[Dynamics of spinal cord changes under the influence of asphyxia sustained in utero].

Changes of the neurons and nerve fibers of the spinal cord at the C4-C5 level were studied in rats exposed to intra-uterine asphyxia and sacrificed at various periods. A gradual increase of the neuronal changes was observed the first 10 days. Although the majority of the neurons were normalized by the 1st-2nd month microscopy showed changed nerve cells in every 100 fields of vision. The abnormal neurons were mainly confined to the areas of colateral circulation. Some of the myelin fibers were vacuolated, and their sheaths--disintegrated.


[Effect of ethanol on the ultrastructure of pancreatic acini].

Histological and ultrastructural changes of the acinar parenchyma of the pancreas under the influence of ethanol were studied on Wistar rats. Lipid inclusions in the cytoplasm of the acinar cells and mitochondrial changes were referred to the functional ones. The opinion of some investigators on the cytotoxic influence of alcohol on the pancreas was not confirmed by the author. The results of this investigation indicate that the leading role of alcohol in the etiology of acute pancreatitis is doubtful.


[Selective dopaminergic regulation of stereotyped forms of aggressive and defensive behavior].

In rabbits the dopaminergic agonists (L-DOPA, apomorphine) induce and control the stereotypic forms of behaviour (thumping, licking and biting) selectively. These forms of behaviour can be used as test-responses for quantitative studies of the neuropsychotropic substances acting on the central dopaminergic synapses, and also for screening the antiparkinsonic agents.


[Immunodepressant action of cyclophosphamide in different strains of mice].

A study was made of the immunodepressive effect of cyclophosphamide (CP) on mice of 3 strains (BALB/c,  CBA, and DBA/2) immunized with sheep red blood cells (SRBC). With the optimal immunizing dose of the antigen (5 X 10(8) SRBC) the most pronounced immunodepression was noted in DBA/2 mice, and with the high dose (6.2 X 10(9))--in DBA/2 and CBA mice. The CP action proved to depend on the dose of the antigen administered; in BALB/c mice a reduction in the number of the antibody-forming cells was the same with both SRBC doses, in DBA/2 mice an increase of the antigen dose led to reduction of immunode pression, and in CBA mice -- to its enhancement (with sufficiently high CP doses). Determination of the rate of oxidative CP hydroxylation by the liver microsomes of mice showed it to be comparatively low in DBA/2 and CBA mice, and much greater in BALB/c mice. It is supposed that the detected differences in the immunodepressive action of CP could be connected with different sensitivity of the target cells and (or) with the peculiarities of its metabolism in mice belonging to different strains.


[Mechanism of tolerance to VI-antigen of Salmonella typhi induced by cyclophosphane].

High dose Vi-antigen treatment and injection of cyclophosphamide 46 to 48 hours later induced in mice a state of immunological unresponsiveness remaining stable in adoptive transfer. Only low amounts of the antigen were revealed in the blood and spleen of tolerant animals 2 to 3 weeks after the tolerogenic treatment. No T-suppressors were found in the spleen of tolerant mice--the cells of tolerant mice failed to suppress the immune response of normal lymphocytes when transferred together to the irradiated recipients, or to induce tolerance in normal mice. Normal spleen cells restored partially the immune responsiveness in tolerant animals. The results obtained suggest that cyclophosphamide tolerance was due to deletion or the long-term inactivation of the immunocompetent cells.


Equilibrium between uptake and elimination of dieldrin by channel catfish, Ictalurus punctatus.

Channel catfish, Ictalurus punctatus, measuring 150 (+/- 5)mm were exposed continuously to dieldrin for 70 days to determine if equilibrium between dieldrin uptake and elimination was reached by muscle tissue. Fish exposed to 13 parts per trillion (pptr) dieldrin reached equilibrium in 56 days, fish exposed to 27 pptr dieldrin were near equilibrium after 56 days, and fish exposed to 49 pptr dieldrin did not reach a balance between uptake and elimination after 70 days of exposure. Evidence of this study showed that balance between uptake and elimination was reached in less time at lower exposure levels than at higher levels.


A Danish twin study of manic-depressive disorders.

The existence of a nation-wide twin register and central psychiatric register has made possible a catamnestic investigation of an unselected and representative sample of twins with manic-depressive disorders. From a total population of 11,288 same-sexed twin pairs born 1870-1920 in Denmark 126 probands from 110 pairs were ascertained. Among the co-twins of 69 monozygotic probands there were found 46 with manic-depressive disorders, and a further 14 had presented other psychoses or marked affective personality disorders or had committed suicide, yielding a proband rate of strict concordance, C1 = 0-67 and of broad, partial concordance, C2 = 0-87. The corresponding direct pairwise concordance rates were 32/55 = 0-58 and 46/55 = 0-84 respectively. For the dizygotic twins the proband concordance rate of C1 was 11/54 = 0-20 and of C2 20/54 = 0-37, and the direct pairwise rates were 9/52 = 0-17 and 18/52 = 0-35 respectively. The differences between the pairwise rates for the monozygotic and dizgotic twins are significant (P less than 0-001 at X2 analysis). This finding is in accordance with previous twin studies of manic-depressive disorders and confirms the evidence of a strong genetic factor. The concordance with respect to unipolar and bipolar forms was not in contradiction to recent evidence of a genetic difference between the bipolar and unipolar form, the latter probably related to the female sex.


Cytogenetic investigation in twins with manic-depressive disorders (22 monozygotic and 27 dizygotic twin pairs).

Chromosome examination was made in 22 monozygotic and 27 dizygotic twin pairs of whom one or both of each pair suffered or had suffered from manic-depressive disorder. We found a significantly higher frequency of chromosome variations among dizygotic twin pairs than was expected from population studies, but not in monozygotic pairs. There was no association between the chromosome variations and manic-depressive disorders. We found no greater intra-pair correlation in monozygotic twins compared with dizygotic twins as regards hypodiploidy, hyperdiploidy and unstable chromosome aberrations, which indicates that the aetiology of such aberrations is mainly of exogenic nature.


Sleep in the rat during food deprivation and subsequent restitution of food.

Continuous telemetric EEG recordings served to determine the vigilance states of the rat during 2 control days, 80 h of food deprivation and 64 h following restitution of food. The recordings were supplemented by measurements of food intake, water intake and motor activity. The following 3 sleep parameters were not significantly changed by food deprivation: the daily amount of the vigilance states, the light-dark distribution of sleep and waking, and the 10 min paradoxical sleep (PS) cycle. During food deprivation, PS was depressed in the dark phase of the diurnal cycle and increased in the light phase. The sleep parameter that was most affected by food deprivation was the duration of sleep episodes. Episodes of slow-wave sleep (SWS) and PS were shortened only in the dark phase of the deprivation days, whereas total sleep episodes were progressively decreased in both diurnal phases. After restitution of food, the episodes of SWS and total sleep were immediately lengthened and tended to exceed the control level. The duration of feeding episodes and meal size were significantly increased in comparison to pre-deprivation values, whereas feeding frequency was decreased. Long episodes of continuous motor activity occurred during the dark phase of the refeeding period, while a fragmented activity pattern was typical for the deprivation nights. It is proposed that the adjustment of the length of behavioral episodes may constitute an important adaptive mechanism for the rat.


Progesterone metabolism in the pineal, brain stem, thalamus and corpus callosum of the female rat.

Specific brain regions, namely, thalamus, tectum, tegmentum, cerebellum, medulla and pineal, from five proestrous rats were incubated for 30 min with [3H]progesterone. After reverse isotopic dilution analysis, the following metabolites were identified in all incubations by purification to constant specific activity, derivative formation and/or gas liquid chromatography trapping: [3H]5alpha-pregnane-3, 20-dione (10-20% of the starting substrate except pineal -- 0.7%), [3H]3alpha-hydroxy-5alpha-pregnan-20-one (1.6-3.8% except for pineal -- 0.5%) and [3H]20alpha-hydroxy-4-pregnen-3-one (0.05-0.11%). Preliminary results from the corpus collosum incubation indicated the presence of the same metabolites. Although some apparent constant specific activities were obtained for 20alpha-hydroxy-5alpha-pregnan-3-one and 5beta-pregnane-3, 20-dione, the low levels of 3H associated with these steroids did not permit a definitive identification. The results indicate the presence of at least delta1-steroid 5alpha-reductase, 3alpha-hydroxysteroid dehydrogenase and 20alpha-hydroxysteroid dehydrogenase activities with progesterone as substrate in the brain regions examined.


Amphetamine-haloperidol interactions in rat striatum: failure to correlate behavioral effects with dopaminergic and cholinergic dynamics.

Previous reports have suggested that the hyperactivity and stereotypy produced by amphetamine (AMP) and the catalepsy produced by haloperidol (HAL) are mediated by striatal dopaminergic mechanisms. In the present study, we have measured the behavioral effects of AMP and HAL, and their effects on striatal dopaminergic function, using both an index of pre-synaptic activity (synaptosomal dopamine (DA) synthesis) and a parameter which we suggest will reflect post-synaptic dopaminergic function (sodium-dependent, high affinity choline uptake). Administration of 2 mg/kg AMP produces hyperactivity and causes a decrease in DA biosynthesis, both of which are blocked by 0.75 mg/kg HAL. AMP (5 mg/kg) produces stereotypy, further decreases DA biosynthesis and causes a decrease in choline uptake, consistent with stimulation of DA receptors. However, while pretreatment with 3 mg/kg HAL completely blocked the stereotypy induced by 5 mg/kg AMP it failed to reverse the effects of this dose on either DA biosynthesis or choline uptake. These data suggest that either 5 mg/kg AMP affects straital dopaminergic and cholinergic parameters by a mechanism independent of HAL sensitive receptors, or the stereotypy produced by high doses of AMP are not related to striatal dopaminergic and cholinergic function.


Interactions of Phytophthora cinnamomi and Trichoderma spp. in relation to propagule production in soil cultures at 26 degrees C1.

Effects of Trichoderma harzianum and T. polysporum on chlamydospore production by two isolates of Phytophthora cinamomi were determined over a 21-day period in nonsterile, sterilized, and sterilized amended soil. Trichoderma was either coinoculated with P. cinnamomi or added to the cultures on day 3 of the incubation period. In nonsterile soil, conversion of mycelial fragments in the P. cinnamomi inoculum to chlamydospores resulted in an initial slight increase in chlamydospore numbers. In cultures where either of the Trichoderma isolates was added on day 3, a significiant reduction in chlamydospore numbers was observed on day 4; however, neither of the Trichoderma isolates at either inoculation time significantly affected clamydospore numbers by day 21. Results from studies with sterilized amended soil indicated that a reduction in P. cinnamomi chlamydospore numbers by either of the Trichoderma isolates was dependent upon availability of simple carbohydrates. In sterilized nonamended soil, the number of chlamydospores was increased in cultures containing either Trichoderma isolate; in sterilized amended soil chlamydospore numbers were initially reduced, followed by a general increase as the nutrients were depleted. Neither of the Trichoderma isolates exhibited significant antagonistic qualities toward P. cinnamomi.


