In contrast to these pleomorphic, clearly mesenchymal tumours reliably produced by 3-M.C., the implantation of pellets of DAB produced fewer tumours which were classified as large, singly occurring hepatocellular carcinomas (H. C. C.). They did not differ in their histological appearance from the description of tumours induced by feeding the carcinogen. Metastases occurred again by implantation in the abdominal cavity. It appears that this is the first experiment in which the reproducible induction of localised H. C. C.s in the liver of the rat has been reported, the only previously reported instance of the induction of H. C. C. by this means having been described some considerable time ago in mice prone to the occurrence of “sponanteous” hepatocellular tumours. This report was disregarded for that reason. The rather low numbers of H. C. C.s induced by DAB implants in the experiments reported here, and their long latency period, suggests a need for further experimental manipulation. None of the usually described associated or precursor phenomena—oval cell proliferation, fasting-resistant glycogen accumulation, nodule formation—were seen either in the parenchyma immediately adjacent to the pellet or in more distant areas. The advantages of this model for further experimentation, and its position in the light of current views of hepatocarcinogenesis, are indicated, while some unsolved problems are underlined. The response of the kidney differed to some extent from that seen in the liver. The implanted pellets of DAB were encapsulated and grdually degraded, but no tumours were induced either in the kidney or, at a distance, in the liver. With pellets of 3-M. C., however, large hydronephrotic cysts with squamous cell metaplasia of the transitional pelvic epithelium progressing to invasive and locally metastasizing squamous cell carcinoma were produced in the kidney of some rats, whereas in others M. F. H.s were the outcome; one of the rats also presented with a mixed tumour. The similarity of the appearance of the 3-M. C.-induced tumours in liver and kidney suggests that the implantation of pellets of 3-M. C. is a reliable tool for the experimental study of M. F. H.s in internal organs.

Notes: Abstract Selective breeding techniques were used to alter allelic frequencies responsible for diazepam sensitivity and resistance. We used the rotarod test to determine the duration of diazepam-induced neurologic deficit in genetically heterogeneous mice. Males were more sensitive than females in the initial population. We then selectively bred for diazepam resistance and sensitivity. A significant difference between the lines was apparent in both sexes after two generations, and divergence has continued over seven generations. Brain benzodiazepine assays indicated that absorption and distribution of diazepam do not differ in the two lines. Differences in brain benzodiazepine concentrations at recovery from ataxia indicated that the two lines differ in central nervous system sensitivity. We found diazepam-induced rotarod impairment to be blocked in a dose-dependent manner by the specific benzodiazepine antagonist Ro 15-1788, indicating that this effect is mediated through BZ receptors. A dose-response curve obtained from generations 6 and 7 indicates a 9- to 14-fold difference in dose required to obtain similar effects in the two lines. These mice are expected to be useful experimental subjects in studies of benzodiazepine mechanisms.

Notes: Summary This report describes the immunolocalization of three monoclonal antibodies along the collecting duct system in rabbit kidney. The antibodies were raised against antigens derived from a membrane fraction of homogenized papillary tissue. Western Blot analysis demonstrated that each of the antibodies recognized a single band of about 190000 (PCD1), 210000 (PCD2) and 50000 (PCD3) daltons. In renal tissue, the antibodies bound specifically to the epithelia of the connecting tubule (CNT), the collecting duct (CD) and the papillary surface epithelium. Differences in the binding patterns of the antisera were limited to the cortex. pCD1 labeled only a few scattered cells in the CNT, and exhibited a heterogeneous binding along the cortical collecting duct (CCD). PCD2 and PCD3 binding patterns were similar. In the CNT, these antibodies bound to the intercalated cells (IC-cells) but not to the CNT-cells proper. In the CCD, both IC-cells and principal cells were labeled. The binding to the medullary collecting duct by all three antisera was identical. The ureter was labeled only by PCD2 and PCD3, and none of the antisera bound to the bladder epithelium. The antibody binding patterns provide information concerning tubular axial heterogeneity and embryogenetic aspects of the CNT and the CCD. These antibodies may be used as differentiation markers in studies of the developing kidney and of renal tissue culture systems.

