Estrous cycle-controlled cell proliferation in the adrenal cortex of female rats.

In a total of 120 rats the number of 3H-Tdr-labelled nuclei of the adrenocortex was counted during various phases of the estrous cycle. In two separate experiments cyclic variations in the number of DNA synthesizing cells were found. The labelling index of the zona glomerulosa showed a significant maximum in proestrus whereas in zona fasciculata a minimum was revealed in estrus. In the glomerulosa the maximum of labelled cells was followed by a mitotic peak in estrus. Due to very low numbers of labelled cells no rhythm was found in the reticularis. The possible endocrine regulations of this endogenous rhythm of adrenocortical cell replication are discussed.


Turnover of tubulin and the N site GTP in Chinese hamster ovary cells.

Radioactively labeled tubulin from Chinese hamster ovary (CHO) cells can be isolated by co-polymerization with nonradioactive porcine brain microtubule protein. 75% of the soluble tubulin in CHO extracts co-polymerizes with the porcine protein through several cycles, without preferential loss of either CHO or porcine subunits. After phosphocellulose chromatography of the co-polymerized microtubules, the CHO tubulin is radiochemically homogeneous, as judged by SDS-polyacrylamide gel electrophoresis. CHO tubulin purified in this way has 1 mole of nucleotide per mole of protein noncovalently bound at the non-exchangeable or N site. This-layer chromatography indicates that the N site nucleotide is entirely ribo-GTP. Label and chase experiments show that the N site GTP exchanges intracellularly with a half-time of 33 hr in growing cells which have a generation time of 17 hr, while the tubulin polypeptides are degraded with a half-time of 48 hr. Intracellular hydrolysis of the gamma-phosphate of the N site nucleotide can be detected but occurs very slowly, with a half-time of 24 hr. These results suggest that the N site nucleotide may function in vivo as a stable structural co-factor of the tubulin molecule and render improbable the possibility that it has a regulatory role in microtubule assembly.


Changes in the mRNA population of chick myoblasts during myogenesis in vitro.

We have analyzed the sequence complexity, frequency distribution and coding capacity of the mRNA populations of primary chick embryo muscle cultures at different stages of myogenesis. Prefusion cultures, fused myofibrillar cultures and cultures blocked for both fusion and myogenesis all contain about 17,000 different mRNA sequences, arranged in three of four abundance classes. The myofibril (96 hr) cultures, however, contain about 2500 sequences in higher concentration and six sequences in exceptionally high concentration, each present in about 15,000 copies per nucleus. These sequences are shown to be 10 times less common in premyogenic (26 hr) cultures and 40 times less common in cultures that have been blocked by BUdR against both fusion and myogenesis. The concentration of these sequences in cultures developing toward myofibril formation correlates well with the capacity of the mRNA to stimulate the cell-free synthesis of muscle-specific proteins. A more direct approach to the identity of the abundant class of myofibril mRNA indicates that it contains the templates for the synthesis of seven polypeptides that are synthesized in particularly large amounts in myogenic cultures, including myosin, actin and tropomyosin. Between 20 and 30% of the abundant mRNA is transcribed from moderately repetitive DNA sequences. The remainder of the abundant, and all of the less-abundant, mRNA is transcribed from single-copy DNA.


Conservative assembly and segregation of nucleosomal histones.

The assembly of new histones into nucleosomes and the segregation of old histones during replication were investigated using a density gradient, sedimentation equilibrium analysis of histones labeled in vivo with dense amino acids. After a 1 hr pulse of dense amino acids and 3H-lysine, nucleosomes were isolated from chick myoblast organ cultures, and the histones were cross-linked to octamers. The octamers were purified from DNA and then banded to equilibrium in cesium-formate guanidinium-HCI density gradients. The cross-linked dense octamers have the same density as the noncross-linked dense histones, and both were significantly heavier than histones synthesized in the presence of light amino acids. This experiment shows that new histone does not mix with old histone in the new nucleosomes, since the labeling protocol allows density labeling of only one histone for every seven preexisting unlabeled histones. Thus the assembly of new histone octamers is conservative. Using essentially the same experimental design, but varying the details of the labeling procedures, we also show that the dense histone octamer is stable over 3-4 generations, that neighboring octamers tend to be synthesized at the same time, and that old and new histone octamers segregate conservatively over 2-3 generations.


Effect of n-butyrate on DNA synthesis in chick fibroblasts and HeLa cells.

n-Butyrate in low concentrations stops reversibly the proliferation of chick embryonic fibroblasts and of HeLa cells, shutting off DNA synthesis. Extensive acetylation of histones is seen at the same time as inhibition of DNA synthesis. Nuclei from n-butyrate-treated HeLa cells remain inactive in control cytosol; control nuclei are strongly inhibited by cytosol from treated cells.


Axial cineangiography in congenital heart disease. Section II. Specific lesions.

The value of axial cineangiography in several forms of congenital heart disease serves as an illustrated supplement to Section I. These techniques visualize defects in the entire ventricular and atrial septum. In persistent atrioventricular (A-V) canal, it is possible to visualize all parts of both septa, status of the A-V valves (two valves versus a common A-V valve), and if a common A-V valve, its degree of override. In tetralogy of Fallot, the bifurcation of the pulmonary trunk, entire ventricular septum and coronary arteries are vividly shown. The presence of true and confluent pulmonary arteries versus systemic or bronchial arteries in pseudotruncus is clearcut. In double outlet right ventricle or in transpositions with or without double outlet right ventricle, the mitral valve-semilunar valve relationships, the left ventricular outflow tract, subpulmonary region and a straddling tricuspid valve are well demonstrated. In asymmetric septal hypertrophy, biventricular angiography may be circumvented.


Radioimmunoassay for human myoglobin. Initial experience in patients with coronary heart disease.

A radioimmunoassay for human myoglobin has been used to study the serum myoglobin level in 13 normal individuals and 68 patients admitted to a Coronary Care Unit because of chest pain. Values in normal individuals ranged from 3 to 75 and averaged 25 +/- 23 (SD) ng/ml. Thirty-two patients with myocardial infarction initially examined within 12 hours of the onset of chest pain all showed clear-cut elevations in serum myoglobin, peak values ranging from 200 to 5500 and averaging 1368 +/- 1357 ng/ml. Seventeen patients with clinically atypical chest pain and no subsequent evidence of myocardial necrosis had myoglobin levels in the normal range, as did 11 of 19 patients with chest pain thought clinically to represent myocardial ischemia but no subsequent evidence of myocardial necrosis by conventional criteria. The final eight patients in the latter group showed mild elevations of serum Mb, peak values ranging from 102 to 280 and averaging 162 +/- 52 ng/ml; the basis for these elevations remains to be clarified.


Guancydine, a new hypotensive agent with complex action.

Guancydine (1-cyano-3-tert-amylguanidine) lowered within normal limits the tensional values in an interval of four hours after its administration in eight out of nine hypertensive patients under experiment. The hypotensive effect of a single oral dose of 500-750 mg persists for about 6-7 hours after its administration. Guancydine does not impair the vasopressor response to angiotensin II but reduces the action of this peptide on the excretion of water, Na, K and Ca through urine. The hypotensive effect of Guancydine is associated with a decrease of platelet adhesiveness and an activation of fibrinolysis. In view of this fact, Guancydine might play a role in the prophylaxis of complications of arterial hypertension - atherosclerosis and trombosis. The increase of venous blood oxygenation after Guancydine could be attributed to the opening of arterio-venous shunts or to the reduction of tissular extraction of oxygen. Guancydine does not seem to be toxic. It produced, in some patients, slight headache and orthostatic hypotension, especially during the first hours after administration.


[Antibiotic prevention using cephalothin].

A single intravenous dose of Cephalotine, given at the start of operations, significantly reduces the incidence of postoperative wound infections. The concentration of Cephalotine was maintained throughout operation. The occurrence of other infections was also reduced. The results of this study imply that operations on patients with higher infectious risk should be accompanied by antibiotic prophylaxis.


Genome analysis of Peromyscus (Rodentia, Cricetidae) VII. Localization of satellite DNA sequences and cytoplasmic poly(A) RNA sequences of P. eremicus on metaphase chromosomes.

A satellite DNA fraction from P. eremicus, having a buoyant density of 1.705 g/ml in neutral CsCl density gradients, was isolated. In situ hybridization experiments, using 3H-RNA complementary to this DNA fraction indicated that the short (heterochromatic) arms of most of the autosomes contained this sequence. Conversely, in situ hybridization using 3H-complementary DNA (cDNA) synthesized from the cytoplasmic poly (A) RNA of P. eremicus (comprising a substantial fraction of total messenger RNA) showed that the number of silver grains in the long arms (euchromatin) was significantly higher than that in the short arms. The X chromosomes showed a distinct localization pattern of both sequences.


The distribution of mitomycin C-induced sister chromatid exchanges in the euchromatin and heterochromatin of the Indian muntjac.

The frequency of sister chromatid exchanges (SCEs) induced by mitomycin C (MMC) in Indian Muntjac chromosomes was determined by the fluorescence plus Giemsa (FPG) technique. Using scanning cytophotometry the relative DNA content of each chromosome was measured with and without acid or alkali pretreatments for C-banding. During acid and alkali treatments, euchromatin lost 20 to 30% of its DNA, while heterochromatin lost less than 5%; an intermediate DNA loss was observed for the short arm of the X chromosome. After growth of cells in the presence of MMC during the first cycle and in the presence of bromodeoxyuridine (BrdU) during the first and second cycles of DNA replication, SCEs in the euchromatin were proportional to DNA content. SCEs at the junctions between the neck of the X chromosome and the long and short arms occurred more frequently than expected. A threshold effect for the induction of SCEs was observed in regions resistant to DNA extraction by acid and alkali treatments (i.e., the neck and short arm of the X chromosome). At high concentrations of MMC, the frequency of SCE at each junction appears to plateau at 0.5.


Androgen-dependent fighting behaviour in male mice.

The influence of testosterone propionate, 17 alpha-methyl-testosterone and cyproterone acetate on isolation induced fighting behaviour of mice was studied in a simple testing procedure. Decreased aggressiveness has been established in mature, sexual experienced and isolated male mice both following castration and administration of the antiandrogen cyproterone acetate, respectively. Replacement therapy with testosterone propionate s.c. and 17 alpha-methyl-testosterone p.o. has been shown to restore the decreased level of aggresiveness after castration.


The effect of cyclic nucleotides on cellular ATP levels and ribosomal RNA synthesis in Ehrlich ascites cells.