Cellular bodies in developing chlemydospores of Thielaviopsis basicola1,2.

The development of chlamydospores of Thielaviopsis basicola from hyphal cells involves the thickening and pigmentation of the cell wall. Electron microscope studies showed that membrane-bound cellular bodies appearing in the cytoplasm of differentiating cells developed from dilated cisternae in the endoplasmic reticulum. Within the membrane-bound bodies, vesicles of up to 0.2-micronm diameter were observed which contained electron-dense particles. Vesicles resembling those seen in the cellular body were also present in the cytoplasm close to the plasmalemma. In newly formed chlamydospore cells where wall thickening was complete, the cellular bodies showed loss of internal organization, and most of the vesicles disappeared, leaving a structure resembling a vacuole. The cellular bodies were not present in undifferentiated hyphae or in mature chlamydospores.


Studies on some characteristics of hydrogen production by cell-free extracts of rumen anaerobic bacteria.

Hydrogen production was studied in the following rumen anaerobes: Bacteroides clostridiiformis, Butyrivibrio fibrisolvens, Enbacterium limosum, Fusobacterium necrophorum, Megasphaera elsdenii, Ruminococcus albus, and Ruminococcus flavefaciens. Clostridium pasteurianum and Escherichia coli were included for comparative purposes. Hydrogen production from dithionite, dithionite-reduced methyl viologen, pyruvate, and formate was determined. All species tested produced hydrogen from dithionite-reduce methyl viologen, but only C. pasteurianum, B. clostridiiformis, E. limosum, and M. elsdenii produced hydrogen from dithionite. All species except E. coli produced hydrogen from pyruvate, but activity was low or absent in extracts of E. limosum, F. necrophorum, R. albus, and R. flavefaciens unless methyl viologen was added. Hydrogen was produced from formate only by E. coli, B. clostridiiformis, E. limosum, F. necrophorum, and R. flavefaciens. Extracts were subjected to ultracentrifugation in an effort to determine the solubility of hydrogenase. The hydrogenase of all species except E. coli appeared to be soluble, although variable amounts of hydrogenase activity were detected in the pellet. Treatment of extracts of the rumen microbial species with DEAE-cellulose resulted in loss ofhydrogen production from pyruvate. Activity was restored by the addition of methyl viologen. It is concluded that hydrogen production in these rumen microorganisms is similar to that in the saccharolytic clostridia.


Angiosarcoma of venous origin arising in radial nerve.

A case of angiosarcoma felt to arise from a vein within the radial nerve is presented. Endovascular angiosarcoma arising in a vein is unusual and only one case of angiosarcoma in a peripheral nerve has been previously reported. Microsurgery followed by radiation has led to a 36-month tumor free survival.


Laser surgery of a digital angiosarcoma: report of a case and six-year follow-up study.

A single digital angiosarcoma was excised by a carbon dioxide laser with small portions of this tumor excised by the scalpel and the high-frequency electrosurgical knife. The patient has been followed for 6 years without any local recurrence or additional lesions. There is a brief summary of investigative studies with lasers in benign and malignant vascular tumors.


Thymomas.

Forty-six thymomas, defined as neoplasms of thymic epithelium, were assessed in respect to histologic type, clinical manifestations and end results. Twenty-three were lymphoepithelial, 12 epithelial, and 11 spindled cell tumors. The sex distribution for all thymomas was approximately equal for males and females but the lymphoepithelial tumors were more common in females and the epithelial tumors more common in males. There was no distribution difference in sex for the spindled cell thymomas which occurred much later in life than the other types. Twenty-five percent of patients were asymptomatic, the tumors being discovered on roentgenograms done on a routine basis or for an unrelated purpose. The most common presenting symptoms were related to myasthenia gravis, and symptoms due to pressure on mediastinal structures were next in frequency. Other endocrine abnormalities were present in 20% of patients. Two male patients also had carcinomas of the thyroid gland. Twenty thymomas were infiltrative tumors having invaded the adjacent mediastinum. There were no examples of extrathoracic spread. Twelve of 16 patients with myasthenia gravis had lymphoepithelial thymomas and four epithelial thymomas. There were no cases of myasthenia gravis in patients with spindled cell thymomas. The crude five-year survival rate was 65%. Only 17% of patients died as a result of recurrent and infiltrative thymomas; the other 18% of patients died from a variety of causes not directly related to the thymomas. The epithelial type of thymoma tended to be more extensive and to pursue a more aggressive course but histologic classification was of little value in predicting final outcome. Poor prognosis is more closely associated with tumors having an infiltrative character. Classification is of importance, however, in alerting the clinician to certain associated diseases.


Action of the vinca alkaloids vincristine, vinblastine, and desacetyl vinblastine amide on axonal fibrillar organelles in vitro.

Axonal ultrastructural changes induced by three Vinca alkaloids, vincristine, vinblastine, and desacetyl vinblastine amide, were studied in vitro at concentrations of 0.01, 0.05, and 0.1 mM in the cat vagus nerve. Disruption of microtubules, appearance of paracrystalline structures, and increase in neurofilaments were induced by all three agents at 0.1 mM. A new type of paracrystal with an electron-dense central core in each subunit was also observed with each drug. Whereas all three compounds affected unmyelinated fibers (vinblastine more so than the other two), only vinblastine significantly damaged the myelinated fibers. The greater effectiveness of vinblastine in causing these in vitro ultrastructural changes contrasts strikingly with the clinical in vivo situation in which vincristine is the most neurotoxic. This suggests that clinical neurotoxicity is associated with additional factors aside from the direct interaction of the Vinca alkaloids with microtubules or tubulin.


Induction of hyperplasia and its suppression by hydrocortisone in organ-cultured rat urinary bladder.

Urinary bladders from Fischer rats organ cultured in a chemically defined medium, Ham's F12, underwent transitional cell hyperplasia which persisted for the duration of the culture period (10 days). The hyperplastic response was initiated at 2 days of culture in basal epithelial cells as evidenced by [3H]thymidine autoradiography. After 2 days, cells classified as intermediate cells were observed replicating DNA in increasing numbers, whereas the frequency of basal cells replicating DNA decreased. The peak periods of basal and intermediate cell DNA replication were at 2 and 5 days, respectively. The total increase in the number of cells in the epithelium during a 10-day culture period was approximately 2.6-fold. The appearance of DNA-replicating cells before the appearance of mitotic figures indicated that the cells of the transitional epithelium are primarily G1 cells, The hyperplastic response in the transitional epithelium was significantly inhibited by hydrocortisone. Epithelia cultured in the presence of hydrocortisone also displayed less atypia than those epithelia cultured in its absence. Hydrocortisone concentrations of 2.1 and 21 micronM inhibited hyperplasia by 75 and 84%, respectively, Cells replicating DNA at 2 days of culture were considerably less sensitive to the hydrocortisone inhibition of [3H]thymidine incorporation into DNA than cells replicating DNA at 4 days of culture. The possibility is discussed that basal and intermediate cells may have different sensitivities to hydrocortisone.


Quantitation with an automated image analyzer of nuclearcytoplasmic changes induced by hydrocortisone in bladder epithelium.

Thirty-eight urinary bladders from Fischer rats were organ cultured for 7 days in Ham's Medium F12 with varying concentrations of hydrocortisone added. Measurements of cell numbers and relative areas of nuclei and cytoplasm (N/C ratios) were made with an automated image analyzer on 1.5-micronm-thick plastic sections of the tissues. Epithelia cultured in the absence of hydrocortisone became hyperplastic and dysplastic and contained approximately 2.5 times as many cells per unit length as uncultured controls. N/C ratios were increased by over 50%, caused mainly by a decrease in the cytoplasmic area. Concentrations of hydrocortisone below 10(-7) M had no significant effect on the hyperplasia or N/C ratios. Increasing concentrations above 10(-7) M resulted in progressive decreases of both the number of cells per unit length and the N/C ratios. Epithelia cultured in 2 X 10(-5) M hydrocortisone were not significantly different from uncultured controls. N/C ratios from 85 fields with a wide range of values were quanititated both with the image analyzer and a manual point-counting technique. Linear regression analysis revealed a close linear relationship and a correlation coefficient of 0.922. The analyzer provided an efficient, accurate tool for quantitating morphological observations.


Effects of metabolism on the binding of polycyclic hydrocarbons to nuclear subfractions of cultured AKR mouse embryo cells.

A high-affinity localization of [2H]methylcholanthrene and/or its metabolites to a specific nuclear fraction (Fraction l) between 4 and 72 hr of exposure is described. Other carcinogenic hydrocarbons, such as benzo(a)pyrene and dibenz(a,h)anthracene, demonstrate a similar markded localization in Fraction l after 24 hr of incubation. The weak carcinogen dibenz(a,c)anthracene, as well as steroid hormones, sho little localization in this fraction. Two  types of binding are measured:and organic solvent: extractable (non-covalent) binding and a nonextractable (covalent) binding. Maximal levels of the combined extractable and nonextractable binding per mass DNA are found at 24 hr of exposure, while at 48 and 72 hr of exposure the binding is reduced. The highest level of the nonextractable binding per mass DNA is also observed at the 24-hr exposure period. However, as the period of exposure increases, the proportion of the total nuclear-bound radioactivity representing the non-extractable type increases. Analysis by high-pressure liquid chromatography of the extractable radioactivity from the fractions indicates that the longer the period of exposure, the greater the extent of metabolism of 3-methylchol-anthrene. When 7,8-benzoflavone (a flavanoid hydroxylase inhibitor) is included in the incubations, practically all metabolic alterations of the parent compound are prevented. In addition, the time-dependent increase in nonextractable radioactivity from all nuclear subfractions is prevented. A metabolic-dependent "covalent" binding of carcinogenic polycyclic aromatic hydrocarbons to the barious nuclear subfractions of chromatin is suggested. This covalent binding is markedly localized in a specific fraction of the chromatin containing rapidly labeled nascent RNA.


Cell surface glycosaminoglycans of cell line MDCK derived from canine kidney.