Notes: Summary The cells of the kidney proximal segment of the migrating arctic lamprey, Lampetra japonica, contain particles of the same size, electron-density and intracellular location as particles identified by others as very low-density lipoproteins (VLDL) in the liver and intestine of teleost fishes and lampreys. These particles are synthesized within the cisternae of the smooth endoplasmic reticulum and elements of the Golgi complex. They are transferred to the lateral intercellular space and lamina propria by way of the Golgi vesicles and an intracellular channel system. Some particles are discharged into the lumina of the sinusoidal capillaries of the lamina propria. Although the physiological role of lipoprotein secretion in the renal proximal segment cells is unknown, the present observations provide morphological evidence that the kidney of the arctic lampreys synthesizes lipoproteins and releases them into the circulation at the time when they are undertaking their anadromous migration.

Notes: Summary Rats were given a lithium-containing diet (40 mmol/kg) to Study the effect of lithium on the structure of collecting ducts from the inner stripe of the outer medulla. The results show that there is a significant increase in the volume density of collecting ducts already after one week on this diet. The volume density of both intercalated and principal cells increases, whereas the volume density of mitochondria in the cytoplasm increases in the intercalated cells only. The increased volume of both principal and intercalated cells seems to be part of a general hyperplasia and hyperactivity of the collecting duct, which may in some way be related to the effects of lithium on vasopressinmediated water transport. The specific changes in the intercalated cells may be a consequence of the effects of lithium on distal nephron potassium and hydrogen ion transport in the distal nephron.

Notes: Summary This investigation provides histochemical evidence for lysosomal storage of sulfated glycosaminoglycans (GAGs) in the interstitial cells of the renal cortex and in macrophage-like cells of the renal medullary zones of rats chronically treated with the drug tilorone. This compound is known to interfere with lysosomal degradation of sulfated GAGs; therefore cells that develop GAG-storage can be assumed to be involved in the turnover of GAGs. In view of this consideration, the most remarkable and still unexplained finding was that the intrinsic interstitial cells in the papilla, which is known to be particularly rich in sulfated GAGs, did not show the cytological symptoms of lysosomal GAG-storage. The present findings may stimulate further studies focused on the cellular sites of turnover of the sulfated GAGs present in the renal medullary interstitium.

Notes: Summary The spatial organization of endoplasmic reticulum (ER) was examined in all segments of rat nephron. Tissues were fixed with glutaraldehyde, impregnated “en bloc” with osmium tetroxide, prepared for and examined by standard (80–100 kV) and high voltage (1 mEV) transmission electron microscopy. In all proximal tubule cells, ER forms a continuous and extensive network of canaliculi and abundant fenestrated saccules which surround mitochondria and cytoplasmic bodies; the cage-like structure of the fenestrated saccules was most evident around the spherical mitochondria of the S3 segment. In the cells of the distal straight and convoluted tubules, the network consists mostly of canaliculi with rare non-fenestrated saccules. The ER network of canaliculi is particularly rich in intercalated cells, in contrast with its rudimentary appearance in the adjacent principal cells of the collecting tubule. In fact, in these cells there are few isolated ER cisternae and they are rarely impregnated. The nuclear envelope is well impregnated in most cells throughout the various segments. Segmental variations in ER organization and its relative abundance are most likely related to the well, established functional heterogeneity of the nephron segments. Moreover, the extensive and unique organization among mitochondria, ER and the basolateral membrane suggests that these three organelles function as a unit which is related to active electrolyte transport. In addition, because of its transepithelial organization, ER may well constitute a transcellular pathway for molecules.

Notes: Summary The nephrons of the freshwater turtles Pseudemys scripta elegans and Mauremys caspica consist of renal corpuscle, neck segment, proximal tubule, intermediate segment, distal tubule and collecting duct. The renal corpuscle has large and scarce capillaries with clear and dark fenestrated endothelial cells containing some rod-shaped bodies, a thin filtration barrier and a well-developed mesangium, the cells of which show secretory, phagocytic and contractile features, and in M. caspica a cilium. The podocytes with a well-developed Golgi apparatus seem to be active secretory cells. Numerous dense bodies similar to lysosomes, but not previously reported in vertebrates, are conspicuous in podocytes of M. caspica. The proximal tubule displays a well-developed brush border with long and densely-packed microvilli and no basal labyrinth; mitochondria are scattered throughout the cytoplasm. Several dense and clear vesicles related to the prominent endocytotic apparatus can be seen. Wavy filament bundles, not previously reported in vertebrate kidneys, can be observed in proximal tubule cells of M. caspica. Three regions can be distinguished in the well-developed intermediate segment as well as in the distal tubule; the latter has a few short microvilli or a smooth luminal surface and lateral interdigitated processes. The collecting duct, the cells of which contain numerous mucous droplets, is similar in both sexes; there is no sexual segment.

