Perinatal events associated with maternal smoking during pregnancy.

To identify components of smoking-related increased perinatal mortality, detailed analyses of data from the Ontario Perinatal Mortality Study (50,000 births, 1,300 deaths, 1960-1961) measured the relationship of maternal smoking to birth weight, gestation, placental complications, and perinatal mortality. Cross-tabulations with other factors and multiple adjustment showed increases with amount smoked of birth weights less than 2500 gm, gestations less than 38 weeks, placenta previa, abruptio placentae, and perinatal mortality. These significant, smoking-related increases were independent of mother's height, weight, hospital status, age-parity group, birthplace, previous pregnancy history, weight gain, time of registration, and sex of child. Maternal smoking had the strongest effect on birthweight in the 8 factor regression, and birth less than 2500 gm increased directly with smoking level from 20% to 340% in 37 data subgroups. Births less than 38 weeks increased 20% and 50% and perinatal mortality increased 20% and 35% for less than 1 pack and 1 + pack smokers, respectively, adjusted for 7 other factors. Placental complications increased consistently with smoking level in all of 37 subgroups except for primiparous less than 1 pack smokers. Adjusted rates increased 25% and 92% for placenta previa, 23% and 86% for abruptions among smokers of less than 1 pack and 1 + packs, respectively. These complications carry high perinatal mortality risk, and account for one-third to one-half of the perinatal deaths attributable to maternal smoking.


The behavior of hypersexual delinquent girls.

The author presents case reports of seven girls with histories of hypersexual behavior associated with delinquency, aggressive behavior, and hypomania. Six of the girls had been exposed to virilizing hormones in utero or in childhood. Most of the girls in the sample were hirsute, tended toward mesomorphic build, and had been young at menarche. The author postulates that exposure to virilizing hormones can result in the kind of hypersexuality and delinquent behavior observed in his sample; he points out the need for adequate medical attention to this type of patient.


The origin, migration and morphology of the primordial germ cells in the chick embryo.

Chick primordial germ cells (PGCs) which separated from the "germinal crescent" entoderm in the period from stages 4 to 8 circulated mostly through the developing blood vessels from stage 10 onward and finally migrated into the gonad. The PGCs making their appearance up to this stage were generally spherical in profile, about 14 mum in diameter. Some of the PGCs in contrast, did not enter the blood vessels but remained in the tissue (mesenchyme) of the embryo proper (tissue PGCs) and possessed pseudopodial processes, suggesting their migration by means of amoeboid movements. The circulating PGCs emerged from blood vessels in the vicinity of developing gonads by three days (gonadal PGCs). The principal mechanism responsible for the subsequent migration of gonadal PGCs is assumed to be amoeboid movements as in the case of tissue PGCs. Notable amounts of PAS-positive glycogen were demonstrated in the cytoplasm of PGCs in all stages obsreved. They also contained yolk and lipids intracytoplasmically, the former dissipating in relatively early stages of development. Electron microscopic observation revealed the electron-opaque, "fragmented nucleolus" in the large nucleus (8 mum in diameter), which represented another prominent feature of chick PGCs. PGCs contained a well-developed Golgi complex and endoplasmic reticulum.


Outpatient evaluation: a new role for the anesthesiologist.

Anesthesiology, although still a relatively young specialty, is expanding beyond the operating room. The author reports material savings in many areas through presurgical interviews in the outpatient department by anesthesiologists.


Factors affecting A-aDo2 after open-heart surgery.

Intrapulmonary shunt, cardiac output, and O2 consumption as factors contributing to alveolar-arterial O2 tension difference (A-aDo2) were tested in 11 patients after open-heart surgery. A-aDo2 was well correlated with intrapulmonary shunt, as expected, but no correlation was found between cardiac index (CI) and intrapulmonary shunt. When total shunt (venous admixture) was above 11%, there was an inverse correlation between A-aDo2 and CI, and it was observed that CI falling below 2.5 L/min/sq m contributed significantly to the enlargement of A-aDo2. Low Pao2 (less than 65 torr, breathing air) was characterized by larger arterial-mixed venous O2 content difference than high Pao2 (is greater than 65 torr, on room air), indicating that A-aDo2 was influenced by cardiac output relative to metabolic demand. Intrapulmonary shunt played a main role in increasing A-aDo2 after open-heart surgery, and systemic factors, including changes in cardiac output and O2 consumption, also contributed to development of hypoxia.


Hepatic clearance of lidocaine during N2O anesthesia in dogs.

Lidocaine catabolism under N2O anesthesia was evaluated in 5 dogs given a lidocaine infusion of 2 mg/kg/min for 20 minutes. Comparison of results with those of a prior similar study with halothane to be significantly faster in the animals given N2O. The extraction ratio for lidocaine, which did not vary with its arterial concentration, was significantly lower with halothane than with N2O. Decreased hepatic catabolism of drugs such as lidocaine should be anticipated in patients anesthetized with potent inhalation agents such as halothane.


The lateral sitting position for neurosurgery.

Over the past 20 years, more than 300 patients have been anesthetized in the lateral sitting position during neurosurgical procedures in the posterior fossa and the cervical and upper thoracic spine. Since the patient can be placed quickly and easily in the horizontal position, the lateral sitting position has a number of advantages over the conventional sitting position, particularly in the treatment of arterial hypotension and venous air embolism. Furthermore, with the patient in the lateral horizontal position, the surgical procedure can be completed satisfactorily.


Inspired air temperature with immersion heater humidifiers.

The temperature of aerosol mixture delivered from 2 types of immersion heater humidifiers was monitored at various sites along the delivery tube. As anticipated, the temperature of the humidified gas decreased as it moved distally from the aerosol generator to the T-adaptor in contact with patient's airway. The immersion heaters with fixed-temperature thermostatic control performed consistently, though most produced inhaled air temperatures higher than desired (greater than 37 degrees C). Immersion heaters with temperature reset features were in general unreliable and invariably gave higher inspired air temperatures with higher settings of the thermostat.


Prolonged neuromuscular blockade with pancuronium in the presence of acute renal failure: a case report.

Prolongation of neuromuscular block following pancuronium administration to anephric patients has been reported. A patient is described whose postoperative course after resection of gangrenous bowel was complicated by acute renal failure and prolonged neuromuscular blockade. A large intraoperative dose of pancuronium was administered without monitoring neuromuscular transmission with a peripheral nerve stimulator. The course was further complicated by administration of gentamicin, an antibiotic known to potentiate neuromuscular blocking drugs. Complete return of neuromuscular transmission occurred 60 hours after administration of last dose of pancuronium and subsequent to peritoneal dialysis. After rapid distribution from plasma, pancuronium, like curare, is eliminated by the renal pathway. The absence of renal excretion considerably increases the duration of action of curare when given in high doses. It has been suggested that doses of pancuronium greater than 3.6 mg/sq m or multiple doses would result in prolonged neuromuscular block in patients without renal function. The case reported illustrates the importance of monitoring neuromuscular transmission during administration of pancuronium in the presence of renal insufficiency to avoid this complication.


Unrelenting hypotension associated with an acute abdomen in a comatose hemophiliac child: a case report.

A comatose 4-year-old hemophiliac presented with an acute abdomen; subsequently he developed unrelenting hypotension. An immediate exploratory laparotomy was required, without time for determining baseline factor VIII (AHF) levels. Despite hypotension and hemorrhage, vigorous intraoperative fluid volume replacement and the administration of fresh frozen plasma and AHF concentrates brought a successful conclusion.


An unusual complication of lumbosacral plexus block: a case report.

Lumbosacral plexus block is a valuable anesthetic technic in the hands of an experineced anesthesiologist. However, as illustrated by this case report, anesthetic dosage must be selected with great care, and patients unable to describe paresthesias should be excluded from use of this procedure.


Prolonged neuromuscular blockade associated with trimethaphan: a case report.

A case of prolonged neuromuscular blockade associated with the administration of trimethaphan to a neurosurgical patient aged 29 is believed to be the possible result of interaction between trimethaphan, a ganglionic-blocking drug, and muscle relaxant. This possibility should be kept in mind when the administration of trimethaphan is being considered.


Anesthetic considerations for pericardectomy in uremic pericardial effusion.

Fifteen patients with impending uremic cardiac tamponade underwent either anterior pericardectomy or pericardiostomy under general anesthesia. Anesthetic agents and technics were selected in the light of physiopathologic derangements involving fluids and electrolyte balance, excretory and cardiovascular disturbances, and pharmacodynamics. Anesthesia was induced with diazepam and maintained with N2O-O2 (70:30) and fentanyl. Pancuronium was used for muscle relaxation. Adequate preoperative assessment of patient, careful monitoring of vital signs, maintenance of the critical balance of fluid and blood replacement, and selection of anesthetic agents for minimal depression of vital multiorgan systems provided excellent results. No postanesthetic morbidity or mortality occurred.


Circulatory responses and halothane concentrations during gastric or gallbladder traction with and without neuromuscular blockade.

Systolic and diastolic blood pressures and heart rates did not change significantly and ventricular dysrhythmias did not occur during gastric or gallbladder traction in patients anesthetized with N2O-halothane, with or without d-tubocurarine or pancuronium neuromuscular blockade. Systolic and diastolic blood pressures and heart rates just before abdominal viscera traction were less (p is less than 0.05) in patients paralyzed with d-tubocurarine than with pancuronium despite similar arterial halothane concentrations. In contrast, these measurements in patients without neuromuscular blockade were similar to those during pancuronium paralysis despite significantly greater arterial halothane concentrations in the unparalyzed patients. The authors conclude that circulatory responses during gastric or gallbladder traction are clinically insignificant during N2O-halothane administration, with or without d-tubocurarine or pancuronium neuromuscular blockade. Intraoperative blood pressures and heart rates were influenced by the neuromuscular blocker chosen. Neuromuscular blockade permitted anesthetic maintenance with lower arterial halothane concentrations than when paralysis was not present.


Methoxyflurane biotransformation and renal function following methoxyflurane administration for vaginal delivery or cesarean section.

Methoxyflurane (MOF) administration for conscious analgesia during vaginal delivery (range 5 to 70 min, mean 23 min) or for anesthesia following delivery of the infant at cesarean section (range 25 to 70 min, mean 44 min) was studied in 18 healthy parturients. Serum ionic fluoride increased significantly in both groups 2 hours after discontinuing MOF with peak concentrations of 11.2 and 14.1 mumol/L in the vaginal delivery and cesarean section groups, respectively. Individual peak serum ionic fluoride levels in the 2 groups of 21 and 25 mumol/L were well below reported levels for subclinical toxicity. Significant ionic fluoride elevations in fetal umbilical venous blood (mean 5.3 mumol/L) were measured in the vaginal-delivery group. Maternal urinary ionic fluoride and oxalate were elevated 24 and 48 hours postpartum. BUN, creatinine, urine volume, and urine osmolality remained within normal range. These data indicate that hazardous elevations of serum ionic fluoride with subsequent renal dysfunction are unlikely following low-dose MOF administration for vaginal delivery or cesarean section.