Morphological observations and biochemical analysis were made on glycosaminoglycans produced by MDCK cells of dog kidney origin growing on a glass surface as a mosaic of epithelium with many multicellular hemishperical vesicles. MDCK cells synthesized glycosaminoglycans, which consisted mainly of heparan sulfate and hyaluronic acid. The majority of the substances were contained in a cell-surface component removable with ethylenediaminetetraacetic acid-trypsin. In the radioautograph of tissue sections, high radioactivity of 35SO4 was observed on the medium-bathed cell surface, where Alcian blue-strained material could be observed. Ultrastructurally, the surface of microvillous processes which were abundant on the cell surface in contact with the medium was stained with ruthenium red. A small amount of chondroitin 4- and 6-sulfates were also synthesized. After 24 hr, the majority of chondrotin [35S] sulfates newly formed were secreted into the cultured medium, whereas haparan [35S] sulfate was released much less, remaining as a cellular component. The biological roles of glyconsaminoglycans produced by epithelial cells are discussed.


Cell density dependence of focus formation in the C3H/10T1/2 transformation assay.

Some of the dynamics of neoplastic transformation in vitro have been studied with the use of benzo(a)pyrene as the carcinogen in the C3H/10T1/2 morphological transformation assay. Experiments that involved the dsipersion of cells into new culture dishes at various times after carcinogen treatment have shown that no change in the fraction of potentially transformed cells occurs while cultures grow to form a confluent monolayer, that little or no change in the fraction of potentially transformed cells occurs for approximately 3 weeks after confluence is attained, and that this fraction increases rapidly some 7 weeks after BP treatment. When confluent benzo(a)pyrene-treated cultures are dispersed in new culture dishes prior to the onset of growth toward focus formation, the formation of transformed foci is suppresssed at high cell densities of seeding. This phenomenon is independent of the total number of divisions undergone by cells after treatment. We suggest that phenotypic expression of morphological transformation is dependent on colony interactions in the C3H/10T1/2 system, which we do not yet understand, but which are independent of time posttreatment either in cell generations or absolute time.


Relationship of nutritional factors to in vitro tumor cell growth and cytotoxicity produced by cytosine arabinoside.

The in vitro relationship between nutritional factors, proliferative status of tumor cells, and the cytotoxic action of cytosine arabinoside (ara-C) was investigated. The reduction in the concentration of only one essential amino acid, L-isoleucine, in the growth medium of A(T1)C1-3 hamster fibrosarcoma cells decreased DNA synthesis in this cell population and slowed the rate of progression of G1 phase cells into S phase of the cell cycle. The complete omission of isoleucine from the growth medium blocked the progression of G1 phase cells into S phase and prevented the cytotoxic action of ara-C. The addition of isoleucine to the isoleucine-deprived cells permitted these cells to enter the S phase and restored their sensitivity to the cytotoxic action of ara-C. When G1 phase cells were placed in a medium containing reduced levels of all the amino acids and vitamins there was a prolongation of the G1 phase. Since medium with low levels of amino acids produced a delay in the entry of G1 phase cells into the S phase, the time interval in which these cells were most sensitive to the cytotoxic action of ara-C was different for G1 phase cells placed in medium with adequate levels of all the amino acids. These in vitro data indicate that nutritional factors can markedly effect the proliferation of tumor cells and the cytotoxic action of ara-C.


The kinetics of contact inhibition in mammalian cells.

To elucidate the process of contact inhibition in mammalian cells, we investigated the kinetics of growth arrest in [3H]thymidine labelled embryonic chinese hamster (Cricetulus griseus L.) cells after the addition of various concentrations of unlabelled cells. It was observed that after the contact inhibition concentration had been reached, the cells grew undisturbed for one more generation. In the following 24 hr the concentration fell back to the level at the beginning of the experiment and stayed there.


Development of the adrenal medullary cells in rats with reference to synaptogenesis.

The adrenal medullae of rats were studied electron microscopically from day 13.5 of gestation. Synapses with thickening of pre-and post-synaptic membranes were first evidenced on the medullary cells of 15.5-day-old fetuses. They increased gradually in number with advancing age. In the medullary cells, a few membrane-limited granules were recognized on day 13.5 of gestation, and thereafter they increased in number. The appearance of the granules was not uniform; some of the granular contents consisted of fine or coarse particles and others contained homogeneous material of varying electron-densities. The population of these granules in the cytoplasm was different from cell to cell. Thus, the discrimination of cell types (NA-and A-cells) was not possible in the prenatal period. Granular discharge of the cells was totally absent in the normal untreated fetuses. However upon the intraperitoneal administration of insulin to the fetus on day 16.5, 18.5, or 21.5 of gestation, discharge by reverse pinocytosis was first evidenced in the medullary cells of the 21.5-day-old fetus.


Studies on protein synthesis during sea urchin oogenesis. I. Synthesis of histone F2b.

Basic proteins were extracted from sea urchin oocytes previously incubated with 3H-lysine and then were analyzed by electrophoresis. A very radioactive band, which showed the same mobility as histone F2b, was analyzed for its amino acid composition. The results show an identity between this protein and histones F2b. In addition, an improved method of isolating large amounts of sea urchin oocytes is described.


The RNA of unfertilized sea urchin eggs is "capped".

It is shown that the RNA of unfertilized sea urchin eggs is active in stimulating protein synthesis in a wheat germ cell free system. This activity is not lowered by conditions that inhibit the methylation processes but is inhibited by a treatment that damages the "cap", A difference in activity in a wheat germ cell free system between the RNA of unfertilized eggs and the RNA from early embryos is described.


Ultracentrifugation of hydrated egg lecithin in benzene solution.

Estimations of molecular weights from sedimentation equilibrium experiments indicate that the degree of aggregation of egg lecithin in benzene solutions increases with increasing addition of water. The molecular weight rises monotopically from about 14,000 in the anhydrous state to about 57,000 at a water/lecithin ratio of 0.31. Sedimentation coefficients were shown to behave in the same manner. Frictional coefficients obtained from the combined results fail to indicate definite differences in asymmetry; however, they seem to indicate that the aggregates in all stages are compact in shape.


A fully automated solid-phase radioimmunoassay for triiodothyronine.

We report the first solid-phase radioimmunoassay for triiodothyronine with use of antibody-coated tubes, which is designed specifically for the fully automated radioimmunoassay instrument, Micromedic Systems' "Concept 4 Automatic Radioassay." Antisera to triiodothyropropionic acid/bovine serum albumin were raised in rabbits, purified by ammonium sulfate precipitation, and coated onto polypropylene tubes. Analytical recovery of exogenous triiodothyronine added to sera from normal men and women and pregnant women was quantitative. Intra-assay and inter-assay coefficients of variation are 4-5 and 6-9%, respectively. Correlation coefficients (r) for comparison of sample values with those obtained by three commercial laboratories were 0.95, 0.84, and 0.91. The sensitivity of the assay is 0.5 microng/liter. The assay can be performed either manually or be fully automated on the "Concept 4."


Effect of sodium sulfate on the hydrolysis of 17-hydroxycorticosteroid- and p-nitrophenyl-glucuronides with beta-glucuronidase preparations from bovine liver.

Previous papers dealt with the discovery that hydrolysis of 17-hydroxycorticosteroid glucuronides in urine with beta-glucuronidase preparations from bovine liver is increased by adding sodium sulfate to the incubation medium. Here, we conclude that sodium sulfate not only increases the activity of bovine liver beta-glucuronidase on 17-hydroxycorticosteroid glucuronides and p-nitrophenyl glucuronides, but also removes the inhibitory activity of substances of high molecular weight in urine and, moreover, inhibits urine putrefaction during the hydrolysis. The net effect is an increased yield of urinary 17-hydroxycorticosteroids. In the incubation with sodium sulfate (Na2SO4, final concentration 80 g/liter, 500 Fishman units of beta-glucuronidase per milliliter of urine, pH 5.0, 48 degrees C, 18 h) the analytical recovery of 17-hydroxycorticosteroid glucuronides added to 12 urine samples proved to be 98 +/- 1.8% (95-100).


Modification of the Pisano method for vanilmandelic acid using high pressure liquid chromatography.

A method for the measurement of urinary vanilmandelic acid (VMA) is described based on a modification of the colorimetric procedure of Pisano. VMA is oxidized to vanillin with subsequent measurement of the vanillin by high pressure liquid chromatography with electrochemical detection (LCEC).  The technique has been successfully applied to a number of human and animal urine samples. Urinary VMA values obtained by the LCEC method are in good agreement with those determined by the method of Pisano. The new method is both more selective and more sensitive than the colorimetric procedure.


A turbidimetric immuno assay (TIA) with automated individual blank compensation.

A simple and sensitive turbidimtric immunoassay (TIA) has been described. The automated method has been based on the use of the LKB reaction rate analyser, an instrument that is already present in many clinical chemical laboratories. The precipitin reaction is accelerated and enhanced by poly-ethylene glycol 6000 at a concentration of 50 g/l. Sample volume 2--100 micronl. Use of antiserum: 3--15 micronl per test. Sampling rate 30--60 samples per hour. The choice and the usefulness if antisera is discussed. The major advantage of the present method is  the individual measurement and compensation for the initial absorbance (340 nm) immediately after the mixing of antigen and antiserum. This enables the determination of very low serum protein concentrations (from 0.005 mg/ml) which is not possible with other methods that require separate blank determinations. The turbidity course is different  in antigen and antibody excess. The The chance of misinterpretation is small. The precision of the method is satisfactory. Within-run precision is 1--2% (C.V.) in the IgA/anti-alpha-system. Between-run-precision in the IgM/anti-micron system: 3.9% (C.V.). The day-to-day precision for IgG, IgA and IgM determinations resulting from a 7-month period of quality control was found to be 6.1% and 10.5%, respectively. RID and TIA methods agree very well. IgM determinations in cord serum from 104 newborns revealed a mean value and standard deviation of 14.6 +/- 6.3 I.U./ml (0.13 +/- 0.06 mg/ml).


A critical evaluation of a procedure for measurement of serum bile acids by radioimmunioassay.

A radioimmunoassay for the quantitative determination of cholic acid conjugates has been developed. Antisera were raised in rabbits injected with cholylglycine coupled by amide linkage to bovine serum albumin. The antibodies reported in this communication were not found to be mono-specific in the strictest sense. This assay is relatively simple, rapid, sensitive, reliable, and most importantly, specific for bile acids; the specificity of each antibody, however, must be thoroughly characterized at the dilution in routine use. The range of normal values for cholic acid conjugates in serum was found to be 0.1--1.6 micronmol/l (mean: 0.62, +/- S.D.: 0.4).


Postmenopausal urinary problems.

