Intensive therapy for hepatic coma.

Intensive therapy for hepatic coma. Of 45 patients observed in the ICU with severe acute hepatic insufficiency, 15 patients were in hepatic coma. All patients received combined treatment consisting of standard conservative methods (drug therapy) and surgical methods for temporary support of liver function (hemodialysis, exchange blood transfusion, pig liver perfusion). Intensive therapy which began during the early phase of hepatic coma enabled us to bring six patients out of the coma, four of whom completely recovered. These studies showed that of the diseases causing hepatic coma, the worst results were obtained with viral hepatitis. This may be explained both by the extensive liver damage and marked metabolic disturbances, which led to failure of other vital organs and systems. At present, the combined therapy contributes to a greater percentage of recovery of patients with hepatic failure and coma.


Circular ribosomal DNA and ribosomal DNA: replication in somatic amphibian cells. Pulse labelled rDNA from cultured somatic cells of Xenopus laevis was examined by electron microscope autoradiography. The pattern of replication closely resembles that of bulk chromosomal DNA and differs considerably from rDNA synthesis during amplification in the oocyte. - About 0.15% of the rDNA molecules in the purified preparations were circular. The presence of interlocked circles of equal size indicates that the circles are not in vitro cyclization artefacts, but may represent free rRNA genes. A low frequency of circles was also seen in Xenopus blood rDNA. Their stability in high concentration of formamide suggests that they too did not arise after DNA extraction.


X-ray induced visible alterations in the giant chromosomes of Phryne cincta (Nematocera, Diptera): relation of radiation sensitivity to pronuclear chromosome structure. In order to induce chromosomal rearrangements, males were exposed to x-rays and then mated to non-irradiated females. The number of each type of structural alteration was determined by examination of the polytene chromosomes of the F1 progeny. -- A comparison of the results with similar studies made on Drosophila revealed a significantly greater sensitivity in Phryne. Parallel to that an extremely high frequency of small inversions was ascertained in Phryne, and the observed ratio of inversions to translocations was the inverse of that which would be expected from purely mathematical considerations based on the lengths of the different chromosomes. These facts allow the conclusion that the paternal pronuclear chromosomes in Phryne are highly spiralized. Besides, the kinetochore-to-translocation-breakpoint distance was measured in both of the chromosomes involved in each reciprocal translocation and the differences (kinetochore-break distance differences) were registered and from them the arrangement of the chromosomes in the pronucleus of Phryne deduced. The data obtained support the assumption of an ordered, polar-field type of orientation. In Drosophila, in contrast, the comparable data showed that the pronuclear chromosomes are not spiralized and are randomly arranged (Bauer, 1939). -- These results seem to indicate that a close correlation exists between the different radiation sensitivities of Drosophila and Phryne and the different states of spiralisation and arrangements of their chromosomes in the pronucleus stage. It is hypothesized that the influence of the maternal genome on the degree of spiralization of the paternal chromosomes could account for differences in the pronuclear chromosome structure of both species.


[Allergies to animal hair in diseases of the upper respiratory tract (author's transl)]. Among 214 patients diagnosed in 1972 as being allergic to animal hair, 79 (48 women) were re-examined later. In almost all cases with a positive personal (33) or family history (26) of allergy there was a particularly rapid sensitization. The largest proportion of pathogenic allergens was from guinea pigs (39), cats (17), golden hamsters (11), rabbits (6), horses, dogs and budgerigars (2 each). 14 patients reacted to the main antigen alone in prick or intracutaneous tests, while in all other cases there were cross-reactions, most frequently to epithelium of sheep, cattle or horse. In interpreting intranasal provocation tests with the chief antigen, the degress of rhinorrhoea was particulary reliable (59 subjects). The most effective treatment was complete withdrawal of allergens: of 65 persons who removed their domestic pet or avoided contact with it, 32 were cured at once and a further 19 within about eight weeks. 14 subjects continued to have symptoms, but in half of these there was also house dust allergy, while others had resumed contact with domestic animals.


Cholinergic control of the nocturnal prolactin surge in the pseudopregnant rat. The possible involvement of cholinergic mechanisms in the control of rhythmic secretion of prolactin has been examined in the pseudopregnant rat. Baseline data were obtained in decapitated animals in which the diuranl surge was observed in the 1430--2030 h range and the nocturnal surge during the 2330-0530 h interval. Atrial cannulation permitted a faithful reproduction of the prolactin pattern seen in decapitated rats if at least 3 days elapsed between the cannulation operation and bleeding, while cardiac puncture, under light ether anesthesia, appeared to suppress the diurnal surge. This latter observation appeared to depend on the time of sampling in relation to the onset of the dark phase of the daily lighting cycle. Atropine (35 or 70 mg/100 g BW) was found to inhibit the nocturnal surge in animals bled by either the cardiac puncture or cannulation techniques --an effect which was reversed by pretreatment with eserine (50 or 100 mug/100 g BW). Nicotine (0.75 mg/100 g BW) was found to delay, but not completely to inhibit, the nocturnal surge. When evaluated in the light of other available information, these observations suggest that a complex cholinergic mechanism is involved in control of the nocturnal rhythm of prolactin in pseudopregnancy and that the two surges, diurnal and nocturnal, are differentially controlled.


The differential effects of diestrous progestogen administration of proestrous gonadotrophin levels. Progesterone or d-norgestrel, a totally synthetic progestogen, administered subcutaneously at 1330 h on diestrus (Day 2) of 4-day cyclic rats, inhibited ovulation and increased the vaginal cycle length in a dose-related manner. d-norgestrel was at least 5 times as potent as inhibitor of ovulation as progesterone. The dose-related inhibition of ovulation was directly related to suppression of the proestrous serum LH surge. Proestrous serum FSH levels were not depressed at the minimum dose of d-norgestrel that produced both a 100% reduction in ovulation (MED100) and a significant decrease in proestrous serum LH. However, progesterone produced a significant decrease in proestrous serum FSH at its anti-ovulatory MED100. Progesterone and d-norgestrel were equipotent (100 mug/100 g BW) with respect to significant suppression of proestrous serum FSH levels. Follicular growth was retarded, but only at the higher doses of either progestogen which suppressed FSH and LH. These data suggest that a) the increase in acute anti-ovulatory potency of a synthetic non-estrogenic progestogen over progesterone lies in its ability to reduced selectively serum LH levels at low doses, b) the progestational block of ovulation takes place via the hypothalamic-pituitary axis and not the ovary, and c) the retardation of follicular growth that accompanies ovulatory inhibition after diestrous administration of high doses of a progestogen takes place only when both serum FSH and LH are significantly reduced.


Sequential patterns of circulating luteinizing hormone and follicle-stimulating hormone in female sheep from early postnatal life through the first estrous cycles. Patterns of circulating LH and FSH were examined during several 6-h periods in developing female sheep from 3 weeks of age through the first estrous cycles (30--40 weeks of age) and early pregnancy. With the onset of pulsatile LH secretion, beginning 11 weeks after birth, circulating LH increased to levels 2- to 5-fold greater than those observed in the adult. Thereafter, high and variable levels of circulating LH persisted through young adulthood when ovarian cyclicity including ovulation became manifest. Although prior to the initial estrus changes in frequency of LH release (number of LH discharges/6-h period) appeared to occur randomly from week to week, after first estrus, changes in frequency became predictable. During the early (day 1) and late (day 15) luteal phases of the first estrous cycles, when circulating progesterone was low (less than 0.6 ng/ml), the frequency of LH release was increased (5--7 discharges/6 h) while during the mid-luteal phase (day 7--12), when circulating progesterone was high (2--4 ng/ml), the frequency was diminished (0--2 discharges/6 h). Massive and sustained discharges of LH which resembled preovulatory surges were observed only shortly before first estrus. In contrast to the pulsatile release pattern of LH, concentrations of circulating FSH within the 6-h periods were relatively constant and within the range found in the adult. These findings lead to the following conclusions: a) changes in negative feedback control are not directly responsible for the onset of ovarian cyclicity in the sheep as indicated by the lack of differences in mean concentrations of circulating LH and FSH before and after first ovulation; b) the onset of the surge mode of gonadotropin secretion occurs only shortly before first ovulation; and c) progesterone may play an inhibitory role in regulating tonic LH secretion during the first estrous cycles.


Localization and mechanism of stimulatory feedback action of estrogen: effect of limbic forebrain implantation of estradiol benzoate on advancement of ovulation. The sites and mechanism of the ovulation-inducing action of estradiol benzoate (EB) were studied by brain implantation of the crystalline steroid through chronically implanted outer cannula at 12:00 on diestrus day 2 in the 5-day cyclic rat. EB implantation in the medial amygdala or the bed nucleus of stria terminalis advanced cyclic changes in vaginal smears, timing of ovulatory LH release, and ovulation by 1 day, resulting in 4-day cycle. When implants in the bed nucleus of stria terminalis were placed for a shorter period of time on diestrus day 2, from 12:00 to 20:00, advancement of these parameters were similarly observed. Serum concentration of FSH and that of prolactin were significantly elevated at 20:00 on the same day in the rats implanted with EB in the medial amygdala or the bed nucleus of stria terminalis, compared with those in the non-treated controls. LH was not affected. The implantation in the arcuate nucleus was also effective to advance ovulation, but the anterior deafferentation prevented the effect. In contrast, EB implantation in the medial septal nucleus, the medial preoptic area, or the medial basal prechiasmatic area was consistently ineffective to advance vaginal cycle and ovulation. Multiunit activity in the arcuate nucleus showed an afternoon elevation on the day of implantation in these areas and as well on the day following, while it did not show such elevation on the day of implantation in the medial preoptic area. It is concluded that EB acts on the medial amygdala and the bed nucleus of stria terminalis in the mid-diestrus in 5-day cycle to stimulate FSH and prolactin release without affecting LH, which changes trigger a chain of reproductive events inducing early release of ovarian steroid responsible for early ovulatory gonadotropin release. The arcuate nucleus in one of the sites of stimulatory action of estrogen, but it requires the neural influence presumably from the medial amygdala and the bed nucleus of stria terminalis via the preoptic area for stimulating the ovulatory hormone release. EB exposure is considered to be endowed with the increase of its responsibility to this neural influence.


Mechanism of action of a new toxin from gonyaulax tamarensis on nerve membranes. The toxin from the dinoflagellate Gonyaulax tamarensis blocks nervous conduction through a selective inhibition of the mechanism whereby the membrane undergoes an increase in permeability to sodium ions. The effect is exerted only from outside of the nerve membrane. These effects are exactly the same as those exerted by tetrodotoxin or saxitoxin.