Amino acid starvation of Ehrlich ascites cells leads to a significant decrease of the intracellular ATP concentration concomitant with a marked decrease in nucleolar RNA polymerase activity. Addition of 8-bromoguanosine 3':5'-monophosphate (br8cGMP) to the amino-acid-deficient culture medium increased the cellular ATP levels and restored the rRNA synthesis capacity of nucleoli to control levels. Exogenous br8cAMP overcame the effects of br8cGMP. Administration of br8cAMP to exponentially growing ascites cells resulted in a shrinkage of ATP levels and in an inhibition of nucleolar RNA synthesis similar to that observed under shift-down conditions. These effects of br8cAMP could be antagonized by exogenous br8cGMP or hypoxanthine. Since the br8cGMP-induced increase in the total adenine nucleotides was abolished in the presence of azaserine (an inhibitor of the amidation of formylglycineamide ribonucleotide) it is concluded that cyclic nucleotides exert at least a part of their regulatory effects on cell proliferation by regulating nucleotide biosynthesis de novo.


Different pools of free myoinositol in chick-embryo cells as indicated by infection with Newcastle-disease virus.

Infection of chicken fibroblasts with Newcastle-disease virus indicates that cellular inositol is compartmented in at least two pools. Only the smaller pool is directly connected with the biosynthesis of phosphatidylinositol. Entrance of exogenous inositol into this pool is inhibited by phlorizin but not by the virus. Three hours after infection Newcastle-disease virus blocks the entrance of inositol from the small pool into one (or more) subsequent larger pool(s). About five hours after infection the virus enhances the catabolism of phosphatidylinositol in chicken cells and about seven hours after infection the permeability of the plasma membrane increases.


The location of secondary structure in histone H4.

A total of eight peptides cleaved from calf thymus histone H4 have been studied at several ionic strengths by circular dichroic, infrared and nuclear magnetic resonance spectroscopies to follow the formation of alpha helix, beta structure and self-aggregates. The results are compared with data obtained previously on three other peptides and on the intact molecule in order to define the location of secondary structure in histone H4. It is concluded that there are two alpha-helical sections, the first from residues 55 to 67 and the second of about 12 residues in the region between residues 70 to 90. beta-Structure formation takes place only in the C-terminal part of the intact H4 molecule. Nuclear magnetic resonance studies of the peptides prove that it is the basic N-terminal regions of histone H4 that remain free when the molecule self-aggregates.


Inhibition of fructose-1,6-bisphosphate aldolase from rabbit muscle and Bacillus stearothermophilus.

Phosphoglycollohydroxamic acid and phosphoglycollamide are inhibitors of rabbit muscle fructose-1,6-bisphosphate aldolase. The binding dissociation constants determined by enzyme inhibition and protein fluorescence quenching suggest that two distinct enzyme inhibitor complexes may be formed. The binding dissociation constants of the two inhibitors to Bacillus stearothermophilus cobalt (II) fructose-1,6-bisphosphate aldolase have also been determined. The hydroxamic acid is an exceptionally potent inhibitor (Ki = 1.2 nM) probably due to direct chelation with Co(II) at the active site. The inhibition, however, is time-dependant and the association and dissociation constants have been estimated. Ethyl phosphoglycollate irreversibly inhibits rabbit muscle fructose-1,6-bisphosphate aldolase in the presence of sodium borohydride, presumably by forming a stable secondary amine through the active-site lysine reside. A new condensation assay for fructose-1,6-bisphosphate aldolases has been developed which is more sensitive than currently used assay procedures.


Analysis of calf-thymus satellite DNA: evidence for specific methylation of cytosine in C-G sequences.

Digestion of purified calf tymus satellite I (phi = 1.714 g/cm3) with a series of restriction enzymes shows that modification in this satellite occurs preferentially in the sequence C-G. This was also shown to be the case in the other satellites and in bulk chromosomal calf thymus DNA. Cloning of purified satellite I DNA in Escherichia coli makes sites, previously modified, available for cutting with certain restriction enzymes. All these 'new sites' contain the sequence C-G. High-resolution mass spectros-copy establishes that the satellites contain a low concentration of 5-methylcytosine. This infers that methylation which inhibits retriction enzyme cutting must occur preferentially in the sequence C-G. Hybridization of cRNA of cloned satellite I DNA with the satellites III (phi = 1.706 g/cm3) and IV (phi = 1.710 g/cm3) shows that there is no or little sequence homology between these satellites. Digestion of calf thymus satellite I DNA with endoR. EcoRI and subsequent hybridization studies with the fragments shows two EcoRI fragments in addition to the usual 1400-base-pair EcoRI repeat unit.


Sugar-specific antibodies reactive towards cell-surface carbohydrates.

Antibodies generated against sugar-azoproteins have been purified by affinity chromatography using elution with monosaccharide. They have high specificity for monosaccharide and they react with terminal, non-reducing sugars on the surfaces of human erythrocytes. Some of them, either native or as Fab fragments, inhibit mitogenic stimulation of lymphocytes by concanavalin A.


Synthesis and transport of myosin in Physarum polycephalum.

Immunological techniques have been used to study the rate of synthesis and intracellular transport of myosin in the slime mould Physarum polycephalum. Quantitative precipitation of myosin in homogenates of Physarum was achieved using an antimyosin antibody produced in rabbit in response to purified Physarum myosin. Dodecylsulphate-gel electrophoresis revealed that about 50% of the precipitated material is myosin. The rates of synthesis of total cellular protein and myosin were measured over the mitotic cycle. Both were found to increase exponentially or linearly between two successive nuclear divisions. Similarly, no difference in the proportion of myosin-synthesising polysomes, assayed by precipitation with antimyosin serum, could be detected between the S phase and G2 phase of the mitotic cycle. Myosin makes up nearly 2% of total plasmodial proteins. Its transport into the nucleus occurs predominantly during the G2 phase.


A cytoplasmic high affinity estrogen-binding protein in the embryonic chicken liver.

1. Embryonic chicken liver contains cytoplasmic estrogen-binding sites with high affinity for estradiol (Kd = 1 X 10(-9) M) and with low capacity (200-1100 binding sites/cell, dependeing on the age of the embryo). 2. Only estrogenic hormones are bound with high affinity. 3. At least the estrogen-bindin moiety of these binding sites is a protein. 4. By agarose gel chromatography an apparent molecular weight of 50000-60000 daltons was determined, very similar to that of the nuclear estrogen-binding protein. 5. In low salt the cytoplasmic binding sites exhibit a strong tendency to aggregate, which is abolished by mild trypsination. 6. A single injection of estradiol into the egg seems to cause a translocation of the cytoplasmic binding protein to the nucleus. 7. During the embryonic development the number of binding sites increases up to the age of 19 days. Thereafter it decreases again reaching a constant low level soon after hatching. 8. Data are presented to show that this low level of binding sites can not be due solely to a degradation or inhibition in vitro by a component eventually present in the liver homogenate of hatched animals.


The replication of ribosomal DNA in Physarum polycephalum.

The DNA coding for ribosomal RNA in Physarum polycephalum exists as a collection of extra-chromosomal molecules of molecular weight 37 X 10(6). We have investigated the replication of rDNA, with the following results. (a) Replication of rDNA is unscheduled. This means that molecules that are replicated at any particular time in one mitotic cycle have an equal probability of replicating again in each time interval in the subsequent cycle. Similarly, in a single cycle, some molecules replicate more than once, and some not at all. (b) Replication forks appear to move bidirectionally from points 45% or 33% from one end of the DNA. Replicating molecules observed by electron microscopy are all linear.


Non-equilibrium method for the radioimmunoassay of clozapine in the presence of metabolites.

Cross-reactions with metabolites are an ever-recurring problem encountered in the use of radioimmunoassay techniques to determine active compounds in biological material. Metabolites may interfere with the assay of the parent drug to a variable extent. Taking 8-chloro-11-(4-methyl-1-piperazinyl)-5H-dibenzo[b,e][1,4]diazepine (clozapine as an example, it was shown that the extent to which the antiserum produced interacts with the parent drug and the metabolites can be estimated by determining the equilibrium constants and the kinetics. In the present case, therefore, it was advantageous to carry out the radioimmunoassay in disequilibrium, i.e. in order to differentiate the metabolites from the parent drug, the sample was incubated with the antiserum for 10 min, after which the labelled antigen was added and the reaction mixture again incubated for a brief, exactly timed interval. It was shown that cross-reactions did not occur in mixtures of clozapine and its N-demethyl and N-oxide metabolites in the propor tions 1:1:2 over a range of concentration of 1.5-48 ng clozapine per 100 microliter human plasma. The equilibrium constants measured with the clozapine goat antiserum were as follows: clozapine 1.2 X 10(8) M-1, the N-demethyl metabolite 4.6 X 10(7) M-1 and the N-oxide metabolite 3.7 X 10(7) M-1 (pH 7.5 and 20 degrees C).


Rotational behavior induced in rats by intranigral application of GABA-related drugs and GABA antagonists.

GABA and GABA-related drugs such as muscimol, gamma-hydroxybutyric acid and baclofen injected unilaterally into the substantia nigra of rats elicited contraversive turning. Unilateral injections of picrotoxin and bicuculline produced either ipsi- or contraversive turning depending on the volume of vehicle. I.p. applied haloperidol did not abolish the muscimol-induced turning. This and the direction of rotational behavior suggests that the turning behavior elicited by GABA-related drugs is not mediated by the nigrostriatal dopaminergic tract.


Effect of 1-deamino-6-carba- (8-arginine)- vasopressin on organ blood flow in the female guinea pig.

An analogue of (8-arginine)-vasopressin, characterized by removal of the Nalpha-amino group and replacement of the disulphide bridge with a thioether bridge (dCAVP), was found to increase blood flow to the brain, kidneys and adrenal glands of female guinea pigs whilst decreasing perfusion of the skin and skeletal muscle. An increase was noted in the proportion of 15 micrometer microspheres trapped in the spleen, probably contingent upon reduced arteriole-venous shunting. These responses were obtained both at estrus and during the luteal phase of the estrous cycle. In contrast, uterine and vaginal blood flows showed a significant reduction only during the luteal phase.


Different behavioural patterns induced by apomorphine: evidence that the method of administration determines the behavioural response to the drug.