Effect of isoproterenol and propranolol on halothane MAD in dogs.

To evaluate the effect of propranolol on anesthetic requirement, changes in minimum alveolar concentration (MAC) of halothane accompanying acute (2 and 10 mg/kg IV) and chronic (200 mg/day orally for 10 days) propranolol administration were determined in dogs. MAC did not change significantly in either case. Beta-adrenergic blockade was achieved with the 2 mg/kg propranolol dose, as indicated by abolition of the pulse rate increase in response to isoproterenol infusion. Isoproterenol itself did not alter MAC. Mean arterial pressure (MAP) and heart rate (HR) at any given level of alveolar halothane were the same before and after chronic propranolol administration. Acute IV administration of propranolol, especially at the 10 mg/kg dose, reduced MAP and increased HR only transiently.


Galactorrhea-amenorrhea syndromes: etiology and treatment.

Fifteen patients with galactorrhea-amenorrhea syndromes were studied before, during, and after treatment with bromergocryptine. Galactorrhea and amenorrhea were noted after pregnancy (6 patients), after oral contraceptive therapy (5 patients), and in association with pituitary adenoma (4 patients). Before treatment prolactin values were elevated ranging from 27 to 125 ng/ml, while luteinizing hormone and progesterone levels failed to show ovulatory peaks or luteal phase progression. Eleven patients had luteinizing hormone-releasing hormone tests before therapy. Response was normal in 8, subnormal in 2 pituitary adenoma, and supranormal in 1 patient with premature ovarian failure. Treatment with bromergocryptine was associated with a lowering of serum prolactin, cessation of lactation in all, and return of ovulatory menses in 14 of 15 patients. All relapsed when therapy was discontinued. Four patients became pregnant while on therapy. Long-term bromergocryptine therapy is effective for all forms of galactorrhea-amenorrhea syndromes studied.


Marital sexual dysfunction: ejaculation disturbances.

The three clinically encountered disturbances of the male orgasmic phase--that is, premature ejaculation, inability to ejaculate, and retrograde ejaculation--are discussed. The problem of defining premature ejaculation is explained and the possible pathogenic mechanisms of its origin are reviewed. The classic premature ejaculator is described. Also included are an elaboration of the newer therapeutic approaches to the problem and a suggested clinical protocol for screening patients. The other two, less common dysfunctions are briefly discussed.


Concordance in physical growth for monozygotic and dizygotic twins.

Heights and weights were analysed for 636 twins who had been repeatedly measured from birth to 4 years. MZ twins were less concordant for birth weight than DZ twins, principally as a result of a few MZ pairs who exhibited very large differences. By one year of age, however, MZ twins had become more concordant for weight (Rmz = 0-87) while DZ twins moved further apart (Rdz = 0-55). For height, at birth, the MZ correlation was 0-58 while the DZ correlation was 0-82, but by 2 years, the MZ correlation reached 0-89 and the DZ correlation regressed to 0-58. The actual size differences within pairs followed a parallel course--for MZ twins, the differences became smaller following birth, whereas DZ differences became larger. The results are discussed in terms of (a) prenatal influences that differntially affect birth size within MZ pairs and DZ pairs, and (b) the rapid convergence of each twin on his genetic growth curve.


A note on the accuracy of recalled age at menarche.

In a longitudinal study, age at menarche for Swedish girls was recorded by school nurses. Some four years after the event the girls were asked to recall the date. The correlation between actual and recalled age at menarche for 339 girls was r = 0-81 +/- 0-05. 63 per cent of the girls recalled the date within +/- 3 months. It is concluded that recalled age is too inaccurate for most purposes.


Seasonality in twin births.

Twin births have occurred seasonally in England and Wales since 1940. The probability of a twin birth is highest in November and lowest in May. The difference between the two probabilities is of the order of seven per cent.


[Lincomycin formation by strain R-367 on a complex medium not containing corn steep].

A new complex medium for biosynthesis of lincomycin by strain R-367 was developed using one-factor experiments and mathematical schemes of planning. A complex of mineral salts containing nitrogen, sulphur, phosphurus, magnesium and trace elements was introduced into the content of the new medium. This provided elimination of corn steep liquor from the medium. The linomycin production level in the fermentation broth with the use of the new medium was at an average 2.5 times higher than that on the initial medium containing corn steep liquor.


Effect of substrate on metabolite production of Alternaria alternata.

Alternariol and alternariol monomethyl ether are commonly associated with weathered grain sorghum. Production of these metabolites and altenuene by isolates of Alternaria alternata was evaluated on various sterile grain substrates. At 35% moisture content and 25 C, metabolite yields were highest on rice, intermediate on sorghums, and lowest on wheat and yellow corn. Fourteen-to 21-day cultures on milled rice were best in terms of ease of metabolite recovery, even though yields were higher on 28-day cultures of rough and brown rice. Metabolite production was reduced when rice was supplemented with yeast extract or yeast extract plus Czapek-Dox broth.


Immunological studies on the proteins of human eccrine sweat.

Proteins present in the eccrine sweat collected from the skin surface of male volunteers were studied. The protein content ranged from 0.154--0.774 g per liter. A total 2282 ml sweat was concentrated until a final protein content of 25 mg/ml was reached. Paper electrophoresis revealed 3 fractions migrating like alpha-, beta-, and gamma-globulins. Eight different proteins were demonstrated by immunoelectrophoretic analyses using rabbit antisera directed against human sweat proteins. Three of these proteins were identified as blood serum proteins. One of the remaining 5 proteins could be of epidermal origin; another one which occurs in comedones as well as in sweat seems to be eluted from the sebum film present on the skin surface.


Effect of methadone addiction on N-demethylation activity of rat liver microsomes.

Adult male rats were addicted to methadone via small dosages in their drinking water. The N-demethylation activity of microsomal mixed function oxidase in liver was determined for partially addicted (25 days of drug consumption) and fully addicted (38 days of drug consumption) rats. Compared to controls which did not receive the drug, enzyme activity expressed on a microsmal protein basis was increased in partially addicted, but not increased in fully addicted animals. Enzyme activity expressed on a liver weight basis was decreased in partially addicted animals but unchanged from contro levels in fully addicted animals. Brief (three-day) exposures to phenobarbital or high dosages of methadone led to an increase in N-demethylation activity.


[Transport and utilization of alditols in the yeast Rhodotorula gracilis glutinis (author's transl)].

The obligate aerobic yeast Rhodoturula gracilis was found to take up the alditols D-glucitol, D-mannitol, ribitol, xylitol, D-arabinitol, L-arabinitol and erythritol by means of a constitutive mobile membrane carrier. This uptake involved active transport, that is, it was dependent on the supply of metabolic energy, leading to the accumulation of alditols inside the cells. The accumulation ratio (intracellular concentration to extracellular concentration, Si/SO) was much lower for alditols than for monosaccharides. As for monosaccharides, this ratio decreased with increasing extracellular concentration, even to values below 1. The kinetic data showed that the carrier system for alditols was identical to that for monosaccharides, though it had a much lower affinity and maximum velocity for alditols. Hence the uptake of alditols was blocked in the presence of monosaccharides. Only ribitol and L-arabinitol were catabolized following enzyme induction. The other alditols were not broken down.


Although alditols reached the interior of the cells of Rhodotorula gracilis by means of the constitutive monosaccharide carrier, a second carrier of pentitols was found to be inducible. Ribitol and L-arabinitol, entering the cells by the constitutive carrier, induced a single catabolic enzyme system, including a carrier of pentitols. This induction depended on de novo protein synthesis. In addition to ribitol and L-arabinitol, the induced catabolic enzyme system was also specific for xylitol and D-arabinitol; these two alditols were not inducers. Hexitols, erythritol and pentoses were not transported or catabolized by the induced system. The induced carrier had a lower half-saturation constant, KT, and higher maximal velocity, V, than the constitutive carrier of penitols. Kinetic studies of induced transport indicated, that this was an active process which appeared to be coupled with the simultaneous uptake of H+-ions (H+-substrate symport). The induced transport of pentitols was strongly inhibited by monosaccharides, although not competitively. The enzyme synthesis induced by ribitol or L-arabinitol was repressed by monosaccharides.


Growth and physiology of Candida utilis NCYC 321 in potassium-limited chemostat culture.

When grown in a defined simple salts medium, plus vitamins, Candida utilis displayed an absolute requirement for potassium. But the potassium content of this yeast was exceedingly variable and, with aerobic chemostat cultures (grown at a dilution rate of 0.1 h-1; 30 degrees C; pH 5.5), was low (less than 0.2%, w/w) when they were potassium-limited and high (greater than 2%, w/w) when glucose-limited. With potassium-limited cultures, the cell-bound potassium content also varied markedly with growth rate, though hardly at all with glucose-limited cultures; magnesium- and phosphate-limited cultures gave intermediate responses. Changes in cell-bound potassium content correlated only weakly with changes in the cellular contents of magnesium, phosphate and RNA, but strongly with changes in both the Yglucose and Y0 values, indicating an involvement of potassium in the generation of energy by oxidative phosphorylation reactions and/or the utilization of this energy for growth processes. Studies with isolated mitochondria revealed that potassium-limited organisms had a changed content of cytochrome b relative to cytochrome a, and lacked coupling at either site 2 or site 3 of the respiratory chain. These results are discussed in relation to the reported functions of potassium in the growth of micro-organisms, and the organizational differences between prokaryotic and eukaryotic cells.


Sporobolomyces salmonicolor var. fischerii, a new yeast.

A new variety of Sporobolomyces salmonicolor, a basidiomycetous yeast isolated from the cerebrospinal fluid of a human case of meningitis has been described. The variety can be differentiated from the species primarily by its ability to assimilate D-galactose and L-arabinose.


Spore germination in Dictyostelium discoideum. II. Effects of dimethyl sulfoxide on post-activation lag as evidence for the multistate model of activation.