Lack of secondary intoxification by red tide poison in the American lobster Homarus americanus. Lobsters are able to feed on shellfish which are toxic with PSP (paralytic shellfish poisoning from Gonyaulax tamarensis) with no apparent harm to themselves, and no measurable assimilation of the poison into their tissues. Lobsters consumed food containing in excess of 1000 mug PSP. There was no impairment of respiration (oxygen consumption) measurable two to three hours after feeding, and no PSP measured in the meat of the claws and tail 48 to 120 hours after feeding. The only PSP was in the guts and contents which were measured 48 hours after feeding began.


Absorption of radioactive lead (210Pb) by different parts of the intestine in young and adult rats. Absorption of radioactive lead (210Pb) was studied in an intestinal preparation from adult and young rats. Absorption was higher in the jejunal parts than in the duodenum or colon. Moreover, absorption in young rats was greater than in adults. No difference in absorption was found after loading with large amounts of lead. Maintaining the animals on a low calcium diet decreased rather than increased lead absorption.


[Latent infection with Corynebacterium kutscheri in mice after peroral inoculation and its provocation by cortisone (author's transl)]. Latent infection with Corynebacterium kutscheri in mice and its provocation by cortisone were studied with a rifampicin-resistant strain of the organism. Mice having been infected perorally began to excrete the organisms in feces within 6 hours, and most of them were found to be carrying the organisms in the intestine, especially in the cecum even 90 days after infection. In such state of latency, however, no organisms were detected in other main organs, and neither visible lesions nor serum agglutinin was detectable. The latent infection with excretion of the organisms in feces after peroral infection was shown to become overt and fatal by cortisone treatment made even 90 days after infection. In infected mice excreting no organisms in feces and having bites on their skin, the wounds became severe ulcers after cortisone treatment resulting in septicemia.


[Genetic relationship of two mutant genes which producing three different syndromes in the mouse (author's transl)]. We determined that leaner gene (la) is located in the linkage group XVIII and closely linked to Es-1, which is known to be located closely to tottering gene (tg). Double heterozygote (la/tg) produced by mating between la heterozygote and tg heterozygote showed an intermediate syndrome between those seen in tottering (tg/tg) and leaner (la/la) mice. Both leaner and tottering mice showed neuromuscular disorders, but their clinical and pathological characteristics were different. Leaner mice were found to represent a so-called cerebellar mutant having the reduced size of cerebellum and severe cytoarchitectonic abnormalities with focal losses of Purkinje and granular layer cells. Tottering was, however, another mutation having epileptiform seizures, and it was characterized pathologically by cellular losses and shrinkage as well as vesiculations of cytoplasmic membranous structures in the cerebellum. The double heterozygote was shown to have both pathologic characteristics seen in each homozygote, and also showed shrinkage of Purkinje cells and vesiculation of the endoplasmic reticulum and Golgi apparatus. These clinical and pathological findings supported the genetic data suggesting that la and tg constitute an allele.


[Sterilization of diets and drinking water in the autoclave in the Laboratory Animal Center, Nippon Veterinary and Zootechnical College (author's transl)]. Efficiency of sterilization was shown to vary according to the type of autoclave, and this would be taken into consideration for preparing sterilized diet and water for laboratory animals. It seems necessary to sterilize diet and water for 60 and 150 minutes, respectively, at 120 degrees C using the autoclave in the Laboratory Animal Center of the Nippon Veterinary and Zootechnical Collge.


[A few characteristics of mouse having high sensibility to anaphylatic shock separated from El mouse (author's transl)]. During an experiment of shaking in El mice, a strain which was less susceptible to convulsion was found out. In this strain (ASK), the mortality to anaphylactic shock and susceptibility to audiogenic seizure were compared with five other strains of mice. Six strains of mice (ASK, EL, IDT, JCC-ICR, dd, C57BL/6J) were sensitized by two subcutaneous injections with 2 mg/head of crystalline egg albumin at five and six weeks of age, and then challenged by the intravenous injection of 0.125, 1, 8, 64, 512 and 4096 mug/head at seven weeks of age. In both sexes of ASK strain, the mortality was the maximum (75--95%) after the challenge of 8--4096 mug egg albumin. The mortality of the female El and IDT and the both sexes of JCL-ICR strains was middle (55--70%) after the challenge with 64 mug egg albumin, while that of other strains (dd and C57BL/6J) was very low (0--20%). The susceptibility of 3-weeks-old mice of ASK, El, IDT, JCL-ICR and dd strains was low (1.9--19.0%), whereas mice of DBA/2 strain showed a very high susceptibility (80%) to 110 phons.


A heritable hyperlipemic rabbit.

A heritable hyperlipemic rabbit. In the course of study on serous cholesterol of rabbit, we found a male mutant with remarkably high cholesterol level (HLR). The characteristics of this mutant could be described as follows: (1) Serous chemical determinations for the mutant's serum revealed extremely high levels of total lipids, and beta-lipoprotein as total cholesterol, being 3011 mg/dl, 10.6 units and 447 mg/dl respectively. High cholesterol level continued these two years during observation, though some fluctuation has been observed. (2) Such hypercholesterolemia was heritable. Determinative gene for hypercholesterolemia seemed to be recessive and not sex-linked, because, while F1 generation did not show so high cholesterolemia, one female rabbit among twelve off-springs born by full-sib mating showed persisting hypercholesterolemia similar to original HLR. (3) HLR may serve as a model of human familial hyperlipemia Type II or III according to the classification by Fredrickson et al.


[Effects of creatinol O-phosphate on the isolated rabbit's atrium and on the rat's phrenic nerve diaphragm preparation]. Creatinol O-phosphate (COP) enhanced the semiisometric contraction of isolated rabbit's atria and those of rat's phrenic - diaphragm preparations immersed in hypocalcic Ringer or Krebs solution. Creatine-phosphate elicited a similar effect.


Parasexual genetic analysis of the cellular slime mold Dictyostelium discoideum A3. Haploid strain A3 of the cellular slime mold Dictyostelium discoideum is valuable for biochemical studies because it is capable of axenic growth. Mutants of A3 temperature-sensitive for growth and resistant to the drugs cycloheximide, acriflavin, or methanol were isolated.--Heterozygous diploid recombinants, formed at low frequency by cell and nuclear fusion, were isolated by selecting temperature-resistant progeny of mixed cultures of two nonallelic temperature-sensitive haploids (LOOMIS 1969). Each drug-resistant mutation was found to be recessive. Two independently isolated methanol-resistant mutants were in one complementation group.--Diploids of A3 heterozygous for drug resistance formed drug-resistant segregants with a frequency of approximately 10(-4). Segregants selected for resistance to a single drug were either haploid or diploid; the fraction which was haploid varied from 0.11 to 0.86, depending on the selected marker. Segregants selected for resistance to two or three drugs were almost all haploid.--Using this parasexual cycle of diploid formation and haploidization, linkage of these temperature-sensitive and drug-resistance mutations to each other and to mutations studied by KATZ and SUSSMAN (1972) and by WILLIAMS, KESSIN and Newell (1974b) was analyzed. The methanol-resistant mutants were found to be partially resistant to acriflavin, and unlinked to the mutant selected for acriflavin resistance, which was methanol-sensitive. Of the expected seven linkage groups in D. discoideum, five, and a possible sixth, have been marked.--Linkage analysis of a mutant abnormal in morphogenesis showed that its phenotype results from two unlinked chromosomal mutations.


Mast cells and immunoglobulin E in inflammatory bowel disease. The numbers of mast cells and of IgE-containing immunocytes in the bowel wall of patients suffering from Crohn's disease of ulcerative colitis have been estimated and the results compared with those found in normal control specimens. In ulcerative colitis there is a slight rise in the number of mast cells and it appears that these participate in the inflammatory process in a non-specific manner; the number of IgE-containing immunocytes is not significantly altered. In Crohn's disease there is an almost total absence of stainable mast cells in affected areas of the bowel, together with a marked decrease in IgE-containing immunocytes. It is suggested that these findings are due to degranulation of mast cells and consumption of IgE as a result of an immediate hypersensitivity reaction in the bowel wall, this being one component of the protein inflammatory and immunological response to the entry of a variety of antigenic material.


Radioiodination of L 1210 cells.

Radioiodination of L 1210 cells. The lymphoid leukaemia L 1210 cells of mice were labelled with 125I. The cell homogenates were fractionated and from the microsomal fraction 90 per cent of the radioactive material could be precipitated with perchloric acid, whereas only 4 per cent was precipitated from the soluble fraction. Papain bound with Enzacryl AH released 31 per cent of radioactivity. It was concluded therefrom that the surface proteins of the cells were labelled. Electrophoretic separation of these proteins in polyacrylamide gel with sodium dodecyl sulphate was performed and 6--8 radioactive fractions of surface peptides were found.


Colonic pseudo-obstruction: a new complication of jejunoileal bypass. Five female patients ranging in age from 25 to 44 years are reported in whom jejunoileal bypass (three end-to-side and two end-to end), performed for morbid obesity, was complicated 1 1/2 to three years later by symptoms of colonic pseudo-obstruction. In each size, the colon was markedly elongated, dilated, and atonic but with no demonstrate organic obstruction. The cause of this complication is not known. Full thickness rectal biopsy in one case showed normal intrinsic nervous plexuses and ganglia. Serum electrolytes were normal. Functional and defunctionalized small bowel were not involved. Symptoms varied from complete colonic paralysis to incapacitating crampy abdominal pain and distention. In the three patients with end-to-side bypass, dilatation affected the entire colon, while, in the two patients with end-to-end bypass, the dilatation was localized to colon distal to the anastomosis with the defunctionalized small bowel. Resection of the affected portion of colon in one case resulted in recurrence distal to the new site of drainage of defunctioned bowel. Treatment with anti-anaerobe antibiotics in two cases produced dramatic but temporary relief of symptoms.


The role of polecats of the genus Putorius Cuvier, 1817 in natural foci of adiaspiromycosis. Adiaspiromycosis provoked by the fungus Emmonsia crescens was diagnosed in two different species of polecats. In total, 76.2% of steppe polecats (Putorius eversmanni) and 35.6% of dark polecats (P. putorius) were infected. The infection was not demonstrated at all in ferrets (P. furo) reared as fur animals. The differences found in the extent and intensity of infection in the two species of wild polecats are explained by different ecological requirements of these related carnivores. The habitat of the exoanthropic P. eversmanni offers more opportunity for a contact with the pathoergont than that of P. putorius; the latter may occasionally pass over to a pronouncedly hemisynanthropic way of life. The synanthropy of the hosts probably has a negative influence on the life cycle of the fungus. Although the inhalation of aleuriospores is the main route of infection, the ingestion of adiaspores also plays an important role. From this aspect are also evaluated the trophic relationships between mustelid carnivores and small rodents: the predatory species gets into contact with the adiasporic phase of the pathoergont localized in the lungs of wild rodents. The extremely high extent and intensity of infection by the elements of E. crescens in polecats probably results from the cumulation of both routes of infection.


