Plasma carnitine levels in children with protein-calorie malnutrition before and after rehabilitation.

Plasma carnitine and albumin levels were measured in children suffering from protein-calorie malnutrition, before and after rehabilitation, and in apparently healthy controls. Both the constituents were lower in malnourished children and improved after treatment. Plasma carnitine showed significant positive correlation with plasma albumin.


Rapid diagnosis of pyruvate and ketoglutarate dehydrogenase deficiencies in platelet-enriched preparations from blood.

Radiochemical methods are described in detail to measure the activities of the pyruvate dehydrogenase complex and of the ketoglutarate dehydrogenase complex in platelet-enriched fractions. Determinations can be completed in one day with as little as 5 ml of venous blood. Activities are proportional to the length of the incubation and the amount of tissue protein added, show appropriate dependence on added cofactors, are stable for up to 2 days at -20 degrees C, and do not appear to be affected by diet. The pyruvate dehydrogenase complex appears to be fully activated (dephosphorylated) in these preparations. Activities were comparable in platelet-enriched fractions from 25 normal subjects and 25 patients with a variety of neurological and psychiatric diagnoses. Mean values (+/- S.E.M.) for these 50 individuals were 169+/-9 pmol/min per mg protein for the pyruvate dehydrogenase complex and 535+/-27 pmol/min per mg protein for the ketoglutarate dehydrogenase complex. These values are comparable to those found in cultured skin fibroblast with similar techniques. Deficient pyruvate dehydrogenase activity (19+/-6 and 11+/-4 pmol/min per mg protein) was demonstrated in platelet-enriched preparations from two brothers whose fibroblasts had previously been shown to be deficient in pyruvate dehydrogenase and who responed to a ketogenic diet. Experimental detail critical to obtaining reproducible results with these methods are stressed (notably the crucial importance of maintaining the purity of the radioactive substrates). These techniques allow identification of patients with pyruvate dehydrogenase deficiencies within one day without requiring liver or muscle biopsy.


A Spanish family with erythrocyte pyruvate kinase deficiency: contribution of various immunologic methods in the study of the mutant enzyme.

Erythrocyte PK deficiency was detected in a 38-year-old man from Catalonia, in Spain. His father and his three children were proven to be heterozygous for the same mutant PK variant. This variant was characterized by low immunologic specific activity, normal (or slightly increased) stability to heat and to urea; normal isoelectric point, increased K0.5 for phosphoenolpyruvate, increased inhibition by ATP and normal activation by 0.35 mM fructose 1,6-diphosphate. The mutant PK variant was antigenically identical with wild enzyme as tested against anti wild erythrocyte PK serum by double immunodiffusion and micro complement fixation. The utility and the significance of the immunologic methods to be used in the study of mutant PK variants are discussed.


beta-Galactosidase in mucopolysaccharidoses and mucolipidoses. Deficiency of GM1 beta-galactosidase in liver and leukocytes.

beta-Galactosidase activities were studied in livers and leukocytes of mucopolysaccharidoses and mucolipidoses (I-cell disease and adult "beta-galactosidase deficiency" with macular cherry-red spots). Marked deficiency of hepatic 4-methylumbelliferyl (4MU) and GM1 beta-galactosidases was demonstrated in these diseases. Leukocyte GM1 beta-galactosidase was also deficient in mucolipidoses. The parents of the patients with I-cell disease and "beta-galactosidase deficiency" had normal beta-galactosidase activity in plasma and leukocytes, compared to the low enzyme activity in heterozygous carriers of GM1-gangliosidosis. The cause of this enzyme deficiency in these diseases is not clear at present. It seems to be affected seondarily by exgenous factors such as unknown stored materials in the cells. Mucopolysaccharides were not increased in the livers of two cases of I-cell disease and a case of "beta-galactosidase deficiency".


Lactosylceramidase assays for diagnosis of globoid cell leukodystrophy and GM1-gangliosidosis.