Mutant spores of Dictyostelium discoideum, strain SG-10, differ from wild type spores in their ability to spontaneously germinate, to be activated with 5% Dimethyl Sulfoxide (DMSO), and to be deactivated with 0.2 M sucrose. Both heat-activated wild type and mutant spores began to swell after a lag of 60-75 min at ambient temperature. Suspension of heat activated spores in 5% DMSO resulted in blockage of spore swelling and a concomitant severe inhibition of respiration; removal of 5% DMSO allowed resumption of respiration and the spores began to swell after a lag of only 15 min. It was concluded that 5% DMSO allowed the early reactions (M) to proceed but blocked the later reactions (R) of post-activation lag. Treatment of one day old spores with 20% DMSO solution for 30-120 min quantitatively activated the population. The post-activation lag time was directly dependent on the time of 20% DMSO treatment. Spores activated with 20% DMSO treatment could be deactivated by incubation at 0 degrees C; the spores most quickly deactivated at 0 degrees C were those within 10 min of swelling. Mitochondrial transport inhibitors such as azide and cyanide caused deactivation in an analogous manner. It is hypothesized that spores proceed to the second portion of the lag phase called (R) before the environment determines if dormancy is reimposed or if germination will proceed. The sensitive strain (SG-10) showed a greater degree of "damage" than the wild type after supraoptimal treatment with 40% DMSO. The spores became more resistant with age to the "damaging" action of 40% DMSO. All the observed effects of DMSO treatment were compatible with our multistate model of activation which suggests that the early portion of the lag phase (M) may involve a relative uncoupling of oxidative phosphorylation while the later portion (R) may require tight coupling.


Studies on metabolic role of 5'-Methylthioadenosine in Ochromonas malhamensis and other microorganisms.

Several compound containing a thiomethyl group were found to replace vitamin B12 in a protozoan, Ochromonas malhamensis. The order of the effectiveness was as follows: 5'-methylthioadenosine is greater than S-adenosylmethionine is greater than 5-methylthioribose is greater then L-methionine. A similar order was obtained with respect to the permeability of these compounds into the protozoan cells, except for S-adenosylmethionine. 5'-Methylthioadenosine and 5-methylthioribose as well as L-methionine markedly increased the intracellular content of L-methionine. The level of S-adenosylmethionine was also increased by them, but to lesser degree. The thiomethyl group of the compounds was established to be incorporated into S-adenosylmethionine. The metabolic fate of the thiomethyl group of 5'-methylthioadenosine cannot be distinguished from that of L-methionine. A high activity of 5'-methylthioadenosine nucleosidase was detected in the cell-free extracts of the protozoan. These results strongly suggest that 5'-methylthioadenosine would be metabolized to L-methionine would be ocnverted to S-adenosylmethionine. Like L-methionine and vitamin B12, 5'-methylthioadenosine and 5-methylthioribose may play an important role in maintenance of the C-1 pool in Ochromonas malhamensis. Neither 5'-methylthioadenosine nor 5-methylthioribose replaced vitamin B12 in some vitamin B12-requiring bacteria. This result is consistent with the fact that neither compounds was significantly taken up by these bacteria.


Effects of polyoxin D on germination, morphological development and biosynthesis of the cell wall of Trichoderma viride.

When polyoxin D is added to a spore suspension of Trichoderma viride at a concentration from 50-100 mug/ml, it inhibits from 40-60% of germination. This percentage increases if dimethylsulfoxide (DMSO) is added. Mycelium growing in the presence of polyoxin D becomes irregular and loses its rigidity, showing several bulges along the hypha. Under the electron microscope the features of the cell wall and cytoplasmic content are apparently normal. Nevertheless, after incubation with different lytic systems or with (14C) glucose, it can be seen that polyoxin D partially inhibits the biosynthesis of beta-(1-3) glucan and the biosynthesis of chitin to a greater extent attaining inhibition of 83% at 100 mug/ml of the antibiotic concentration. Rgenerating protoplasts are less affected by polyoxin D. They do regenerate slower but the percentage of regeneration is more than 80%. Aberrant tubes synthesized by these protoplasts are not affected, they manifest their usual morphology and lack of chitin is confirmed in their composition.


Influence of WA-335, a factor which blocks serotonin receptors, on neuroleptic-induced catalepsy.

In a previous study, anticataleptic action of cyproheptadine was reported. The present investigation deals with the influence of WA-335 (9,10-dihydro-10-(1-methyl-4-piperidylidene)-9-anthrol), another antagonist of serotonin, on catalepsy induced in rats with spiroperidol, pimozide, fluphenazine and reserpine. WA-335 antagonized catalepsy induced by these neuroleptics (the effect on reserpine-induced catalepsy was weakest). Joint administration of WA-335 and L-DOPA with an inhibitor of peripheral decarboxylase, or WA-335 and amantadine produced a stronger antagonistic effect (spiroperidol catalepsy) than either of these substances separately. WA-335 did not prevent catalepsy induced with physostigmine.


The action of compound WA-335 on the central nervous system.

Compound WA-335 (9,10-dihydro-10-(-1-methyl-4-piperidylidene)9-anthrol) was studied with regard to its antidepressant and central antiserotonin action in mice and rats. WA-335 depresses reserpine-induced hypothermia, particularly in mice, but does not affect ptosis induced with this neuroleptic. The compound diminishes spontaneous motility in mice and rats, including mice stimulated with amphetamine. WA-335 acts synergistically with amphetamine in which it potentiates stereotyping and enhances motility. The compound has no influence on the action of L-DOPA and does not alter hypothermia or increase motility induced with this amino acid. WA-335 does not affect the drop in body temperature or stereotype induced with apomorphine. In mice, WA-335 inhibits motility elicited with L-5-hydroxytryptophan, and in rats as well as mice prevents occurrence of head-twitches due to activation of serotonin neurons. The compound has no effect on the righting reflex abolished with fenfluramine in frogs, and given together with reserpine slightly counteracts abolition of this reflex. WA-335 raises the level of serotonin in the brains of rats, and lowers the level of 5-hydroxyindoleacetic acid. In addition, WA-335 exhibits cholinolytic activity and abolishes the symptoms elicited with oxotremorin in mice and rats.


Behavioral effects of phenoxybenzamine administered intraventricularly in rats.

Phenoxybenzamine (PNB) injected into the lateral ventricle of the brain weakens spontaneous motility of rats and antagonizes hyperactivity induced with noradrenaline, amphetamine and caffeine. Body temperature rises briefly, sleep induced with chloral hydrate is shortened, and high doses enhance amphetamine-induced stereotypy. PNB injected intraventricularly intensifies turnover of noradrenaline but it has no effect on turnover of dopamine in rat brains. The results confirm a blocking influence of PNB on the cerebral noradrenaline receptors.


[Hematopoiesis in the yolk sac of several representatives of the Sauropsida].

Hemopoiesis in the yolk sac of chicken and crocodile embryos was studied at different developmental stages. Primary intravascular erythropoiesis is closely related to the formation of sanguineous islands appearing in the zone of the yolk growth in the visceral mesoderm. With the development of folds enlarging the surface of the yolk resorbtion, around the vessels running within the folds there appear foci of primary granulopoiesis. The process begins in the paravasal mesenchyma which is gradually disguised by hemopoietic cells (in chicken embryos--at the stage of 8 days, in crocodiles--23 days of incubation). The granulopoiesis continues in chicken embryos during 1/3 of the incubation period, in crocodiles--during 2/3 of the incubation time. The leukopoiesis foci are developed more intensively in crocodiles as well at the size of the yolk sac folds. Leukocytic accumulations disappear in the crocodile sac after hatching (38-40 cm). The change of primary erythropoiesis for the secondary one in the chicken is preceded by the appearance of megaloblastic forms of erythrocytes with the compact homogeneous cytoplasm. They are absent from crocodiles.


[Malignant vascular tumors of the lung (cytologic and histologic parallels)].

Analysis of 78 observations (including 6 of the author's own) of primary malignant vascular tumours of the lung, showed that these tumours had no characteristic clinicoroentgenological features; in this connection the question of their preoperative cytological diagnosis is of importance. The retrospective collations of cytological and histological data in 8 patients made it possible to establish criteria of cytological diagnosis of similar tumours. For the first time in the Soviet literature a suggestion was put forward that basing on the data of cytological investigations it is possible in a number of cases to give a detailed verification of tumours of the lung with presumable indication to hemangioendothelioma, angiosarcoma, or hemangiopericytoma. Metastases of malignant vascular tumours of other localizations in the lung are of analogous cytological pattern. correct diagnosis of primary of metastatic tumour can be made only if all the complex of clinico-roentgenological and cytological data are taken into account.


[Ultrastructural localization of lysosomal and nuclear beta-glycerophosphatase activity in the urethral discharge cells of gonorrhea].

Activity of acid phosphotase in the urethra discharge in patients with various forms of gonorrhea (acute, torpid, and chronic) was studied with the use of electron-microscopy and biochemical methods. A positive reaction of nuclei of the epithelial cells to acid phosphotase in lysosomes and perichromatin granules was demonstrated. In polymorphononuclear leucocytes the positive reaction to acid phosphotase could be also sometimes observed in granules of the cytoplasm. The electronograms presented testify to heterogeneity and high activity of acid phosphotase in lysosomes. Neither electron-microscopy nor biochemical methods could help reveal any differences in the activity of acid phosphotase in various forms of gonorrhea.


Optic nerve colobomas of autosomal-dominant heredity.

Fifteen members of a family had congenital colobomas of the optic nerves, all of which were bilateral. The bolobomas appeared as enlargement of the physiologic cup with severely affected eyes having huge cavities in the site of the disc. A variable amount of glial tissue was present within the colobomas. Either macular or extramacular serous detachment or sequelae were present in 21 of the 30 affected eyes, and often led to total blindness. Radioisotope cisternography in one patient failed to implicate cerebrospinal fluid as the source of the subretinal fluid. The disorder is apparently inherited as an autosomal dominant.


Galactose metabolism in regenerating rat liver.

1. Rats trained on a controlled lighting and feeding schedule were subjected to partial hepatectomy or sham operation. 2. After a large period of about 6h the activity of UDP-galactose 4-epimerase increased threefold, reaching a maximum 4 days after partial hepatectomy, and returned to normal values within a fortnight. 3. The enzyme pattern of the UDP-galactose-glycoprotein galactosyltransferase was biphasic, one peak appearing at 20 h, the second at 72 h after partial hepatectomy. 4. The rise in enzyme activities could be blocked by the injection of actinomycin D, and the Km values for UDP-glucose and UDP-galactose were nearly identical in regenerating and adult liver. It is therefore concluded that the increase in enzyme activity is due to synthesis de novo of enzyme protein.


B-scan ultrasonography in idiopathic hypertrophic subaortic stenosis. Study of left ventricular outflow tract and mechanism of obstruction.