Detailed objective assessment of urinary symptoms provides information from which satisfactory treatment can be given. The fundamental investigations are history and clinical examination, endoscopy and two-channel cystometry (using several provocative tests). Detrusor dysfunction is a commoner cause of urinary symptoms than is generally realized and it cannot be diagnosed easily on clinical assessment. Initial treatment of all urinary problems should be conservative as the majority of patients benefit, and surgery should be carried out only after careful selection of patients and operative techniques.


A case of deficiency of N-hydroxylation of amobarbital.

It has been shown recently that the overall metabolism of amobarbital in man is essentially under genetic control. The drug normally undergoes two hydroxylation reactions, leading to 3'-hydroxyamobarbital (C-OH) and N-hydroxyamobarbital (N-OH). This paper describes a sibship in which two mothers who are identical twins show a gross deficiency on N-OH elimination in urine. The whole set of sibship data suggests that this deficiency represents a recessive trait controlled by a single pair of allelic autosomal genes which regulate N-OH formation. Several methodical approaches to assess an individual's capacity for N-OH formation are illustrated. There was no evidence of compensatory or concordant regulation of the two hydroxylation reactions. The case of this family illustrates that the functional lack of a biotransformation reaction is almost certain to be overlooked if one measures only the disappearance of a multimetabolized drug and not the appearance of metabolites.


Development of a radioimmunoassay for theophylline. Application to studies in premature infants.

The use of theophylline in premature infants has necessitated the development of an assay applicable to samples under 0.1 ml. To accomplish this we have developed a radioimmunoassay for theophylline. The antiserum to theophylline has been characterized and found to be highly specific for theophylline and sensitive to 2 ng. The affinity of our antibody and the titer developed in the goat allowed the assay to be performed routinely with the use of a plasma sample size of only 2 to 5 micronl. Theophylline levels of 6 to 20 microng/ml were found 2 hr after maintenance doses of aminophylline, 4 mg/kg per rectum in 12 premature infants. Terminal serum half-lives ranged from 12 to 54 hr.


Further investigations on the pathophysiology of the compartmental syndrome.

A model compartmental syndrome is described in rabbits in which the intracompartmental pressure may be accurately controlled to investigate the pathophysiologic changes resulting from increased intracompartmental pressure. Oxygenation in the tibialis anterior muscle was measured using a medical mass spectrometer. The Po2 declined with increasing intracompartmental pressure from a control value of 10.8 mmHg to a minumum of 2.8 mmHg at a pressure of 90 mmHg. The functional integrity of the peroneal nerve and compartmental muscle was tested by direct electrical stimulation. Functional deficits were first noted when an intracompartmental pressure of 40 mmHg was exerted for 6 hours. The incidence of functional losses increased with increasing pressures and durations of pressure application. All animals subjected to 100 mmHg for eight or more hours lost both nerve and muscle function. These investigations demonstrate that increased intracompartmental pressure alone, without other associated vascular injury, may produce muscle hypoxia and loss of neuromuscular function. The continuous monitoring of intracompartmental pressures may, therefore, be a useful clinical adjunct in the management of patients at risk for a compartmental syndrome.


Control of the yeast Torulopsis glabrata in the stomachs of glucose fed lambs by oral dosing with nystatin and amphotericin "B".

In the absence of specific dietary lipids, large amounts of ethanol (ethyl alcohol) are produced from glucose in the stomachs of ruminant neonates by the resident yeast Torulopsis glabrata. Using new born lambs, oral dosing with the antifungal antibiotics Nystatin and Amphotericin "B" was shown in samples of stomach contents, to control the ethanol production entirely, and to suppress the yeast numbers partially but not permanently. Of 11 lambs receiving heavy and continous doses orally of the pure antibiotic powders, 7 died, of these 5 had developed adhesive peritonitis.


Septicemia and meningoencephalitis in pastured cattle caused by a Haemophilus-like organism ("Haemophilus somnus").

Septicemia and meningoencephalitis developed in 10 pastured cattle 7 months to 3 years of age. Two unrelated herds were involved. Necropsy findings were similar to those previously reported in cattle infected with a Haemophilus-like organism, including multifocal intramuscular hemorrhages, suppurative polyarthritis, and multifocal hemorrhagic thrombi in the brain. A Haemophilus-like organism was isolated from one animal. It was characterized by growth on blood agar or tryptose agar plus a feeder streak under raised carbon dioxide tension, and lack of response to Haemophilus growth factors X and V.


The structure of mesokaryote chromosome.

Condensed and dispersed forms of the chromosomes of the dinoflagellate, Prorocentrum micans, deposited on grids by the microcentrifugation technique were studied by electron microscopy. In the normally condensed form, the chromosomes appear as banded rods surrounded by a peripheral cloud of partially dispersed fibers. Single fibers in these and in extensively dispersed preparations appear as smooth threads of uniform diameter (55-65 A). The chromosome fibers are contrasted by positive-group-specific stains indicating the presence of cationic moieties associated with the DNA. Occasionally Y-shaped chromosomes are seen; these may be replicating structures. These observations are in general agreement with studies of dinoflagellate chromosomes by other techniques, and provide support for the suggestion that these organisms possess a genome organization whose structure is typical of neither prokaryotes nor eukaryotes, and hence may be intermediate forms.


Observations on the mitosis and on the chromosome evolution during the lifecycle of Oodinium, a parasitic dinoflagellate.

The life cycle of the dinoflagellate Oodinium alternates between an ectoparasitic trophic phase and a phase of multiplication as free-living flagellates. The nucleus of the young ectoparasite has rod-like chromosomes similar to those of free-living dinoflagellates. As growth of the trophout proceeds the nucleus becomes increasingly homogeneous. When Oodinium leaves its host, nuclear reorganization processes occur rapidly; they correspond to a peculiar prophase of the first sporogenetic division. The following division stages are similar. A conspicuous fusorial system appears between two archoplasmic areas which are responsible for daughter-chromosome segregation. The nuclear envelope remains intact while the fusorial microtubules are attached at distinct, kinetochore-like structures onto the nucleus. As the chromosomes become more condensed the kinetochore-like formations disappear.


The differential effects of exposure to tobacco smoke on the secretion of luteinizing hormone and prolactin in the proestrous rat.

The acute effects of tobacco smoke inhalation on the spontaneous proestrous rise in serum luteinizing hormone and prolactin have been investigated in the female Wistar rat. It was found that the surge of LH normally seen throughout the afternoon of proestrus was delayed by inhalation of tobacco smoke and that the delay was dose-related to the nicotine content of the cigarettes used in  the experiment. In the case of prolactin neither the timing nor the magnitude of the surge was altered when compared with controls. These results suggest that under certain well-defined conditions inhalation of tobacco smoke of known nicotine content is capable of exerting profound influences on the hypothalamic-pituitary-gonadal axis.


Fourth expansion and glucose perturbation of the Dictyostelium kinetic model.

The kinetic model of carbohydrate metabolism has been expanded to include: (a) the accumulation of alpha and beta-cellulose, insoluble cell-wall glycogen and mucopolysaccharide; (b) the role of RNA turnover as a source of carbon for end-product synthesis and as a buffer regulating the level of uridine nucleotides in this metabolic network; and (c) the role of purine-nucleoside phosphorylase, 5'-AMP nucleotidase, nucleosidediphosphate kinase and polynucleotide phosphorylase. One of many predictions based on this model is that cells differentiating in the presence of glucose will produce sorocarps with an abnormally high trehalose to cellulose ratio. External perturbation of either the model or of developing cells by glucose increases the levels of sorocarp trehalose and glycogen, 5-fold and 6-fold respectively. Evaluation of the experimental data and the simulation analyses have allowed several predictions to be made concerning the compartmentation of metabolites and the permeability of cells to glucose during differentiation.


Characterization of foldback sequences in Physarum polycephalum nuclear DNA using the electron microscope.

An examination of the foldback fraction of nuclear DNA from Physarum polycephalum has been carried out using the electron microscope. Results show that the inverted repeat sequences responsible for the formation of foldback DNA range from 150-3000 bases in length, with a number-average size of 340 bases. About one-half of the inverted sequences form looped structures with loop sizes averaging 1200 bases in length. The distance between adjacent foldback sequences is estimated to be in the range 100-1500 bases.


Studies of glutamate dehydrogenase. Analysis of quaternary structure and contact areas between the polypeptide chains.

Cross-linking of the unimer of glutamate dehydrogenase from beef liver (consisting of six polypeptide chains each having a molecular weight of 56000) with dimethyladipimidate and subsequent analysis by sodium dodecylsulfate electrophoresis shows predominantly the trimeric species (molecular weight 168000). Treatment with dimethylimidates of other chain length yields significantly less trimeric species indicating that the amino groups being cross-linked are within a distance of about 0.85 nm. Comparison of the molar amount of incorporated [14C]dimethyladipimidate with the number of modified amino groups (determined with trinitrobenzenesulfonic acid) shows that although 8-9 of the 34 amino groups have reacted, only 2-3 of them are involved in cross-links. Reaction with dimethylimidates inactivates the enzyme. The loss of the activity is partly concomitant to cross-linking to the trimeric species and not simply due to the modification of essential lysine residues. This is supported by the fact that, although more lysine residues react with mono-functional methylimidates, the loss of activity is reduced. Purified chymotryptic and tryptic peptides of the radioactive-labeled trimeric species were subjected to sequence analysis. Six peptides containing 75% of the total label were identified: one involves the amino-terminal residue alanine-1 and the others involve lysine-105, lysine-154, lysine-269, lysine-358 and lysine-399. Quantitative analysis of the specific radioactivity of each peptide/mol lysine leads to the conclusion that only lysine-105, lysine-154, lysine-269 and lysine-358 participate in cross-links, lysine-269 and lysine-358, respectively, being at isologous and lysine-105 cross-linked with lysine-154 at heterologous contact domains of the enzyme. A model for the planar arrangement of the trimeric species in the quaternary structure of glutamate dehydrogenase is discussed. It includes both isologous and heterologous contact areas between the polypeptide chains.


Changes in sensitivity to operant effects of dopaminergic and cholinergic agents following morphine withdrawal in rats.

Rats trained to bar-press on either a FI 15 sec or FR 30 schedule for water reinforcement were administered various doses of appomorphine, haloperidol, pilocarpine and scopolamine both before and 1--4 months after a 3-day period of continuous morphine administration. All drugs monotonically depressed response rates on both schedules with increasing dose. Chronic morphrine administration produced a persistent increase in sensitivity to apomorphine and pilocarpine and a persistent decrease in sensitivity to haloperidol and scopolamine. The results suggest that both dopaminergic and cholinergic supersensitivity are likely to be involved in protracted abstinence phenomena.


[Histological changes of various organs in aged SD-JCL rats (author's transl)].