The behavioural effects of s.c. injected apomorphine was studied on habituated rats in a test-box designed to measure 8 different components of behaviour. Apomorphine, 1 mg/kg, induced two different behaviours: The "G-type" of behaviour characterized by compulsive gnawing and the "LS-type" of behaviour characterized by increased locomotion, sniffing and repetitive head and limb movements. G-type behaviour was induced when apomorphine, dissolved by heating, was injected s.c. into the flank of the animal. LS-type behaviour was induced both when apomorphine, dissolved by heating, was injected s.c. into the neck and when it was dissolved by heating together with a high concentration of ascorbic acid (1 mg/ml) and injected s.c. into the flank. G-type behaviour could not be elicited by changing the dose which induced LS-type behaviour or vice versa. We therefore conclude that these different behavioural effects of apomorphine were not dose--response effects but were elicited by at least two different synaptic mechanisms in the brain. Experimentally induced changes from one of these apomorphine-induced behaviours to another can therefore not merely be interpreted as a change in the intensity of the behavioural response as is done in e.g. commonly used stereotypy rating scales.


The role of oxygen diffusivity in biochemical reactions.

It has been shown that increasing protein concentrations can decrease oxygen diffusion in 3 in vitro systems. We postulate that it is possible, and in some circumstances probable, that diffusion might be a rate limiting step in both in vitro and in vivo biological systems.


Microspectrography of formaldehyde and fluorescamine-induced fluorescence in rabbit pulmonary neuroepithelial bodies: demonstration of a new, probably polypeptide intracytoplasmic substance.

The microspectrographic analysis of the fluorescence emitted by NEB's in gaseous formaldehyde-fixed lung tissue, posttreated with fluorescamine, revealed the presence of numerous primary amino groups which are clearly different from the serotonin identified in our earlier studies and correspond to a new, probably a polypeptide intracytoplasmic substance.


A bio-implantable solid-state camera for long-term video-recording of abdominal organs in situ.

A new solid-state camera was constructed and implanted in sheep. It permits continuous or selective video-recording of the ovary in the unrestrained animal during an entire oestrous cycle (16 days).


[Effect of amantadine on the emotional reactivity and agressiveness of rats and mice].

The effect of amantadine on various models of aggressiveness was investigated in experiments on male albino rats and mice. Amantadine (30-75 mg/kg) inhibited emotional reactivity and aggressiveness in models of shock- and apomorphine-induced aggression reproduced in rats. The drug also inhibited the isolation-induced aggressiveness in mice. In rats with bilaterally damaged caudate nucleus no inhibitory effect of amantadine on emotional behavior was in evidence, while in those with destroyed midbrain raphe nuclei this effect of amantadine was less ostensible by comparison with controls (sham-operated animals). It is suggested that the antiaggressive effect of amantadine is due to its stimulating action on the strial serotoninergic system.


Treatment of hyperprolactinemia-anovulation syndrome.

Twelve patients with amenorrhea-galactorrhea and hyperprolactinemia are presented. The mean serum prolactin level was 175 ng/ml (range, 37 to 575 ng/ml). Basal gonadotropin levels were normal in all patients. Serum estradiol levels were normal in three women and reduced in nine. The response to luteinizing hormone-releasing hormone was normal in 10 patients and the response to clomiphene citrate was reduced in all women. Radiologic evaluations of the sella turcica and neurologic examinations were performed in all cases. Patients were treated with bromocryptine (2-bromo-alpha-ergocryptine, CB-154), 2.5 to 10 mg/day, for 5 to 35 weeks. In 10 patients normalization of the menstrual cycle was restored, and 9 patients were ovulatory. The galactorrhea ceased or was improved in all cases. Four patients who were treated for infertility became pregnant after one to three treatment cycles. In all cases prolactin levels were normalized (mean level, 10 ng/ml). Side effects were slight and were experienced only on initiation of therapy. The role of prolactin and the significance of normalization of plasma prolactin levels are discussed. Lowering prolactin secretion with bromocryptine allows resumption of normal gonadal function.


[Effect of phosphemide and forpurine on antibody formation].

Tests conducted on mice of the CBA line immunized with sheep erythrocytes have brought evidence that, when introduced to the animals 24 and 48 hours after immunization, phosphemide appreciably reduces the number of cells producing the IgM- and IgG-anibodies. The maximum immunodepressive effect of phopurine was observed when this was introduced simultaneously with the antigen, and also 24 and 48 hours following immunization. Both compounds produced a change in the mass of the immuno-competent organs and reduced the number of nuclei-containing cells in the spleen throughout the entire period of investigation.


[Change in the functional state and tissue oxidative processes of the pancreas under the influence of alcoholic intoxication].

Experiments conducted with albino rats demonstrated that intragastrically introduced ethyl alcohol in a dose of 3 g/kg intensifies the external-secretary function of the pancreas and also increases the activity of the pancreatic juice amylase in the blood, this activity continuing to be elevated 24 hours after a single introduction of the alcohol. The action of the latter reduces the oxygen tension and the redox potential in the glandular tissue, lessens the activity of NAD-dependent dehydrogenases occurring against the background of the succinate-cytochrome-c-oxydoreductase complex activity, which bears evidence to the development of "hypoxy-like" shifts in the metabolic stade of the pancreas. The author concludes that the toxic action of alcohol on the pancreas is associated with disturbances in the redox processes and with the damage of cellular membranes.


Effect of short-term cyclic administration of cyproterone acetate on pituitary-ovarian function in the human.

Short courses of cyproterone acetate, a compound with progestational and antiandrogenic activities, were administered to normally menstruating women during different phases of the menstrual cycle to suppress growth and maturation of the follicles and corpus luteum function. Postovulatory administration of 20 mg of the drug daily for 8 days to two women delayed menstruation by 4 to 6 days, followed by prolonged bleeding and short post-treatment cycles. Plasma levels of progesterone were suppressed temporarily during therapy, but increased immediately after cessation of treatment. Administration of 10 mg of the drug for 8 days during the early follicular phase to two women resulted in irregular bleeding, short cycles, and decreased plasma levels of progesterone throughout the cycle. Reduction of the dose to 2.5 mg during the early follicular phase in two other women also resulted in irregular cycles. When the 2.5-mg dose was administered to three women from the 8th to the 15th days of the cycle, vaginal bleeding and cycle length were normal. Plasma levels of luteinizing hormone and progesterone were suppressed during therapy. In one subject, cervical mucus was found to be hostile to sperm penetration in all three treatment cycles. The results indicate that, with cyclic administration of low doses of cyproterone acetate to women during the late follicular phase, it may be possible to interrupt pituitary-ovarian function, as well as sperm transport through the cervical mucus.


[Immunotropic activity of adapiprazine].

It is shown that the replacement of the ethyl or methyl radical in the molecules of aethaperazine or metherazine with the adamantyl one results in the appearance of a singular immunotropic activity. Such a drug (adapiprazine) exerts an immunodepressive action both on the primary and secondary response of the mice immunized with sheep erythrocytes. The immunodepressive action of adapiprazine is manifest with a low-dosage optimum introduced before immunization. In analogous conditions aethaperazine and metherazine fail to display any immunodepressive action. Adapiprazine arrests the rejection of a skin allotrasplant in mice differing from the donors in the presence of a strong histocompatibility locus. In in vitro tests adapiprazine in a concentration of 10(-5) mol and less has no effect on the spontaneous migration of leucocytes in the capillary, nor upon the process of immune roset-formation.


[Effect of chloroquin on the experimental sensitization with penicillin].

In penicilline-sensitized mice it was possible to decrease the titers of haemagglutinating antibodies by cholroquine.


Plasma hormone profile of threatened abortion and its prognosis.

Hormone levels in patients with threatened abortion were compared with those of women with normal pregnancies to determine the relationship between these hormone patterns and patient prognoses. Plasma progesterone, plasma unconjugated estradiol-17beta (estradiol), and plasma human chorionic somato-mammotropin (HCS) were measured serially in 27 patients with threatened abortion. Twelve of these patients were able to continue their pregnancies, and 15 patients aborted. Of the 3 indices tested, 2 (plasma estradiol and HCS levels) showed marked differences in women who were able to continue their pregnancies and women whose pregnancies ended in abortions; moreover, these findings indicated that these 2 levels could be used as indices for the evaluation of a pregnant patient's prognosis.


Case Report: epignathus-clinical, radiologic, and pathologic considerations.

In retrospect, a diagram could hace been made from the antenatal radiological examination of the case of epignathus presented here. The features of the epignathus are considered with respect to antenatal diagnosis and subsequent reparative surgery. Specific features such as axial organization, maturity of all tissues, and identical ossification points between host and parasite-all of which are evident in this case-favor the malformative origin of epignathi, thus distinguishing them from teratomata.


An overview of IUD research and implications for the future.

The limitations of nonmedicated intrauterine devices (IUDs) and the potential for improving IUD performance through the addition of pharmacologic agents are discussed. The mechanisms of action of IUDs, particularly of the copper and the progesterone-releasing devices, are reviewed. The development of improved intrauterine contraceptive devices will depend on an understanding of these mechanisms of action. In order to determine the optimal design of intrauterine contraceptives, the effects of the pharmacologic agents will also have to be evaluated, both separately and in combination with the carrier (vector).


Identification of an androgen receptor in the cytosol of the female Mastomys prostate.

An in vivo and in vitro study was carried out on the prostate from the female Praomys (Mastomys) natalensis to identify and characterize the binding of androgens within the cytoplasm. The labelled cytosol was prepared and subjected to gel exclusion chromatography and density gradient centrifugation. A macromolecular protein associated with the radioactivity was isolated on Sephadex G-200. Subsequent analysis of the steroid receptor complex showed that the major part of the radioactive steroid (64 percent) was dihydrotestosterone. This binding was inhibited by unlabelled testosterone and could not be demonstrated in liver cytosol. Characterization of this dihydrotestosterone receptor complex revealed a sedimentation coefficient of 4.6 s in the presence of a high salt solution (0.4 M KCl). The complex aggregated in the absence of 0.4 M KCl and sedimented preferentially from 5.6-7.4 s together with polydisperse aggregates of higher sedimentation coefficients. The use of this animal as an experimental model for hormonal studies on the prostate is suggested.


Androgen metabolism in relation to growth stimulation by a uterine cell line.