Studies were made with standard time motion and B-scan echocardiography on 48 patients including 5 with idiopathic hypertrophic subaortic stenosis (hypertrophic obstructive cardiomyopathy), undergoing diagnostic cardiac catheterization. The dimensions of the left ventricular outflow (O) and inflow (I) tracts were measured on the B-scan images. The outflow tract was significantly narrowed in idiopathic hypertrophic subaortic stenosis at both end-systole (1-1+/-0-1 cm) and end-diastole (1-3+/-0-1 cm) when compared with the average width in other patients (2-6+/-0-1 and 3-0+/-0-1 cm, at end-systole and end-diastole, respectively) (P less than 0-001) or normal subjects (2-4+/-0-3 and 2-9+/-0-2 cm) (P less than 0-01). Furthermore, the O/I ratio differed significantly in idiopathic hypertrophic subaortic stenosis (0-5+/-0-1 at end-systole and 0-6+/-0-1 at end-diastole) from that in all other groups (1-4+/-0-1 at both end-systole and end-diastole) (P less than 0-005). There was no appreciable change in the width of the outflow tract from mid- to end-systole in the two patients in whom this was examined. The data support the contention that the anterior leaflet of the mitral valve assumes an abnormally anterior position in idiopathic hypertrophic subaortic stenosis. Though the systolic anterior movement of the tip of the anterior leaflet of the mitral valve shown by M-mode echocardiography could not readily be confirmed with B-scans, we believe that the narrowed outflow tract found in the present investigation contributes to the obstruction that occurs in this disease. We suggest that this outflow tract narrowing is probably caused by hypertrophy of the ventricular septum which in itself contributes to the narrowing, but which also displaces the papillary muscles and thus produces abnormal traction on the mitral valve and striking anterior displacement of the valve apparatus.


Influence of labour on the lecithin, lecithin/sphingomyelin (L/S) ratio and palmitic acid values in the amniotic fluid.

The lecithin/sphingomyelin (L/S) ratio, lecithin concentrations and palmitic acid levels in amniotic fluid were studied during labour and compared with values found before labour. All were significantly higher in women in labour. The explanation and significance of this finding are briefly discussed.


The effect of internal urethrotomy and urethral dilatation on the postoperative course of patients undergoing surgery for stress incontinence.

Three years' experience of a total of 128 patients undergoing surgery for stress incontinence is described. Forty-three patients had an internal urethrotomy performed routinely at the end of the operation, 26 had routine urethral dilatation and 59 had neither procedure. Internal urethrotomy, and to a lesser extent urethral dilatation, resulted in prompt return of spontaneous and efficient micturition following removal of the Foley catheter on the third postoperative day. The postoperative stay averaged 6-4 days in the urethrotomy group of patients, 7-6 days in those who had urethral dilatation and 10-6 days in the others. The two-year cure rate in the patients who had neither procedure performed was 85 per cent and as yet no failures have occurred in the patients who had internal urethrotomy or urethral dilatation performed.


Flexibility in the specificity site of serine proteases.

The statistical availability of tryptophan and tyrosine residues with one ring face fully exposed to solvent was examined for two serine proteases and their derivatives by investigating the formation of charge transfer (CT) complexes between the aromatic donor residues of the protein and the acceptor 1-methylnicotinamide chloride. The availability of the ring face of one of the two exposed tryptophan residues in trypsin has been previously shown to be pH dependent and to parallel the acid side of the pH-activity profile of the enzyme. The present results indicate that, in diisopropylphosphoryl-trypsin (DIP-trypsin), this residue [which was identified as Trp-215 in native trypsin (chymotrypsin numbering)] is locked in a relatively rigid, pH-independent conformation with one ring face rotated out toward the solvent. In the zymogen and DIP-zymogen, the ring face is essentially unavailable. Chymotrypsin, like trypsin, has a pH-depent tryptophan residue available for complexation with the CT acceptor, but unlike trypsin, the pH dependence is apparently associated with dimerization of the enzyme. These and other data suggest this residue is the same as in the homologous trypsin structure, i.e., Trp 215, and that the ring face is mostly buried in the zymogen. Comparison of the crystal structure models of chymotrypsin and chymotrypsinogen shows that, as the specificity pocket opens up from its collapsed structure upon zymogen activation, the ring face of Trp-215 moves out and rotates relative to the surface of the enzyme in such a fashion as to become more accessible to solvent. These observations are in accord with the present CT results and provide additional support for the assignment of changes in Trp-215 availability to parallel changes in the conformation of the specificity pocket of these serine proteases. The present investigation also shows that, although a tryptophan ring face is partly exposed in DIP-chymotrypsin, its statistical availability more closely resembles that of the zymogen than the native enzyme. The reverse appears to be true for DIP-trypsin, which suggests the possibility that the specificity pocket in DIP-chymotrypsin may be partially collapsed while the catalytic residues are frozen in the conformation of the acyl-enzyme.


Physical studies on the H3/H4 histone tetramer.

High-resolution proton magnetic resonance spectroscopy (270 MHz), circular dichroism, and infrared spectroscopies and ultracentrifugation studies have been carried out on the salt-extracted (H3/H4)2 tetramer from calf thymus. The tetramer contains about 29% alpha helix and no beta structure. It is denatured in 6 M urea but can be renatured simply by dialysis to water. The proton spectrum shows a number of perturbed resonances which are not observed in the spectra of either H3 or H4 alone. The observation of these resonances demonstrates that the tetramer contains some elements of tertiary structure. The overall appearance of the spectrum however is close to that of a partially denatured protein. Sedimentation velocity studies show the tetramer to have a frictional ratio of 1.99 in 50 mM acetate/50 mM bisulfite and thus to be hydrodynamically quite different from a globular protein. Two possible structural models compatible with the data are discussed.


Phospholipase A2 complexes with gadolinium (III) and interaction of the enzyme-metal ion complex with monomeric and micellar alkylphosphorylcholines. Water proton nuclear magnetic relaxation studies.

Gadolinium (III) binds competitively with calcium(II) to porcine pancreatic phospholipase A2 (EC 3.1.1.4) and its zymogen. The enzyme-Gd3+ complex exhibits 4% of the hydrolytic activity of the corresponding Ca2+ complex toward a dispersion of dioctanoyllecithin. Dissociation constants for the Gd3+ complex of enzyme and proenzyme were evaluated from water proton relaxation rate (PRR) titrations. At pH 5.8, the dissociation constants for the Gd3+ complexes of enzyme and zymogen are 0.50 and 0.18 mM, respectively. Dissociation constants for the complexes of enzyme with Ca2+, Eu3+, and Tb3+ were evaluated in PRR titrations by competition of these cations with Gd3+ binding. PRR enhancement factors for the Gd3+ complexes of enzyme and proenzyme are 16.4 and 5.8, respectively, at 22 degrees C and 24.3 MHz. Binding of a homologous series of n-alkylphosphorylcholines to the enzyme-Gd3+ complex was investigated through the influence of monomeric and micellar forms of these amphiphiles on the PRR enhancement factor for the enzyme-bound Gd3+. Separate monomer and micelle binding regions were observed in titrations using n-alkylphosphorylcholines with critical micelle concentrations ranging from 15 muM to 13 mM. In every case, the enhancement factors for the enzyme-Gd3+ complexes were significantly greater than that for the tenary complex, enzyme-Gd3+ -monomer. Morever, a synergism was observed in the binding of Gd3+ and micelles to the enzyme. The magnitudes of the PRR enhancement factors for the enzyme-Gd3+ complexes with micelles of n-alkylphosphorylcholines indicate that the bound Gd3+ is freely accessible to the bulk solvent. These results suggest a model for the enzyme-micelle complex in which the active site is spatially removed from the enzyme-micelle interface.


Radioprotective effect of superoxide dismutase on model phospholipid membranes.

1. Hydroperoxide formation in model membranes was measured via the net increase in absorbance at 232 nm after exposure to X-rays or 137Cs gamma rays in the presence and absence of bovine superoxide dismutase and other radical scavengers. 2. Membranes X-irradiated in air to 4200 rad at 210 rad/min exhibited a large increase in absorbance, a major portion of which was O2--mediated since active superoxide dismutase at 1 mug/ml reduced it by more than 80% to the level observed in N2O. In N2 the change in absorbance was smaller than in N2O but not in proportion to the halving in OH production. 3. The net absorbance of membranes exposed to a constant dose from 137Cs increased with decreasing dose rate. A minor component of this effect was due to exposure protraction with decreasing dose rates while the major component was attributed to long chain reactions initiated by ionizing radiation. A corollary effect was also observed, namely, that with reducing dose rate the dose required to elicit a constant absorbance change decreased. Both aspects were abolished by superoxide dismutase at 1 mug/ml. 4. The enzyme protected membranes after an acute exposure and from low level radiation at natural background while its inactivated form sensitized.


Further observations on the photooxidation of DNA in the presence of riboflavin.

Illumination of DNA in the presence of riboflavin results in an increase in buoyant density and a decrease in the temperature of the thermal helix-coil transition (Tm). The increase in buoyant density is maintained even after thermal denaturation, which indicates that it reflects a chemical alteration of a DNA base (presumably guanine). beta-Carotene, a quencher of singlet oxygen, inhibits the increase in buoyant density but it prevents only partially the decrease in Tm. This is taken as an indication that the photo-induced alteration of the DNA structure is due to more than one reaction. Illumination of DNA in the presence of methylene blue also causes an increase in buoyant density , but this increase is not retained upon thermal denaturation.


Developmental changes in plasma membrane fluidity in chick embryo heart.

1. Decreases in the rate of transport of sugars (facilitated transport), amino acids (active transport), and urea (simple diffusion) occur in chick embryo heart during development. This work considers the possibility that changes in the plasma membrane fluidity during development contribute to the observed changes in transport activities. 2. Technics were developed for subcellar fractionation of chick embryos and adult chickens. 3. The depolarization of the fluorescence of 1,6-diphenylhexatriene was used to estimate the fluidity of the lipid portion of plasma membrane enriched fractions of hearts from chick embryos at various stages of development and from adult hearts. 4. There is a pattern of decreasing membrane viscosity as development proceeds. Between 5-6 days and 10 days of embryonic life a 20% decrease in viscosity of the plasma membrane-enriched fraction occurs. Between 10 and 20 days of embryonic life there is no significant change in viscosity. Between 20 days of development (1 day before hatching) and adulthood there is a further 55% decrease in plasma membrane viscosity. 5. It is proposed that the changes in membrane fluidity observed may contribute to developmental changes in membrane transport activities, but other factors must also be involved.


Multiple thermotropic state transitions in erythrocyte membranes. A laser-Raman study of the CH-stretching and acoustical regions.