Age-related histological changes were studied in various organs from SD-JCL rats reared throughout their lifespan. In aged-male rats examined at 5 to 36 months of age were frequently observed nephropathy, periarteritis, skeletal muscle degeneration, pigmentation of the follicular epithelium in the thyroid, fibrosis of the pancreatic islets, hyperplasia of the parathyroid epithelium, and changes of the acini in the extraorbital lacrymal gland. In aged-female rats sinusoid dilatation of the adrenal, and atrophy of the ovary were also noted. Perilobular fat deposition of the liver, dilatation of the gastric gland and severe hemosiderosis of the spleen were observed similar frequency in the both sexes. The nephropathy and cardiovasculopathy were major factors to cause death for males, while the main cause of death for females was tumors, especially of the mammary and pituitary glands. Enlargement of the parathyroid gland, bone resorption and metastatic calcification in the solf tissues were found in rats with severe nephropathy.


[Spontaneous tumors in aged SD-JCL rats (author's transl)].

Incidence of spontaneous tumors was examined in 77 male and 73 female SD-JCL rats reared throughout their lifespan of 5 to 36 months. Total tumor incidences were 60% and 95% for males and females, respectively. The sex difference was mostly attributable to a high incidence of mammary tumor in females. The pituitary tumor was found in 30% of males and 66% of females aged 5 to 36 months while islet cell tumor of the pancreas and thyroid tumor occurred later and the incidence of latter was particularly high in rats lived more than 2 years. Myeloid leukemia appeared to occur as early as 5 months of age.


A simplified method using commercial milk powder for hand-rearing of the Caesarean-derived infant rabbits (author's transl).

An efficient and simplified method for hand-rearing of Caesarean-derived infant rabbits under gnotobiotic condition was devised. The Caesarean-derived infant Dutch or Japanese-White rabbits and their hybrids (F1; Japanese-White female x Dutch male) were reared in sterilized vinyl-isolators by hand-feeding with two kinds of milk diets, A and B, consisted mainly ofa commercial milk powder for dogs and cats (Esbilac) supplemented with several minor components (Table 2) and administered intragastrically once a day through a Nelaton's catheter. Bacteriological examinations revealed that feces and urine were sterile for the first three days. On the third day, Escherichia coli, Staphylococcus epidermidis, Streptococcus faecalis, Bacillus subtilis, Enterococcus, and Bacteroides sp. were given with milk diet and the infant rabbits were reared until 10 to 12 weeks of age. The weaning rate at 5 weeks of age and the raising rate at 3 months of age were approximately 78% and 77%, respectively (Table 3), indicating that feeding once-a-day with a milk diet mainly composed of Esbilac is suitable for the hand-rearing of infant rabbits. There was, however, no significant difference milk diet A and B on the weaning rate.


Breeding of a rabbit strain of hyperlipidemia and characteristic of this strain (author's transl).

We attempted the breeding experiment in order to establish an inbred strain of disease model animal derived from a spontaneously hyperlipidemic rabbit (HLR). The clinical, pathological features and breeding results of HLR strain were as follows; 1) A prominent clinical feature of this strain was conspicuous elevation of serous lipid ingredients including cholesterol (S-ch), beta-lipoprotein (beta-LP). The S-ch level was 400 +/- 70 mg/dl, the beta-LP level 9.1 +/- 1.5 unit and the total lipid 1146 +/- 270 mg/dl in the HLR strain. These values were approximately 9, 10 and 4 times higher than those of normal rabbits respectively. 2) After sacrifice, aterosclerotic lesions were evident extensively in the aorta and renal arteries. Nodular of plane xanthomas were conspicuous over the articuli digitorum of manus and pedis. 3) HLR strain indicated a striking resemblance of the condition to type III hyperlipidemia in the Frederickson's classification of hyperlipidemia in man. 4) Inbreeding was carried out in two lines, viz. sister-brother mating, parent-offspring mating and back crossing. As the result, the second filial generation showed an incidence of HLR of 15.0%, the third generation 50.0%, and the fourth generation 56.1% respectively. In the fourth generation, the coefficients of inbreeding and relationship were 50% and 72.3% respectively.


[Importance of biochemical marker genes in inbred strains of mice. Some problems on checking genetic purity of inbred strains (author's transl)].

In order to check genetic purity of inbred strains of mice, we surveyed five biochemical variants in certain 69 strains, some of which had established from Japanese fancy mice. The loci examined were Hemoglobin beta-chain, Malic enzyme (supernatant form), Isocitrate dehydrogenase (supernatant form), Serum esterase-1, and Serum esterase-2. These loci were all in homozygous states as far as examined, however some subline divergences were found in a few strains, that is to say, sublines were found to be fixed with different alleles as some loci, Important of biochemical marker genes to check genetic purity of strains and several derived problems are mentioned.


[Rearing and management of chimpanzees for experimental infection with hepatitis B virus (author's transl)].

For the purpose of experimental infection with human hepatitis B virus, 14 chimpanzees (Pan troglodytes) were delivered to the Division of Animal Research, Faculty of Medicine, University of Tokyo, Tokyo. These chimps, 11 males and 3 females, born in the West Africa, had been reared for two to six months. Several days after delivery, they were anesthetized with Ketalar in order to make clinical, bacteriological and parasitological examinations; It was found that one of them was in malnutrition, and that another had dislocation of the shoulder joint and the associating abscess. All of them were negative for tuberculin test. In the bacteriological examination, Shigella sonnei was detected in the feces from one of them. Mycoplasma sp. was detected in the materials from the oral cavity of four head. As intestinal parasites, Ascaris sp. were detected in two head, Enterobius vermicularis in eight, Strongloides sp. in two, Oesophagostomum sp. in nine, tape worms in four, and Entamoeba coli in twelve. Microfilaria as blood parasite was detected in 11 of them. The laboratory used for the experimental infection was a room occupying about 42 m2, which had been built by renovating a part of our division building. Each of the cages used for rearing the chimps was contained in the isolation box made of stainless steel. The contaminated air in the isolation box was discharged forcedly into the exhausting duct with a fan, and further passed through the HEPA filter and the Miraceram honeycomb heater, and was then conducted to the already existing ventilation duct of the division building. Each chimp was fed on a ration of 200-350 g of the imported "Purina Monkey Chow 25" and also one grapefruit and one banana daily for the supply of vitamin C. The chimps weighed 16.1 kg on the average on delivery, but gained an average weight of 4.2 kg during the following four months.


[Influence of nystatine, a polyene antibiotic, on the formation of complex-bound sterols and the formation of conidia by Coprinus cinereus (author's transl)].

In contrary to the wild strain, a monocaryotic mutant of Coprinus cinereus (basidiomycete) does not contain hydrosoluble, complex-bound sterols. Under nystatine-treatment, however, the mutant is induced to produce this kind of stereols in abundance. Furthermore, nystatine highly increases sporulation of the mutant and decreases sporulation of the wild strain.


[2-o-Iodotyrosine]-oxytocin and [2-o-methyltyrosine]-oxytocin: basic pharmacology and comments on their potential use in binding studies.

Both [2-o-iodotyrosine]-oxytocin and [2-o-methyltyrosine]-oxytocin display only weak vasopressor and antidiuretic effects on rats. They inhibit the in vitro uterotonic action of oxytocin; this inhibition is not fully competitive. It is concluded that they are not suitable as markers for studies of uterine receptor for oxytocin.


[Effect of neuroleptics, midantan and DOPA on apomorphine sterotypy in rats].

The influence of differently structured neuroleptics on the stereotypyd attitude of rats, induced by apomorphine, was inquired into. Most of the known neuroleptics manifested an inhibitory effect on the apomorphine-induced stereotypy. Clozapine and azabutyrone were effective only in a dose of 25 mg/kg, while carbidine and promazine failed to exert any marked influence on the stereotypy. It seems that the antiapomorphine effect does not correlate with the antipsychotic activity of the neuroleptics.


[Effect of lithium chloride on several indices of cardiac function].

Used in a dose of 100 mg/kg lithium chloride modifies the ECG patterns in rats, rabbits and cats. It increases the voltage of P and R waves, the S-T interval and slows down the rhythm of cardiac contractions. In doses of 50-300 mg/kg the compound raises the coronary circulation rate in narcotized cats. The compound exerts an antiarrhythmic action in chlorocalcium arrhythmia in rats, in strophanthin-induced one in cats and frogs and the intensity of this action is not inferior to that produced by potassium chloride, quinidine, novacainamide, izoptin and even exceeds it.


[Conjugation components in alpha-adrenergic reactions in smooth muscles].

Changes in the electric and mechanical activity conditioned by norepinephrine in the deferent duct, musculus rectococcygeus, aortic band and the portal vein in rats, as well as influence exerted on these changes by the replacement of Na+ ions with NH4+ ones, by the action of novocain (1-10(-3) g/ml), strophanthin (1-10(-5) g/ml), excess (25 mM) and deficiency (0.25 mM) of Ca2+ ions and by that of manganese chloride (2 mM) and lanthanum (1 mM) were analyzed. As evidenced, the basic mode of conjugation in alpha-adrenergic reactions is an intensified supply of Ca2+ ions to the cells, unattended by any changes in the degree of the membrane potential at rest. Depolarization and quickened generation of action potentials occur in musculus rectococcygeus and in the portal vein, in the lastly named case -- under the effect of high norephrine concentrations (10(-6)-10(-5) g/ml).


[Effect of ethyl alcohol on oxidation-reduction processes in the myocardium].

Evidence is given of disturbances of aerobic oxidative process in the myocardial mitochondria occurring in acute and chronic alcoholic intoxication. A single introduction of alcohol in a dose of 1 and 3 g/kg to rats and dogs produces "hypoxyform" changes in the myocardium, viz. a fall of PO2, a reduced activity of most NAD-independent enzymes, activation of succinate- and lactate-dehydrogenases in a moderate dissociation of oxidative phosphorylation reactions. The damaging effect of the alcohol persists for 24 hours after its one-time introduction. Following a 2-month long alcoholic intoxication there was seen a depressed respiratory activity of the mitochondria and of the study enzymes, except for cytochrome-C-oxidase. It is assumed that the cardiotoxic effect of alcohol is due to the damage of the energy-producing system of the mitochondria.


An in vitro bioassay for 1,25-dihydroxyvitamin D3 and other antirachitic agents.

Duodenal tissue from chicken embryos (19 days in ovo) has been maintained in organ culture for 48 hours. The incubation medium and culture conditions were chosen to produce a linear relationship between calcium binding protein production and the logarithm of the dose of vitamin D3 steroid in the medium. The dose-response curves are parallel for vitamin D3 and both its major metabolites, 25-hydroxyvitamin D3 and 1,25-dihydroxyVITAMIN D3. The minimum concentration of 1,25-dihydroxyvitamin D3 capable of inducing calcium-binding-protein synthesis was 1 X 10(-9) M.