Notes: Summary A monoclonal antibody against an antigen (PCD2) derived from the rabbit renal papilla recognized principal and intercalated cells of the collecting duct system in the adult rabbit kidney. Intercalated cells were heterogeneous in the connecting tubule and the cortical collecting duct, where immunoreactive and unreactive cells were shown to coexist. In the outer medullary collecting duct, all intercalated cells exhibited PCD2-immunoreactivity. Connecting tubule cells proper were not recognized by the antibody, whereas all principal cells of the collecting duct revealed specific immunoreactivity. The immunocytochemical heterogeneity of the intercalated cells is discussed in terms of a functional heterogeneity. Cytologically, the immunogold labeling of principal and intercalated cells was shown to occur along the plasmalemma, in the intracellular membrane structures and along the Golgi transport route. This pattern suggests that the antigenic determinant, which is ubiquitous in both principal and reactive intercalated cells, belongs to a membrane protein.

Notes: Summary The medullary pyramid of renculi in kidneys of ringed seals (Phoca hispida) is enclosed by a basket composed of ribbons of stromal tissue continuous with the wall of the calyx. Branched smooth muscle cells with well-developed Golgi complexes and rough endoplasmic reticulum and only an incomplete external lamina are the principal cells in sites near the origin of the ribbons from the calycal wall. Deeper in the corticomedullary junctional region, smooth muscle is progressively replaced with stellate or spindle-shaped cells exhibiting structural characteristics intermediate between those of fibroblasts and smooth muscle fibers. These myofibroblast-like cells contain arrays of parallel microfilaments 6–8 nm thick with associated focal densities and subplasmalemmal dense plaques, caveolae, elongate, often deeply wrinkled nuclei, and well-developed Golgi complexes and rough endoplasmic reticulum. Material resembling external lamina is associated with parts of the surfaces of most myofibroblast-like cells and intermediate junctions are present. Fibroblasts lacking arrays of parallel microfilaments are a minority at any level in the stromal ribbons. Interstitial cells in the vicinity of the corticomedullary junction show similar myofibroblast-like characteristics. The smooth muscle and myofibroblast-like cells presumably assist expression of urine from the papilla and calyx, and possibly participate as pacemakers for the urinary tract.

Notes: Abstract Two methods of glomerular filtration rate estimation have been evaluated, based on the intravenous administration of 99mTc-DTPA and the measurement of renal time activity curves by means of a computer linked gamma camera. A single 20 min plasma sample was also required. These methods were designed to minimize the component of error arising from decay statistics. One method was based on using a constant fraction of the cardiac activity in lieu of a perirenal region of interest for the background correction, the other was based on deconvolution by a constrained least squares technique. The first method, based on modifying the background correction, led to poor results (residual standard deviation 18.9 ml/min when compared with the plasma clearance method). The second method, based on constrained least squares deconvolution, worked as well as previously reported methods (residual standard deviation 14.5 ml/min) and appears suitable for clinical use.

Notes: Abstract It is now known that cardiac atria play an important role in blood pressure and volume regulation. Mechanical distension of the atria results in the release of a potent diuretic and natriuretic agent or agents termed the atrial natriuretic factor (ANF). Several structurally related forms of ANF exist in man and it is thought that these represent precursory forms of a single optimally active molecule and/or the presence of more than one form of active ANF. The chemical structure of ANF between different mammalian species is similar. ANF receptors have been identified in kidney, brain, vascular endothelial and smooth muscle cells, tracheal and bronchial smooth muscle and the adrenal glands of many mammalian species, including man. This would suggest that ANF influences blood pressure and volume homoeostasis by affecting any one of a number of biochemical or physiological mechanisms via different target tissues. ANF is now considered a potentially valuable therapeutic agent for the treatment of hypertension and congestive heart failure. Synthesis of potent receptor antagonists could be extremely useful in the treatment of various clinical situations which are produced or complicated by endogenously produced ANF, such as chronic orthostatic hypotension.