1. We have examined the Raman scattering from erythrocyte ghosts at 2700 cm-1 -3000 cm-1 (CH-stretching region), that at low frequencies due to acoustical vibrations, as well as that due to v (-C=C-) and v (=C-C=) of beta-carotene, in the temperature range of -30 degrees C to  +30 degrees C. For comparison we have also evaluated the Rama spectra of liposomes composed of lecithins+/-cholesterol. 2. Plots vs. temperature of the intensities (I) of the approx. 2880 cm-1 and 2932 cm-1 bands relative to the intensity of the thermally stable 2850 cm-1 band, i.e. the (I2880/I2850) and (I2932/I2850) ratios, revealed discontinuities in both erythrocyte membranes and liposomes. 3. In erythrocyte membranes, plots of (I approximately 2880/I2850) and (I approximately 2932/I 2850) reveal several discontinuities. Those reported by (I approximately 2880/I 2850) occur at +17 degrees C and approx. -4 degrees C. The ratio (I approximately 2932/I 2850) reveals a transition between -20 degrees C and +10 degrees C, 1/2 maximal at -8 degrees C. The ratio  (I 1527/I 1158), representing the relative intensity of v (-C=C-) vs. v (=C-C=) of membrane-bound beta-carotene, changes discontinuously with the 17 degrees C transition, but is stable over the rest of the temperature change studied. 4. The data indicate that a major membrane-state transition takes place below 0 degrees C but that an additional transition occurs at approx. 17 degrees C.


[A study of the protein composition of the zona pellucida of mammalian oocytes and zygotes by a method of microelectrophoresis in polyacrylamide gel].

Z. pellucide from oocytes and zygotes of CBA mice and albino rats were lysed in a microvolume of 1% buffered sodium dodecylsulphate (SDS). The microlysate (about 1,0--2,0-10(-7) g protein) was fractionated by means microdiscelectrophoretic technique, using 10%- and 15%-polyacrylamide gel containing 0.1% SDS. The new proteins with increased anodic mobility was found among zonal proteins after fertilization in CBA mice and rats. Homogenous fraction of glycoprotein (approximately molecular weight 44 000) is found in z. pellucida of rat oocytes. This glycoprotein fraction from rat zygote coats produces 3 components moving to anode under electrophoresis. The data obtained support the hypothesis that the mechanism of "zonal fraction" under fertilization in mammals can be due to partial lysis of coat proteins by proteinase of cortical granules.


Glucose disappearance rate, insulin response and growth hormone response in the small for gestational age and premature infant of very low birth weight.

The purpose of the present investigation was to study the disappearance rate of intravenously administered glucose, plasma insulin, plasma growth hormone (GH) and their responses to an intravenous glucose load in 29 infants who were small for gestational age (SGA). The findings were compared to those in 19 premature babies of similar birth weight who were appropriate for gestational age (AGA); in all the weights were below 2,000 g. The babies were divided in two groups according to their age when tested, at less than 24 and 25-48 h. SGA infants cleared glucose more rapidly than AGA infants (p less than 0.01) whose clearance rates were similar to those previously reported in normal full-size and full-term newborns. Arterial insulin values before glucose loading were similar in all groups. The insulin response to glucose was poor in both SGA and AGA infants, but there was no significant difference between these two groups of infants. The response in the two groups at 24 and 25-48 h was similar. In 8 SGA babies insulin response was studied shortly after glucose injection to determine beta-cell release, but there was no significant difference of insulin level before and after glucose administration. In 5 AGA and 8 SGA babies insulin response was studied simultaneously in portal vein and aorta. In AGA babies the insulin response was completely different in the two vessels with a high response in the vein; on the contrary, in SGA infants response was variable. Growth hormone levels were high before test in SGA and AGA; after a glucose load the plasma GH levels increase, but the response was identical in SGA and AGA.


The regulatory effect of estrogens on fetal growth. IV. Brain development in growth accelerated fetuses in rabbits.

Brain weights and DNA, RNA, and total protein concentrations were determined on 31-day-old fetuses whose growth had been accelerated incidental to estrogen deficiency following maternal oophorectomy. Compared to controls, there were no differences in the DNA, RNA, or total protein concentrations, although the mean brain weight was increased. The higher mean brain weight in the growth accelerated fetuses was related to the increased mean fetal weight, and did not represent an independent response to the estrogen deficiency. A comparison of the influence of fetal weight versus gestational age on the weight of the fetal brain shows that gestational age exerts an effect apart from that of fetal weight.


Automation in cervical cancer screening. Part 1: fixed cell scanning systems.

Cytology automation is highly desirable if a mass cancer screening system is to be effective, and has been the subject of widespread research and development effort. Several experimental systems have been developed based on fixed cell scanning or flow techniques, but clinical trials have produced disappointing results. The major problem with these early systems has been that of false positive "abnormal cell" signals caused by cell overlaps and artifacts. Most current research is devoted to the development of better techniques for cell presentation, to the improvement of pattern recognition techniques, and to the exploitation of novel cell parameters in flow systems.


Nanosecond time-resolved emission spectroscopy of a fluorescence probe adsorbed to L-alpha-egg lecithin vesicles.

Nanosecond time-resolved emission spectra (TRES) are fluorescence emission spectra obtained at discrete times during the fluorescence decay. The complete data-set obtainable is a surface representing the intensity at all wavelengths and times during the emission decay time. When 2-p-toluidinonaphthalene-6-sulfonate (2,6 p-TNS) is adsorbed to egg lecithin vesicles, an excited-state reaction associated with energetic changes of the emitting species occurs on the nanosecond time scale. Convolution of the fluorescence decay with the excitation response introduces an artifact in the time-dependent spectra. A precedure is described by which this artifact can be eliminated. The data for the generation of time-resolved emission spectra are obtained with a computer-interfaced instrument based on the single-photon counting method.


[The tranquilizing properties of sodium hydroxybutyrate].

Experiments were conducted on rats. It was shown that sodium oxybutyrate depressed the escape response to punishment of the partner and motivated aggression in much lower doses than the defence conditioned reflex and unmotivated aggression, prevented negative consequences of fear and rage reactions; it also normalized conditioned reflexes in neurotic rats, this pointing to its marked tranquillizing action.


[Factors determining prolonged chimerism of lymphoid tissue in mice treated with cyclophosphamide].

Conditions of long-lasting chimerism induction in adult CBA mice treated with cyclophosphamide and spleen cells (CBAXC57BL/6)F1 were investigated. The necessary requirements are high cell doses (over 50X10(6)), cyclophosphamide dose equal or over 200 mg/kg and a short interval (3-6hr) between their injections. The data obtained were interpreted from the point of view of the capacity of cyclophosphamide to induce reversible lymphocyte DNA damages. After the interaction of lymphocytes with the antigen these damages are fixed with the resultant elimination of the corresponding cell clone.


[The relationship between the number of antibody forming cells in the spleens of rats immunized with Sal. typhi O-antigen and the number of background cells producing antibodies to that antigen].

It was shown on 5 series of rats by the method of local passive hemolysis in gel that the number of antibody-forming cells (AFC) in the spleen after the immunization with O-antigen from Sal. typhi correlated with the number of background AFC to the given antigen in the animals of the same series. This, possibly, pointed to the fact that the number of spontaneous AFC reflected the number of B-lymphocyte immunocompetent to the given antigen and could be used - to some extent - for the assessment of the immunological reactivity to this antigen before the immunization.


[The influence of low-molecular extracts from heterologous thymus, epiphysis and hypothalamus on the immune response in mice].

Extracts from bovine tissue were prepared by the acetic acid treatement and purified by gel filtration. All the preparations were injected into CBA mice for 10 days. Extracts from the thymus and the epiphysis proved to increase the number of antibody-forming cells generated in the spleen and the level of serum hemagglutinins in response to the immunization with sheep red cells. As to the hypothalamic extract, it depressed the immune response.


[The adjuvant effect of IgG molecule fragments present in the serum of partially hepatectomized rabbits].

A F (ab)2 -like fragment of IgG isolated from the serum of partially hepatectomized rabbits was capable of enhanceing an immune response in rabbits xenogenous erythrocytes. This enhancing effect was revealed by an increase of both the number of antibody-forming cells in the spleen and the titres of 19S and 7S hemagglutinins.


[Sex ratios in mouse embryos during the postimplantation period of development].

Sex ratio in the embryos if the CBA mice was studied on the 10th and 18th days of gestation to analyze the relationship between the mortality and sex of the embryo. Spontaneous mortality of embryos was shown to be independent of sex of the embryos , and sex ratio of the embryos at different stages of embryogenesis was no different from 1 : 1.


[Experimental neuropathological syndromes during the development of hyperactive determinant dispatch stations in the caudate nuclei].

Hyperactive determinant dispatch stations (DDS), representing generators of pathologically increased excitation, were created in the rostral part of both caudate nuclei in rats with the aid of tetanus toxin, which disturbed various kinds of inhibition. Formation of such DDS generators in the indicated structures induced stereotypes, impulsive convulsive movements and catalepsy.


[Effect of dilysosomal macrophages on the development and outcome of pneumonia induced by the causative agent of enzootic abortion of sheep].

Repeated intranasal injections of increasing doses of bacterial lipopolysaccharide pyrogenal resulted in accumulation of numerous macrophages in the upper and middle parts of the mouse lungs; these macrophages were filled with leukocyte lysosomes. In these animals dilysosomal macrophages increased local resistance to the agent of enzootic ewe abortion and thus could change the lethal form of induced pneumonia into the nonlethal one.


The binding of various mercurial compounds to serum proteins.

Binding study of 203Hg-labeled Hg2+, PMA, MMC and EMC tp serum albumin of six mammalian species, bovine hemoglobin and bovine lambda-globulin is presented. Both MMC and EMC bound only weakly to serum albumin and lambda-globulin and more strongly to hemoglobin; Hg2+ bound very strongly to both albumin and hemoglobin and weakly to lambda-globulin; and PMA bound most strongly to albumin, next to hemoglobin and the least, to lambda-globulin. The available binding sites varied from one to five per molecule of protein. Human serum albumin has the lowest association constants with all four mercurial compounds, indicating that it was not as tightly bound to mercurial compounds as found with serum albumins from other species.


Reduction mammaplasty with a vertical bipedicle and transverse scar: a follow-up.

A technique for reduction mammaplasty has been developed. The method has been used in 174 patients and the complications and final results have been analysed. If a transverse scar can be accepted the method gives an attractive, predictable shape, with very few complications and very little morbidity.


A twin study of human red blood cell catechol-o-methyl transferase.

Significant sibling-sibling and within-family correlations of human red blood cell catechol-o-methyl transferase activity have suggested a high degree of genetic control over levels of activity of this catecholamine-related enzyme. However, family studies do not disentangle genetic from environmental similarities as causative of within-family correlations. We therefore undertook a human twin study using the monozygotic-dizygotic comparison method. Twelve pairs of monozygotic twins had an intraclass correlation of 90 in red blood cell catechol-o-methyl transferase activity, and seven pairs of dizygotic twins had an intraclass correlations of 37. Heritability of the enzyme activity was estimated by different methods as between 68-100 per cent.


Transposition of the spleen in rats with portal hypertension.