A myogenic cell line with altered serum requirements for differentiation.

Dfferentiation properties of a cell line, L84, which originated from a non-fusing clone isolated from the myogenic line L8, are described. In nutritional medium supplemented with 10% serum used routinely with L8 cells, L84 cells continue to proliferate to very high densities and fail to form multinucleated fibres. When grown in medium supplemented with 2% horse serum of 2% horse serum plus 0.1% microng/ml insulin, L84 cells behave very similarly to L8 cells grown in medium supplemented with 10% horse serum: when the cultures reach confluency, proliferation decreases and cells start to fuse and form a dense network of fibres. Large increases in creatine kinase activity and synthesis of myosin are associated with cell fusion. Under conditions in which L84 cells do not fuse the increase in these synthetic activities is not observed, even after extremely high cell densities are reached. The data show that L84 cells retain the programme for their differentiation into muscle fibres. The difference between L84 and its progenitor line L8 lies in the sensitivity to the environmental conditions which trigger the expression of this programme.


Scanning electron microscopy (SEM) of the chick embryo primitive streak.

The structure of the cells forming the primitive streak was examined by SEM in a series of embryos at Hamburger and Hamilton's stages 2--5. Specimens were prepared by stripping the endoderm from fresh embryos in New Culture and by fracturing whole fixed embryos along and at right angles to the primitive streak. At all stages of examination the SEM appearance of cells within the privitive streak was quite different from that of ectodermal, endodermal or mesodermal cells away from the streak. Streak cells were closely packed, lay with their long axes directed from ectoderm to endoderm and possessed many flat leaf-like processes. By contrast the ectoderm formed a columnar epithelium, the endoderm a flat epithelium and the mesoderm was a layer of loosely arrangedcells with long. thin processes. Within the streak SEM did not show any differences between cells that could identify them specifically as future endoderm or mesoderm cells. It was concluded that during gastrulation all the cells migrating through the primitive streak have the same appearance regardless of their eventual destination in the embryo. This structure may be attributable to the type of movement made by cells during invagination.


Developmental pattern of small intestinal enterokinase and disaccharidase activities in the human fetus.

The concomitant appearance of enterokinase (EK) and trypsin activities in the human intestinal mucosa is indicative of the importance of EK as an activator of trypsinogen and therefore as the key enzyme in protein digestion. Enterokinase can be detected in fetal mucosa from the 26th week of gestation on, paralleling appearance of tryptic activity in meconium. The developmental pattern of EK activity increases with age. Between 26 to 30 weeks of gestation, the EK activity is only 6% and full term babies (40 weeks) 20% of that found in older children. In contrast, lactase studies during development show a lactase activity of only 30% in human fetuses between 26 to 34 weeks of gestation as compared to full term babies. During the same gestational period, sucrase and maltase activities reach 70% of the full term. In addition, the distributional pattern of EK differs from the disaccharidases, showing the highest activity in duodenum and the lowest in ileum, whereas disaccharidases are highest in jejunum with lower activity in duodenum and ileum. Differences in topographical distribution and time of appearance of EK and disaccharidases may be attributed to differences in orgin as well as subcellular localization of these enzymes. It is conceivable that the premature infant, between 26 to 30 weeks of gestation, is better equipped to deal with hydrolysis of alpha-glucosides than of lactose.


Kayser-Fleischer-like rings in patients without Wilson's disease.

Three patients, one with cryptogenic cirrhosis, one with active chronic hepatitis and one with neonatal hepatitis, were found to have corneal pigmentation rings indistinguishable from early Kayser-Fleischer rings on slit lamp examination. They did not have the clinical features of Wilson's disease and their serum copper and ceruloplasmin concentrations were normal. Urinary copper excretion rates and hepatic concentrations were only slightly raised but were below the range found in symptomatic Wilson's disease. It is concluded that the Kayser-Fleischer ring would no longer be considered as pathognomonic of Wilson's disease.


[Disordant monozygotic twins with Mayer Rokitansky Kütser syndrome (author's transl)].

Two patients with vaginal agenesis are reported. Each had a normal twin. In one case, a Mayer Rokitansky Küster syndrome could be diagnosed by laparoscopy. In the other case it was probably the same syndrome. Monozygocy of the two pairs of twins was shown by examination of features of serum proteins, serum enzymes and erythrocytes with a probability of 99,8% and 99,1%. Only three other cases of discordant twins with MRK syndrome were reported in the literature, at least two of them monozygotic. Extremely rare cases of familial MRK syndrome were also found. Discordance of monozygotic twins makes a genetic etiology unlikely.


[Severe infections in our obstetric-gynecologic case material including nosocomial infections (1972--1976) (author's transl)].

A consideral by increased incidence of severe infections in 1975 provided the impetus for a critical analysis of such cases (n = 29) during the last four years. In the group of infections acquired outside the hospital setting, 13 were adnexal infections and 2 were amniotic infections. The group of true hospital infections (n = 14) consisted of 4 entoplastic cases, 7 cases with contaminated infusions and 3 cases of amniotic infections. A shift in the pathogenic spectrum in favor of gram-negative bacteria, particularly Klebsiella, Pseudomonas and anaerobes, was observed in those infections acquired within the hospital setting-- The mortality rate for the whole group was 10% (!).-- In addition to the usual hygienic measures used in hospitals to successfully combat endemic diseases resulting from severe nosocomial infections, a short-term perioperative prophylactic antibiotic, the advantages and disadvantages of which were mentioned, was integrated in the resistance dispositive. In severe adnexal inflammation (2 times in connection with an intrauterine device), the method of choice for surgical therapy was adnexectomy or, if necessary, hysterectomy and drainage. Septicemic diseases in connection with venipuncture material (entoplastic) can be avoided, for the most part, by carefully selecting the type and nature of the material as well as through preventive hygienic measures. The infusions contaminated exclusively by Klebsiella led to particularly serious infectious disease.


[Spontaneous rupture of gravid rudimentary horn of an uterus bicornis unicollis without connection with the cervical channel (author's transl)].

It is reported on a 24-year-old nullipara, who came to be operated with the diagnosis of perforated extrauterine gravidity. It was the question of the perforation of a gravid uterus bicornis with missing connection of the gravid horn to the cervical channel. The importance of an early diagnosis of uterus malformations is pointed out. The modus of implantation in the atresic uterus horn is placed under discussion.


Effect of iron and protein deficiency on the expulsion of Nippostrongylus brasiliensis from the small intestine of the rat.

The relationship between iron deficiency and protein deficiency and infestation of the rat with the nematode Nippostrongylus brasiliensis was investigated. There was a significant delay in the expulsion of N. brasiliensis from the small intestine of both iron deficient and protein deficient animals and those with a combined deficiency of iron and protein. Iron repletion returned the time of worm expulsion to normal and this would appear to be related to iron deficiency per se rather than to anaemia. Antibody initiated damage to worms was normal in the control animals and in animals with nutritional deficiencies. This suggests that the defect in worm expulsion occurs either in the cell-mediated immune system or in one of the other mediators of expulsion. Extrapolation to the human situation has important therapeutic implications in that iron and protein deficiency may play an important role in the perpetuation of helminth infestations. Thus, to be successful antihelminth therapy should be accompanied by iron and protein supplementation.


[Common virus infections of the skin. Molluscum contagiosum, herpes zoster, herpes simplex, condylomata acuminata, verrucae vulgares].

The clinical diagnosis, etiology and predisposing factors of these common viral skin diseases are discussed with special emphasis on new therapeutic approaches.


[New transplantation materials for reconstructive arterial surgery].

For reconstruction of femoral artery occlusion autologous vena saphena is the best material. In all cases of reocclusion after autologous vein bypass or in cases without adequate vena saphena the heterologous Solco-Collagen-Graft and the new alloplastic Gore-Tex-Teflon arterial prosthesis are two alternative materials. First clinical experiences with the Gore-Tex-Graft in 45 patients and clinical experiences with the bovine Solco-Graft in 54 patients are presented. Both the bovine Solco-Graft and the alloplastic Gore-Tex-Graft provided a patency rate of 75% in patients with chronic arterial diseases state III-IV after 1 year. The special indications for either heterologous bovine Solco-Graft or alloplastic Gore-Tex-prosthesis are demonstrated.


[Experiences with mastodynon in the treatment of the premenstrual syndrome].

Experiences with Mastodynon in the therapy of premenstrual syndrome are reported. The results of the phytohormon Mastodynon were evaluated in 52 cases by measurement of the basal temperature.


Psychophysiologic test performance in normal twins and in a pair of identical twins with essential tremor that is suppressed by alcohol.

In a sample of 52 adults male twin pairs (26 MZ, 26DZ) performance in a series of psychophysiologic tests was measured. The intraindividual variability was determined by repetitive tests in the same subjects. In a number of cases the measured parameters proved relatively stable within the same peron; identical twins, however, were no more similar to one another than fraternal twins. In a pair of identical twins with autosomal-dominant essential tremor the degree of the tremor could be effectively reduced by alcohol application.


Gonads of Carassius auratus (teleosts) in organ culture: a new technique and the effects of testosterone.

The use of a semi-natural medium (Eagle's MEM supplemented with chicken embryo extract) appears to be satisfactory for ovarian and testicular fragment cultures of Carassius auratus. The male germ cells show a normal degeneration which might be attributed to a lack of pituitary hormones: gonadotropic and, possibly, somatotropic hormone. The ovarian fragments maintain a normal organization during 21 days of culture. Only the oocytes in late vitellogenesis massively atresiate at the beginning of culture. The addition of fetal bovine serum to this medium produces surprising effects on the testicular explants, particularly on the spermatogonial mitoses. This might be attributed to the presence of STH and androgen in the serum. The modifications of ovarian explants, on the other hand, are identical to those observed with Eagle's MEM supplemented with chicken embryo extract. M 199 has been tested as a synthetic medium; the preliminary results are encouraging. Testosterone, when added to the semi-natural medium, permits complete spermatogenesis in the testicular fragments during the 21 days of culture, but does not produce any effect on the ovarian explants.


[Indications for the transfusion of erythrocyte concentrates].

Whole blood transfusions as usual are often not effective. The high fluid level overloads the circulatory system and the share of red blood corpuscles improves the hematocrit inconsiderably. On the contrary after application of erythrocyte suspension which has reducted volume the hematocrit increases and risks of the circulatory system will be prevented. Risks by means of whole-blood transfusions; advantages, indications, and production of erythrocyte concentrations are outlined.