An assay procedure was developed for accurate estimation of lactosylceramidase II in the presence of relatively high activity of lactosylceramidase I. The procedure involves determination of lactosylceramide-cleaving activities under two different assay conditions, and lactosylceramidase II activity is calculated by the difference. Applicability of the procedure was evaluated with separated soluble fractions of the two beta-galactosidases from normal human brains, and with whole homogenates of gray and white matter, liver and cultured fibroblasts from control individuals and from patients with globoid cell leukodystrophy or GM1-gangliosidosis. The use of the lactosylceramidase I assay procedure developed by Wenger, D.A., Sattler, M., Clark, C. and McKelvey, H. ((1974) Clin. Chim. Acta 56, 199-206) and of the present procedure permits accurate diagnosis of both globoid cell leukodystrophy and GM1-gangliosidosis with one natural substrate, lactosylceramide, irrespective of the relative proportion of the two beta-galactosidases in the tissue.


Some properties of monoclonal cryoglobulin M appearing in the course of malignant lymphoma with macroglobulinemia.

Monoclonal cryoglobulin IgM was separated from the serum of a patient with malignant lymphoma and macroglobulinemia. The purified cryoglobulin in immunoelectrophoresis formed the precipitin lines with anti-mu and anti-kappa serum and showed an abnormal electrophoretic mobility due to large aggregate formation. In comparison to normal IgM preparations a significantly lower content of hydroxyproline was found in the cryoglobulin studied. The possible role of hydroxyproline-containing peptides in the formation of the oligomeric structure of the IgM molecule is discussed.


Effect of stilboesterol therapy on thyrotrophin-releasing hormone (TRH) responsiveness in males.

TRH responsiveness was investigated in twenty-four males (aged 66-83 years) undergoing stilboesterol therapy for carcinoma of the prostrate. Twelve were taking 2-6 mg daily by mouth for periods of 5 days to 12 months (mean 5 months) (Group A) and the remainder were on daily i.v. doses of 250-500 mg for periods of 3-10 days (mean 8 days) (Group B). In the former group TRH responsiveness was enhanced compared to age matched controls. A contrary effect was observed in the latter group in that the group mean TRH response was significantly lower than that of Group A or of the control group. Basal serum TSH values were unchanged in Group A compared to controls, but a significant elevation in the basal mean value was observed in Group B. The mechanism of action of stilboesterol in the subjects studied remains to be elucidated. Our data are in keeping with either a hypothalamic or pituitary locus for such actions.


Thyroid function and antibody studies in pernicious anaemia.

Thyroid antibodies were demonstrated in 57% of thirty pernicious anaemia patients without overt thyroid disease. Elevated basal thyroid stimulating hormone (TSH) levels and an enhaced TSH response to thyrotrophin releasing hormone (TRH) only occurred in thyroid antibody positive subjects; by contrast the thyroid antibody negative subjects in the older age group frequently and undetecable basal TSH levels and an impaired TRH response. Thyroid hormone concentrations provided no absolute evidence of hypothyroidism in any of the patients.


A human immunoglobulin in myeloma protein with anti-gastric parietal cell autoantigen activity.

A previously characterised IgA Kappa myeloma protein was isolated and purified. The (Fab) alpha fragments of this immunoglobulin were obtained. The IgA and Fab fragments reacted in vitro with gastric parietal cells (GPC) using animal gastric sections and with smooth cytoplasmic membranes obtained from rabbit fundus (gastric) mucosa. This was seen macroscopically and by light and electron microscopy. Evidence is thus provided which supports the concept that this homogeneous immunoglobulin is a typical monoclonal autoantibody.


Effects of a gold salt on lymphocyte responses.

The effects of sodium aurothiomalate (SATM) on certain lymphocyte functions in vitro were tested. Lymphocytes were obtained from both healthy donors and patients with rheumatoid arthritis (RA). SATM depressed tritiated thymidine uptake in lymphocyte cultures stimulated by phytohaemagglutinin, and in a mixed lymphocyte response assay. Cytotoxic effector cell generation assayed by percentage specific 51Cr release was also depressed by SATM using lymphocytes from healthy donors, and from patients with RA. These effects imply that SATM inhibits both exogenous thymidine uptake, and blastogenesis, and suggest that gold salts may act in RA by interference with T-cell dependent functions.