Portal hypertension was induced in rats by progressive occlusion of the portal vein or by dimethylnitrosamine (DMNA) cirrhosis. Portasystemic venous connections did not develop in relation to the spleen when this organ was intraperitoneal, but after subcutaneous transposition most of the collaterals were parasplenic and the portal venous pressure fell.


Increase in striatal acetylcholine by picrotoxin in the rat: evidence for a gabergic-dopaminergic-cholinergic link.

Picrotoxin, 2 mg/kg i.p., a GABA receptor blocking agent, increased rat striatal acetylcholine content by approximately 70% without altering the levels of this amine in the cerebral hemispheres, mesencephalon, diencephalon, hippocampus and cerebellum. Striatal choline levels were concomitantly decreased by about 25%. This dose of picrotoxin also increased striatal homovanillic acid levels by about 30%, an effect which was not antagonized by pretreatment with the dopamine receptor stimulating agent, piribedil. Picrotoxin did not affect striatal choline-O-acetyltransferase or cholinesterase activity after in vitro incubation. The action of picrotoxin on striatal acetylcholine levels was partially antagonized by pimozide and completely blocked by alpha-methyl-para-tyrosine pretreatment while the intraventricular injection of 6-hydroxydopamine was without effect. Convulsions were not prevented by any of these treatments. The results are interpreted as follows: picrotoxin released dopamine through disinhibition of the dopaminergic neurons as a result of blockade of gabergic receptors. The increased dopaminergic activity inhibited cholinergic neurons and lead to an increase in acetylcholine content. The data thus provide evidence for a possible gabergic (inhibitory)--dopaminergic (inhibitory)-cholinergic link terminating in the striatum.


Quantitative differences between sympathetic nerve terminals.

The fluorescence histochemical method for catecholamines shows differences in both innervation density and fluorescence intensity between sympathetic nerve terminals in different organs. Quantitative electron microscopy shows differences in vesicle density in varicosities of nerve terminals in heart, spleen and vas deferens of the rat. The electron-dense core of the small noradrenergic vesicles has a different mean volume and a different core-size distribution in different nerve terminals. The significance of these findings is discussed.


Dietary chloride requirement of broiler chicks fed on a wheat-soya bean diet.

1. A diet based on soybean meal and wheat contained insufficient chloride to support normal growth in chicks. 2. Ammonium chloride and sodium chloride were equally good sources of supplementary chloride for the chick. 3. When seven diets providing increasing proportions of chloride from 608 to 1808 mg/kg, were fed to chicks, maximal growth at 21 d was obtained at about 1200 mg chloride/kg diet.


Evidence of a novel role for monovalent cations in pyruvate kinase catalysis.

In the pyruvate kinase (EC 2.7.1.40) reaction, the complete enzyme-products complex consists of enzyme, pyruvate, ATP, a divalent cation, and a monovalent cation, usually K+. The dissociation of this complex can be slow permitting reversible enolization of pyruvate in this complex during the course of the forward reaction. High concentrations of each component in the enzyme-products complex inhibits the forward reaction, but only elevated concentrations of the monovalent cation decrease the net rate of product dissociation relative to that of pyruvate enolization. This result indicates that the monovalent cation can be the first component released from the enzyme-products complex and that the presence of an inhibiting monovalent cation concentration leads to re-formation of the complete complex, which is necessary for pyruvate enolization. The evidence suggests that the monovalent cation can bind and release with each turnover of the enzyme. While the data do not permit the conclusion that first release of monovalent cation is the exclusive pathway for dissociation of the enzyme-products complex, no other component once released can reassociate rapidly enough to form a complete complex during the forward reaction. Inhibition by these components must be attributed to the formation of abortive complexes.


Ultrastructure of melanin formation in Verticillium dahliae with (+)-scytalone as a biosynthetic intermediate.

Transmission and scanning electron microscopy showed that melanin of wild-type Verticillium dahliae occurred as granules in microsclerotial cell walls and in a fibrillar network encapsulating the walls. An albino microsclerotial mutant and a brown microsclerotial mutant of V. dahliae did not form melanin granules. When albino microsclerotia were treated with (+)-scytalone (a metabolite that the brown mutant accumulates), they formed melanin granules and turned black. These granules were similar in appearance and distribution to those in the wild type. Melanin granules of the wild-type isolate and the scytalone-treated albino mutant were formed at a maximum rate in microsclerotia from 5- to 8-day-old cultures. These observations suggest that scytalone is a natural intermediate of melanin synthesis in V. dahliae.


Use of mutants to establish (+)-scytalone as an intermediate in melanin biosynthesis by Verticillium dahliae.

Melanin biosynthesis in Verticillium dahliae Kleb, was studied with mutants deficient for normal black melanin or for production of microsclerotia. Seven genetically different mutants had apparent blocks in melanin biosynthesis. Four mutants (brm-1 to -4) produced brown microsclerotia and extruded pigments into media; three (alm-1 to -3) produced albino microsclerotia. Other mutants produced no microsclerotia (nms) or had greatly reduced numbers of microsclerotia (rms). Mutation alm-1 was due to a single recessive gene; the other melanin-deficient characters were recessive but their genetic bases were not determined. Cultures of the brown mutants brm-1 and -3 extruded and accumulated a metabolite that blackened the albino microslerotia of alm-1 to -3. The metabolite was identified as (+)-scytalone (3,4-dihydro-3,6,8-trihydroxy-1(2H)naphthalenone). Pigment formed by alm-1 microsclerotia from (+)-scytalone had chemical and physical properties identical with those of melanin in the wild-type fungus. (+)-Scytalone was produced and converted to melanin by microsclerotia but not by conidia or hyphae. Conversion of (+)-scytalone to melanin appeared to involve two or more enzymes and probably involved conversions to 1,3,8,-trihydroxynaphthalene and 1,8-dihydroxynaphthalene. Albino mutants of Thielaviopsis basicola, Drechslera sorokiniana, Pleospora infectoria (Alternaria), Ulocladium sp., and Curvularia sp. also converted scytalone to pigments indistinguishable from the melanins found in their respective wild types. Scytalone melanin may be common in fungi with dark brown or black pigments.


Lysogeny in Streptococcus bovis.

Forty-eight strains of Streptococcus bovis were tested for induction with mitomycin C. Eleven inducible strains were found and the lysates of all 11 contained bacteriophage-like particles. The lysate of one strain also contained large numbers of particles which closely resembled polyheads. Eight of the lysates produced zones of inhibition on indicator strains and viable bacteriophages were detected in six lysates. The sources of the S. bovis were ovine and bovine rumen fluid (23 strains), ovine and bovine feces (5 strains), blood cultures from human endocarditis (8 strains), and a wide variety of other sources (12 strains). Only 1 of 23 rumen S. bovis strains was lysogenic, which suggests that lysogenic S. bovis strains are not an important source of bacteriophages in the rumen.


Bilateral pneumonia and inappropriate secretion of antidiuretic hormone in a premature infant.

A 6-week-old infant born prematurely had severe hyponatremia and other features of the syndrome of inappropriate secretion of antidiuretic hormone (SIADH). This disturbance was believed to be secondary to extensive bilateral pneumonia with collapse of the right upper lobe. Although this association has been recognized in adults, this is the first report of its occurrence in an infant. SIADH must be considered in the differential diagnosis of hyponatremia in association with pneumonia in an infant.


Importance of mammary gland DNA synthesis on carcinogen-induced mammary tumorigenesis in rats.

DNA synthesis in mammary gland estimated by [3H]thymidine incorporation was significantly higher on the day of proestrus than on the second day of diestrus in 50-day-old female Sprague-Dawley rats. The percentage of progressive mammary tumors, tumor growth rate, and the number and the weight of tumors per tumor-bearing rat were significantly higher in the animals given a single i.v. injection of 5 mg 7,12-dimethylbenz(a)anthracene at proestrus than in the animals given it at diestrus. Inhibition of DNA synthesis at proestrus by 2-bromo-alpha-ergocryptine also suppressed mammary tumorigenesis by the carcinogen. In 90-day-old rats in which little difference was found in mammary gland DNA synthesis between proestrus and diestrus, there was no difference in mammary tumorigenesis between animals given the carcinogen at proestrus and animals given it at diestrus. On the other hand, the prestimulation of mammary gland DNA synthesis by prolactin increased the growth, the number, and the weight of carcinogen-induced mammary tumors. These results demonstrate the importance of mammary DNA synthesis at the time when a carcinogen acts on the glands in mammary tumorigenesis.


Synthesis of 4-hydroperoxy derivatives of ifosfamide and trofosfamide by direct ozonation and preliminary antitumor evaluation in vivo.

A one-step synthesis of 4-hydroperoxyifosfamide and 4-hydroperoxytrofosfamide is described. The method involves direct ozonation of ifosfamide and trofosfamide and offers improved yields in comparison with Fenton oxidation and greater convenience in comparison with ozonation of the appropriate 3-butenyl phosphorodiamidate. Evaluation of the 4-hydroperoxy derivatives of cyclophosphamide, ifosfamide, and trofosfamide against leukemia L1210 in vivo suggests a superior effect for the ifosfamide derivatie.


Two stable Fenton oxidation products of cyclophosphamide (NSC-26271) as precursors of 4-hydroxycyclophosphamide (NSC-196562) under physiologic conditions.

Cytostatic and chemical properties of two cyclophosphamide (CP) 4-peroxides, 4-hydroperoxycyclophosphamide and 4-hydroxycyclophosphamide anhydro-dimer, that were earlier isolated and identified after Fenton oxidation of CP, are summarized and discussed. Their cytostatic toxicities, which are quantitatively comparable to those of the primary metabolite of CP, 4-hydroxycyclophosphamide, are explained from their apparent spontaneous conversion to mono-oxidized forms of CP, both in vitro and in vivo. The conversion is shown to proceed via autocatalyzed reactions, which are thought to imply the occurrence of free radicals during deoxygenation. The point is raised whether any qualitative difference in cytotoxicity might be caused by the action of these free radicals. Current dose-effect studies of residual colony-forming ability after treatment of 3T6 mouse fibroblasts and Yoshida sarcoma cells in suspension culture are introduced. Further studies on the local toxic action of the radicals under in vivo conditions are necessary if the peroxides are to be introduced in local or regional clinical treatments.


Synthesis and structure-activity relationships of pre-activated analogs of cyclophosphamide (NSC-26271).

The synthesis of approximately 80 cyclic or acyclic analogs of cyclophosphamide, isophosphamide, triphosphamide, and phosphoramide mustard is outlined briefly. All of the analogs contain at least one 2-chloroethyl or bis(2-chloroethyl)-amino group bound to phosphorus. Representatives of some of the various classes of analogs were evaluated against murine L1210 leukemia in vivo. Some of the cyclic oxidized derivatives and a phosphorodiamidic chloride yielded long-term survivors. 4-Peroxycyclophosphamide was also active against ic implanted L1210 leukemia and against two other murine tumors, Lewis lung carcinoma and C3H mammary tumor.