[Effect of alfalfa meal on experimental hyperlipidemia]. The influence of alfalfa meal on hyperlipidemia induced by dietary cholesterol was examined and changes in serum total cholesterol (TC), triglycerides (TG) and non-esterified fatty acid (NEFA) in rabbits were recorded. Serum lipid levels of groups treated with alfalfa meal (R-1) as well as those not treated (R-2) were found to be elevated. TC of R-1 was lower than that of R-2. The inhibition effect of alfalfa meal on elevation of TC was apparent to a certain extent. The inhibition effect of alfalfa meal on elevation of TG and NEFA was greater than that of TC, thus it is suggested that alfalfa meal can be successfully utilized for experiments with lipids, when the rabbit is the experimental animal of choice. Effect of riboflavine-2,3,4,5-tetranicotinate (RN-4) on changes of TC, TG and NEFA in rats fed 1% cholesterol diet was also examined. RN-4 was mixed in the diet (0.5, 0.25 and 0.125%). TG levels in groups treated with RN-4 (Rt-2, Rt-3 and Rt-4) were lower than previously observed.


[Morphine dependence and preference. (I). Morphine preference in naive and morphine-experienced rats]. The spontnaeous morphine intake ratio (M-SIR) under free access to morphine-admixed food and quinine-admixed food conditions was measured for 3 weeks in naive and morphine-experienced rats. In the case of morphine (0.5 mg/g of food) vs. quinine (0.5 mg/g of food), naive rats gradually increased M-SIR from 17% to 77%. Using a higher level of morphine- and quinine-admixed food (1 mg/g vs. 1 mg/g of food), M-SIR was more rapidly increased than that in the lower group. Thus while on the 10 approximately 60 mg/kg/day dose range, the M-SIR was gradually increased dose dependently in naive rats due mainly to the positive reinforcing properties of morphine. Morphine-experienced rats showed a significant increase in M-SIR for the first 4 days specifically as compared with naive rats. Morphine dependent rats thus obtained morphine in sufficient amounts to maintain dependent states only after the first 2 approximately 3 days. This choice behavior revealed the psychological aspects of morphine dependence in rats and the preference for morphine was also observed after withdrawal for more than 2 weeks as secondary abstinence syndrome.


[Characteristics of aggressive behavior induced by chronic administration of theophylline in rats]. In previous studies we reported that chronic administration of theophylline induced affective aggressive behavior in rats. The present study was designed to clarify characteristics of the aggressive behavior observed, and compare it with that in the olfactory bulbectomized and isolated rats. To measure aggressive behavior, the reflexive fighting test, the test for killing response to various objects and the aggressiveness rating test were used, and in addition, the open field test was employed to objectively measure emotionality. It was only the theophyllinized rats that assumed the fighting position not only during the electric shock period but also during the adaptation and sound shock periods. The theophyllinized pairs fought quicker and more frequently than did the other treated pairs. During a 3 min observation the percentage of mouse-killers was much higher among the theophyllinized group (62.5%) than among the other treated groups (the olfactory bulbectomized and the isolated groups increased mouse killing to 87.8% and 62.5%, respectively, though the theophyllinized group remained at 62.5%. The theophyllinized killers attacked constantly rat-pups and dead as well as live mice. In the open field test, rearing and ambulatory activities increased more in the olfactory bulbectomized group and decreased more in the isolated group than in controls, but no change was observed in the theophyllinized group. Defecation which has a negative correlation with aggressiveness decreased more only in the theophyllinized group than in the controls. Prior to treatment there was a positive correlation between the frequency of ambulation and that of rearing (r = 0.535, n=32, p less than 0.01).


[Effect of carbon tetrachloride on biliary excretion of 3,4-benzpyrene in rats]. 14C-labelled 3,4-benzpyrene (14C-BP) in dose of 100 mug/animal was injected into female Sprague-Dawley rats under urethane anesthesia. The rats were cannulated at the duct and the output of metabolites in the bile was determined. Non-treated rats excreted 35.5% of the given dose within one hr. The main metabolites excreted in bile were fractionated chromatographically into three. One of these was a metabolite which was hydrolyzed by beta-glucuronidase, the other two were unknown substances. Unchanged BP was not detected. In rats pretreated with CCl4 (0.5 ml/kg, p.o.) 24 hr before experiment, the biliary excretion of BP-metabolites was found to be reduced, that is only 10% of the given dose was excreted within one hr. It was noted that the output within the first 90 min after injection of BP was significantly reduced. In the CCl4-treated rats, bile flow was found to be lowered. However, the reduced output of BP-metabolites was considered not as the result of lowered bile flow, but as the result of disturbance of BP-metabolism in the liver. Thus it is suggested that impaired hepatic junction as the result of CCl4 intoxication may last for more than two weeks, when the hepatic junction is evaluated in terms of biliary excretion of BP.


[Pharmacological analysis on the mode of action of mandelamidine, a long-acting hypotensive agent (1). Comparison with guanethidine]. MA produced a temporary and long lasting hypotension in anesthetized animals. MA like guanethindine caused relaxation of the nictitating membrane in unanesthetized cats and inhibited the contraction of the nictitating membrane elicited by electrical stimulation at various frequencies of preganglionic cervical sympathetic nerve in anesthetized cats. The mode of neuron blocking action of MA differed from guanethidine on the contraction of the nictitating membrane elicited by stimulation at high frequencies. Neuron blocking action of MA was also shown on rat inferior eyelid, dog carotid occlusion, rabbit jejunum and guinea-pig vas deferens. Moreover that of MA like guanethidine on the cat nictitating membrane was reversed by a following administration of methamphetamine. MA slightly potentiated the effects of norepinephrine, epinephrine and reduced the effects of tyramine on the responses of the cat nictitating membrane and blood pressure, but did not potentiate the effects of norepinephrine on the guinea-pig vas deferens and rabbit pulmonary artery. MA produced a contraction of the cat nictitating membrane and this contraction was inhibited by pretreatment with reserpine or phentolamine. However, MA did not produce a rise in blood pressure in spinal cats nor did it reduce the contractile force and heart rate in guinea-pig atria. These findings suggest that sympathomimetic action and uptake inhibition of the catecholamine of MA was weaker than that of guanethidine.


[Phamacological studies on bamboo grass (1). Acute toxicity and anti-inflammatory and antiulcerogenic activities of water-soluble fraction(Folin) extracted from Sasa albomarginata Makino et Shibata]. Folin showed no significant toxic effects in mice. The tail pressure test in mice revealed a potent analgesic effect with a hypothermic effect in rats. A significant decrease was also seen in rat dextran or carrageenin edema. There were no inhibitory effects on the formation of granulation tissue in rats. In addition, Folin administration revealed a tendency to a decrease in pylorus-ligated ulcer of the rat by inhibiting the gastric secretion. Some protective effects from aspirin or caffeine-induced gastric ulcers of the pylorus-ligated rats were also obtained in this fraction.


[Effects of penfluridol, a psychotropic agent, on operant behavior of rats]. Effects of penfluridol, a diphenylbutylpiperidine type psychotropic agent, on operant behavior were investigated and compared with those of chlorpromazine and haloperidol in rats trained on the following 5 schedules. Fixed ratio (FR 30) of food reinforcement and differential water reinforcement of low rate (DRL 20 sec) schedules were used for positively reinforced behaviors. Continuous (Sidman-type) and discriminated avoidance schedules were used for negatively reinforced behaviors. Conditioned suppression of response (CER) under FR 30 developed by stimulus presentation with electric shock was also applied. When 1 mg/kg of penfluridol was given orally, no change was observed in respondings of all the performances. At higher doses (2 approximately 8 mg/kg, p.o.), the respondings were inhibited in proportion to the dosage except in DRL performance, in which only correct response rate decreased at 8 mg/kg. These inhibitory effects were observed more apparently in the negatively reinforced behaviors than in the positively reinforced ones. Furthermore, a clear dose-effect relationship was obtained in the former. CER was not all attenuated by 2 approximately 8 mg/kg of penfluridol, thus a diazepam-like effect was not confirmed. These behavioral effects suggested that penfluridol has neuroleptic properties similar to those observed with chlorpromazine or haloperidol. However, in general, the inhibitory effects reached the maximum level approximately at 16 hr and lasted for 2 approximately 3 days after oral administration of the drug. Intensity of the effect of penfluridol was estimated to be about 1/8 approximately 1/10 that of haloperidol, according to the results obtained in the avoidance performances.


[Application of group psychology of animals into pharmacological studies. 12. Specificities of actions of psychotropic drugs on emotional and exploratory behaviors of a single mouse or groups of 2 or 3 animals]. The exploratory movement recorder of Tokyo University of Education type was utilized to assess the exploratory motor activity of groups of 1, 2 and 3 mice of the dd-strain. As the animals were observed during 15 min periods to move spontaneously, psychotropic drugs were administered and the specific actions of these agents were determined. Each drug was given i.p. in a dose which proved non-effective in a single mouse. Observations were as follows: The ratio of exploratory motor activity of each pair of mice as compared to a single mouse was not always different from that of control animals. When a 3rd mouse was added to the pair, the motor activity either increased or decreased considerably. The rate of increase in motor activity was calculated 45 min after caffeine, pentetrazol, ephedrine and DL-amphetamine and was found to be less than in the control animals. After administration of chlordiazepoxide, diazepam, imipramine, chlorpromaxine or mescaline, the rate of increase in motor activity was considerably greater than in the controls. With dimorphoramine or hexobarbital, the rate was much the same as control. After beta-oxy gamma-aminobutyric acid, the rate was slightly increased and with diphenhydramine the rate was slightly less than control. Thus the exploratory motor activity of 3 mice as compared to that of 2, decreased considerably after administration of psychotropic stimulants, while after psychotropic depressants, an even greater increase was observed. We have already reported on the mutual relation of mice in fighting episodes and E1-mice convulsions. Those relations were, however, responses elicited by the physical stimulation. In contrast, the mutual relation in this paper is more of a natural nature. In view of observations in this paper, it is considered that CNS stimulants increase the mutual, emotional, checking behavior in groups of 3 animals while CNS depressants decrease the checking behavior. Such behavior is one example of the emotional conduct when a 3rd party is added to a 2 member group.