Metabolism of intravenously administered dipeptides in rats: effects on amino acid pools, glucose concentration and insulin and glucagon secretion.

1. Studies were performed to investigate the metabolic fate of dipeptides when administered intravenously in rats. Glycyl-leucine, glycylglycine or glycylsarcosine was injected into the jugular vein. The plasma disappearance rate after the peak plasma concentrations was most rapid for glycyl-leucine and least rapid for glycylsarcosine. 2. During urine collection for 40 min, trace amounts of glycyl-leucine and glycylglycine and 13% of the injected glycylsarcosine were excreted. 3. Neither glycylglycine nor glycyl-leucine was detected in the liver, muscle, intestinal mucosa or renal cortex, but concentrations of glycine or leucine, or both, in these tissues were increased after each injection. In contrast, glycylsarcosine was recovered in all these tissues with concentrations in the renal cortex being far greater than in any other tissue, but sarcosine was found only in the renal cortex and intestinal mucosa. 4. The changes in plasma concentrations of free amino acids, glucose and glucagon, and tissue concentrations of free amino acids, were similar after the intravenous administration of glycyl-leucine and an equimolar mixture of free glycine and leucine. However, the amount of insulin secreted during the 40 min after glycyl-leucine injection was 1-6 times that produced after the injection of the corresponding amino acid mixture. 5. Results show that, within the present experimental conditions, the intravenous administration of dipeptides is as effective as that of the corresponding free amino acids in enriching the tissue pools of amino acids. It is suggested that efficient hydrolysis by cellular enzymes prohibits accumulation of intact dipeptides in body tissues.


Spatial disorientation in the monkey.

The incidence of misdirected movements of the hand during tactile discrimination performance in the dark has been tabulated for monkeys with bilateral removals of parieto-prestriate cortex. The observations support the existence of a spatial disorientation (in addition to their inaccuracies of reaching) in these animals.


Correlation of the response of the flushing device to compression with the clinical picture in the evaluation of the functional status of the shunting system.

The functional status of cerebrospinal fluid shunting systems provided with flushing devices has commonly been assessed by the response of the flushing device to digital compression and a review of the clinical status of the patient. In order to study the reliability of the response of the flushing device as an indicator to shunt function, 47 hydrocephalic children with implanted shunts were studied. In 40% of these children, the flushing device was pumping well at the time of shunt obstruction, and in 50% it was not, although the patient's clinical status was perfectly normal. Accordingly, we conclude that the flushing device is a very poor indicator of shunt functioning.


Focal seizure disorder secondary to multiple shunt revisions. A case report.

A case of focal seizure disorder secondary to multiple ventricular shunt revision is presented. The literature reviewed indicates this complication of shunting is probably due to low grade infections or bleeding at the puncture site at the time of shunt revisions.


Acute schizophrenia: an efficacious outpatient treatment approach as an alternative to full-time hospitalization.

A research study was designed to test the hypothesis that acutely schizophrenic patients treated for a few minutes a day in a specially designed, city-county hospital outpatient clinic could remit as frequently, rapidly, and economically as a similar group managed on the ward of the same hospital. The results of the study permitted acceptance of the hypothesis. To wit, 90% of the clinic patients remitted in a median time of 12.5 days, as opposed to 70% of the ward patients remitting in a median time of 19.5 days and for approximately six times the cost.


Thyrotropin-releasing hormone in rat brain: nyctohemeral variations.

The concentration of thyrotropin-releasing hormone (TRH) was measured by specific radioimmunoassay in three different brain regions of rats sacrificed every 4 h over a 24 h period. TRH concentration reached a zenith at 1200 h both in the hypothalamus and  the amygdala but did not vary in the forebrain. These results indicate that a nyctohemaral variation of TRH levels exists in hypothalamic and extrahypothalamic brain areas.