The metabolism of radioactive testosterone, 5alpha-dihydrotestosterone, 4-androstene-3beta,17beta-diol or 4-androstene-3alpha,17beta-diol by the human cell line NHIK 3025, derived from a carcinoma of the uterine cervix, was studied. The cells were grown in Eagle's minimal essential medium (MEM) with a steriod concentration of 10-(7) M for 4 days. Androgen metabolism by this cell line is essentially the same as for other androgen-responsive cells. The most interesting testosterone metabolite in this system is 4-androstene-3beta,17beta-diol, and the separation of this compound from 4-androstene-3alpha,17beta-diol and the two corresponding 5alpha-reduced diols is described. Since 4-androsterone-3beta,17beta-diol is a more potent growth factor for these cells than testosterone, the small conversion of testosterone to 4-androstene-3beta, 17beta-diol observed could be responsible for the growth stimulation by testosterone.


[The surgical treatment of discrete subaortic stenosis (author's transl)].

The AA. examine their experience concerning 23 patients operated on for a discrete subaortic stenosis since 1969. Immediate results are satisfactory, even if there have been postoperative complications in 26% of the patients. Late results are less satisfactory, above all because of the progression of aortic insufficiency far from the operation. Date of literature are nor allowing valuable indications about the natural history of the patient after surgical treatment, because of the scarcity of postoperative hemodynamic controls and, especially, of serial examinations. The AA. believe, on the ground of considered data and personal experience, that it is prudent, awaiting for further critical evaluation of late results, to limit surgical indications, in asymptomatic patients, to cases with left ventricular-aortic gradient superior to 70 mmHg.


[Subaortic stenosis due to antero-lateral muscular band, associated with ventricular septal defect and underdevelopment of the aortic arch. Anatomical features in three cases (author's transl)].

The pathological findings in three cases with ventricular septal defect and severe coarctation or tubular hypoplasia of the aortic arch are reported. The left ventricular outflow tract was partially obstructed by muscle situated on its antero-lateral wall. The haemodynamic relationship between this type of subaortic stenosis and aortic arch underdevelopment, due to unequal partitioning of the blood flow between the ascending aorta and the pulmonary artery during foetal life, is emphasized. The normal anatomy of the left ventricular infundibulum is described and some embryological hypotheses to explain this antero-lateral muscular band are forwarded.


[Supravalvular aortic stenosis. Report of twelve cases (author's transl)].

Twelve patients having supravalvular aortic stenosis were operated. Indication to the surgery was set up with a pressure gradient across the stenosis greater than 50 Hg millimeters and/or in a presence of important symptoms and/or with electrocardiographic signs of left ventricular hypertrophy. The technique and results of operation are described. The indication criteria to surgery on the basis of clinical results are discussed.


Stimulation of adenylate cyclase in homogenates of isolated intestinal epithelial cells from hamsters. Effects of gastrointestinal hormones, prostaglandins, and deoxycholic and ricinoleic acids.

The mechanism by which intestinal secretagogues evoke fluid secretion in the small bowel and colon has been suggested to involve mucosal adenylate cyclase. Adenylate cyclase activity was assayed by conversion of [32P]ATP to [32P]cyclic AMP in a system of pure epithelial cells isolated from the small intestine of the hamster by vibration in buffer. Several gastrointestinal hormones were tested for their capacity to stimulate adenylate cyclase; vasoactive intestinal peptide and impure cholecystokinin-pancreozymin (but not the 99% pure preparation or pure cholecystokinin octapeptide) were potent stimuli, but pentagastrin, glucagon, secretin, and gastric inhibitory peptide were impotent. Two prostaglandins, PGE1 and PGE2, were potent stimuli of adenylate cyclase. Two other compounds that provoke intestinal secretion of fluid, deoxycholic acid and ricinoleic acid (castor oil), were ineffective stimuli of adenylate cyclase. These experiments do not support a clear-cut relationship between a compound's ability to stimulate adenylate cylase and its activity as an intestinal secretagogue.


Segregation of cytoplasmic incompatibility properties in Culex pipiens fatigans.

Maternally inherited variants, which arose within a laboratory colony of Culex pipiens fatigans, have been studied by rearing cultures from single egg rafts. Segregation, i.e, variation of cytoplasmic incompatibility properties between the male progeny of individual females, was demonstrated. Also, from the daughters of individual females, sub-lines were derived within which all the males showed the same incompatibility or compatibility properties. Among the descendants of tetracycline-treated individuals were lines which superficially simulated these phenomena, but theses lines ultimately reverted to the cytoplasmic compatibility type of the strain which was submitted to the treatment. The types of variation s in cytoplasmic incompatibility properties that have been studied are discussed.


["Methamphetamine-stereotypies" and brain dopamine levels of rats treated with single or repeated doses of alpha-methyl-para-tyrosine (author's transl)].

Male albino Wistar rats were injected intraperitoneally with saline solution (1.0 ml/kg) or a-methyl-paratyrosine (50 mg/kg) once or daily X 7 and methamphetamine (10 mg/kg) was injected 3 hr or 24 hr after the last injection of these drugs. Stereotyped licking and biting activity were scored during 120 min after methamphetamine injection and, in parallel brain dopamine levels were measured. Methamphetamine given 3 hr after single or repeated administrations of alpha-methyl-para-tyrosine was not effective for stereotyped behavior and brain dopamine levels in similarly treated rats decreased markedly in comparison with saline-treated rats. Methamphetamine 24 hr after repeated administration of alpha-methyl-para-tyrosine clearly induced stereotypies and brain dopamine levels in these rats did not differ from saline-treated rats. The results suggest a possible role of brain dopamine on "methamphetamine-stereotypies".


[Effect of chronic oral administration of clonidine on the endogenous catecholamine metabolism in spontaneously hypertensive rats (author's transl)].

The present experiments were undertaken to elucidate the effect of chronic oral administration of clonidine on the endogenous catecholamine metabolism in spontaneously hypertensive rats (SHR). A three month administration of clonidine (20 microgram/kg/day p.o.) produced a slight decrease in blood pressure and heart rate. No significant change was observed in the body weight. A one month administration of clonidine (300 microgram/kg/day p.o.) produced hypotension and weight loss. No significant effect was observed in heart rate. After a two month administration of clonidine (300 microgram/kg/day p.o.), the catecholamine content in the brainstem showed a significant decrease as compared with that in control. 3H-norepinephrine (NE) turnover rate after intraventr culari injection of 3H-NE tended to decrease in the brainstem of SHR while no significant difference was recognized from a view point of uptake, biological half life, elimination rate constant of 3H-NE between control and clonidine administered rats. Under the same experimental conditions, a marked rebound hypertension occurred when medication is discontinued in some cases of this group of SHR.


Ultrastructural alterations in collagen associated with microscopic foci of human emphysema.

Postmortem and biopsy lung samples were examined with a dissecting microscope and classified into three categories: no overdistended air spaces visible, mild destruction and some fenestration of the septa with minimal confluence of air spaces, and more advanced breakdown of alveolar walls with at least a sixfold increase in air space to a 1300 mu diameter. Prominent ultrastructural tissue alterations were found within the focal regions where fenestration of the alveolar wall and some confluence of air spaces were identified. Edema and rearrangement of the supportive tissues were noted in the alveolar septa adjacent to foci having the greatest alveolar enlargement. Randomized electron dense collagen fibrils were found only in these areas, whereas the normal alveolar septa contained uniformly parallel collagen fibrils. Many of the randomized fibrils had sufficient swelling to display prominent internal spiraling. Other changes in structure and cell numbers were consistently found in association with the altered collagen. These included: separation of the type I epithelial cells from the basal lamina and some autolysis of the cytoplasmic organelles, numerous flattened type II epithelial cells, and aggregations of pulmonary macrophages.


A new case of XX-male (XX/XXY mosaic).

We report a 10-year-old male patients with the predominant karyotype 46,XX. In only one cell of the testis culture a metaphase with 47,XX+G was found. The theories of etiology of XX-males and the reported cases of XX/XXY-mosaics in the literature were discussed.


The cytology of spontaneous neoplastic transformation in culture.

Thirteen cell lines derived from embryos of the mouse, rat and hamster were sampled at closely spaced intervals for cytology and spontaneous neoplastic transformation. Transformation, determined by growth of the cells as malignant neoplasms in compatible hosts, occurred in eight lines and appeared to be unrelated to time in culture, passage number, proliferative activity of the cells or type of serum used to supplement the culture medium. In an effort to establish cytologic criteria for malignancy of rodent fibroblasts, cultures were examined for 19 cytologic abnormalities. The percentage of cells or colonies showing each of the abnormal properties was recorded. Five of the 19 were found to correlate with neoplastic transformation in all cell lines. These were cytoplasmic basophilia, reduced cytoplasmic spreading on substrate, cording, high nuclear: cytoplasmic ratio and clumping. Each of these properties is amenable to quantitation or computerized image analysis. On the basis of the five cytologic criteria, the percentage of colonies diagnosed neoplastic, borderline and non-neoplastic was determined for each line. Although these percentages varied from one transplant generation to another, the percentage of neoplastic colonies tended to increase with time in all lines which became malignant, and the borderline colonies tended to decrease, suggesting a progression from borderline to neoplastic state.


Clonal growth of chinese hamster cell lines in protein-free media.

A protein-free synthetic medium, MCDB 301, has been developed for clonal growth of Chinese hamster ovary cell lines. Medium F12 was developed originally for that purpose, but later failed to support good growth without small amounts of serum protein. Growth was restored by the addition of nonphysiological amounts of commercially prepared thyroxine or smaller amounts of the trace element selenium. The thyroxine preparation was shown to contain sufficient selenium to account for all of its growth-promoting activity. MCDB 301 contains increased concentrations of calcium chloride and glutamine, and a smaller amount of cysteine than medium F12. It also has been supplemented with 19 inorganic ions, in addition to selenium and those in medium F12, in order to insure against possible future deficiencies as chemicals are purified further. A Chinese hamster lung line which will not grow in MCDB 301 alone will grow when the medium is supplemented either with methylcellulose or with insulin. The growth-promoting activity is thought to be an impurity shared in common by both substances. The probable "essential" role of impurities in cellular growth in most synthetic media and the problems involved in attempting to develop a truly "defined" medium are discussed.


Toxicity of nystatin and its methyl ester toward parental and hybrid mammalian cells.