[Analysis of the mode of action of ifenprodil on cerebral and peripheral circulation in dogs]. Pharmacological properties of ifenprodil, a vasodilating agent similar in chemical structure to isoxsuprine were investigated. In anesthetized dogs and cats, ifenprodil (0.3 approximately 1 mg/kg, i.v.) caused a temporary prominent depressor response with an increment in heart rate and an occasional enlargement of pulse pressure followed by a mildly durable hypotension. These two phases of hypotension of ifenprodil could not be demonstrated in spinal rats and cats. Ifenprodil also decreased the pressor response of norepinephrine and reversed the pressor effect of epinephrine to a depressor one. The lowering effect of ifenprodil on the constant-rate perfused pressure of vertebral artery in dogs was half as potent as isoxsuprine. Ifenprodil significantly reduced the contraction to norepinephrine in the isolated guinea-pig vas deferens. This was greater than the constrictor response produced by preganglionic sympathetic nerve stimulation. The inhibitory effect of ifenprodil was less potent than that of phentolamine on the responses to norepinephrine in isolated rabbit ascending aorta strips. In the isolated guinea-pig right atrium, ifenprodil (10(-5) g/ml) showed slightly negative inotropic and chronotropic effects, and reduced the responses to isoproterenol. It is suggested that the two phases of lowering effects of ifenprodil on the constant-rate perfusion pressure of either vertebral or femoral artery in dogs are due to a possible neurogenic and an alpha-adrenolytic action, but the precise mechanism is not clear. Ifenprodil like isoxsuprine has a weak beta-sympathetic effect.


[Effects of adrenergic agents on secretion of salivary kallikrein in the submandibular gland of the dog]. The effects of sympathetic nerve stimulation and administrations of sympathomimetics on the secretion of salivary kallikrein induced by stimulation of Chorda tympani were examined quantitatively and qualitatively in the dog submandibular gland. Both sympathetic stimulation and administrations of sympathomimetics such as norepinephrine, epinephrine, and isoproterenol caused increased secretion of salivary kallikrein. Among these effects, those of the sympathetic stimulation, norepinephrine, and epinephrine were not completely abolished by pretreatment with phenoxybenzamine or propranolol, but the effect of isoproterenol was abolished by pretreatment with propranolol, and was hardly influenced by pretreatment with phenoxybenzamine. From these results, it would appear that secretion of salivary kallikrein may be mediated through adrenergic alpha- and beta-receptors. On the other hand, in contrast with the activities of the salivary kallikrein secreted by Chorda tympani stimulation or administration of isoproterenol which were not inhibited by SBTI, the activities of the salivary kallikrein secreted by sympathetic stimulation as well as administration of norepinephrine or epinephrine were markedly inhibited by SBTI. From these results, it is concluded that secretion of glandular kallikrein is due either to Chorda tympani stimulation or is mediated through adrenergic beta-receptor, while secretion of plasma kallikrein is mediated through adrenergic alpha-receptor.


[Effects of nialamide on brain monoamine consumption and liver glycogen content in rats chronically treated with insulin]. Male rats weighing about 50 g were fed a normal-balanced diet for 100 days during which time i.p. injections of saline solution (2 ml/kg) or insulin (4 IU/kg) and/or nialamide (30 mg/kg) were given every two days. The animals were sacrificed 16 hr after the last injection and the brains and livers were extracted for the measurement of tyramine consumption and liver glycogen deposition. It was found that the decrease of glycogen deposition and tyramine consumption induced by nialamide was inhibited by a simultaneous administration of insulin.


[Anti-inflammatory effects of ketoprofen - with special reference to inhibition of leukocyte migration]. It was observed that ketoprofen inhibited the increased vascular permeability induced by US-PF and in vitro leukocyte migration. The suppressive effect of ketoprofen on increased vascular permeability induced by US-PF was found to be the same as that of indomethacin. The effect of ketoprofen on the in vitro chemotaxis of rabbit peritoneal neutrophils was found to be the same as that of oxyphenbutazone and less than indomethacin. If the side effects of this drug are proven to be mild or not serious, and effective results are seen in clinical trials, ketoprofen may then be classified as a potent non-steroidal anti-inflammatory agent wifh analgesic actions in dysergic inflammation.


[Pharmacological analysis of the mode of action of mandelamidine with sustained antihypertensive effect. (II). Mechanism of transient hypotensive action of mandelamidine]. The mode of a transient hypotensive action of dl-Mandelamidine (MA) was studied in both anesthetized and unanesthetized animals. In the blood pressure of unanesthetized rats, a transient hypotensive action of MA (1 approximately 30 mg/kg i.v.) like papaverine (1 approximately 10 mg/kg i.v.) was much less predominant than in rats anesthetized with urethane (1.5 g/kg s.c.). The transient hypotensive action of MA (10 mg/kg i.v.) in rats anesthetized with urethane (1.5 g/kg s.c.) was not reduced when MA was injected continuously. Moreover C6 (5 mg/kg i.v.), propranolol (1 mg/kg i.v.), diphenhydramine (10 mg/kg i.v.) and atropine (2 mg/kg i.v.) did not block this transient hypotensive action. MA blocked the pressor action of epinephrine (3 mug/kg i.v.) in three minutes, but then potentiated it. MA (10 mug/kg i.a. approximately 1 mg/kg i.a.) caused a temporary vasodilation on the perfused leg artery and vertebral artery in dogs anesthetized with sodium pentobarbital (30 mg/kg i.v.). In the perfused leg artery, atropine (2 mg/kg i.v.) and propranolol (1 mg/kg i.v.) did not block the vasodilation of MA, and MA showed no marked blocking effect of the vasoconstriction of norepinephrine (0.2 mug/kg i.a.). In the dog heart-lung preparation and the guinea-pig heart, MA showed inhibiting effects. On the contraction of the cat nictitating membrane elicited by the stimulation of the postganglionic cervical sympathetic nerve, the blocking action of MA (1 mug/kg i.a. approximately 1 mg/kg i.a.) was much less predominant than that of phentolamine (10 mug/kg i.a.). In the rabbit descending aorta, MA (3 X 10(-4) g/ml) antagonized non-competitively the contraction elicited by norepinephrine. These findings suggest that a transient hypotensive action of MA depends upon the inhibiting effects on the cardiovascular system, and the adrenergic alpha-blocking effect of MA may be very weak.


[Anti-inflammatory action of 2-(2-fluoro-4-biphenylyl) propionic acid (flubiprofen]. 2-(2-fluoro-4-biphenylyl) propionic acid (flurbipofen, FP-70), a newly synthesized anti-inflammatory agent, was examined for its effects and then compared with the actions of other standard drugs. Inhibition of capillary permeability in mice: the activity of FP-70 was 7.8 times as potent as ibuprofen (IP), 61 times that of acetylsalicylic acid (AS), 5.7 times that of indomethacin (IM) and 25 times that of phenylbutazone (PB). Inhibition of carrageenin-induced edema in rat paw: the activity of FP-70 was 52 times as potent as IP, 250 times that of potent as AS, 14 times that of potent as IM and 110 times that of potent as PB. Inhibition of heat-induced coagulation of BSA: the activity of FP-70 showed the marked effects much the same as IP. In adrenalectomized rat, FP-70 was also effective. Inhibition of the ultra-violet erythema in guinea-pigs, the activity of FP-70 was 26 times as potent as IP, 790 times that of AS, 25 times that of IM and 68 times that of PB. Regarding the inhibition of CMC induced pouch in rats, the potency of FP-70 was to the same degree as that of predonisolone. In adjuvant induced arthritis in rats, FP-70 reduced and cured inflammatory syndromes and activity was the same or more effective than that of IM. From these results, FP-70 appears to be a most potent nonsteroidal anti-inflammatory agent.


[Phamacolofical studies of ifenprodil].

[Phamacolofical studies of ifenprodil]. Pharmacological effects of ifenprodil (IP), 4-benzyl-alpha-(p-hydroxyphenyl)-beta-methyl-1-piperidineethanol-L-(+)-tartrate monohydrate (Funai Pharmaceutical), a potential vasodilator, were studied in the dog, rabbit, guinea-pig and mouse. 1) Intravenous administration of IP (0.1 approximately 1 mg/kg) exhibited a continuous fall in blood pressure, an increase in heart rate and an increase in cardiac contractile force in the dog. The depressor and chronotropic effects of IP appeared likewise in the pithed dog, and the depressor responses were slightly more evident in the rabbit. The cardiovascular responses to IP were not affected by pretreatment with atropine or diphenhydramine, however, the chronotropic and inotropic effects were completely removed and the depressor response was depressed to some extent by propranolol. On the contrary, a pressor response to adrenaline was reversed to a depressor one after IP in the dog. 2) In heart-lung preparation of dogs, IP brought about slight cardiac stimulating effects, such as an increase in contractile force and a rise in aortic pressure. 3) IP induced a transient slight excitation in in situ movements of the rabbit intestine, and a similar excitation followed with a inhibition in movements of isolated intestine (10(-6), 2.5 X 10(-5) g/ml. This slight excitation was not affected by pretreatment with atropine, whereas the inhibition was depressed to a certain degree by simultaneous pretreatment with phenoxybenzamine and propranolol. IP produced a slight tonic increase in the guinea-pig isolated intestine and inhibited markedly the excitatory effect of histamine in the intestine. 4) IP did not affect the bronchial muscle but inhibited prominently a contractile respose of the muscle to histamine in in situ experiments on guinea-pig. 5) Locomotor activity was reduced dose-dependently and thiopental hypnosis was potentiated similarly after subcutaneous administration of IP (5 approximately 40 mg/kg), while skeletal muscle relaxant effects were not observed. 6) Survival time under a condition of acute anoxia was not changed with a dose of 0.5 and 1 mg/kg, but shortened with a dose of 5 mg/kg. The results suggest that IP has a alpha-blocking, a beta-stimulating and a moderate antihistaminic effects, and shows some cardiovascular effects, such as continuous depressor and chronotropic effects, and a moderate central inhibition.