Effect of methotrexate on the cell cycle of L1210 leukemia.

The influence of methotrexate (MTX) on the proliferative activity of cells in different phases of cell cycle has been studied. MTX (5 mg/kg) was injected i.p. 3 days after the inoculation of 5 X 10(6) leukemia cells, into F1 (DBA X C57 BL) mice. It was shown that MTX causes degeneration of cells, being in G1- as well as in S-phase at the time of drug injection. Incorporation of 3H-TdR was suppressed for a period ranging from 2 to 12 hr after MTX administration, which is demonstrated by the decrease in the number of grains per cell. The number of cells labeled after 3H-TdR injection was also sharply decreased during this period. For a period of 3 until 15 hr after MTX administration the mitotic index decreased significantly as a result of inhibition of DNA synthesis. The blocking of the G1-S transition was evident during 4 hr after MTX. Thereafter the G1-S transition proceeds at a rate which is practically equal to that for nontreated controls. MTX did not inhibit transition to mitosis of cells being in G2-phase and in a very late S-phase at the time of drug injection. The sensitivity of G1-cells to the cytocidal effect of MTX shows that for L1210 leukemia cells MTX can be classified as a cycle-specific drug killing both G1-and S-cells rather than S-phase specific agent with self-limitation.


Cell differentiation during early development of Nassarius reticulatus L. (Gastropoda, Prosobranchia). II. Morphological changes of nuclei and nucleoi.

In Nassarius reticulatus the nuclei and nucleoli undergo important morphological changes from the zygote to the 16-cell stage. In the zygote and in the trefoil (2-cell) and 4-cell stage, several agranular, fibrillar nucleolus-like bodies 1 mum diameter are present in the interphase nucleus. Granular nucleoli first appear at the 8-cell stage. These nucleoli have fibrillar and  granular regions. The granular regions are made up predominantly of ribosome-liek osmiophilic granules. From the 16- and 32-cell stage onwards, the one or two spherical nucleoli of each nucleus measure 2.5 mum in diameter and show a concentric organization with a very dense central region surrounded by a broad peripheral zone containing numerous granules, possibly of ribonucleoprotein. At the same time the number of ribosome-like particles increase in the karyoplasm and that of ribosomes in the cytoplasm. These findings are surprising because in eggs with radial cleavage, which have been subjected to more detailed analysis, the first granular nucleoli appear after the end of the cleavage, at the blastula or gastrula stage. The early appearance of granular nucleoli which seem to be characteristic of several eggs with spiral cleavage is discussed in connection with biochemical data on RNA-synthesis.


Ultrastructural study on the hypothalamic-hypophysial-adrenal axis in fetal rats.

The adrenal glands of decapitated and encephalectomized fetal rats were investigated electron microscopically and compared to those of normal intact fetal rats. Although the adrenal cortices did not show three zones (zona glomerulosa, fasciculata, and reticularis) on the 16.5th day of gestation when the decapitation or encephalectomy was carried out in utero, the zonation was recognized in fetuses operated on the 21.5th day of gestation. The same was true for normal control fetuses. However, cytoplasmic characteristics suggesting steroidogenesis in the cortical cells were reduced to various degrees in the encephalectomized or decapitated fetuses, especially in the latter ones. The change in cytoplasmic appearance was more conspicuous in the inner portion of the cortex. This result suggests that for the maintenance of normal adrenocortical function the hypothalamus may be indispensable even during the prenatal life of rats.


Mass spectrometric analysis of permethylated glycosphingolipids II. Comparative studies on different blood-group active and related erythrocyte membrane glycosphingolipids.

Three isomeric ceramide tetrasaccharides--P blood-group active globoside, lacto-N-neotetraosyl ceramide as ABH blood-group precursor, both isolated from human erythrocytes and "asialo ganglioside" from human brain as reference standard--and two ceramide pentasaccharides--H blood-group active glycosphingolipid, obtained from blood-group B active ceramide hexasaccharide of human B erythrocytes after alpha-galactosidase treatment and ceramide pentasaccharide from rabbit erythrocytes with B-like blood-group activity--were investigated by mass spectrometry after permethylation. The carbohydrate moiety exhibits differences not only concerning the sugar sequence but also with regard to the position of some glycosidic linkages: Oligosaccharides containing N-acetylhexosamine substituted at position 4 produce spectra that are distinctly different from those containing C-3 substituted N-acetylhexosamines, thus allowing the differentation between type 1 and type 2 carbohydrate chains. Moreover, oligosaccharide ions with a hexose at the cleavage site exhibit a fragmentation pattern different from those with a N-acetylhexosamine at the "reducing terminal". The intensity ratio between parent ion and parent ion -32 mass units is Q greater than or equal to 3 in the first case, whereas in the latter case Q is less than 1. The Q-values are given for 14 oligosaccharide ions. Differences in the composition of the ceramide residues can also be deduced from the mass spectra.


Allergy and parasites: the measurement of total and specific IgE levels in urban and rural communities in Rhodesia.

Eighty adult asthmatics living in an African city had a significantly higher serum IgE level (799 u/ml) than the control group (350 u/ml). A high proportion (78.7%) of the asthmatics had demonstrable circulating mite-specific IgE antibodies. The rural population of a filariasis endemic region was investigated and although no allergic subjects were identified, the group had a significantly higher IgE level (1613 u/ml) than the asthmatics and also showed a relatively high incidence of grass pollen-specific IgE antibodies (35%). The discrepancy between clinical history and laboratory results supports the mast cell saturation hypothesis and suggests: (a) an explanation for the susceptibility to allergy of African and Asian immigrants to Great Britain, and (b) a practical approach for preventing allergic reactions in vivo.


Rapid radioimmunoassay for methotrexate in biological fluids.

We report a radioimmunoassay for the cytostatic folic acid antagonist methotrexate. Antiserum was raised in rabbits immunized with a conjugate of methotrexate and methylated bovine serum albumin. The calibration curve of the assay is linear from 1 to 100 ng methotrexate per 100-mul sample, 0.1 ng methotrexate still inhibiting the binding of the tracer by 20%. The coefficient of variation ranged from 4.04% to 11.58%. The reaction of the antiserum proved to be specific for methotrexate, as only insignificant interference could be demonstrated with folic acid, folinic acid (leucovorin), or tetrahydrofolic acid. With the method, results are available about 20 h after blood sampling at a daily capacity of at least 48 samples (i.e., about 15 determinations), depending on the equipment. Concentrations of methotrexate in plasma and erythrocytes were determined in five patients who were on continuous treatment with the drug. In one patient the concentration/time curves were followed in plasma and erythrocytes by frequent determinations for 24 h after a parenteral dose. The results indicate that the procedure is well suited for clinical investigations.


Estimates of the heritability of serum lipoprotein and lipid concentrations.

Heritability (h2) of a serum lipid or a serum lipoprotein concentration is the proportion of variability in that concentration, among individuals, which is associated with heredity. We derive a familiar formula for estimating heritability (h2) from a simple linear model for inheritance. The formula was implied by Fisher (1918) and by Wright (1921); it was derived from a consideration of variances by Jensen (1967), and from a consideration of path coefficients by Rao et al. (1974). The estimate of heritability (ĥ2) is ĥ2 = 2(rMZ - rDZ) for twin studies, where rMZ and rDZ are the sample intraclass correlation coefficients among monozygous and dizygous twins, respectively. We calculate ĥ2 for some published twin data (Heiberg 1974) on serum lipoprotein and serum lipid concentrations.


Anti-actin specificity of human smooth muscle antibodies in chronic active hepatitis.

Thirty sera reacting by IFL technique in titres greater than or equal to 100 with smooth muscle fibres of rat stomach, rat renal glomeruli, and with the membrane region of thyroid cells were randomly chosen among sera sent in for routine testing of tissue antibodies. All sera but one were found to be derived from patients with chronic active hepatitis. The smooth muscle and other relevant cell staining were abolished after absorption of sera with actin, prepared from rabbit skeletal muscle and found to be homogeneous by SDS gel-electrophoresis and by electron microscopy. The actin anti-bodies were purified by precipitation of sera with F-actin and elution of the precipitates at acid pH. The purified antibodies stained all tissues in the same way as the original sera. In double immunodiffusion tests all thirty sera gave precipitation with actin. Thus, it was concluded that these broad-reacting SMA are directed against actin. The finding of high-titred SMA is of diagnostic value and supports the clinical diagnosis of active chronic hepatitis. In addition, anti-actin antibodies eluted from human sera are a suitable tool for studying actin-containing cellular structures.


Contraposed curve patterns in monozygotic twins.

Monozygotic twins with Marfan's disease presented with opposing lumbar curve patterns. Differences in degrees of curvature correspond to the levels of severity of other clinical manifestations of the disease.


Pulmonary infection due to Torulopsis glabrata; report of a case treated with flucytosine.

A case of pulmonary infection due to Torulopsis glabrata in a 43-year-old man without other known pulmonary or systemic disease is described. The diagnosis was made from multiple positive sputum cultures. After treatment with flucytosine for 21 months, there was marked symptomatic improvement and radiographic clearing, but sputum cultures continued to yield a few colonies of T glabrata.


Failure of estrogens and androgens to inhibit bone resorption in tissue culture.

The effects of estrogens, androgens, and glucocorticoids on bone resorption were compared in organ culture. At a concentration of 10(-5) M, corticosterone, testosterone, dihydrotestosterone, dehydroepiandosterone, estradiol, ethinyl estradiol and estrone did not inhibit the release of 45Ca from 19-day fetal rat long bone shafts in control or para-thyroid hormone (PTH)-treated cultures after 2 or 5 days. Cortisol inhibited both control and PTH-stimulated resorption at 10(-5) M and 10(-6) M. 17beta-Estradiol was inhibitory at 3 X 10(4) M. However, it was less effective than the estrogenically inactive epimer 17alpha-estradiol, in 8 day control cultures or in cultures containing low concentrations of PTH. At 10(-5) m, neither 17beta-estradiol nor 17alpha-estradiol inhibited resorption stimulated by prostaglandin E2 or by osteoclast activating factor.


Hypothalamic regulation of pituitary FSH secretion.