Nystatin methyl ester (NME), the methyl ester derivative of the polyene macrolide antibiotic nystatin, is known to be effective against fungi and is now found to be relatively less toxic than the parent antibiotic nystatin (NYS) to animal cells in culture as measured by 51Cr release, cell survival at different posttreatment periods and cell growth. NYS and NME were tested on TK- mouse (B82) and hamster (B1) cells, HGPRT- mouse (RAG) cells, and on lysolecithin-fused cells selected in HAT medium and confirmed as B82-RAG and B1-RAG hybrids by chromosomal analysis plus polyacrylamide gel electrophoresis of lactate dehydrogenase. NME was less toxic and caused less immediate membrane damage than NYS when tested in all five cell systems. However, differences in innate polyene sensitivity were evident between the three parental cell types. B82 and B1 cells were more resistant than RAG cells to NYS and NME. B82-RAG hybrids reflected the higher level resistance of B82 parental cells, and B1-RAG hybrids reflected the higher level resistance of B1 cells. Where one parental cell type is relatively more polyene sensitive, the use of polyenes in the future may be applicable as selective agents in cell hybridization.


Comparison of in vivo and in vitro ribosomal RNA synthesis in nucleolar mutants of Xenopus laevis.

Cells of embryos carrying a lethal nucleolar mutation have been maintained in vitro for extended periods of time. Normally these mutants live only 9 to 12 days after fertilization but their cells in culture will survive for more than 3 months. The extent of ribosomal RNA (rRNA) synthesis was determined in primary cultures prepared from normal embryos and nucleolar mutants having different numbers of ribosomal RNA genes. We found that the accumulation of radioactivity into rRNA for normal and mutant embryos was similar in vivo and in vitro. In primary cultures of normal embryos which have two nucleoli per cell and mutant embryos which have only one nucleolus per cell, the incorporation of radioactivity into rRNA was similar even though the normal cells have twice as many rRNA genes. Thus the mechanism which regulates dosage compensation of the rRNA genes operates both in vivo and in vitro.


The development of chick spinal cord in tissue culture. I. Fragment cultures from embryos of various developmental stages.

Explants from neural tube and spinal cord of chick embryos at developmental stages 8 through 36 were cultured on collagen-coated cover glasses for 21 days. The cultures of neural tube at stages 10 to 14 contained many neuronal precursor cells which gave rise to mature neurons. This was verified by cumulative labeling of cultures with tritiated thymidine. Explants from spinal cords of stages 26 and 27 contained fewer precursor cells, and at stage 36, only 7% of mature neurons were labeled. Regardless of the stage of development at which explants were made (stages 8 through 36), all cultures had a similar appearance after 21 days, indicating that cells from explants taken from earlier developmental stages (before neurons were formed) "caught up" with the explants from later developmental stages, which already had formed neurons at the time of explantation.


Human lymphocyte subpopulations: rosette formation with sheep, human and horse red blood cells.

Rosette formation between human lymphocytes and horse red blood cells could be promoted by a low pH medium, overnight incubation and a temperature of 4 degrees C. The percent of sheep, horse and human rosette-forming cells in the peripheral blood were 71.7 +/- 1.8, 30.5 +/- 2.8 and 28.3 +/- 3.4 respectively. However, their percentages in thymuses were 97.1 +/- 1.1, 91.4 +/- 2.4 and 89.0 +/- 3.4. Using preparations of isolated subpopulations, it was observed that the horse and human red cell rosette-forming cells were probably also "early" sheep red cell rosette-forming cells. Rosette formation with all three types of red blood cells were inhibited by a preparation of Fetuin-glycopeptide.


Clones producing antibodies of more than one class.

Clones producing antibodies directed against oligo-D-alanine (D-ala) determinants have been demonstrated in X-irradiated recipient mice after transfer of limited numbers of spleen and thymus cells. Seven out of eighty-one recipients were found to have donor allotype clones, all of which produced IgG1 antibodies to D-ala; two of these clones also produced small quantities of IgG2a antibodies. In addition, each of the seven responder mice contained IgM antibodies to D-ala. Isoelectric focusing (IEF) patterns of anti-D-ala antibodies from each of the seven responder mice were restricted and compatible with one to three antibody types. Most of these IEF patterns were significiantly different from each other. Five mice have been used for a comparison of IgM and IgG1 combining sites by affinity and specifity measurements. In each case affinity and specificity of IgM and IgG1 antibodies were alike. No similarity was found when comparing IgM from one clone with IgG1 from another one. These data led us to conclude that a single precursor cell can give rise to a clone producing antibodies of the IgM class and different IgG subclasses all of which share combininig sites.


Lymphocyte activation by various Mycoplasma strains and species.

The capacity of various mycoplasma strains and species to induce lymphocyte transformation in vitro was studied. Of six strains of Mycoplasma pulmonis studied, five displayed mitogenic activity with rat lymphocytes. Among those M. pulmonis strains, our MP15 isolate and the Negroni strain exhibited particularly potent mitogenic capacity. The murine mycoplasmas M. neurolyticum and M. arthritidis shared this mitogenicity for rat lymphocytes. However, the human mycoplasmas M. fermentans, M. pneumoniae, M. hominis, M. orale, and Acholeplasma laidlawii did not activate rat lymphocytes. Lymphocytes obtained from germfree rats were activated to the same extent as those from animals bred under conventional conditions. The mitogenic potency exhibited by mycoplasma was not restricted to infective microorganisms, and preparations of killed mycoplasma particles exerted an extensive lymphocyte transformation. The data show that the mitogenic activity of mycoplasmas is not confined to a single mycoplasma isolate and that it acts in a nonspecific manner.


Mycoplasma-dependent activation of normal lymphocytes: induction of a lymphocyte-mediated cytotoxicity for allogeneic and syngeneic mouse target cells.

Mycoplasma arthritidis, M. hominis, and M. arginini were tested for their ability to induce a cytotoxic response from normal CBA mouse lymphocytes against 51Cr-labeled allogeneic target cells. In most cases, the mycoplasmas alone were not toxic for the target cells. Furthermore, the mycoplasmas did not result in decreased lymphocyte viability but, in fact, contributed to enhanced lymphocyte survival. In the absence of normal CBA lymphocytes, mycoplasmas alone did not induce a significant amount of cell damage in either the allogeneic or the syngeneic target cells. Strains of M. arthritidis and M. hominis, when added to the lymphocyte-target cell mixtures, induced statistically significant increases in 51Cr release from both target cell types at each assay period after 6 h. The release of 51Cr was taken as a measure of cell death. M. arginini induced only low levels of cytotoxicity or none at all. Both arthritogenic and non-arthritogenic strains of M. arthritidis induced the cytotoxic response. The degree of cytotoxicity produced was directly related to the size of the initial inoculum. The presence or absence of serum in the culture medium did not contribute significantly to the cytotoxicity response.


Mycoplasma-dependent activation of normal lymphocytes: role of arginine and nonviable mycoplasma antigen in the induction of lymphocyte-mediated cytotoxicity for syngeneic mouse target cells.

The ability of arginine supplementation to abrogate or potentiate Mycoplasma arthritidis-mediated cytotoxicity induction by normal CBA mouse lymphocytes was investigated. Also, nonviable antigens of M. arthritidis were assayed for their ability to induce a cytotoxic response. The presence of additional arginine in the culture medium had no significant effect on the amount of cytotoxicity produced. In some cases, an increase in cytotoxicity was seen in the presence of additional arginine. Amino acid analysis of the culture supernatants showed that the arginine in the nonsupplemented medium was exhausted during the assay, whereas ample amounts of arginine were present in the supplemented medium. However in both cases the amount of cytotoxicity was comparable. Nonviable antigens of M. arthritidis were unable to induce a significant cytotoxic response in the presence of normal lymphocytes. The same antigens exhibited a suppressive response when tested for blastogenesis in the presence of normal lymphocytes. A discussion of lymphocyte involvement is presented.


Cell-mediated immune response in Mycobacterium leprae infected mice.

Positive skin reactions to homologous antigens in mice infected with Mycobacterium leprae were observed at the time that the bacillary multiplication was nearing the plateua level. Once developed, the skin reactivity persisted for a considerable length of time. Lymphocyte transformation studies indicated that the lymphocytes from infected animals were unable to respond to specific antigenic stimuli in the early phase of infection. Possible explanation for this failure could be that the lymphocytes although sensitized, were not capable of responding to the stimuli due to a rather low level of sensitization. The elevated lymphocyte stimulation index in the later stages of infection, coupled with skin reactivity indicates the production and proliferation of antigen-reactive lymphocytes which in turn govern the activity of mononuclear phagocytic cells through the production of lymphokines. Occurrence of all the events appears to coincide with the time closer to the plateauing of M. leprae in the foot pad of the mouse. It is suggested that the cell-mediated immune response in the M. leprae infected mouse is basically a T cell rather than a B cell response.


Progressive nerve lesion in a disease-arrested leprosy patient. An electron microscopic study.

An ulnar nerve biopsy from a patient with purely neural leprosy of the borderline tuberculoid group, who developed ulnar and median paralysis after 2.5 years of DDS therapy, was examined using light and electron microscopes. The nerve parenchyma was largely replaced by collagen fibrils. There were many onion bulbs similar to those seen in hypertrophic neuropathies. Bizarre fibroblasts such as those seen in hereditary sensory neuropathy were also demonstrated. A few Schwann cells contained M. leprae. It is suggested that considerable proliferation of fibrous tissue may be a reactive phenomenon in response to the continued presence of fragmented M. leprae and their products. Ischemia following a marked progressive increase of intraneural collagen is an important cause of atrophy of Schwann cells followed by segmental demyelination and necrosis of the axons in this healed leprosy patient.


HLA antigen and susceptibility to leprosy.

Fifty-nine leprosy patients (31 tuberculoid, 28 lepromatous) have been HLA typed and compared to 125 healthy individuals who have had no known contact with leprosy patients in the past. HLA antigens were determined by the microdroplet lymphocyte toxicity method developed by Prof. Terasaki of UCLA. In order to detect 31 HLA antigens, a total of 58 antisera were used in a No. T-7 research tray. AW24 antigen showed the most remarkable deviation. The frequency of this antigen in leprosy patients was 25.4% and that in normal subjects of the control group was 63.2%. The difference in frequency between the two groups was statistically significant when the p value was corrected for the number of antigens being tested. Uncorrected p values of the BW39, A9, and B8 antigens were also statistically significant. Frequencies of the majority of HLA antigens in tuberculoid leprosy patients were similar to those of corresponding HLA antigens in lepromatous leprosy patients.


Correlation between clinical and histopathologic classification in leprosy.

A detailed histopathologic study, utilizing hematoxylin and eosin stain, was done in 95 fresh uncomplicated cases of leprosy. The microscopic features were classified according to Ridley's (1974) definition, while the clinical grouping was done using the criteria of Ridley and Jopling (1962). The disparity between them on comparison was explicit. In one third of the cases the two were in consonance with each other. In many there was a shift of one step either towards the tuberculoid or lepromatous end of the spectrum. The remaining were classified as indeterminate because of early histopathologic changes. This disparity was expected because the parameters used for the histopathologic classification were precise and also took into account the immunologic aspect. The histologic definitions seem practical and may prove useful in assessment of the status of the disease with or without treatment.