[Effects of 2-(4-benzyl-piperidino)-1-(4-hydroxyphenyl)-1-propanol (ifenprodil) on the cardiovascular system in vivo]. Effects of ifenprodil (2-(4-benzyl-piperidino)-1-(4-hydroxyphenyl)-1-propanol) tartrate on the cardiovascular system were studied in mongrel dogs anesthetized with pentobarbital sodium and with alpha-chloralose and in rabbits. Ifenprodil (12.5 to 5,000 mug/kg i.v.) lowered systemic blood pressure in a dose-dependent manner in association with increased heart rate and augmented myocardial contractile force in dogs. Fall in blood pressure and tachycardia caused by ifenprodil were inhibited by pretreatment with phenoxybenzamine (5 mg/kg). The pressor responses to adrenaline and bilateral carotid occlusion were moderately reduced by pretreatment with ifenprodil (5 mg/kg), while the response to noradrenaline was attenuated only slightly. Ifenprodil (less than 1 mg/kg i.v.) decreased the total peripheral resistance and resistances of common carotid, internal carotid, vertebral, coronary, superior mesenteric, renal and femoral vascular beds in dogs. Ifenprodil increased the vertebral and femoral blood flow consistently (93.6 and 174.0%, respectively) but decreased the renal blood flow. Intra-arterial injection of ifenprodil (0.01 to 5 mug/kg) produced a dose-dependent increase in blood flow of vascular beds tested. The femoral and vertebral arterial blood flows were in particular consistently increased. Increases in the vertebral and femoral blood flows following i.v. and i.a. injections of ifenprodil were only slightly attenuated by pretreatment with phenoxybenzamine. Intravenous injections of ifenprodil (12.5 to 5,000 mug/kg) produced tachycardia in conscious rabbits but caused bradycardia after pretreatment with reserpine. From these results it was revealed that ifenprodil causes a fall in systemic blood pressure and an increase in the blood flow particularly in vertebral and femoral vasculatures following the vasodilating action directly on vascular smooth muscle. Blocking action of alpha-adrenoceptors in vessels was also demonstrated.


[Fluorohistochmical studies on localization of 5-hydroxytryptamine neurons in the rat brain following administration of thiopental sodium]. The localization of 5-hydroxytryptamine (5-HT) neurons in the rat brain was studied by the fluorescence histochemical method of Falck and Hillarp. A model experiment and tissue experiment were used to measure the characteristics of wave length of 5-HT fluorophor and we decided 420 mmu for excitation maxima and 520 mmu for emission maxima. The 5-HT neurons both in frontal and sagittal sections of the brain from reserpine-nialamide-treated rats were observed fluorohistochemically. The brain was divided into 5 portions; medulla oblonga, pons, midbrain, diencephalon and telencephalon. In frontal sections, from B1 to B8 cell groups were localized in raphe nuclei, in the region of 200 mu width on both sides from the central plane, excluding most of B2 cells and cells of B9 group which were situated on both side apart from the central plane. Microphotographs taken from about 34 areas of a sagittal section were composed to determine the interrelationship between B cell groups of 5-HT neurons. The frontal sections untreated with reserpine-nialamide showed a faint fluorescence of 5-HT neurons, however, after the administration of thiopental sodium 80 mg/kg i.p. the fluorescence of 5-HT neurons was increased in intensity and the observable cells were increased in number, especially in the B7 cell group. The yellow fluorescence of 5-HT nerve terminals in nuc suprachiasmaticus in the sagittal section was increased in intensity and varicosities of the terminals were readily observed. These results verified the increase of 5-HT in 5-HT neurons after the administration of thiopental and suggest that the 5-HT neuron may play a role in the mechanism of the central action of a thiopental.


[Cesarean section. 5-year study of results at a medium size department for obstetrics and gynecology]. A report about the results of caesarean section from 1970-1974 is given. The rate of caesarean section was 4,2%. The list of indication is divided into vital indication, preventive indication before onset, at onset and in the first stage of labor. The morbidity was 5%, mortality 11%. The perinatal mortality was 4,4%. It is emphasized that caesarean section is still the most dangerous method of delivery.


[Selective kidney angiography. Indispensable diagnostic method in urogenital tuberculosis]. Selective angiography of the kidneys was performed in 119 patients with urogenital tuberculosis. The diagnosis was confirmed by bacteriological and/or histological examinations. Diagnostic value. Under optimal technical conditions changes of the small renal arteries even in the parenchymal stage I of renal tuberculosis can be seen. These cannot be diagnosed with other methods. In stage II and III the degree of destruction and the localisation of the specific process can be demonstrated more efficiently by angiography than by urogram. In stage III/3 (kidney without function) the differential diagnosis between tuberculosis and the late form of polycystic renal disease is possible. Therefore the selective renal angiography ought to be performed more frequently for intensive diagnosis and also to reach a definite diagnosis before surgical procedures. Special surgical planning is more precise and knowledge of vascular anatomy reduces the operative risk.


[Studies on tissue cultures from calf eyes]. The growth of lens epithelium and iris tissue was studied by biomicroscopy and electronmicroscopy, its velocity measured. With the Warburg-method it was possible to show the oxygen-uptake of both tissues without and after addition of different substances. Pilocarpine lowered the O2-uptake of the epithelia to less than 1/3 of its normal value, whilst it raised that of the iris to the double amount. Napthochinone inhibited the O2-uptake of both tissues to little more than half of the normal level. The least differences as compared to the controls showed methyl-prednisolone: it lowered scarcely the O2-uptake of the epithelium, and the O2-uptake of the iris to almost half the normal rate. The author concludes that cataract developes, if oxygen-uptake of lens epithelium and iris tissue are reduced in the same way without any possibility of compensation. Under certain conditions this cell-biological method may be employed to detect cataractogenous substances.


[On the toxicity of CT-1341 evoked by long-term administration. III. Subacute toxicity of alphadolone acetate and solvent contained in CT-1341 (author's transl)]. Subacute toxicity of solvent adjuvant, alphadolone acetate and solvent, 20 per cent Cremophor solution contained in CT-1341 was studied by using rats of both sexes. Alphadolone acetate and Cremophor solution were intraperitoneally injected every day for a period of one month. Total rats tolerated to daily administration of 60 mg/kg of alphadolone acetate or of 20 ml/kg of 20 per cent Cremophor solution, without showing significant changes in body weight curves and food intake. No change was observed in blood cells and in biochemical data of blood and urine as compared with control. Rats subjected to daily administration of 60 mg/kg of alphadolone acetate presented slight patho-histological changes such as swelling of cells of the liver and kidneys, and also cell infiltration of pericapillary tissues of the lung.


[Effect of acute cold exposure on blood composition in rats treated with 6-Hydroxydopamine (author's transl)]. Rats injected with 6-hydroxydopamine (6-OHDA) into the lateral ventricle showed greater elevation in plasma FFA and beta-hydroxybutyrate and significant decrease in blood sugar and lactate following acute cold exposure at -10 degrees C for 2 hours. Changes in the blood composition induced by the cold exposure in rats treated with 6-OHDA intravenously were not different from those observed in control rats. Adrenomedullectomized rats treated with 6-OHDA responded to the cold exposure with rather marked elevation in plasma FFA and beta-hydroxybutyrate. These results were discussed from the view-point of compensatory changes in metabolic regulation in the absence of norepinephrine.


[Effects of thinner fibers on spinal neurones in the cat (author's transl)]. The aim of the present experiment is to investigate the effects of thinner nerve fibers on spinal nerve cells by using differential blocking method of thicker nerve fibers. 1. As to the differential blocking of a thicker alpha motor fibers of gastrocnemius nerve by applying trapezoid wave current, significant correlation was obtained between the decreasing amplitude of alpha-volleys and the increasing latency of the volley. In single fiber analysis, the relation between the strength of the blocking current and conduction velocity was summarized on 30 alpha and 41 gamma fibers. The blocking current of each single fiber was shown to be inversely proportional to 2.01 power of conduction velocity (correlation coefficient -- 0.71)...


[Diurnal variations and the effect of cold exposure on the tissue histamine content in the rat (author's transl)]. Diurnal variations and the effect of cold exposure on histamine content in the liver, pinna and interscapular brown adipose tissue were investigated in the rat. A significant circadian rhythm of the histamine content was observed in the pinna and liver. In these tissues the content showed the peak at 5:00 a.m. and was maintained at low levels during the daytime. No significant variation was seen in the histamine content of brown adipose tissue. Acute cold exposure at -10 degrees C for 60 min caused an increase in the histamine content of the liver, while in the other two tissues the content was not affected by the cold exposure. Chronic cold exposure at 5 degrees C for 2 weeks caused a marked increase in the histamine content in brown adipose tissue and a moderate increase in the liver, while the content decreased in the pinna.


Effect of osmotic pressure on neurogenesis in cultures of chich embryo spinal cords. The osmotic pressure of the medium in stoppered, roller tube cultures increased by an average of 17 +/- 6 mOsM per kg of water during 3 days of incubation at 37 degrees C irrespective of the initial osmolality (280 to 340 mOsM) of the medium. The increase was apparently due to evaporation of water from the medium into the gas phase of the roller tube. This observation led us to study the effect of osmotic pressure on neuronal differentiation in cultures of chick embryo spinal cords. Spinal cords were excised from stage 16 to 19 (2.5 to 3 days of incubation) or stage 36 (10 days) chick embryos and cultured as fragments on collagen-coated cover slips in roller tubes at 37 degrees C for 21 days. The medium was adjusted to 283 +/- 3,300 +/- 3,323 +/- 3, or 342 +/- 3 mOsM per kg with saturated choline chloride solution or distilled water. The results indicate that the nature of the neuronal differentiation in vitro was not altered by the osmolality of the medium. The proportion of cultures containing neurons was influenced by osmolality. In the 300 +/- 3 mOsM medium, 75% of all the stage 36 cultures initiated contained neurons, and 52% of all the stage 16 to 19 cultures initiated contained neurons. In the other media the proportion of neuron-containing cultures was lower. Two conclusions were drawn. Neurogenesis in cultures of embryonic chick spinal cord fragments is sensitive to an increase in the initial osmotic pressure of the medium as small as 20 mOsM above the optimal 300 mOsM. As a result of the 17 mOsM increase which always occurred in the culture medium between feedings, the optimum osmolality for neuronal development is in fact a range, from 300 to 317 mOsM.


Incorporation of labeled glucosamine into glycoproteins by organ cultures of hamster trachea: adverse effects of HEPES buffer. Hamster tracheas were cultured in serum-free CMRL 1066 medium buffered with either NaHCO3 alone or HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulphonic acid) plus NaHCO3 in an atmosphere of 95% O2 and 5% CO2. Afther 2 days in culture, tracheas maintained in HEPES plus NaHCO3-buffered medium showed an altered surface morphology. Histiological examination after 6 days of culture with HEPES plus NaHCO3 showed the presence of only scattered clumps of cilia. Incorporation of [14C]-glucosamine into intracellular glycoproteins was reduced by 75% and into secreted glycoproteins by 54% in cultures buffered with HEPES plus NaHCO3, compared to NaHCO3-buffered cultures. Incorporation of [H]fucose into intracellular glycoproteins was also reduced, although no effect was observed on secreted glycoproteins.