Notes: Summary Oxazepam has two opposing actions on behavior: a responsedecreasing or depressant action and a response-increasing or disinhibitory action. The course of the two actions in chronic dosing was determined in rats in a test in which punished and unpunished schedules of reinforcement were alternated. The depressant action (measured by a decrease in the rate of unpunished behavior) was observed to undergo tolerance after 3–4 doses, while the disinhibitory action (measured by an increase in the rate of punished behavior) failed to show tolerance and even increased throughout the chronic series. The selective tolerance of the depressant action is probably due to neuronal adaptation, but changes in metabolism also may be involved. The increase in the rate of punished behavior is attributed, at least in part, to a progressive unmasking of the disinhibitory action as tolerance to the depressive action develops.

Notes: Summary Theoretical models for sample selection by diagnosis using one, one-out-of-two, two, two-out-of-three, and three concurring opinions are illustrated. These models are then applied to two categories, schizophrenia and depressive disorders, for an actual sample of patients diagnosed by multiple observers. The conclusion is reached that sample selection by a single opinion is not sufficiently reliable for research studies. The choice of alternative methods of selection depends on the needs of a particular study. Samples chosen on the basis of two-out-of-three concurring opinions provide the least total error. When erroneous inclusions are particularly undesirable, higher degrees of consensus—e.g. agreeing pairs or triplets—are necessary.

Notes: Summary The results of the interaction of imipramine, physostigmine and d/l amphetamine in various intravenously administered dosages were studied on the cortical and subcortical electrical activity of cats. Imipramine was shown to cause a cortical synchronization, hippocampal desynchronization, rise in the threshold of electrocortical arousal and limbic convulsive activity which was potentiated by amphetamine. Imipramine increased the amount of physostigmine, but decreased the amount of amphetamine needed for cortical desynchronization. The possible significance of these findings for the neuropsychopharmacological action of imipramine is briefly discussed.

Notes: Summary CMA (p-chloro-N-methylamphetamine) lowers the cerebral 5-HT concentration in test animals but has virtually no effect on the catecholamine concentrations. Moreover, this compound was found to behave in depressive patients like an antidepressive drug, not like a central stimulant of the amphetmine type. The study described was conducted in order to establish whether CMA influences the overall metabolism of indoleamines in man. Such an influence was clearly demonstrable. Our findings are consistent with the hypothesis that CMA releases 5-HT from its depots. It has not been explained why a considerable proportion of the released 5-HT is excreted unchanged and why the increase in 5-HIAA excretion is so small: the overall activity of MAO was found not to be inhibited. No indications of abnormal 5-HT degradation were found at this time. Patients with vital depressions who improved on CMA medication showed a lower 5-HIAA excretion before treatment than did patients who were refractory to CMA treatment. This is consistent with earlier observations. The possible cause of this phenomenon is discussed. Among the various possibilities considered, an abnormal 5-HT metabolism is regarded as the most plausible. Pertinent investigations are being continued.

Notes: Summary Rats were kept on isotonic saline as drinking fluid for a period of 4 weeks. Glomerular filtration rate and proximal transit time of Lissamine green remained unchanged. The intrinsic reabsorptive capacity of the proximal tubular epithelium—measured by the “shrinking-droplet-method”—decreased significantly. As a result, fractional proximal reabsorption of sodium and water decreased from 51 to 38 per cent of the filtered load. Sodium excretion rose from 0.06 (controls) to 1,25 per cent of the filtered load. These changes were reversible by injection of d-aldosterone. The results were confirmed by measuring the TF/P ratio of inulin at the end of the proximal convolution. The results indicate that in rats chronically loaded with salt, the inhibition of fractional proximal reabsorption is due to a decreased secretion of mineralocorticoids, and is the cause of the higher sodium excretion. Apparently, the decreased proximal reabsorption is partly compensated in more distal parts of the nephron.

Notes: Summary To investigate the influence of plasma urea levels on the concentrating power of the kidney, the blood circulation of rats was connected to a dialyser; with dialysis against a urea-free cleaning solution, the urea level of the blood was lowered for three hours. During this, the urine flow of the animals was kept constant by a regulating arrangement. This control was achieved by removing fluid from the blood circulation in the case of too high urine flow and by putting in fluid in the case of too low urine flow. It was found that the concentration of urea in the urine had a direct linear relationship with the plasma urea level. The total electrolyte concentration fell only slightly compared to the plasma urea level falls. If the urea could have been completely removed, high concentrating of the electrolyte, to more than double isotonic, would still have been possible. The simplest way to interpret the results of this research is by the hypothesis that two different concentrating mechanisms are at work in the rat kidney, the one essentially concentrating electrolytes, and the other principally urea. The concentrating mechanism for electrolytes could be localised in the outer medulla, and for urea in the inner medulla.