Development of a health and safety program at the Louisiana State University School of Veterinary Medicine.

An accident prevention and zoonotic disease control program at the School of Veterinary Medicine, Louisiana State University, was developed by a Health and Safety Committee comprised of members from all instructional departments. The program incorporated safety educational input during the basic, preclinical, and clinical years; a standardized accident reporting procedure; rabies vaccination, with serologic evaluation; a blood serum bank; and recommendations for future control procedures.


Allosteric regulation of beef liver arginase activity by L-ornithine.

Inhibition of beef liver arginase by L-ornithine was investigated with two sets of independent experiments. Progress curves of the production of urea were simulated with two integrated Michaelis-Menten equations for competitive and non-competitive inhibition by ornithine. Both fitted the curves well, but failed to correctly predict the inhibition when the reaction was started with ornithine already present. Measurement of initial rates of reaction enabled an allosteric model to be built in accordance to Monod-Wyman-Changeux: arginine preferentially binds to the active state R and ornithine preferentially binds to the inactive state T. In the absence of both ligands, the R in equilibrium T equilibrium slightly favours the active state and both states bind ornithine more strongly than arginine. No great variation was observed in the 6 parameters of the model by assuming the enzyme to be a trimer or a tetramer. The model was able to predict not only the initial rate curves, from which it was derived, but also the progress curves independently obtained.


Lymphocytic choriomeningitis in laboratory personnel exposed to hamsters inadvertently infected with LCM virus.

Seven of 27 students and staff oa a university laboratory became ill with lymphocytic choriomeningitis (LCM) following exposure over an 18-month period to hamsters implanted with tumors. The clinical symptoms were compatible with LCM infection and all the patients' sera had fluorescent antibody to LCM virus; 5 of them also had complement-fixing (CF) antibodies. Complement-fixing antibodies were found in 9 of 9 hamsters with tumors and 16 of 31 without tumors (excluding 6 hamsters with anticomplementary sera) and in 8 of 14 rabbits, but in none of 49 rats. Lymphocytic choriomeningitis virus was isolated from 4 pools of liver and spleen, 2 each from hamsters with and without tumors, and from a frozen stored sample of the tumor cells. Interlaboratory transmission may have occurred via infected tumor cells. Control was achieved by destroying all animals in the project room as well as all hamsters in the building. The possibility that many tumor cell lines carried in hamsters may be infected with LCM virus has important implications for laboratory safety as well as for the validity of the research results obtained, inasmuch as the virus enhances the growth of some tumors and retards others.


Malignant catarrhal fever in bison.

Malignant catarrhal fever was diagnosed in 3 herds of American bison (Bison bison) in South Dakota from 1973 to 1976. Clinical signs included depression, nasal and ocular discharge, conjunctivitis and keratitis, and diarrhea. Herd morbidity ranged from 3 to 53.8%, and mortality was 100%. At necropsy, ulcerative lesions were found throughout the alimentary tract, trachea, and bronchi. Microscopically, necrotizing vasculitis without thrombosis was found in virtually every organ examined.


Influence of flavonoid-copper complexes on cross linking in elastin.

A protective and curative effect of some flavonoids on collagen maturation in lathyrism has been previously demonstrated. This action appeared to involve cross linking of collagen. This paper deals with the effect in vitro of flavonoids and flavonoid-copper complexes on the oxidative deamination of lysine epsilon-amino groups in [4,5-3H]-lysine-labelled elastin. Flavonoids alone do not affect the reaction but it is evident that some flavonoid copper complexes are strongly effective: the aldehyde groups that are quickly formed then enable cross linking to occur. The lysine epsilon-amino groups of elastin are specifically concerned: in fact no effect was observed on free [4,5-3H]-lysine or on [4,5-3H]-lysine-labelled proteins obtained from mouse liver. The lysyl oxidase seems to interfere with the flavonoid-copper complees to regulate the oxidative deamination of lysine epsilon-amino groups.


Microdetermination of tryptophan in compounded poultry rations.

A reliable method for the microdetermination of tryptophan in compounded poultry rations has been developed. Samples are hydrolyzed with Ba (OH)2, and the hydrolysate is passed through a Dowex-50 column to remove the salts and soluble carbohydrates. N-1-Naphthylethylenediamine is added to the purified feed hydrolysate, and color is read at 530 nm.


Trypsin as a catalyst for peptide synthesis.

Trypsin-catalyzed syntheses of peptides were performed using various N-acylated amino acid or peptide esters as donors and amino acid derivatives, peptides, or their derivatives as acceptors. The synthesis was almost quantitative under optimal conditions. Considerably more enzyme and a more alkaline pH were necessary for synthesis than hydrolysis. Another very important condition was the concentration of the starting materials; higher concentrations resulted in much better product yields. The nucleophile specificity for synthesis was also important; hydrophobic or bulky amino acid residues were most efficient at the P1' position, and L-proline as well as D-amino acid residues were the worst choices. The synthesis was also dependent on the solubility of the products synthesized; the yield was higher with products of lower solubility. As donor esters, good substrates were all useful. Accordingly, fragment condensation was possible using N-acylated peptide esters and various peptides. The present study suggests that trypsin may become a useful tool for peptide synthesis.


Kinetics for the secretion of procollagen by freshly isolated tendon cells.

Fibroblasts isolated by enzymic digestion of chick embryo tendons were incubated for several hours in suspension under conditions in which they were in steady state in terms of the synthesis and secretion of procollagen. Under these conditions, the cells synthesized and secreted about 630 microgram of procollagen/10(9) cells/h. The cells were labeled with [14C]proline for 15 to 120 min and then the kinetics of secretion were followed by chasing the label and assaying the 14C-peptides digestible by collagenase in the cells and in the medium. The results demonstrated that secretion of collagenase-digestible peptides did not follow the kinetics of a single first order process but suggested at least two pseudo-first order process with half-times of 14 and 115 min. The [14C]procollagen secreted during 0 to 30 min and 90 to 120 min of chase was the same in terms of the ratio of pro-alpha1 to pro-alpha2 chains, the size of the pro-alpha chains, the extent of interchain disulfide bonding, the extent of prolyl hydroxylation, and the degree of helicity as tested by resistance to pepsin digestion. Addition of ascorbic acid to the incubation medium increased slightly the extent of prolyl hydroxylation but did not alter the kinetics of secretion. The results suggested that the kinetics of secretion are influenced by a two-compartment system in which at least one metabolic pool contributing to the secretory process is present as a "side pocket."


Developmentally regulated proteases from the basidiomycete Schizophyllum commune.

The basidiomycete Schizophyllum commune produces three chromatographically distinguishable proteases which are capable of attack on a variety of other enzymes from S. commune and other sources. These proteases, which are produced during a specific phase of the development cycle, exhibit typical enzyme kinetic patterns, are active in the neutral to weakly alkaline pH range and are inhibited by phenylmethylsulfonyl fluoride, soybean trypsin inhibitor, and ovomucoid. No pattern of specificity toward the test enzymes could be discerned. The proteases co-purify with the activity which causes the increase in cold lability of S. commune phosphoglucomutase reported previously. In addition, one of the protease enzymes could be purified to the point where it had no significant ability to release trichloroacetic acid products from denatured substrates at pH 3 or pH 7. When undenatured hemoglobin was used as a substrate, the purified protease releases a relatively large molecular weight nonheme peptide. Relatively large peptides are also formed after proteolysis of rabbit muscle phosphoglucomutase. These results suggest that the protease carries out only limited proteolysis.


Covalent structure of the membranous segment of horse cytochrome b5. Chemical cleavage of the native hemoprotein.

The complete covalent structure of the membranous segment of horse liver cytochrome b5 has been determined. This peptide spans residues 91 to 133 of the cytochrome molecule, and contains the segment responsible for the association of the hemoprotein with microsomal or synthetic vesicles. Two peptides, residues 91 to 127 and 128 to 133, comprising the entire membranous moiety were isolated from a tryptic digest of urea-denatured apoprotein. The membranous segment (residues 91 to 127) could be separated from all other tryptic peptides by a single gel filtration step. Trypsin digestion of succinylated cytochrome produced similar peptides, residues 89 to 127 and 128 to 133. The covalent structures for residues 89 to 127 and 128 to 133 were derived from automated sequenator analysis of tryptic peptides. Chemical cleavage at tryptophanyl, or methionyl residues, or both, by the method of Ozols and Gerard ((1977) J. Biol. Chem. 252, 5986-5989) provided the overlapping peptides from which the following unique sequence was deduced: (formula: see text).


Crystallization and preliminary crystal data of porcine pepsinogen.

Single crystals of porcine pepsinogen, suitable for x-ray diffraction studies, have been grown with lithium sulfate as the precipitant. These pepsinogen crystals were dissolved, activated, and assayed for proteolytic activity. The specific enzymic activity of the dissolved crystalline protein was nearly twice that of the commerical pepsinogen from which the crystals were grown. Incubation at pH 8 before assay demonstrated that the crystals are free of pepsin. This crystal form of pepsinogen belongs to the monoclinic space group C2 with 4 molecules in the unit cell. The unit cell dimensions are a = 104.8 +/- 0.5 A, b = 43.1 +/- 0.1 A, c = 88.4 +/- 0.3 A, and beta = 91.3 degrees.


Familial idiopathic hyperphosphatasia: a study of two young siblings treated with porcine calcitonin.

A study of two siblings with a severe infantile form of familial idiopathic hyperphosphatasia is reported. A girl aged one year was followed for two years while receiving intermittent treatment with porcine calcitonin. This induced a clinical remission, a reduction of both the high serum level of alkaline phosphatase and the raised urinary excretion of hydroxyproline, and a remarkable improvement in bone structure as seen radiologically. Her sister aged two months received porcine calcitonin for three weeks, during which clinical improvement, no change in the serum level of alkaline phosphatase and a marked decrease of the excretion of hydroxyproline were recorded.


Motility of triton-demembranated sea urchin sperm flagella during digestion by trypsin.