Improved methods for reducing calcium and magnesium concentrations in tissue culture medium: application to studies of lymphoblast proliferation in vitro. We have compared several methods for reducing calcium and magnesium concentrations in tissue culture medium, with the objective of producing selective deficiency effects on the growth of mouse (L5178Y) and human (P1R) lymphoblasts. In experiments in which calcium- and magnesium- "free" McCoy's medium was supplemented with 15% horse or fetal calf serum, enough calcium and magnesium was provided by serum to support normal lymphoblast growth rate. Either dialysis or chelating-resin treatment of horse or fetal calf serum reduced calcium and magnesium contents approximately 100-fold. Use of dialyzed sera resulted in reduced growth rate, although in most cases the reduction in growth could be attributed to other effects of dialysis on serum, inasmuch as growth in those experiments was not restored to normal by the addition of calcium and magnesium to the medium. In contrast, the reduction of lymphoblast growth rate that occurred when resin-treated serum was used was always attributable to removal of calcium and magnesium, as normal growth always occurred in cultures to which calcium and magnesium were added. To demostrate a growth-inhibiting effect on either mouse or human lymphoblasts by severe reduction of either calcium or magnesium in the presence of normal amounts of the alternative cation, it was necessary to (a) expose McCoy's Ca-Mg-"free" medium to chelating-resin to reduce further the residual cation concentrations; (b) wash cells from stock cultures in a medium devoid of calcium and magnesium prior to inoculation into experimental cultures; (c) reduce the proportion of serum in the final medium from 15 to 5%; and (d) add 100 muM EGTA to cultures. Under these conditions, growth of both cell types was completely abolished in the presence of normal magnesium but in the absence of added calcium, and markedly reduced in the presence of normal calcium but in the absence of magnesium. These modifications did not compromise growth in cultures containing normal concentrations of both ions.


Local and systemic immune responses following oral immunization of foetal lambs. Foetal lambs were immunized orally 6-15 days before birth by introducing horse spleen ferritin into the amniotic fluid. Immunized and non-immunized lambs were killed at birth, usually before they had suckled, blood and intestinal contents were collected and single cell suspensions were prepared from spleen, mesenteric lymph nodes and jejunum. Specific antibody was detected in serum and intestinal contents of all immunized lambs which had not suckled. Specific antibody was usually not detected in samples from non-immunized lambs. In immunized lambs antibody activity in serum was associated with IgM and in intestinal contents with IgA and IgM. In agreement with these findings, the levels of IgM and IgA in serum and intestinal contents of immunized lambs were relatively high. Generally, immunoglobulins were not detected in samples from non-immunized lambs. Relatively high proportions of cells secreting specific antibody were present in the tissues of immunized but not non-immunized lambs. In the spleen most of the cells were secreting IgM antibody, in mesenteris lymph nodes IgM cells predominated and small numbers of IgA cells were detected, and in the jejunum approximately equal numbers of IgA and IgM cells were secreting specific antibody.


Biological activities of endotoxins detoxified by alkylation. It has been previously observed that lipopolysaccharides can be detoxified by alkylation and yet retain their adjuvant activity. Our present findings confirm these results and show, moreover, that these derivatives did not lose their capacity to protect mice against lethal irradiation and lost only partially their ability to interrupt pregnancy or to induce blast transformation of murine B-lymphocytes. However, in contrast with lipopolysaccharides, these alkylated preparations did not enhance the nonspecific resistance of mice to a bacterial infection. The various bilogical functions of endotoxins can therefore be separated and are not uniformly related to their toxicity.


Comparative infectivity of trachoma organisms in HeLa 229 cells and egg cultures. Thirty-two trachoma strains were simultaneously titrated in the yolk sac of embryonated chicken eggs and in HeLa 229 cells pretreated with diethylaminoethyl-dextran to determine the relative sensitivity of the two culture systems for detection of the infection. The strains tested were both yolk sac- and HeLa cell-established isolates from the eye and the genital tract. The study showed that the cell culture was of equal or greater sensitivity than yolk sac culture and that ocular and genital isolates differ in their ability to propagate in egg and cell culture: genital strains grow better in cell culture than in eggs and ocular strains grow only slightly better in cell culture.


Selective reduction and alkylation of the COOH-terminal disulfide bridge in bovine growth hormone. Conditions leading to the cleavage of both disulfide bridges in human growth hormone caused the reduction of only one disulfide bond in bovine growth hormone. Partially reduced and alkylated derivatives of bovine growth hormone were prepared and characterized. It was shown that the reduction and alkylation modified the COOH-terminal disulfide bond, however, this modification does not result in the dissociation of the dimeric form of bovine growth hormone or cause a significant loss of growth-promoting activity.


Clinical, electrophysiological, quantitative, histologic and ultrastructural studies of the index branch of the radial cutaneous nerve in leprosy. I. Preliminary report. The index branch of the radial cutaneous nerve has been demonstrated as a constant nerve which can readily biopsied under local anesthesia and yields a nerve which is of suitable size for quantitative and qualitative studies both by light and electron microscopy. It supplies a limited but constant area where the sensory loss does not disturb the patient. Definite ultrastructural changes have been demonstrated in the clinically normal nerves of leprosy patients. These nerves have also revealed a loss in the small myelinated and unmyelinated fibers, which correlated with the common clinical findings of the absence of sweating and dissociated sensory loss in this disease. Gross damage has been encountered in nerves which showed only early signs of clinical damage even with refined methods of sensory testing. These nerves would have passed as normal on routine testing by No. 5 nylon. Regeneration of small fibers was noted following loss of large size fibers. Nerve conduction velocity may be a useful tool in early diagnosis.


Spinal cord stimulation versus detrusor stimulation. A comparative study in six "acute" dogs. Spinal cord stimulation and direct bladder stimulation were evaluated in an "acute" comparative study of six dogs. Both means were effective in inducing strong detrusor contraction and rise in intravesical pressure. Spinal cord stimulation, however, induced greater rise in outflow resistance than did direct bladder stimulation. Effective net intravesical pressure was higher with direct stimulation and thus more successful in achieving bladder emptying.


Biophysical comparisons of calicivirus serotypes isolated from pinnipeds. Biophysical properties of three new San Miquel sea lion virus isolates from pinnipeds were examined and compared with those of previously characterized serotypes. The caliciviruses showed an identical sedimentation rate of 183S in 5-20% sucrose. Buoyant densities in CsCl were in the range of 1.35-1.39 g/ml, with differences among the serotypes.


Amyloidosis in leprosy.

Amyloidosis in leprosy. Histopathologic studies were performed on systemic amyloidosis of leprosy patients in two Japanese leprosaria. Amyloid deposits were observed in 23 of 50 cases (46%) autopsied from 1955 to 1959 (Group A), and in 17 of 110 cases (15.4%) from 1962 to 1971 (Group B). These incidences are higher than those of the pre-chemotherapeutic period in Japan. The average age of amyloidal cases was significantly lower than that of nonamyloidal cases in both groups. There was no predilection for either type of leprosy in Group B. Pyelonephritis was frequently observed in both groups; the incidence of amyloidosis in Group B correlated with pyelonephritis. Amyloid deposits may be related to renal disorders associated with lepra reactions or inflammatory lesions. Amyloidosis has been classified into primary and secondary type, but the criteria seem uncertain. We referred to a classification based on immunologic finginds. In the severely affected lesion of the kidney of one case, we observed foci of foreign body type giant cells, suggesting a possibility of resorptive process in human renal amyloidosis.


[First results of parenteral treatment of male fertility disorders using kallikrein: oligozoospermia]. 57 andrological patients with oligozoospermia were treated with kallikrein--a kinin-liberating proteinase--over a period of 7 weeks. The statistical analysis of the different semen parameters showed that during parenteral application of 3 X 40 KU Kallikrein per week a significant increase of the sperm motility (total motility and progressive motility) occurred. Improvement of sperm motility did exist during the observation period up to 3 months after withdrawl of the therapy. 2, 3 and 5 months after beginning of the treatment a significant increase of the sperm count was found. A significant increase of the total sperm output was observed 3 and 5 months after initiation of the medication. Finally, the percentage of normal shaped spermatozoa was increased 2 and 5 months after beginning the kallikrein application. The conception rate was 28%.


Leprosy of the larynx.

Leprosy of the larynx. Five instances of lepromatous leprosy involving lesions of the larynx were encountered among a series of 280 laryngeal lesions. These are briefly described as involving the epiglottis in all cases, vocal cords in two, and extension into the pyriform fossa in one instance.


Effect of methionine on cephalosporin C and penicillin N production by a mutant of Cephalosporium acremonium. A mutant with enhanced potential to utilize sulfate for cephalosporin C production was isolated from a strain of Cephalosporium acremonium. The mutant displayed potency levels more than twofold that of the parent in the presence of sulfate but its productivity was severely inhibited by more than 0.5% of methionine which gave high cephalosporin C production with the parent. In a complex medium norleucine stimulated cephalosporin C production by the mutant in the presence of sulfate, whereas it showed no effect on the parent. In an incubation  system with sulfur-starved cells of the mutant, L-methionine, but not the D-isomer, gave lower cephalosporin C production and a delayed production of penicillin N. However, it exhibited a stimulatory effect in the presence of valine or alpha-aminoadipic acid, the constituent amino acids of the antibiotic. Norleucine showed a similar effect to that of L-methionine in the presence of sulfate. On the basis of these results, characteristics of the mutant are discussed in connection with the effect of methionine.


Stimulation of cephalosporin production by methionine peptides in a mutant blocked in reverse transsulfuration. The previously reported inability of methionine to stimulate cephalosporin C production in a cysteine auxotroph is due to cysteine interference with methionine uptake. In such a cse, "illicit transport" of alanylmethionine can be used to demonstrate the efficacy of methionine in such mutants blocked in the path from methionine to cysteine. This result supports the hypothesis that the stimulatory activity of methionine is not due to its ability to donate sulfur to the caphalosporin C molecule.


Evidence for a sex hormone in Dictyostelium discoideum. A dialyzable sex hormone is produced by one strain of Dictyostelium discoideum (NC-4) that induces macrocyst formation (sexual stage) in its opposite mating type (V-12).