Notes: Summary To investigate the mutual interdependence of urea and sodium in producing concentrated urine, solutions of different concentrations of sodium and urea were infused into the stomach of rats. After two days' infusion, the urine and tissues from the different zones of the kidneys were analysed for sodium, potassium and urea content. It was found that the efficiency of the rat kidney in producing urine with high sodium content was diminished if urea was infused together with the sodium chloride. Under all conditions sodium and urea concentrations increased from the cortex to the papilla of the kidneys. When pure sodium chloride solutions were infused, sodium and urea concentrations in the urine were nearly equal to the corresponding concentrations in the papilla. Infusion of solutions containing NaCl and urea caused production of urine with a higher sodium concentration and lower urea concentration than in the papilla. These results show that the high concentration of urea in the medulla of the kidney does not arise by passives diffusion of urea from the collecting ducts into the medullary tissue. The possibility that the concentration of urea in the inner medulla is produced by counter current diffusion of urea in the vasa recta is discussed. The concentration effect necessary for this counter current multiplication could be created by an osmotic flow of water from the vasa recta into loops of Henle, if the urine in the loops becomes hypertonic by taking up sodium chloride from the collecting ducts.

Notes: Summary Light- and electronmicroscopic evaluation of proximal convolutions of the rat kidney were made following injection with either mineral oil or castor oil. The following observations were made: 1. Injection of mineral oil does not result in a complete blockade of the tubular lumen and leads to morphological lesions of epithelial cells. 2. Injection of the more viscous castor oil, in contrast, leads to a dilatation of the tubular lumen and a compression of the brush border of proximal tubular cells. Thus a better blockade of longitudinal flow is achieved. A toxic effect of castor oil on the tubular epithelium could not be detected. Mechanical lesions, similar to those observed after mineral oil injection, were less frequent. They consisted in a penetration of oil into the cells, perhaps as consequence of the tubular dilatation. 3. In oil-injected proximal tubular segments the following cytologic phenomena were observed: a) a cessation of the normal pinocytosis, which is induced in the presence of tubular fluid, b) simultaneously the “large resorption vacuoles” disappear, indicating that they originate from confluence of pinocytotic vesicles, c) in dilated tubules protein resorption vacuoles are found to have ruptured towards the tubular lumen suggesting a high hydrostatic pressure within the vacuoles.

Notes: Summary Four weeks after clamping one renal artery, the perfusion pressure of the clamped kidney was normal whereas the mean arterial pressure had increased to 188 mm Hg. Urinary excretion of sodium and water of the clamped kidney had not changed while that of the untouched kidney increased considerably. Glomerular filtration rate (per g kidney), intratubular pressure, tubular diameter, transit time of Lissamine green and fractional sodium and water reabsorption of the proximal tubule were normal in both the clamped and the untouched kidney. In the untouched kidney, the transit time through Henle's loop was shortened and the sodium and water reabsorption in it was significantly reduced. These changes, although partly compensated by higher reabsorption in the distal tubule and collecting duct, are responsible for the increased sodium and water excretion of the untouched kidney. They are probably due to an increased medullary blood flow. In the clamped kidney, fractional sodium and water reabsorption of the whole nephron did not differ significantly from normal values. However, sodium reabsorption was lower in the distal tubule and higher in the collecting duct. Sodium transport in the distal tubule seemed to be impaired; throughout its length sodium TF/P ratio averaged 0.6. These results may explain the different urinary excretion found in human hypertension following unilateral renal artery stenosis (Howard-Test).

Notes: Summary Osmotic gradients of 150 mosmol/l between intratubular fluid and blood in the kidney of Rana ridibunda induce a fluid movement down the osmotic gradient which is stronger into the tubule than out of it. In either case Na+ is reabsorbed “dry”. If the active Na+-transport is blocked by Furosemid an osmotically downhill movement of isotonic fluid out of the tubule obtains. Reversal of the osmotic gradient induces a reversal of flow but the inward moving fluid is diluted. The tubular cells seem to inhibit the backwash of Na+. If the osmotic gradient induces an outflow of fluid out of the tubule no influx of Na+ occurs even in the presence of a concentration gradient for Na+ in the opposite direction. The Na+-concentration in the urine falls considerably below that of the perfusion fluid. This finding indicates Na+-reabsorption against a concentration gradient. Upon blocking of the Na+-transport by Furosemid water enters the tubules. The results of the experiments in which the Na+-transport was blocked by Furosemid prove that the tubules are impermeable for Na+ in the direction of secretion. The “dilution” of the tubular fluid as a result of an osmotic gradient which induces an influx of water isnot due to a reabsorption outbalancing “secretion”. If instead of the easily penetrating NaCl the slowly penetrating Na+-cyclohexanesulphamate is used, less Na+ is transported. However, in every other respect the results obtained are similar to those described above.