[Manufacture and transfusion of leukocyte and thrombocyte concentrates with special reference to hematological problems].

Great quantities of granulocytes or platelets can be removed from a single donor by following methods: 1) Continuous flow cellseparation 2) Intermittent flow cellseparation 3) Filtration leukapheresis. Using one of the methods mentioned above 1.0 X 10(10)--5.0 X 10(10) granulocytes can be obtained during a four hours procedure. The yield of platelets is more than 5.0 X 10(11) cells. The procedure itself is well tolerated and no severe side effects were seen.


[Manufacture and transfusion of leukocyte and thrombocyte concentrates with special reference to organizational problems].

One year-long experience with the Haemonetics blood cell separator (model 30) in a blood bank is presented. The collection, indications for transfusion and transfusion itself of 62 leucocytes and 96 platelets concentrates, as well as organisatory problems are discussed in detail.


[The use of human albumin in infusion therapy].

A survey on physiologic functions of albumin is given. Furthermore the two main-indications of albumin, the treatment of blood-volume-deficiency, including the use of albumin for the iso-volemic-hemodilution and the substitution in hyproteinemia are discussed. A comparison of albumin and plasmaproteinsolution follows. Finally limiting factors in albumin therapy due to the lack of coagulation factors are mentioned.


[The use of stroma-free hemoglobin solutions as blood substitute].

Solutions of stroma free human hemoglobin and of polymerized hemoglobin were used for perfusion of isolated organs of the experimental animal (Wistar rats). Both preparations proved to be suited for maintainance of sufficient oxygen transport. Following intravenous infusion of greater amounts of hemoglobin or polymerized hemoglobin in the experimental animals (3.0 g/kg body weight), the renal losses amounted 30% and less than 10% respectively. Half live was only two hours in the case of hemoglobin and 13 hours with the polymerized hemoglobin, despite molecular weight was only doubled in the latter preparation. Since the minor amount of the different hemoglobin preparation was excreted by the kidneys, the greater amount was stored in the organism or metabolized respectively. The intravenous infusion did not cause chemically demonstrable signs of liver toxicity. Neither bilirubin concentration nor enzyme activity showed significant alterations. Additionally, all animals survived the high dosed intravenous infusions. However, histological evaluations showed distinct alterations caused by the hemoglobin preparations. Hemoglobin was found inside the liver parenchymal cells. The liver cells and kidney cells showed signs of toxic effects. The polymerized hemoglobin was not found in liver parenchymal cells. In contrast to hemoglobin this preparation was stored inside the Kupffer's cells. In contrast to hemoglobin the polymerized form behaves like erythrocytes. The rapid elimination of hemoglobin (half live only two hours) renders this substance unsuited for blood substitution. However, half life of polymerized hemoglobin is 13 hours, and from this point of view the polymerized form is suited for blood substitution. Considering the histologically demonstrable alterations additional experiments are required before hemoglobin solutions are used in human subjects.


[Changes in body composition during various reducing diets].

A method of measuring body density is presented which allows much greater precision in calculating the fat content of the human body than the usual methods of under-water weighing. Using this method, body fat was determined for three groups: normal weight, moderately overweight, and obese subjects. In spite of vast amounts of energy stored in the obese patient, no advantages for the pre- or post-surgical phase result. The explanation for this paradoxical situation is seen in two factors: 1. The cardiovascular system is not capable of delivering more energy because mobilisation of fat depends on corresponding oxygen intake. 2. Aside from this, a metabolic limiting factor exists in the mobilisation of protein reserves for the production of glucose, which leads to a considerable protein loss the first few weeks of a period of abstinence. In posttraumatic or post-operative situations, this protein mobilisation is particularly strong, especially when connected with a high metabolic intake. The high nitrogen loss and/or the high metabolic rate, is illustrated on the basis of severe skull and brain traumas. The differences between absolute fasting and a 200kcal-protein formula diet were experimentally investigated. The nitrogen loss under the protein diet was decisively less. An anabolic phase even reappeared after three weeks of the diet. The 200kcal-diet therefore represents a means of pre-operative weight reduction without protein loss. The alterations in body composition are almost exclusively the results of reduction in excess fat. This is demonstrated by means of clinical examples.


[Biokinetic characterization of human glycerin utilization].

23 patients received controlled infusions of 10% glycerol solution using an "Infusomat". During various rates of infusion biokinetic parameters and renal excretion were measured. The concentration of glycerol in the serum rises over-proportional with increasing rates of infusion. The extrapolated maximal metabolic turnover capacity is 54 micronmol.kg-1.min-1. Halb-maximal turnover rate is reached at a serum level of 0,56 micronmol.ml-1. After an infusion of 0,3 g.kg-1.min-1 glycerol disappears from the blood with an elimination constant of 0,024 min-1 and a half life of 28,9 minutes. Renal excretion increases with the dose but remains below 10% of the dose in the investigated range. From the data it can be concluded that glycerol cannot be applicated in higher doses than other polyols. No adverse reactions have been observed in the range up to 0,3 g.kg-1.min-1.


[Dangers of a wrong diet in patients with renal insufficiency in the predialytic state or during chronic hemodialysis].

Patients with chronic renal failure and on chronic intermittent hemodialysis have several metabolic risks such as retention of urea and other products of nitrogen metabolism, catabolism, acidosis, edema, dehydration, hyper- and hypotension, hyperkalemia, renal osteopathy, and renal anemia. Uremic coma is usually avoided by balanced nutritional therapy and treatment with hemodialysis. A dietary regimen containing protein with high amounts of essential amino acids is an important part of treatment. In patients on chronic hemodialysis disorders of protein and amino acid metabolism are caused by either deficiency of essential ingredients of the food or by the metabolic defects due to chronic uremia.


[Nutritional habits and protein status of surgical patients during the preclinical phase].

In this study the importance of nutrition during the preoperative period has to be find out. Three methods are helpful in it: 1. anamnesis of nutrition; 2. status of the subcutaneous fat tissue by measuring the skinfolds with a Harpenden-Caliper; 3. proteinstatus. 24 patients after gastric surgery and 20 patients of a control group were investigated. There was found a loss of weight inabout half of these 24 "gastric patients" preoperatively. These special patients showed a malnutrition in their anamnesis, thin skinfolds, low serum albumin and total protein concentration and a decreased concentration of immunoglobulins. They got six times more infectious complicationes than the patients without a preoperative loss of weight. It is proposed to develop an adequate plan for preoperative and post-operative nutrition.


[Risks to donor and recipient in blood collection and blood transfusion].

A summary is given on the local and general risks of blood donation for the donor and especially on the recipient's risks to blood transfusion itself, including massive transfusion. The transfusion reactions are grouped in (1) risks in connection with the technique and physics of transfusion (cooling, air embolism, microaggregation, circulatory overload), (2) biochemical-metabolic risks (citrate intoxication, acidity, coagulation deficiencies), (3) immunological-serological risks (allergic and hemolytic reaction, addendum: pyrogenic reaction), (4) dangers of infection (bacteria: septic-toxic reaction, protozoae: malaria, viruses: herpes group, cytomegaly, hepatitis). In order to cut down the overall frequency of transfusion reactions, the use of blood derivatives instead of whole blood has been described.


[The manufacture and transfusion of deep frozen preserved blood].

For the past ten years we have been evaluating the preservation of human red cells by freezing. A comparative study of several techniques revealed no major differences as far as clinical effectiveness is concerned. The recovery percentage varies from 90-95%. Due to the washing procedure, necessary for removing the intracellular cryophylactic substance glycerol, no side effects from these ghosts have been observed. The maximum storage time in the post-thaw period should not exceed 24 hours, because the so-called "closed system" is interrupted. Since the preparations and storage of these cells is laborious and expensive, frozen blood should be used in selected clinical situations, i.e. rare blood types, autologous transfusions.


[Blood transfusion and cross matching].

To avoid hemolytic transfusion reactions different methods of cross match techniques are described and the relation between the recipient's and the donor's blood quantities are discussed in this regard.


[Forensic problems in blood transfusion].

According to the Austrian Criminal Law the physician is made responsible for damages caused by blood-transfusion, because of criminal carelessness, unless, no serious guilt is proven and the sequences of the transfusion--incident are so insignificant that neither a damage of health nor any inability to carry on a profession longer than fourteen days has been caused. The physician is obliged to make the patient aware of the dangers of blood-transfusion in the limits of this comfort of such procedure and is not allowed to proceed the transfusion against the declaired will of an adult even though according to medical conception this would be necessary. In urgent indicated cases the physician has the possibility to ignore the wish of a non-adult person and decide against the will of the guardian without exposing himself to the danger of criminal pursuit. Consequences of Civil Law are not considered.


[Transfusion problems relative to the HL-A system].

The importance of the HL-A system for blood transfusions (febrile reactions, substitution of leukocytes and platelets, prevention of anti-HL-A immunisation) is described. Furthermore, the problem of finding an HL-A identical donor for a given recipient which arises from the polymorphism of the HL-A system is discussed.


Ureaplasmal epithelial lesions related to ammonia.

The pathological effects of ureaplasmas on oviductal epithelium (ciliostasis and deciliation) were duplicated by adding ammonia to the medium as ammonium sulfate or by adding jack bean urease, which hydrolyzed the urea in the medium.


Effect of maternal immunization on the antibody response of low responder rats.

The offspring of low responder F344 female rats that were immunized with poly(Glu52Lys33Tyr15) aggregated with MeBSA prior to mating showed a higher antibody response to the polypeptide antigen than did the offspring of unimmunized females. Immunization of the mothers with unaggregated polypeptide, or with DNP-BGG, did not affect the antibody response of the offspring even when high doses of antigen were used. When the polypeptide used to immunize the mothers was given as an aggregate, some crossed the placenta to the fetus. The antigen was first detected in the placenta, blood and liver of the fetus at 15 days of gestational age. After birth, it was in the liver and spleen up to 6 weeks af age, and thereafter it was present only in the bone marrow.


Immunoglobulins in human cervico-vaginal secretions.

The concentration of IgG, IgA and IgM has been measured in the cervico-vaginal secretions of 8 women with a normal menstrual cycle, 52 pregnant women, 6 post-menopausal women and 12 women with total hysterectomy. No significant difference in immunoglobulin levels was found in the cervico-vaginal secretions of women with a normal cycle as compared to those of post-menopausal or pregnant women. A significant decrease of the IgG/IgA ratio was noticed during ovulation as a consequence of increased IgA secretion. In patients with hysterectomy, the secretions are of vaginal origin only and contain negligible quantities of IgA. Secretory IgA is found essentially in the superior genital tract. IgM is present in trace amounts in all secretions and does not vary considerably. The secretion of immunoglobulins may be under hormonal control: in addition to ovarian hormones, corticosteroids seem also to be involved. The local application of a fluorinated corticosteroid into the vagina has produced a significant decrease of secretory IgA production. The therapeutic possibilities of administering a fluorinated corticosteroid in cases of sterility which are due to the production of anti-spermatozoal antibodies are discussed.