Suppression of prolactin and thyrotropin secretion in the rat by antiserum to thyrotropin-releasing hormone.

Administration of antiserum to synthetic thyrotropin-releasing hormone (TRH) to male and female rats cause a 50% and a 70% suppression in serum levels of prolactin and thyrotropin, respectively, as compared with controls injected with normal rabbit serum. The degree of suppression was similar in diestrous and proestrous female rats and in male rats. These findings support the view that, in addition to its original designation, TRH also has a physiological role in regulating release of pituitary prolactin.


5.9-S RNA, a new RNA characterized in several mammalian cell lines.

A new species of RNA has been isolated from several different cell lines, both oncornavirus producing and non-producing. This RNA, which we designate 5.9-S RNA is present in the cellular cytoplasmic fraction at very low concentration (approximately 1% of the quantity of 4-S RNA), but it accumulates to much higher levels in two murine oncornaviruses, Moloney murine sarcoma leukemia virus complex and Gross leukemia virus, where it represents as much as 10% of the low-molecular-weight RNA fraction associated with the 70-S RNA genome. The electrophoretic mobility and fingerprint analysis of T1 RNase digest products show that this species of RNA is approximately 160-165-residues long, and can be unequivocally distinguished from all previously described species of RNA in this size range.


Comparative analysis of three guinea pig satellite DNA's by restriction nucleases.

The structures of guinea pig satellite DNAs I, II, and III have been analyzed by digestion with seven restriction nucleases. From the cleavage patterns it is obvious that the long-range periodicities in these three satellites differ rather characteristically Satellite I is fairly resistant to six nucleases and gives only a number of weak discrete bands which do not show a simple regularity. By the restriction nuclease from Arthrobacter luteus, however, it is cleaved extensively and yields very heterogeneous breakdown products. This is consistent with the high extent of divergence previously found for this satellite, e. g. by sequence analysis. Satellite II is almost completely resistant to all nucleases, indicative of a high degree of sequence homogeneity of this satellite. Satellite III is completely broken by the restriction nuclease from Bacillus subtilis into fragments which form a novel, highly regular series of bands in gel electrophoresis. The patterns show that the satellite is composed of tandem repeats ofapproximately 215 nucleotide pairs length, each repeat unit containing two cleavage sites for this nuclease. The data are consistent with the assumption that 30--40% of all cleavage sites have been eliminated by a random process. Satellite III DNA yields weak degradation patterns of the same periodicity with a number of other restriction nucleases. Cleavage sites for these nuclease are clustered on separatesmall segments of the satellite DNA. In this respect, the satellite is similar to others, notably the mouse satellite DNA. The three guinea pig satellites are examples of more general types of satellite structures also found in othe organisms. Similarities and differences to other satellites are discussed with special consideration to theories on the evolution of this class of DNA.


Studies on a carboxypeptidase Y mutant of yeast and evidence for a second carboxypeptidase Activity.

Immunological studies on the carboxypeptidase Y mutant prcl-l of Saccharomyces cerevisiae revealed the origin of mutation in the structural gene of carboxypeptidase Y. The absence of carboxypeptidase Y has no effect on growth, even after drastic changes of growth conditions. A double mutant (prc 1- leu2-) lacking carboxypeptidase Y and auxotrophic for leucine is able to grow on the peptide benzyloxycarbonylglycylleucine (Cbz-Gly-Leu) as sole nitrogen source, indicating the existence of a second carboxypeptidase. Using a new peptidase test, the existence of this second enzyme, called carboxypeptidase S, was confirmed biochemically.


Cortisone cataract in children with nephrotic syndrome.

In a group of 16 children with idiopathic nephrotic syndrome treated with corticosteroids for longer than 12 months, 9 developed a posterior subcapsular cataract (PSC). No correlation between the frequency of PSC and the duration of treatment and the total dose of treatment with steroids was demonstrable. However, the patients with PSC had received considerably higher average daily doses than those without PSC. Two patients with normal ophthalmologic findings at the end of treatment showed PSC 6 and 9 months later respectively. Only one patient acquired a significant impairment of visus.