Notes: Summary Four weeks after constricting one renal artery distal tubular function was studied using the micropuncture technique. In the untouched kidney tubular fluid remained hypotonic up to 90% distal tubular length. We believe this to be the result of the high inflow of tubular fluid from Henle's loop. A reduced permeability for water could not be demonstrated. Except for the early part of the distal tubule there was a marked increase in sodium concentration in the clamped kidney as compared to values obtained in the untouched one and in kindneys of control animals. In addition no potassium secretion could be observed in the clamped kidney. The distal steadystate concentration for sodium was increased in the clamped kidney and normal in the untouched one. Reabsorptive half-time, measured by the split droplet technique, was prolonged in the clamped kidney and shortened in the untouched one. Early distal urea concentration was lower in the untouched kidney than in the clamped one and in control kidneys, while at the end of the distal tubule urea concentrations did not differ significantly. Both kidneys had normal glomerular filtration rates and proximal tubular transit times, whereas a shortened transit time through Henle's loop of the untouched kidney was found. The results indicate that active sodium transport in the distal tubule of the clamped kidney differs from that of the untouched one, possibly due to the different renin concentration in both kidneys.

Notes: Summary At different times after injection into the renal artery of131I-albumin, the distribution of the tracer in kidneys of the rabbit and the dog was illustrated by a series of autoradiograms. In order to determine the mean circulation time for albumin in the kidney, the radioactivity over the kidney was simultaneously registered by external monitoring. A short mean circulation time was measured for albumin (plasma) passing cortex, and a longer one for that fraction of the albumin that passed through medulla, papillae, and capsule. The investigation rendered probable that the mean circulation time for blood through the kidney determined by the external technique applies to blood passing cortex. The circulation times for blood through medulla, papillae, and capsule are not included in the calculation of the mean circulation time.

Notes: Summary Restitution of skeletal damage caused by strontium in growing Sprague Dawley rats is accompanied by an increase in the dialysable serum fractions of strontium and calcium. The results suggest that the renal manipulation of cations is influenced in bone lesions caused by foreign metals.

Notes: Summary This experiment investigated the possibility that chlordiazepoxide (CDP) has unique properties that account for state dependent learning, and prevention of conflict-induced behavior fixations. One group of rats were given a discrimination problem on a Lashley jumping stand, but on even days all responses were punished. Another group were treated the same way except than on even days all responses were rewarded. Each of these groups were subdivided, half of the Ss were given CDP on even days, the other half no drug. The results showed that punishment on even days for the response to be learned disrupted learning more than reward for responses that were to be avoided. CDP practically eliminated these disruptive effects and aided learning for the punishment group, but led to a slower rate of learning for the reward group. These findings confirmed the hypothesis that CDP attenuates the effects of negative incentives, and that this property accounts for the drug's cue value in discrimination learning and for its fixation prevention characteristics.

Notes: Summary Atropine or scopolamine improved conditioned avoidance behavior for most rats which performed poorly, in spite of extensive training, in a shuttle-box procedure. As previously reported, d-amphetamine also improved performance in many of these animals, but there was no particular relationship between a rat's responses to the cholinergic blocking agents and to d-amphetamine. The effect of any one of the 3 agents was, for the most part, reversible after the drug effect had dissipated. Physostigmine was quite potent in disrupting avoidance behavior in rats that performed well in the shuttle-box, even in animals that were overtrained. This impairment was antagonized by atropine or scopolamine, partly antagonized by d-amphetamine, and not antagonized by methyl atropine. Poor performers were found to be very sensitive to the disruptive effects of physostigmine, losing much of their escape behavior after relatively small doses. The results are interpreted as evidence for a central cholinergic system with inhibitory influences for modulating stimulus-response patterns. Under normal circumstances this inhibitory system probably functions in an integrated manner with the adrenergic mobilizing system for the central control of learned behavior. Centrally-active anticholinergic drugs of the muscarinic type appear to influence behavioral responses by inducing a response disinhibition.