Temporal changes in plasma LH and FSH concentrations were monitored during the afternoon of proestrus in controls and in rats in which the spontaneous LH/FSH surges were blocked with Nembutal. These values were compared with those obtained following electrochemical stimulation (ECS) of either the medial preoptic area (MPOA) or the dorsal anterior hypothalamic area (DAHA) in similar Nembutal-blocked animals. Whereas MPOA-ECS (60 muA/60 sec) elicited a release of both FSH and LH, similar unilateral stimulation of the DAHA resulted in a pronounced increase in plasma FSH and only a slight elevation in plasma LH. Increasing the amount of DAHA tissue stimulated (100 muA/60 sec) caused a significantly greater release of FSH but not LH. Bilateral DAHA-ECS (60 muA/60 sec) failed to produce a greater release of FSH than that observed after unilateral 100 muA/60 sec ECS but resulted in increased concentrations of LH in plasma. Surgical separation of the MPOA from the DAHA, leaving the preopticotuberal fibers intact, did not alter the spontaneous temporal patterns of discharge of FSH or LH 19-21 days post-operatively, although peak LH concentrations were reduced. Further, unilateral ECS (60 muA/60 sec) of the MPOA in such preparations elicited a release of FSH and LH similar to that  observed in intact MPOA-ECS rats. In contrast, unilateral DAHA ECS (60 muA/60 sec) in rats with transected hypothalami, caused no release of LH and an attenuated FSH discharge when compared with intact DAHA-ECS rats (peak valued 189 +/- 8 ng/ml vs 274 +/- 11 ng/ml). These studies suggest the existence of specific cell bodies in the DAHA which can cause selective release of FSH when activated. Coexisting with this system is that level of control which is believed to be responsible for the cyclic discharge of both FSH and LH of which the MPOA is a component part.


Hormonal control of bone collagen synthesis in vitro: effects of parathyroid hormone and calcitonin.

The effects of parathyroid hormone (PTH) on bone collagen synthesis were assessed in organ cultures of fetal rat calvaria by measuring the incorporation of [3H]proline into collagenase-digestible (CDP) and non-collagen protein (NCP) using purified bacterial collagenase. 1) PTH decreased the incorporation of labeled proline into CDP at concentrations similar to those which stimulate bone resorption in vitro. 2) This effect was observed in bones treated for 6 h, but not for 3 h; it was maximal at 24 h and was maintained for 96 h. Bones treated with PTH for 48 h and transferred to control media for 48 h showed recovery of CDP labeling to control values. 3) the effect was specific for bone collagen. There was little alteration in the incorporation of proline into NCP, and incorporation into collagen was not inhibited. 4) The effect could be ascribed to decreased collagen synthesis and not to changes in amino acid uptake, precursor pool size, or degradation of newly synthesized CDP. In 3 hour experiments, PTH did increase the labeling of CDP and NCP, but only at tracer concentration of proline in the medium, compatible with an early stimulation of amino acid uptake. 5) Similar inhibition was observed with purified bovine (1-84) PTH and synthetic bovine PTH (1-34) as well as with crude homologous PTH obtained from rat parathyroid gland culture fluid. Human (hCT) and salmon (sCT) calcitonin did not inhibit the effect of PTH on the labeling of CDP nor did they stimulate CDP labeling directly at concentrations which inhibited bone resorption. Dibutyryl cyclic-3',5'-adenosine monophosphate (D3cAMP) inhibited labeling of CDP at concentrations of .03 to .3 mM, thus mimicking the action of PTH. However, in this system DBcAMP inhibited 45Ca release, thus mimicking CT. We conclude that the direct effect of PTH on bone collagen synthesis is a slow reversible inhibition, not opposed by CT. This effect may be mediated by cAMP formation in bone cells.


Intracellular localization of calcium in the chromaffin cells of the rat adrenal medulla.

The ulstrastructural localization of calcium in the chromaffin cells of the adrenal medulla was carried out by using potassium pyroantimonate during fixation. Calcium-containing deposits were either diffuse within the cytoplasm or associated with membrane-bounded organelles. Variable amounts of precipitates were found within the nucleus and in the Golgi complex. However, the major sites of calcium antimonate deposits were the secretory granules and the mitochondria. The morphological identification of calcium-storing organelles in the adreno-medullary cells may be useful in evaluating the involvement of such intracellular compartments during the secretory process.


Pulsatile release of LH and secretion of ovarian steroids in sheep during the luteal phase of the estrous cycle.

The concentration of LH and progesterone in jugular venous plasma and the secretion of steroids by the ovary were measured every 10 minutes for 2 hours on days 12, 14, and 16 of the estrous cycle in 5 ewes with utero-ovarian autotransplants. A pulse of LH occurred about once every 2 hours, when the concentration rose from a basal level of 0.57 +/- 0.08 ng/ml to a peak of 2.97 +/- 0.57 ng/ml. Within 5 minutes of the pulse of LH, the secretion of estradiol (an exclusive product of the follicle) rose rapidly from a basal level of 0.75 +/- 0.12 ng/min to reach a peak value of 2.16 +/- 0.33 ng/min in about 30 minutes. In contrast, the secretion of progesterone from the corpus luteum, and the concentration of progesterone in the peripheral plasma changed very little following the pulse of LH. The secretion of androstenedione, which arises from the follicle and corpus luteum, increased from 3.03 +/-0.75 ng/min to 7.85 +/- 1.78 ng/min by 30 minutes after the pulse of LH. These findings indicate that the follicle, and possibly the stroma, respond rapidly to episodic fluctuations in the concentration of LH and are probably involved in the negative feedback loop between the ovary and the hypothalamic pituitary system. The fluctuations in the secretion of progesterone from the corpus luteum, on the other hand, are unrelated to pulses of LH.


Twenty-four-hour resting prolactin levels in male rats: the effect of septal lesions and order of sacrifice.

Diurnal variation in resting prolactin levels was studied in male rats that were normal, sham-operated, or lesioned in the septal nuclei. Separate groups of rats were sampled by decapitation every 3 or 4 hours in two separate studies during a 24 h period consisting of 12 h light and 12 h dark. Male rats did not exhibit a significant diurnal variation in resting prolactin levels. A septal lesion had no effect on resting prolactin levels. Prolactin levels were found to be highly responsive to environmental disturbances and were subject to the effects of sequential sacrificing procedures. The possibility was raised that prolactin exhibits a diurnal variation in sensitivity to stress which varies as a function of circulating levels of corticosterone.


The diurnal rhythm of plasma aldosterone, plasma renin activity, plasma cortisol and serum growth hormone and subnormal responsiveness of aldosterone to angiotensin-II in the patients with normotensive acromegaly.

The diurnal rhythm of plasma aldosterone concentration (PA), plasma renin activity (PRA), plasma cortisol (PC) and serum growth hormone (GH) were examined in 5 cases of normotensive acromegaly and the results were compared with the observations in normal subjects. Moreover, the response of PA to angiotensin-II infusion was studied in 6 cases of normotensive acromegaly. A normal diurnal rhythm with the lowest values in the evening or midnight and the highest values in the morning was observed in 3 of 5 cases in PA and 3 of 4 cases in PC. On the other hand, no apparent rhythm of GH was observed in any cases and that of PRA in 4 of 5 cases. Although there was a significant positive correlation between PA and PC, no significant correlation was demonstrated between PA and PRA. The response of PA to angiotensin-II fusion was significantly suppressed in normotensive acromegaly as compared to the normal subjects in spite of normal levels of PRA except for 1 case. The above observations were interpreted to suggest that the aldosterone regulation system is slightly altered in a certain number of patients with normotensive acromegaly in contrast to the normal subjects in which PRA is the main contributing factor. The low PA and suppressed response of PA toangiotensin-II infusion may suggest the defective action of angiotensin-II infusion on the adrenal gland.


Secretion of gastrin and calcitonin after ingestion of meat extract in patients with peptic ulcer.

Serum gastrin and calcitonin were determined after administration of meat extract to patients with peptic ulcer in order to investigate what role gastrin and calcitionin are playing in peptic ulcer. Serum gastrin levels were elevated after ingestion of meat extract in 9 out of 11 patients with peptic ulcer. Serum calcitonin (CT) levels were elevated rather gradually in 7 subjects. It was suggested that increased endogenous gastrin levels may have stimulated the release of CT. When salmon calcitonin (SCT) was admininstered intravenously at a dose of 100 MRCu before administration of meat extract, the rise of gastrin and CT was suppressed in 67% of cases as to gastrin and in 43% as to CT.


Inhibition of transfer ribonucleic acid nucleotidyl transferase (EC 2.7.7.25) from Ehrlich tumor cells by proflavine sulfate and ethidium bromide.

Transfer RNA nucleotidyl transferase from Ehrlich tumor cells was inhibited by proflavine sulfate and ethidium bromide but not by actinomycin D, chromomycin, rifamycin SV, rifampin, daunomycin, or alpha-amanitin. Complete inhibition of nucleotidyl transferase activity was attained at a final concentration of 1 mmol/1 proflavine sulfate, while 2 mmol/1 ethidium bromide was required to completely inhibit the enzyme. CMP incorporation into tRNA was more sensitive to both proflavine sulfate and ethidium bromide than was AMP incorporation.


Haptoglobin--haemoglobin interaction. Heterogeneity of the haptoglobin 1-1 molecule in its binding affinity for horse R/E-hemoglobin.

The formation of two different complexes when haptoglobin (Hp) and haemoglobin (Hb) are mixed in a 1:1 molar ratio is demonstrated by isoelectrofocusing. In these two complexes, the affinity of Hp for Hb is shown to be different, since Hb can be displaced only from one of the complexes, by a further addition of Hp. This is confirmed by a quantitative study of the reaction stoichiometry, when [Hp]/[Hb] = 1 and [Hp]/[Hb] greater than 1, which allows an evaluation of the amount of each complex formed. All these data cannot be explained other than by the existence of two forms of Hp molecule and a reaction scheme which fits these experiments is proposed.


On the heterogeneity of native ribosomal subunits in Ehrlich-ascites-tumor cells cultured in vitro.

Native small ribosomal subunits in cultured Ehrlich ascites tumor cells, analyzed by high-resolution CsCl isopycnic centrifugation, consist of at least five classes of particles. These particles with buoyant densities of 1.39, 1.42, 1.45, and 1.51 g/cm3 were designated as SI, SII, SIII, SIV and SV, respectively. They were different from the ribosome-derived 40-S subunits which have a density of 1.54 g/cm3. Native large ribosomal subunits consist of at least two classes of particles with densities of 1.57 g/cm3 (LI) and 1.59 g/cm3 (LII), respectively. The ribosome-derived 60-S subunits have the same buoyant density as LII particles. Labeling of Ehrlich ascites cells with radioactive uridine and a subsequent chase in the presence of RNA-synthesis inhibitors shows that radioactivity was incorporated first in precursor particles with a density of 1.48 g/cm3 and then subsequently appeared in SIII and SII particles. Met-tRNAf is found exclusively on native 40-S particles with densities of 1.42 and 1.49 g/cm3. This result has been observed in cells labeled with [35S] methionine in vivo as well as with tRNA charged in vitro. The possibility that SII particles contain 40-S initiation complexes is discussed.