The survival curves for a population of reactivated spermatozoa exposed to digestion by trypsin indicate that a large number of trypsin-sensitive targets must be digested before the flagellum disintegrates. Changes in flagellar movement during trypsin digestion can be very small, especially when the spermatozoa are reactivated at 0.25 M KCl. They are not the changes which would be expected if elastic resistance of the trypsin-sensitive structures responsible for maintaining the integrity of the axoneme is a significant determinant of flagellar bend amplitude. By carrying out trypsin digestion under a variety of conditions, at least six distinct effects of trypsin digestion on parameters of flagellar movement have been detected. These include a gradual increase in the rate of sliding between tubules, gradual and abrupt changes in beat frequency accompanied by reciprocal decreases in bend angle, changes in the symmetry and planarity of bending, and selective interference with mechanisms for bend initiation and bend propagation.


Structural analysis of human neutrophil migration. Centriole, microtubule, and microfilament orientation and function during chemotaxis.

Orientation of nucleus, centriole, microtubules, and microfilaments within human neutrophils in a gradient of chemoattractant (5 percent Escherichia coli endotoxin-activated serum) was evaluated by electron microscopy. Purified neutropils (hypaque-Ficoll) were placed in the upper compartment of chemotactic chambers. Use of small pore (0.45 mum) micropore filters permitted pseudopod penetration, but impeded migration. Under conditions of chemotaxis with activated serum beneath the filter, the neutrophil population oriented at the filter surface with nuclei located away from the stimulus, centrioles and associated radial array of microtubules beneath the nuclei, and microfilament-rich pseudopods penetrating the filter pores. Reversal of the direction of the gradient of the stimulus (activated serum above cells) resulted in a reorientation of internal structure which preceded pseudopod formation toward the activated serum and migration off the filter. Coordinated orientation of the entire neutrophil population did not occur in buffer (random migration) or in a uniform concentration of activated serum (activated random migration). Conditions of activated random migration resulted in increased numbers of cells with locomotory morphology, i.e. cellular asymmetry with linear alignment of nucleus, centriole, microtubule array, and pseudopods. Thus, activated serum increased the number of neutrophils exhibiting locomotory morphology, and a gradient of activated serum induced the alignment of neutrophils such that this locomotory morphology was uniform in the observed neutrophil populayion. In related studies, cytochalasin B and colchicines were used to explore the role of microfilaments and microtubules in the neutrophil orientation and migration response to activated serum. Cytochalasin B (3.0 mug/ml) prevented migration and decreased the microfilaments seen, but allowed normal orientation of neutrophil structures. In an activated serum gradient, colchicines, but not lumicolchicine, decreased the orientation of nuclei and centrioles, and caused a decrease in centriole-associated microtubules in concentrations as low as 10(-8) to 10(-7) M. These colchicines effects were associated with the rounding of cells and impairment of pseudopod formation. The impaired pseudopod formation was characterized by an inability to form pseudopods in the absence of a solid substrate, a formation of narrow pseudopods within a substrate, and a defect in pseudopod orientation in an activated serum gradient. Functional studies of migration showed that colchicines, but not lumicolchicine, minimally decreased activated random migration and markedly inhibited directed migration, but had not effect on random migration. These studies show that, although functioning microfilaments are probably necessary for neutrophil migration, intact microtubules are essential for normal pseudopod formation and orientation, and maximal unidirectional migration during chemotaxis.


Quantitative analysis of tubulin and microtubule compartments in isolated rat hepatocytes.

A combined morphometric and biochemical approach has been used to identify and quantitate microtubules and tubulin in isolated hepatocytes. The total soluble pool of microtubule protein was estimated by specific high affinity binding to radiolabeled colchicine. Scatchard analysis of the data identified two populations of binding sites: high affinity-low capacity sites resembling tubulin and low affinity-high capacity sites believed to represent nonspecific colchicine-binding sites. Data from these studies indicate that tubulin represents 1% of the soluble protein of the cell, that 9.0 X 10(-14) dimers of tubulin are present per microgram soluble hepatocyte protein, and that the average hepatocyte contains 3.1 X 10(7) tubulin dimers. Our calculations suggest that this amount of tubulin would form a microtubule 1.9 cm in length if totally assembled. However, stereological measurements indicate that the actual length of microtubules in the cytosolic compartment of the average hepatocyte is only 0.28 cm. Thus, these experiments suggest that only 15% of the available tubulin in hepatocytes of postabsorptive rats is assembled in the form of microtubules.


A study of communication specificity between cells in culture.

We have examined the specificity of communication between cells in culture by co-culturing cells derived from mammalian, avian, and arthropod organisms. Both mammalian and avian culture cells have similar gap junctional phenotypes, while the insect (arthropod) cell lines have a significantly different gap junctional structure. Electrophysiological and ultrastructural methods were used to examine ionic coupling and junctional interactions between homologous and heterologous cell types. In homologous cell systems, gap junctions and ionic coupling are present at a high incidence. Also, heterologous vertebrate cells in co-culture can communicate readily. By contrast, practically no coupling (0-8%) is detectable between heterologous insect cell lines (Homopteran or Lepidopteran) and vertebrate cells (mammalian myocardial or 3T3 cells). No gap junctions have been observed between arthropod and vertebrate cell types, even though the heterologous cells may be separated by less than 10 nm. In additional studies, a low incidence of coupling was found between heterologous insect cell lines derived from different arthropod orders. However, extensive coupling was detected between insect cell lines that are derived from the same order (Homoptera). These observations suggest that there is little or no apparent specificity for communication between vertebrate cells in culture that express the same gap junctional phenotype, while there is a definite communication specificity that exists between arthropod cells in culture.


Isolation and characterization of plasma membrane-associated cortical granules from sea urchin eggs.

Cortical granules, which are specialized secretory organelles found in ova of many organisms, have been isolated from the eggs of the sea urchins Arbacia punctulata and Strongylocentrtus pupuratus by a simple, rapid procedure. Electron micropscope examination of cortical granules prepared by this procedure reveals that they are tightly attached to large segments of the plasma membrane and its associated vitelline layer. Further evidence that he cortical granules were associated with these cell surface layers was obtained by (125)I-labeling techniques. The cortical granule preparations were found to be rich in proteoesterase, which was purified 32-fold over that detected in a crude homogenate. Similarly, the specific radioactivity of a (125)I-labeled, surface glycoprotein was increased 40-fold. These facts, coupled with electron microscope observations, indicate the isolation procedure yields a preparation in which both the cortical granules and the plasma membrane-vitelline layer are purified to the same extent. Gel electrophoresis of the membrane-associated cortical granule preparation reveals the presence of at least eight polypeptides. The major polypeptide, which is a glycotprotein of apparent mol wt of 100,000, contains most of the radioactivity introduced by (125)I-labeling of the intact eggs. Lysis of the cortical granules is observed under hypotonic conditions, or under isotonic conditions if Ca(2+) ion is present. When lysis is under isotonic conditions is induced by addition of Ca(2+) ion, the electron-dense contents of the granules remain insoluble. In contrast, hypotonic lysis results in release of the contents of the granule in a soluble form. However, in both cases the (125)I-labeled glycoprotein remains insoluble, presumably because it is a component of either the plasma membrane or the vitelline layer. All these findings indicate that, using this purified preparation, it should be possible to carry out in vitro studies to better define some of the initial, surface-related events observed in vivo upon fertilization.


Histogenesis of mouse cerebellum in microwell cultures. Cell reaggregation and migration, fiber and synapse formation.

A microwell culture system was developed for analysis of cell movements and interactions during nervous system histogenesis. Cells from trypsinized 7-day-old mouse cerebellum reaggregated within hours into clusters which later developed interconnections consisting of either sheets of migrating cells and cell processes or cables of fiber bundles with cells migrating along their surfaces. Granule cells in several stages of differentiation, basket and/or stellate neurons, some larger neurons, and two types of neuroglial cells were identified in reproducible, nonrandom patterns by scanning and transmission electron microscopy. Axonal and dendritic processes, both with growth cones, and numerous synapses were generated in vitro.


Assembly of smooth muscle myosin into side-polar filaments.

The in vitro assembly of myosin purified from calf aorta muscle has been studied by electron microscopy. Two types of filament are formed: short bipolar filament similar to those formed from skeletal muscle myosin, and longer "side-polar" filaments having cross bridges with a single polarity along the entire length of one side and the opposite polarity along the other side. Unlike the case with skeletal myosin filaments, antiparallel interactions between myosin molecules occur along the whole length of side-polar filaments. The side-polar structure may be related to the in vivo form of myosin in vertebrate smooth muscle.


Some clues as to the formation of protrusions by Fundulus deep cells.

One of the ways in which Fundulus deep cells move in vivo is by putting out long, fingerlike protrusions. This involves a change in the shape of the cell as a whole, with cytoplasmic flow, and is not just a local phenomenon. Moreover, particles on the cell surface move toward a protrusion as it is forming, suggesting surface flow. The role of surface flow is discussed both on a grown level and in respect to molecular fluidity. Long, stable protrusions can be pulled from cells by the application of negative pressure at a constant rate and these behave in a similar way to those formed during cell locomotion. Such long protrusions must be structured. The importance of contractile properties of the cytoplasm in the formation of protrusions was studied by treating cells with media that modify cellular contractility.


Synchrony in the rhythm of the contraction-relaxation cycle in two plasmodial strands of Physarum polycephalum.

Rhythmicity of contraction of plasmodial strands of Physarum polycephalum was studied, by measuring the isometric tension exerted by isolated segments of the strands. When 2 strands were connected by way of a plasmodial mass, the contraction-relaxation cycle of the 2 strands synchronized. Such cycle activity of the strand was found to be well coordinated with shuttle streaming in the plasmodial mass which had been in connection with the strand. The presence of a control system which induces synchrony of periodic activity over the entire body of a plasmodium is discussed.


Synaptonemal complex karyotyping in Melanoplus differentialis.

The chromosomal axes of the spermatocytes of the grasshopper Melanoplus differentialis have been studied with a modification of the microspreading procedure used previously. The whole complement of synaptonemal complexes (SCs) and the axis of the X chromosome have been described and measured. The relative length of each SC is characteristic and constant and permits the construction of an idiogram. Relative lengths of SCs are almost equal to the relative lengths of mitotic chromosomes of spermatogonia (with the exception of the X chromosome), thus extending to an invertebrate the relationship between SCs and mitotic chromosomes that has been demonstrated in mammals. All the SCs except the 3 smallest (which are apparently telocentric) show a small short arm beyond the kinetochore. The progression of changes in the chromosomal axes during meiotic prophase has been staged by centriolar behaviour. During leptotene, axes are first formed near the nuclear envelope at a special (polar) region. SCs also begin to appear in the polar region and extend towards the nuclear interior. The beginning and completion of synapsis is not synchronous among bivalents. The X-axis is formed in midzygotene and shows a characteristic sequence of changes in shape during pachytene. Cells in post-synaptic stages show whole chromosome complements with characteristic chiasmatic configurations. Kinetochores are prominent and bipartite during diplotene-diakinesis.