Human anaphylatoxin (C3a) from the third component of complement. Primary structure. C3a anaphylatoxin is derived from the third component (C3) of the blood complement system. Selective proteolysis of C3 by activated proenzymes indigenous to blood generates the C3a fragment. Human C3a was isolated from inulin-activated serum containing 6-aminohexanoic acid, according to recently published procedures (Hugli, T. E., Vallota, E., and Müller-Eberhard, H. J. (1975) J. Biol. Chem. 250, 1472-1498). The human C3a fragment is a highly cationic molecule exhibiting an approximate molecular weight of 9000 and composed of 77 amino acid residues. It consists of a single polypeptide chain containing 8% cysteine and lacks both tryptophan and carbohydrate. A tentative primary structure for the human C3a molecule, deduced from overlapping peptides obtained after cyanogen bromide cleavage, tryptic and chymotryptic digestion, is: See article. Two cystelhylcysteine sequences were established at positions 22, 23 and 56, 57 in human C3a. The 6 half-cystine residues in C3a are all interconnected through three disulfide linkages intersecting in a disulfide knot. The functionally amino acid residues distributed among 14 residues at the COOH-terminal end of C3a. This unusually cationic COOH-terminal region of C3a is presumed to play an important role in the interaction of this protein molecule with cellular receptors. A comparison between the linear sequence of human C3a and the NH2-terminal sequences of light and heavy chains of human immunoglobulin indicates that limited identity exists.


Human prothrombin activation.

Human prothrombin activation. Human prothrombin has been purified from American Red Cross Factor IX concentrates. Studies of the activation of the human prothrombin with the use of sodium dodecyl sulfate electrophoretic analysis of activation products indicated that human prothrombin activation is similar to bovine prothrombin activation. Molecular weight analysis of human prothrombin and intermediated by sodium dodecyl sulfate co-electrophoresis with bovine prothrombin and its intermediates resulted in molecular weights of 70,000 for prothrombin, 51,000 for intermediate 1, 41,000 for intermediate 2, 23,000 for intermediate 3, and 13,000 for intermediate 4. Amino acid compositions of human prothrombin and intermediates are similar to those for bovine prothrombin and intermediates. NH2-terminal sequence studies of human prothrombin, intermediates, and alpha-thrombin A and B chains placed the intermediates in the parent human prothrombin molecule as described for the bovine system. Intermediate 3 is the NH2-terminal of prothrombin, and intermediate 1 is the COOH-terminal segment of the zymogen. Intermediate 4 is the NH2-terminal of intermediate 1. Intermediate 2', the immediate precursor of alpha-thrombin, is the COOH-terminal segment of intermediate 1. In general, a high degree of homology in the primary structure of prothrombin and intermediates was observed between the human and bovine system. The NH2-terminal sequences of human intermediate 2' and alpha-thrombin A chain are identical. However, human intermediate 2' isolated in a manner identical with that used for the isolation of bovine intermediate 2 is homologous with bovine intermediate 2, beginning with residue 14.


Mass spectrometric study of the enzymatic conversion of cholesterol to (22R)-22-hydroxycholesterol, (20R,22R)-20,22-dihydroxycholesterol, and pregnenolone, and of (22R)-22-hydroxycholesterol to the lgycol and pregnenolone in bovine adrenocortical preparations. Mode of oxygen incorporation. Incubation of cholesterol with a bovine adrenocortical mitochondrial acetone-dried powder preparation yielded (22R)-22-hydroxycholesterol (I), (20R,22R)-20,22-dihydroxycholesterol(II), and pregnenolone (III) which were conclusively identified by combined gas chromatography-mass spectrometry. Incubations with [4-14C]cholesterol yielded I, II, and III with specific activities (determined from partial mass-spectral scans) not significantly different from those of the used substrate or the cholesterol reisolated after the incubation, demonstrating that the isolated compounds arose mostly, if not entirely, from the substrate cholesterol. Incubations in an 18O-enriched atmosphere yielded I, II, and III with 18O at position C-22, C-20 and C-22, and C-20, respectively, providing evidence that the hydroxyl groups of the side chain of I and II and the C-20 oxygen atom of III originated from molecular oxygen. The distribution of the oxygen atoms in II after incubation with 18O2 and 16O2 (devoid of 16O18O) proved that the hydroxyl groups of the side chain of II were introduced from two different molecules of oxygen, consistent with a sequential hydroxylation of cholesterol. No (20S)-20-hydroxycholesterol was found. Incubation of I in an 18O-enriched atmosphere afforded II and III with 18O at C-20.


Effects by heme, insulin, and serum albumin on heme and protein synthesis in chick embryo liver cells cultured in a chemically defined medium, and a spectrofluorometric assay for porphyrin composition. Primary chick embryo liver cells, which had been previously cultured in Eagle's medium containing 10% fetal bovine serum, had the same characteristics (inducibility of delta-aminolevulinic acid synthetase and synthesis of plasma proteins) when cultured in a completely defined Ham F-12 medium containing insulin. Insulin was active in the physiological range; 2 to 3 nM were sufficient to increase the induced delta-aminolevulinic acid synthetase to 50% of the maximum effect obtained with a saturating amount of insulin (30 nM). Serum albumin added to the Ham-insulin medium caused protoporphyrin but not uroporphyrin, generated in the cultured liver cells, to be transferred to the medium. As little as 10 mug of human serum albumin per ml caused the transfer of one-half of the protoporphyrin. Bovine serum albumin was only about 1/30 as effective. A spectrofluorometric method and calculation procedure are described for quantitation, in the nanomolar range, of total porphyrin and the percentage of this that is protoporphyrin or uroporphyrin plus coproporphyrin. The method is satisfactory for the measurement of porphyrins generated by 1 mg wet weight of cells in culture in 20 hours. Heme (0.1 to 0.3 muM), when added to the medium as hemin, human hemoglobin, or chicken hemoglobin, specifically inhibited the induction of delta-aminolevulinic acid synthetase by one-half. This high sensitivity for heme was observed under conditions in which the defined medium was free of serum and where a chelator of iron was added to the medium to diminish the synthesis of endogenous heme. Heme endogenously generated from exogenous delta-aminolevulinic acid also inhibited the induction; chelators of iron prevented this inhibition. The migration of heme from the mitochondria to other portions of the cell is discussed in terms of the affinities of different proteins for heme. A hypothesis of a steady state of liver heme metabolism, controlled by the concentration of "free" heme, is presented. The different effects of heme on the synthesis of a number of proteins are summarized.


A requirement for reversible binding between aggregating embryonic cells before stable adhesion. Chick embryonic liver and neural retina cells aggregate in a two-step process. Initially, cells formed a loose association in a step that apparently did not require metabolic energy. Cells bound in this manner were dissociable by mild shear forces or by simple dilution. The results of the dilution experiments suggest a readily reversible binding of single cells to form these types of aggregates. In a second step, which required metabolic energy, the cells became firmly, or stably attached. The formation of both types of bond was temperature-dependent. Kinetic studies indicated that the formation of reversible bonds between cells was required before the cells could become stably attached, and that reversibly bound cells were converted directly into stably bound cells.


Amino acid transport by aggregates of cultured chicken heart cells. Effect of insulin. Single heart cells dissociated from 14-day-old chicken embryos were reagregated into spheroidal clusters on a gyratory shaker and centrifuged to form cohesive discs of approximately 400 aggregates. These cultured cells accumulated 2-amino[1-14C]isobutyric acid against a gradient. When incubated for 3 hours in a protein-free, buffered, balanced salt solution, concentrative transport decreased to a stable basal level. Incubation in the presence of sodium bovine insulin prevented this fall in transport activity and resulted in increased 2-aminoisobutyric acid uptake to concentrations 40 time sthat in the medium during a subsequent 3-hour transport assay. Intracellular accumulation of 2-aminoisobutyric acid was linear during the initial 15 min of transport in the presence and absence of added insulin. Basal transport of 2-aminoisobutyric acid was temperature-dependent, requied extracellular sodium greater than 125 meg/liter, and demonstrated saturation with an apparent Vmax of 3.4 mmol/liter/10 min and an apparent Km of 2.6 mM. Basal transport activity was not reduced by cycloheximide or puromycin even after 3 hours of exposure...


Scanning microscopy of dissociated tissue cells. A method is described for studying by scanning electron microscopy (SEM) all the surfaces of fully differentiated cells from intact tissues. Thus, cell faces normally hidden from view are exposed and made available for SEM examination. This is achieved by fixing the tissue in OSO4 and then soaking it in a 1% solution (in water) of boric acid. After different periods of time, varied according to particular tissue, slight mechanical pressure will cause the fixed tissue to dissociate into its component cells. These are then made to adhere to a substrate and are taken through critical point drying, etc., for examination. Observations are reported on the topography of whole hepatocytes, adsorptive cells of the intestinal epithelium, proximal tubule cells of the rat kidney, mammary tumor cells of the mouse, and rat sarcoma cells. Several other tissues are reported to dissociate when similarly treated, but for each the procedure must be slightly modified.


Differential effect of hormones on macromolecular synthesis and mitosis in chick embryo cells. Exposure of serum-deprived confluent monolayers of chick embryo cells to fresh serum results in maximal stimulation of synthesis of RNA and protein followed by increased DNA synthesis and mitosis. The addition of insulin to quiescent cultures effects a similar acceleration of synthesis of RNA and protein, but little stimulation of DNA synthesis and mitosis is evident. However, the simultaneous addition of insulin and hydrocortisone to resting cells causes a significant increase in the rate of DNA synthesis although the level reached is considerably lower than that obtained with serum and still no mitosis occurs. Unexpectedly, insulin plus hydrocortisone prevents maximal synthesis of RNA and protein in contrast to insulin-treated cultures. Nuclear autoradiography and percent mitosis of cells incubated with various regulatory factors indicate that cell heterogeneity exists and is reflected in the metabolic responses of subpopulations to specific regulatory signals.


Microtubular origin of mitotic spindle form birefringence. Demonstration of the applicability of Wiener's equation. Meiosis I metaphase spindles were isolated from oocytes of the sea-star Pisaster ochraceus by a method that produced no detectable net loss in spindle birefringence. Some of the spindles were fixed immediately and embedded and sectioned for electron microscopy. Others were laminated between gelatine pellicles in a perfusion chamber, then fixed and sequentially and reversibly imbibed with a series of media of increasing refractive indices. Electron microscopy showed little else besides microtubules in the isolates, and no other component present could account for the observed form birefringence. An Ambronn plot of the birefringent retardation measured during imbibition was a good least squares fit to a computer generated theoretical curve based on the Bragg-Pippard rederivation of the Wiener curve for form birefringence. The data were best fit by the curve for rodlet index (n1) = 1.512, rodlet volume fraction (f) = 0.0206, and coefficient of intrinsic birefringence = 4.7 X 10(-5). The value obtained for n1 is unequivocal and is virtually as good as the refractometer determinations of imbibing medium index on which it is based. The optically interactive volume of the microtubule subunit, calculated from our electron microscope determination of spindle microtubule distribution (106/mum2), 13 protofilaments per microtubules, an 8 nm repeat distance and our best value for f, is compatible with known subunit dimensions as determined by other means. We also report curves fitted to the results of Ambronn imbibition of Bouin's-fixed Lytechinus spindles and to the Noll and Weber muscle imbibition data.