Respiration and ATP level in BHK21/13S cells during the earlist stages of rubella virus replication.

This study is concerned with the effect of virus infection on the energy balance of the cell and the possible involvement of mitochondria in virus replication. The process of rubella virus adsorption and penetration is immediately associated with a stimulation of O2 uptake. Shortly afterwards, ATP levels decline, possible because of increased ATP utilization. Virus infection appears to protect mitochondria against the uncoupling effect of dinitrophenol.


Detection of double-stranded RNA in Semliki Forest virus-infected cells by an indirect solid phase radioimmunoassay: an assay for interferon.

An indirect solid phase radioimmunoassay using rabbit antibodies to the synthetic double-stranded RNA poly rI-rC and 125I-labelled sheep anti-rabbit IgG could detect 1 ng of poly rI-rC. This assay could also detect the presence of double-stranded RNA in Semliki Forest virus-infected chick fibroblasts and can be used as an assay for interferon activity.


Lyophilized combination pools of enterovirus equine antisera: preparation and test procedures for the identification of field strains of 19 group A coxsackievirus serotypes.

This paper describes the preparation of seven combination pools of equine antisera, designated J though P, for identification of 19 coxsackievirus A immunotypes. Each pool is composed of 4 to 6 antisera; the serotypes included are A1-6, 8, 10-15, and 17-22. These pools, unlike the previously prepared A-H enterovirus pools, were lyophilized from volumes of 0.5 ml dispensed into 5-ml vials, and when rehydrated with 5 ml of diluent provide 50-antibody-unit material ready for use in identification tests without further dilution. Procedures for using the antiserum pools are given, and guidance is provided for interpreting the results of serum neutralization tests in identifying field isolates.


Radioimmunoassay of urinary free cortisol.

The radioimmunoassay for urinary free cortisol described in this paper is simple, rapid, and reproducible. The method uses a commercially available antibody preparation and is performed in two steps. The first step includes an extraction and a column purification to remove materials antigenically similar to cortisol from the urine. The second step is the radioimmunoassay using dextran-coated charcoal to separate bound and unbound cortisol. 3H-cortisol is added prior to any mechanical manipulation to allow calculation of analytical recovery for the purification procedure. The coefficient of variation for interassay determinations was a maximum of 10.3% and for intraassay determinations a maximum of 5.7%. Analytical recovery averaged 97.7%. One technician can analyze 100 samples per week.


The location of nuclei of different labelling intensities in autoradiographs of the anterior forebrain of postnatial mice injected with [3H]thymidine on the eleventh and twelfth days post-conception.

The location of neuron nuclei of different labelling intensities in autoradiographs of the anterior forebrain of two 22 day old mice which had been injected with [3H]thymidine at 11 and 12 days post-conception respectively was charted on photocollages of sections enlarges 175 times. The pattern of distribution of the heavily labelled nuclei, i.e. those nuclei belonging to cells most likely to have been born shortly after the time of [3H]thymidine injection, indicated that the inner two thirds of the neocortex is laid down along a ventro-dorsal gradient, i.e. the lateral neocortex starts to form before the dorsal; and that cells born at a particular time lie in cortical layer VI at the dorsal edge of the gradient is traced ventrally. Progressively more weakly labelled cells formed intermediate steps in this migration. A model or cortical growth fitting these findings is presented. Some inferences are also made about the possible role of the ganglionic eminences in providing cortical cells, at least during the initial stages of cortical histogenesis.


Survey of finfish and shellfish for volatile N-nitrosamines.

A variety of finfish and shellfish obtained on the local retail market were analyzed for 14 volatile N-nitrosamines by using a gas-liquid chromatographic method demonstrated to be sensitive to 10 ppb (microng/kg). A total of 78 samples (fresh, frozen, and canned), representing 26 varieties of fish, were analyzed. No nitrosamines were found.


Direct plating versus dilution plating in qualitatively determining the mold flora of dried beans and soybeans.

Two methods presently used for examining whole foods and feeds for viable molds were evaluated for their relative effectiveness in the qualitative determination of the total number of mold species present in soybeans and dried beans: the direct plating method and the serial dilution method. Sixty-nine soybean samples and 40 dried bean samples were examined. Although the quantitative results of this study were inconclusive, the qualitative results show that the direct plating method was substantially more effective in detecting individual mold species. An average of 12.9 and 10.9 species was detected by the direct plating method in whole soybean and dried bean samples, respectively. An average of 4.4 and 2.8 species was detected by the dilution method in ground soybean and dried bean samples, respectively. A total of at least 37 mold species were found in the study, including 10 toxicogenic species. With few exceptions, detection rates of the 37 individual species were substantially greater among the samples examined by direct plating than those examined by serial dilution.


Negative chemotaxis in cellular slime molds.

This study confirms the suggestion of earlier workers that the vegetative amoebae of Dictyostelium repel each other while those of Polysphondylium violaceum do not. When Dictyostelium amoebae were placed in drops on thin and thick agar, the cells moved out faster on the thin agar, presumably because the repellent was more concentrated. This did not occur with Polysphondylium amoebae. Also, if 2 drops of cells were placed side by side, or a single drop was placed near an edge, in Dictyostelium there were fewer cells emerging between the drops (or near an edge) than on the far side. Polysphondylium showed no such difference. However, Polysphondylium amoebae were repelled by Dictyostelium cells (but not vice versa) when drops of each were placed beside one another. Finally, if Dictyostelium discoideum cells were placed in drops over thick and thin agar, but separated from the agar by a dialysis membrane, the cells again spread farther on the thin agar, indicating that the repellent is a dialyzable molecule.


Kinetic studies dealing with immobilized reversible enzyme system. Experimental evidence for shift of apparent equilibrium constant within bienzyme system.

Two chemostat methods for studying the immobilized enzyme kinetics are presented. The methods are applied to glucose-6-phosphate isomerase immobilized within a proteic membrane. The first method is based on the steady state behavior of a Continuously Stirred Tank Reactor (C.S.T.R.) and the second one on a diffusion-reaction process. The apparent equilibrium ratio between glucose 6-phosphate and fructose 6-phosphate is measured for the immobilized enzyme system. No modification is observed with a monoenzyme membrane. In the presence of a glucose-6-phosphate dehydrogenase activity the product of the first reaction is trapped by the second one and due to the local intramembrane concentrations an apparent modification of the equilibrium ratio is observed. The apparent modification is studied with the bienzyme membrane as a function of the concentration of NADP co-substrate of the second enzyme. A "Michaelian" relationship is observed.


Synthesis of myelin glycosphingolipids (galactosylceramide and galactosyl(3-O-sulfate)ceramide (sulfatide)) by cloned cell lines derived from mouse neurotumors.

Clonal cell lines derived from both spontaneous and chemically induced rat and mouse brain tumors were screened for their ability to incorporate H232SO4 into galactosyl(3-O-sulfate)ceramide (sulfatide). High levels of 35SO4 incorporation into sulfatide were found only in two of the mouse cell lines studied (G26-20 and -24). Tumors produced by subcutaneous injection of these cell lines into C57BL/6 mice were also unique in that they contained high levels of both sulfatide and galactosylceramide. The synthesis of large amounts of sulfatide and galactosylceramide by a clonal cell line of neurological origin suggests that the original tumor was of oligodendrocyte or Schwann cell origin. In common with a large number of mouse and rat astrocyte cell strains and their derived tumors, these glial cells lacked the ability to synthesize gangliosides such as monosialotetraglycosylceramide and disialotetraglycosylceramide (as judged by analytical and [3H]GlcNH2 incorporation studies). This appears to be a unique characteristic of neuroblastoma-derived cell strains such as N18, NB2a, and NB41A.


Immobilization of enzymes on slowly soluble carriers.

The preparation and some properties of microspheres composed of oxidized polysaccharides and some vinyl polymers are described. The microspheres contain immobilized enzyme and can be slowly solubilized in water solutions, thereby releasing active a enzyme into the surrounding medium. The kinetic characteristics of the immobilized enzyme bound with a fragment of matrix after complete solubilization are unchanged, but the enzyme exhibits high thermostability. These preparations could have a wide range of medical applications, e.g., to form a drug "depot" directly in an affected organ.


Microtubule and microfilament rearrangements during capping of concanavalin A receptors on cultured ovarian granulosa cells.

Thin-section electron microscope analysis of rat and rabbit-cultured granulosa cells treated with concanavalin A (Con A) at 37 degrees C revealed coordinated changes in the cytoplasmic disposition of microfilaments, thick filaments, and microtubules during cap formation and internalization of lectin-receptor complexes. Con A-receptor clustering is accompanied by an accumulation of subplasmalemmal microfilaments which assemble into a loosely woven ring as patches of receptor move centrally on the cell surface. Periodic densities appear in the microfilament ring which becomes reduced in diameter as patches coalesce to form a single central cap. Microtubules and thick filaments emerge associated with the capped membrane. Capping is followed by endocytosis of the con A-receptor complexes. During this process, the microfilament ring is displaced basally into the cytoplasm and endocytic vesicles are transported to the paranuclear Golgi complex along microtubules and thick filaments. Eventually, these vesicles aggregate near the cell center where they are embedded in a dense meshwork of thick filaments. Freeze-fracture analysis of Con A-capped granulosa cells revealed no alteration in the arrangement of peripheral intramembrane particles but large, smooth domains were conspicuous in the capped region of the plasma membrane. The data are discussed with reference to the participation of microtubules and microfilaments in the capping process.


Muscle actin filaments bind pituitary secretory granules in vitro.

Hog anterior pituitary secretory granules sediment at 3,000 g. When rat or rabbit skeletal muscle actin filaments are present with the granules, the sedimentation decreases markedly. Depolymerized actin or viscous solutions of Ficoll and collagen have no effect on granule sedimentation. With this assay, actin filaments bind secretory granules (consisting of the proteinaceous core plus limiting membrane), secretory granule membranes, mitochondria, artificial lecithin liposomes, and styrene-butadiene microspheres, but have little or no interaction with membrane-free secretory granule cores and albumin microspheres. A secretory granule-actin complex sedimentable between 3,000 g and 25,000 g can be isolated. Metal ions, nucleotides, salts, dithiothreitol, or pretreatment of the granules with trypsin do not destroy the binding, which appears to be a lipophilic interaction.