Notes: Summary We have studied the interactions of reserpine and the monoamine oxidase inhibitors, nialamide and isocarboxazid, in rabbits. EEG recordings were made from animals with acute and chronically implanted electrodes. Gross behavioral observations were made in freely moving rabbits and brain amine concentrations of norepinephrine and serotonin were also determined. Emphasis was placed on observing drug effects over periods of time ranging up to 13 days. Rabbits given isocarboxazid or nialamide alone exhibit a phenomenon rarely seen in the control animals, namely partial activation, an EEG pattern in which slow waves of high amplitude are maintained in the cortex while fast activity appears in subcortical structures. This effect was also observed in rabbits treated with nialamide and reserpine in combination. By appropriate dosage schedules in which reserpine was administered to animals pretreated with monoamine oxidase inhibitors it was possible to maintain extended periods of frank arousal or EEG activation over a period of several days. The absolute concentrations of brain amines bore little or no relationship to the EEG effects observed, however, increased ratios of serotonin to norepinephrine were observed in conjunction with EEG activation and signs of behavioral excitement.

Notes: Summary The rotarod test of motor coordination in mice was modified by increasing the rotation speed every 30 sec until the animals fell off. This procedure yielded a stable, proficient level of performance within four brief trials; the approximately normal distribution of performance times provided an equivalent measure of either improvement or impairment caused by drugs and permitted the use of parametric statistical tests. A total of 240 mice were assigned to 20 different groups of 12 each, administered oral doses of placebo or different drugs, prior to trial 4. The use of a ratio score (performance time in trial 4 divided by the same animal's time in trial 3) provided a measure of drug-induced changes, controlling for individual differences among animals in over-all level of performance. Two phenothiazines (chlorpromazine and perphenazine) impaired performance at low doses, with a progressively greater decrement at increasing doses (4, 8, 16 mg/kg); 2 barbiturates (pentobarbital and amobarbital) showed an all-or-none effect, with no significant decrement at the lower doses (20 and 40 mg/kg) but almost complete incapacitation at the highest dose of 80 mg/kg. Performance superior to the placebo condition was found with the 2 lower doses of pentobarbital and with 3 d-amphet-amine doses (4, 8, 16 mg/kg). An analysis of individual differences gave evidence that the animals which were inferior in prior performance were more susceptible to both improvement and impairment of performance under the influence of drugs.

Notes: Summary The offspring of female mice treated with 2.5 mg/kg DDT during the second or third trimester of pregnancy showed a delayed acquisition of the conditioned avoidance response. If DDT was given in the first trimester, or if 3 mg/kg of Parathion was given during the pregnancy, no effect on the conditioned avoidance response of the off-spring was seen.

Notes: Summary In rats micro-injections in corpus striatum of quaternary chlorpromazine and related drugs give rise to highly characteristic neuroleptic effects: antagonism of amphetamine-induced stereotyped behaviour and development of catalepsy. There is no effect of injections in hippocampus or septum. Dopaminergic mechanisms in corpus striatum seem to play a central role in neuroleptic action.

Notes: Summary 1. The effects of amphetamine on the sleep-wakefulness cycle were studied in 48 kittens during the growth period of 1–28 days of age. Recordings of EEG, EMG of the posterior neck muscles and respiratory rhythms were made as well as observations of gross behavior. 2. Percent time of wakefulness was increased with amphetamine (0.3 mg/kg). This effect became more marked with age, i.e., being more significant after 16 to 18 days of age (P〈0.005) than before (P〈0.05). These increases in the intensity of the effect during growth may be related to the postnatal development of the structures responsible for wakefulness. 3. Percent time of “activated” sleep was diminished significantly at all ages including the newborns (P〈0.005). This lack of the influence of age suggests that the structures responsible for “activated” sleep are well developed at birth. 4. The increase of percent time of slow wave sleep was observed. This increase is regarded as a “passive” manifestation of time left available after a severe diminution of “activated” sleep time and relatively slight increase of wakefulness time during amphetamine medication.

Notes: Summary Rats of 3 strains were observed at regular intervals and their activity was recorded using four categories of behaviour-rearing, moving, grooming and immobile. Strain differences in control activity were found. Nicotine and physostigmine reduced the activity of the more active rats and increased that of the less active animals. Rearing behaviour was particularly susceptible to depression by both drugs. The similarity of effect of the two drugs supports the hypothesis that one of the actions of nicotine in the brain is the release of acetylcholine.