Gametic differentiation in Chlamydomonas reinhardtii. III. Cell wall lysis and microfilament-associated mating structure activation in wild-type and mutant strains. Cell fusion between mating type plus (mt+) and minus (mt-) gametes of Chlamydomonas reinhardtii is analyzed structurally and subjected to experimental manipulation. Cell wall lysis, a necessary prelude to fusion, is shown to require flagellar agglutination between competent gametes; glutaraldehyde-fixed gametes ("corpses") of one mating type will elicit both agglutination and cell wall lysis in the opposite mating type, whereas nonagglutinating impotent (imp) mutant strains are without effect. The fusion process is mediated by a narrow fertilization tubule which extends from the mt+ gamete and establishes contact with the mt- gamete. Formation of the tubule requires the "activation" of a specialized mating structure associated with the ml+ cell membrane; activation causes microfilaments to polymerize from the mating structure into the growing fertilization tubule. Mating structure activation is shown to depend on gametic flagellar agglutination; isoagglutination mediated by the lectin concanavalin A has no effect. Gametes carrying the imp-l mt+ mutation are able to agglutinate but not fuse with mt- cells; the imp-l gametes are shown to have structurally defective mating structures that do not generate microfilaments in response to gametic agglutination.


Temperature dependence of anaphase chromosome velocity and microtubule depolymerization. The time course of chromosome movement and decay of half-spindle birefringence retardation in anaphase have been precisely determined in the endosperm cell of a plant Tilia americana and in the egg of an animal Asterias forbesi. For each species, the anaphase retardation decay rate constant and chromosome velocity are similar exponential functions of temperature. Over the temperature range at which these cells can complete anaphase, chromosome velocity and retardation rate constant yield a positive linear relationship when plotted against each other. At the higher temperatures where the chromosomes move faster, the spindle retardation decays faster, even though the absolute spindle retardation is greater. Chromosome velocity thus parallels the anaphase spindle retardation decay rate, or rate of spindle microtubule depolymerization, rather than absolute spindle retardation, or the amount of microtubules in the spindle. These observations suggest that a common mechanism exists for mitosis in plant and animal cells. The rate of anaphase chromosome movement is associated with an apparent first-order process of spindle fiber disassembly. This process irreversibly prevents spindle fiber subunits from participating in the polymerization equilibrium and removes microtubular subunits from chromosomal spindle fibers.


A rapid phytohemagglutinin induced alteration in lymphocyte potassium permeability. The exposure of rat and human lymphoid cells to mitogenic concentrations of phytohemagglutinin resulted in an apparent decrease in cellular K+ without a significant change in cellular Na+ when the cells were washed with isotonic Hepes buffered choline chloride prior to cation determination. The apparent reduction in total cellular Na+ plus K+ concentration, however, was not accompanied by a change in cell volume. We inferred that the constant cell volume could occur only if the lost intracellular K+ was exchanged for an external cation during the washing procedure used to prepare cells for Na+ and K+ measurement. This inference was supported by the quantitative recovery of lost cellular K+ in the choline chloride washing solution and the demonstration that a comparable proportion of 86Rb+ (K+ analogue) 42K+ was lost from prelabelled cells during choline chloride washing. Use of medium 199 with Hanks salts, 150 mM NaCl, or 100 mM MgCl2 as the washing solution did not prevent K+ exchange although exchange was less in the presence of MgCl2. These findings indicate that phytohemagglutinin produces a rapid alteration in lymphocyte plasma membranes so as to allow abnormal K+ exchange. This observation is of importance because investigators who measure intracellular solutes in phytohemagglutinin-treated lymphocytes must consider the possibility of lossduring preparative washes. Also, changes in membrane permeability following phytohemagglutinin treatment may modulate mitogenesis and/or permit the transmission of chemical messages between cells.


Influence of Concanavalin A, wheat germ agglutinin, and soybean agglutinin on the fusion of myoblasts in vitro. Although muscle cell fusion was shown to be an energy-requiring process, release of myoblasts from an EGTA fusion block could be accomplished with Earle's balanced salt solution (containing 1.8 mM Ca++) free of glucose or any other energy-produced metabolite. The effect of concanavalin A, abrin, and the lectins from wheat germ, soybean, and Lens culinaris on myoblast fusion was examined with synchronized myoblast cultures upon release from fusion block. At a concentration of 15 mug/ml, these lectins were found to inhibit the fusion process to the extent of 62%, 41%, 32%, 8%, and 19%, respectively. Concanavalin A inhibition could be prevented by alpha-methyl-D-mannoside. The inhibitory effect of all the lectins except abrin could be reversed by changing to the normal, serum-containing medium. The number of binding sites was 3.4 X 10(7), 6.1 X 10(7), and 1.7 X 10(6), respectively. Although myoblasts were found to have about twice as many binding sites for wheat germ agglutinin as for concanavalin A, concanavalin A was determined to be twice as effective as wheat germ agglutinin as an inhibitor of myoblast fusion. These findngs raise the possibility that specific cell surface glycoproteins may be an important factor in this process.


Spontaneous and lectin-induced redistribution of cell surface receptors on embryonic chick neural retina cells. The mobility of plant lectin receptors in the plane of the membrane is examined for cells prepared from embryonic chick neural retinas by a variety of procedures. Cells liberated from the intact tissue by trypsin treatment followed by mechanical dissociation are able to redistribute their receptors into 'caps' both spontaneously and in the presence of a multivalent lectin. These cells, dispersed by trypsinization, upon repair in culture for a suitable period of time lose their ability to redistribute lectin receptors. Cells dispersed by mechanical means without prior trypsin treatment are unable to undergo 'cap' formation. In addition, cells within intact tissues are also unable to redistribute their lectin receptors into 'caps.' Based on these observations we propose that within solid tissues which have assumed their characteristic architecture, cell surfaces are immobilized, and that this phenomenon may be a critical parameter in determining the potential of a cell to undergo morphogenetic rearrangements.


Determinants of experimental neurosis in cats. In order to assess the relative effectiveness of Masserman's contingent method and Wolpe's noncontingent method of inducing experimental neurosis in cats, cats in four independent groups were trained to a uniform criterion to eat in a standard apparatus and then shocked either contingently upon eating or noncontingently upon eating or noncontingently. Cats shocked noncontingently were matched with those shocked contingently for the number of shocks administered. Ss then were assessed for persistence and/or generalization of the inhibition of eating in the same or in a discriminably-different setting. Masserman's contingent method induced significantly greater inhibition at the 0.5 level. This finding is consistent with prediction that would be made on the basis of the delay-of-reinforcement gradient. It is suggested that the contrasting results obtained by Wolpe and Smart may be attributable to confounding variables introduced unintentionally.


The septum: neural systems involved in eating, drinking, irritability, nuricide, copulation, and activity in rats. Eating, drinking, irritability, muricide, and copulation were measured in 45 male rats before and after one of three coronal knife cuts in the septal region or a control operation; in addition, activity was measured postoperatively. (a) Cuts posteroventral to the septum resulted in slight hypophagia, marked hyperdipsia, irritability, asexuality, and decrease in activity. (b) Cuts of the fornix resulted only in slight decrease in activity. (c) Cuts through the anterior septum resulted in irritability, slight hyposexuality, and slight decrease in activity. No significant correlations between the various behavioral effects were found. It was suggested that the neural pathways mediating eating, drinking, irritability, muricide, copulation, and activity are relatively distinct and that neural activity in the septal region probably does not influence behavior in any unitary fashion.


Conspecific aggression in the laboratory rat. Attacks by dominant colony males of a domesticated rat strain on conspecific strangers introduced into the colony include both a full range of threat displays and actual biting attack. These dominant males attack and bite both anesthetized and unanesthetized strangers, but threat displays and bites to anesthetized rats are limited. Lesions resulting from these bites are nonrandomly distributed, with most damage to the head and upper back and very few bites to ventral surfaces. When introduced strangers were left in the colonies for several days, 66% were killed, indicating that domesticated rats are capable of lethal attacks on conspecifics without external provocation.


Cortical evoked potential, personality, and intelligence. The averaged cortical evoked potential (CEP) was measured from 296 male subjects in two studies and was correlated with scores on intelligence and personality tests. For both studies, auditory stimuli were used with bipolar electrode placement to the Cz and T3 scalp positions. In the first study, with 84 subjects, stimulus intensity was 95 dB (all dB readings re 20 muN/m2). Intelligence was measured with the Mill Hill Vocabulary Scale and personality with H. J. Eysenck's PEN questionnaire. The second study, with 212 subjects, used additional stimuli at 75 dB and 55 dB. Intelligence was measured with the Ravens Progressive Matrices. State anxiety and other state measures at the time of testing were also taken. Neither study found any relation between the CEP and IQ or personality. However, more bored subjects showed larger CEP amplitudes. The results are discussed in terms of Ertl's neural efficiency hypothesis and Eysenck's arousal theory of personality.


Male hamster preference for odors of female hamster vaginal discharges: studies of experiential and hormonal determinants. Male hamsters approach sources of odors from female hamster vaginal discharges and spend significantly more time around these odor sources than around control locations in the test box. This preference for female hamster vaginal odors appears in sexually inexperienced as well as experienced males, even in individuals isolated from females since the time of weaning. Castration significantly reduces the sex odor preference, and treatment with testosterone propionate partially restores it.


Isolation-induced aggression in newly hatched ducklings. Newly hatched Khaki Campbell ducklings (Anas platyrhynchos domesticus) were initially housed in pairs and subsequently transferred to isolated housing conditions. Subjects living with another bird displayed filial behavior and little aggression upon encountering another duckling in a test arena. In contrast, ducklings housed in isolation exhibited aggressive pecking in addition to filial behavior when another duckling was subsequently encountered. In Experiment 2, ducklings were housed with an imprinting object (i.e., an object that elicits attachment behavior), but they were otherwise isolated from other birds. These subjects displayed little aggression when they were subsequently reunited with a conspecific, indicating that the aggression-precluding effects of social housing are not limited to the particular social stimulus with which the ducklings are housed. In Experiment 3, ducklings were again housed with an imprinting object, but this time the object was behind glass, thereby precluding tactile contact with it. Since these birds also exhibited little aggression when reunited with a conspecific, it is apparent that visual stimulation from an imprinting object is sufficient in itself to preclude subsequent aggression. These findings suggest that stimulation that elicits attachment behavior is the critical factor mediating isolation-induced aggression.