In vitro transcription of RNA in nuclei, nucleoli and chromatin from physarum polycephalum.

Methods for isolating nuclei, nucleoli and chromatin from Physarum polycephalum which retain high levels of endogenous RNA polymerase activity are described. Under carefully controlled conditions with respect to mono- and divalent cation concentrations RNA synthesis in nuclei displayed linear kinetics for at least 30 min and the RNA products had a similar size distribution to nuclear RNA synthesis observed in vivo. Chromatin showed 60% of the nuclear transcriptional activity but no conditions were found where faithful transcription of the template occurred. Isolated nucleoli were 5-fold more active than nuclei and the endogenous RNA polymerase activity was insensitive to alpha-amanitin. Under carefully controlled conditions, the nucleoli appeared to support the accurate transcription, re-initiation and processing of rRNA chains in vitro.


Visualization of interphase chromosomes.

Using a modified Giemsa-banding technique we have observed what appear to be chromosomes during interphase in nuclei from Allium cepa root tips and Chinese hamster cells (CHO line). During telophase through G1 chromosomes progressively uncoil and decondense. During S chromosomes are comparatively decondensed, but some segments have structure similar to chromosomes in G1 and G2. During G2 the chromosomes progressively recondense and coil in apparent preparation for prophase. Although specific structural modifications of chromosomes occur in G1, Sand G2 nuclei, chromosomes appear never to decondense to the point that they lose their 3-dimensional integrity, but remain in distinct domains throughout interphase.


Electron-microscopic autoradiography of tritiated testosterone in rat testis.

The feasibility of a technique for autoradiography of diffusible substances has been further tested by analysing the localization of steroids in rat testes with the light-and electron-microscope. Testes of rats were perfused with tritiated testosterone (3 min) followed by 15-min perfusion with buffer containing a 100-fold of unlabelled testosterone. Tissue samples were frozen, freeze dried, fixed in osmium vapour and embedded in Epon. To exclude extraction of steroids, contact with water and other solvents was prevented during cutting of thin sections on an ultracryotome and further treatments for autoradiography. Light- and electron-microscopic observations indicate that the highest concentration of labelled testosterone was present within the basal parts of the Sertoli cell cytoplasm and in lipid inclusions of Sertoli cells within the seminiferous tubules. This is the first account of autoradiography of steroids at the electron-microscope level.


[Free intra-peritoneal endometrioma associated with ileal and appendicular endometriosis (author's transl)].

The authors report three rare localisations of endometriosis: ileal, appendicular and free endometrioma of the pouch of Douglas and study the diagnostic difficulties of digestive endometriosis and the interest of routine histological examination of structures found apparently free in the pouch of Douglas.


[Effects of stress and highly selective vagotomy on mast cells in the wall of the rat stomach (author's transl)].

Quantitative changes in the gastric mast cells in ulcer diathesis and especially in the development of acute ulceration have already been studied. Qualitative changes may be asse-sed by Alcian Safranin blue stain which permits one to distinguish two sub groups: the blue mast cells and the red mast cells, which alone undergo degranulation. This report describes the modifications in number and aspect of the mast cells of the stomach of the white rat after stress due to restraint at variable intervals after highly selective vagotomy. The density of the mast cells and their appearance depend partly on vagal nerve supply. After highly selective vagotomy, their density increases while the initially unbalanced proportion of red and blue mast cells becomes normal in the long term. After stress, there is a reduction in the number of mast cells and, above all, disappearance of the red forms whether the rats are vagotomised or not. This study takes into consideration multifactorial problems which intervene in the etiology of stress ulcers and the necessity of continuing the study of the consequences of vagotomy.


Decrease of prolactin by methysergide in amenorrheic hyperprolactinemic women.

The effect of methysergide (MES), a serotoninergic antagonist on prolactin (PRL) blood levels was studied in five hyperprolactinemic amenorrheic women. The drug was administered for five days at a daily dosage of 11.2 mg. MES decreased significantly the PRL blood levels in all subjects (p less than 0.01). Since the MES has been shown to have antiserotoninergic effects and since serotonin has been thought to be involved in the control of PRL release, the effects of MES in lowering PRL might be due to a decrease of serotonin tone.


Galactorrhea and hyperprolactinemia associated with chest wall injury.

A 48 year old premenopausal woman presented with galactorrhea and amenorrhea associated with chest wall burns. Basal serum prolactin levels were raised, and were further elevated by the administration of L-dopa, chlorpromazine and TRH. Intercostal nerve block and bromocryptine treatment reduced prolactin levels to normal, but did not noticably reduce milk secretion.


Periodicity and self-actualization in women.

Studied the relationship between menstrual cycle phase and self-actualization as measured by the Personal Orientation Inventory (POI). In classroom testing sessions, 160 high school students completed the POI and personal data sheets, which gave information about their menstrual cycle phase. Premenstrual, menstrual, and midcycle groups of 20 Ss each were formed from those girls eligible for inclusion in each condition. Three one-by-three analyses of variance were run using the TC, I, and TC + I scores of the POI, which were selected as the best overall measures of self-actualization. A significant (p less than .05) difference on TC + I was found; subsequent t-tests revealed that the menstrual group's TC + I score was significantly (p less than .02) higher than that of the midcycle group. The difference between menstrual and premenstrual Ss on TC + I, though not significant, lay in the same direction and was of the same order of magnitude. Possible explanations for the results and implications for future testing that involves females are presented briefly.


3H estradiol in catecholamine neurons of rat brain stem: combined localization by autoradiography and formaldehyde-induced fluorescence.

3H setradion-17beta was localized in nuclei of catecholamine (CA) neurons of the rat lower brain stem, i.e., midbrain, pons and medulla, by employing a combined formaldehyde-induced fluorescence (FIF)-autoradiography approach which permits the direct visualization of 3H estradiol uptake in relation to CA-neuron perikarya and terminals. In the medulla, noradrenalin (NA)-containing neurons in and adjacent to the nucleus (n.) reticularis lateralis (group A1) and in the vicinity of the n. tractus solitarii (group A2) concentrated 3H estradiol. In the pons, NA neurons dorsal to the n. olivaris superior (group A5), in the locus ceruleus (group A6) and ventral to the pedunculus cerebellaris superior (group A7) displayed 3H estradiol uptake. In the midbrain, dopamine (DA)-containing neurons in the n. cuneiformis (group A8), in the zona compacta of the substantia nigra (group A9) and adjacent to the n. interpeduncularis (group A10), were not seen to concentrate 3H estradiol. This is in contrast to DA neurons in the n. arcuatus of the hypothalamus (group A12), previously reported to concentrate 3H estradiol. In addition, certain non-CA-containing neurons with nuclear uptake of 3H estradiol were observed to be surrounded by CA terminals in such areas as substantia grisea centralis, n. tractus solitarii and n. tractus spinalis nervi trigemini. This suggests a second mode of interaction between CA neurons and estrogen target neurons. The estradiol-CA neuron relationships described may provide morphological substrates underlying functional interactions between estradiol and CA neurons in the rat brain.


Changing meal patterns and suppression of feed intake with increasing amounts of dietary nonprotein nitrogen in ruminants.

Goats were injected intraruminally during spontaneous meals with ammonium chloride, urea, ammonium lactate, or sodium lactate arranged in a Latin square experimental design randomized for order of treatments. Urea and ammonium injections shortened meal length by 20 to 30%. Rate of eating and meal frequency were reduced. Sodium lactate injections reduced meal size. In cows, meal length and meal size also were measured. Grain concentrate, corn silage, and chopped hay were fed as complete mixed rations. In the concentrates 58+ of the nitrogen was either from soybean meal or urea. Length of the first meal after feeding was reduced from 24.3 min with soybean meal to 12.4 with urea. Meal size was reduced from 3.2 kg to 1.8 kg when urea was fed. Total feed intake was similar, 12.0 kg/day (soybean meal) and 11.6 kg/day (urea) since spontaneous meal number and size were 17 and .30 kg for soybean meal but increased to 23 and .36 kg for urea. The physiological basis for the limit on meal length with urea rations is unknown but is an important factor in successful feeding of urea when eating time is limited for cows.


Vitamin A transport in plasma of the non-mammalian vertebrates: isolation and partial characterization of piscine retinol-binding protein.

Studies were conducted to explore vitamin A transport in the non-mammalian vertebrates, especially Pisces, Amphibia, and Reptilia, and to isolate and partially characterize piscine retinol-binding protein. Retinol-containing proteins in fresh plasma obtained from bullfrogs and a turtle exhibited similar properties to those found in mammalian and chicken plasma: i.e., molecular weight of about 60,000-80,000 as estimated by gel filtration and binding affinity to prealbumin on human prealbumin-Sepharose affinity chromatography. In sharp contrast, vitamin A-containing proteins in plasma from larvae of bullfrogs as well as three fishes (carp, blue sharks, and young yellowtails) appeared to be present in plasma as monomeric retinol-binding proteins without any affinity to human prealbumin. On the other hand, plasma vitamin A in the lamprey (Cyclostomes) was found to exist exclusively as an ester form in association with the lipoproteins of hydrated density less than 1.21 g/ml. Piscine retinol-binding protein was isolated from pooled plasma of young yellowtails and was converted (1000-fold purification) to a homogeneous component by a procedural sequence that included gel filtration on Sephadex G-100, chromatography on SP-Sephadex, gel isoelectric focusing, and, finally, polyacrylamide gel electrophoresis. Purified piscine retinol-binding protein showed physico-chemical properties distinctly different from the mammalian and chicken retinol-binding proteins examined, i.e., a smaller molecular weight of approximately 16,000, a lower isoelectric point of 4.3, a prealbumin mobility on analytical polyacrylamide gel electrophoresis, and a lack of binding affinity for human prealbumin; however, it displayed similar characteristics in two ways: a 1:1 molar complex with retinol, and a high content of tryptophan (four residues). These results strongly suggest that the piscine retinol-binding protein is a prototype of the specific vitamin A-transporting protein in plasma of the vertebrates, being modified later in evolution, during phylogenetic development of the vertebrates, to acquire a binding site for prealbumin on the molecule.