Pharmacologic response to pentobarbital in actively immunized mice.

Mice were placed on an immunization schedule known to result in the production of antibodies directed against a variety of barbiturates (antibody binding capacity = 2.71 pmole 3H-phenobarbital bound/ml undiluted serum). The pharmacologic response to barbiturate in these actively immunized mice and suitably treated controls was investigates using rotarod apparatus for monitoring CNS depression. It was found that the response to pentobarbital in actively immunized mice was decreased, as reflected by an increase in the time the mice remained on the rotarod and a shift of the dose--response curve to the right. The decreased pharmacologic response to pentobarbital was not the result of changes in levels of the hepatic drug metabolism components. Furthermore, the alteration of pharmacologic response in actively immunized mice was selective for barbiturate and did not modify the ataxia produced by administration of another depressant agent, ethanol.


Nitroglycerin and propranolol on myocardial O2 consumption during myocardial ischemia.

The effects of i.v. and intracoronary (i.c.) nitroglycerin, i.v. propranolol, and the combination of propranolol and nitroglycerin on myocardial oxygen consumption (MVO2) and lactate utilization were studied in situ ischemic working dog hearts. I.v. nitroglycerin reduced MVO2 9% which was associated with the peripheral vasodilatory actions of the drug. I.c. nitroglycerin which had no detectable effects on the peripheral vasculature had no significant effect on MVO2. I.v. propranolol caused an 8% reduction in MVO2 and this action was associated with a negative chronotropic and a slight negative inotropic effect. However, the combination of propranolol and nitroglycerin which was more effective than either drug alone, reduced MVO2 18% thereby indicating that the effects of the two drugs on oxygen consumption are additive. These results support the concept that the reduction of MVO2 seen with nitroglycerin is principally due to peripheral vasodilatory actions of the drug while that seen with propranolol is due to that drug's effect on the heart and the effects of the two agents are complementary.


Short-term benefits and costs of U.S. family planning programs, 1970-1975.

Between 1969 and 1975, the U.S. family planning program helped its patients avert 1.1 million unwanted and mistimed births. These averted births resulted in short-term savings to the government for health and social welfare services of $1.1 billion, compared to a federal investment in family planning of $584 million--a benefit/cost ratio of $1.80 for every federal dollar invested.


Iatrogenic disease, drug metabolism, and cell injury: lethal synthesis in man.

One important consideration to be appreciated from the examples of "lethal synthesis" presented in this paper is that the mechanisms of toxic substances must be further explored, a true understanding of the complex modalities of cellular injury achieved, and the significance of cellular changes determined before we can preestablish the hazards of chemical or environmental contaminants. The identification of compounds that may modify the intracellular milieu by adduction of marcromolecules and those that may be activated to produce proximate toxins could be accomplished on the basis of their distinguishing structure and metabolism. Assaying for the levels and functions of xenobiotic systems may help us to preduct the potential risk to individuals. It is incumbent upon us now to become more critical of the side-effects of "therapy", to weigh the jeopardy against the benefits involved. And to do so, we must attain a better comprehension of the complicated mechanisms of cellular injury.


Histology of the baboon endometrium during the menstrual cycle and pregnancy.

The histologic characteristics of baboon endometrium during the menstrual cycle and pregnancy were studied and compared with those of human endometrium. Eight phases of endometrial change during the menstrual cycle are described. The samples were dated on the basis of sex skin changes. The basic histology of the baboon endometrium is similar to that of human endometrium, but some differences were observed. The growth of baboon endometrium is more sluggish, secretion is less intense, and many of the changes, especially in the stroma, are localized, less diffuse than they are in human endometrium. During pregnancy, decidual transformation in the baboon is much less intense than that in the human. No differences were observed between endometrium in association with preimplantation embryos and that from nonpregnant animals at comparable times following ovulation.