Notes: Summary Measures of skilled motor performances, both of a task-oriented (tests of eye-hand coordination) and incidental (control of facial and ocular muscles) nature were recorded for a sample of 20 healthy young adults before and after single administrations of perphenazine, opipramol, imipramine and placebo at doselevels commonly supposed to produce mood or behavioral effects. It was anticipated that such performances would be sensitive even to slight changes in the subjects' physiological and psychological state; the aim was to test the power of tests of subtle skills in providing indices of slight to moderate behavioral effects. The performance measures remained surprisingly little affected by all drugs, despite their sensitivity to drug-independent improvement in performance throughout the experimental day, and despite evidences of drug-related effects, especially for imipramine and opipramol, in simple objective physiological measures, and for imipramine alone in subjective measures taken concurrently. There may be a class of skilled sensory-motor acts, particularly those related to well-learned daily activities, which, rather than being vulnerable to adverse effect, remain efficient even in the presence of signs of disturbance of bodily function.

Notes: Summary Using climbing rope and bar-pressing behavior methods, rats were rendered tolerant to Δ 9-THC, cannabis extract, mescaline and LSD-25. Cross-tolerance experiments showed that rats refractory to Δ 9-THC and cannabis extract were still sensitive to mescaline and LSD-25, and vice-versa. These results suggest that, in spite of the similarity of the clinical symptoms produced in man by the 3 drugs, Δ 9-THC may have its psychotomimetic effects produced by different mechanisms from those of LSD-25 and mescaline.

Notes: Summary In stop flow experiments during infusion of 1,2-3H-18-aldosterone glucuronide there was no proximal tubular secretion of this metabolite in contrary to experiments during infusion of 1,2-3H-aldosterone. The discrepancy must be explained by formation of 1,2-3H-aldosterone glucuronide out of3H-aldosterone in proximal tubular cells, while the 1,2-3H-18-aldosterone glucuronide in the plasma of the renal artery blood is only filtered in the glomerulum.

Notes: Summary After an intravenous injection of 1,2-3H-aldosterone the ratios of 1,23H-tetrahydroaldosterone glucuronide to 1,2-3H-aldosterone glucuronide in the blood plasma of three normal and two nephrectomized persons were compared. In the three normal persons the ratios were of the same magnitude as in the two nephrectomized persons. On the basis of glomerular filtration of tetrahydroaldosterone glucuronide, as previously demonstrated by renal clearance studies in man [13, 15] and stop flow experiments in dogs [6], we made the following deductions: 1. 18aldosterone glucuronide in blood plasma is only filtered in the glomerulum. 2. The renal clearance of 18-aldosterone glucuronide, as investigated during aldosterone infusion in man [13, 15], therefore must be explained by glomerular filtration of only one fifth and by formation of four fifth of 18-aldosterone glucuronide by proximal [15] tubular cells. 3. This means that about 80% of 18-aldosterone glucuronide excreted in urine is formed by the kidneys.

Notes: Summary The arteriovenous differences of total and individual free fatty acids (FFA) have been determined in the myocardium, in the electrically stimulated skeletal muscle (gastrocnemius) and in the kidney of closed chest anesthetized dogs. The concentration of total FFA was determined by titration, their composition was determined by gaschromatography. When the arterial concentration of total FFA was increased by norepinephrine infusions the percentage of olic and of palmitic acid increased and the percentage of stearic and linolic acid decreased. When the concentration of total FFA was decreased by glucose infusions the percentage of olic and palmitolic acid decreased and the percentage of palmitic and stearic acid increased. Under control conditions in the myocardium 44% of the total FFA extraction were related to olic and 24% to palmitic acid and in the skeletal muscle 43% were related to olic and 29% to palmitic acid. During norepinephrine induced increases of arterial FFA concentration myocardium and skeletal muscle extracted only olic, palmitic and palmitolic acid. At the same time in the myocardium stearic and linolic acid and in the skeletal muscle stearic acid was produced. In the kidney under control conditions 40% of the total FFA extraction were related to palmitic and 40% to olic acid. Since the extraction of olic, palmitic and palmitolic acid was higher than the extraction of other FFA at comparable arterial concentrations, it is concluded, that these three FFA are used in preference to other FFA as energy sources. Norepinephrine causes, probably via activation of lipase, in the myocardium and in the working skeletal muscle a breakdown of stored or in plasma transported triglycerides.