Condyloma-like lesions possibly caused by Neisseria gonorrhoeae. Report of a case.

A 2-year old girl suffered from continuous pyogenic inflammation in the anogenital region probably enhanced by neurological alterations. In one episode, the inflammatory process presented multiple moist papillomatous growths on the labia majora and along the anal margin which are thought to have been caused by gonococcal infection.


Marked instability of character of cloned epithelial cells.

Cell lines derived from 7,12-dimethylbenzanthracene-induced rat mammary carcinomas show a marked drift in properties when cultured in vitro. The following characteristics alter progressively with the number of passages: cloning efficiency, requirement for hydrocortisone, requirement for fetal sera, growth response to conditioning factor(s) and ease of detachment from the culture vessel by trypsinization.


Investigation of insulin sensitivity in treated subjects with ketosis-prone diabetes mellitus.

Two similar intravenous infucion techniques have been utilized to investigate insulin sensitivity in young subjects with recent-onset ketosis-prone diabetes mellitus. All subjects presented initially with mild to moderately severe ketoacidosis and had been treated with daily insulin therapy for two to eight weeks at the time of study. Six diabetics and 10 normal subjects (group A) received intravenous infusions of glucose (6 mg./kg./min), insulin (80 mU./min.) for 150 minutes. Steady-state plasma glucose (SSPG) and insulin levels (SSPI) were reached by 90 minutes and maintained through the end of the study. As all subjects achieve simolas SSPI while simultaneously receiving similar glucose loads, the SSPG can be used to measure individual insulin sensitivity. Under these conditions the diabetics in group A had a mean SSPG (+/-S.E.) of 99+/-26 mg./100ml., which was not different from the level of 98 +/-14mg./100 ml. for their control subjects. Six diabetics and six normal subjects  (group B) received infusions of only glucose (6 mg./kg./min.) and insulin (80 mU./min.). Similar SSPI levels were attained in both the diabetic and control subjects, and their mean SSPG (+/-S.E.) levels were not significantly different (83+/- 13 vs. 61 +/-6 mg./100 ml.). One diabetic in group A and two diabetics in group B had SSPG levels above the highest values measured in their control groups. However, all three had markedly elevated fasting plasma glucose levels on the day of study. In contrast, nine well-controlled diabetics had normal insulin sensitivity. These results suggest that well-controlled subjects with ketosis-prone diabetes mellitus have normal sensitivity to insulin.


Nosological problems of myoclonic epilepsies in childhood and adolescence.

An attempt is made to clarify the confusion over the classification of the myoclonic epilepsies of childhood. Six types of seizure are described, the classification being based on clinical and electroencephalographic findings. The types of myoclonic epilepsy are infantile spasms, myoclonic astatic epilepsy, myoclonic absence, myoclonic epilepsy of childhood, myoclonic epilepsy of adolescence and photomyoclonic epilepsy, the last being subdivided into myoclonic jerks and eyelid myoclonia. Synonyms are given for all types, based on the major papers in the literature over the last 16 years. The similarity of infantile spasms and myoclonic astatic epilepsy is discussed, as are the so-called 'Lennox' and 'Lennox-Gastaut' syndromes.


Treatment of generalized epilepsies of childhood and adolescence with sodium valproate ("epilim").

A total of 142 patients (84 per cent aged less than 20 years) with various forms of generalized epilepsy have been treated with sodium valproate alone or in combination with other drugs. The mean duration of symptoms was six years, and half the patients had daily seizures. Nine patients had typical absences, 33 had absences with automatisms, 28 had tonic-clonic seizures with or without photosensitivity, and 72 had various forms of myoclonic epilepsy. Dosage varied from 23 to 54mg/kg and twice-daily administration was usual. Estimation of serum levels did not assist in management. Fits ceased in 63 per cent of all cases and a further 18 per cent showed improvement greater than 50 per cent. Of the 69 with 3c/sec spike-and-wave discharges, 81 per cent became free from all fits, as did 77 percent of those with myoclonic jerks. Fits ceased in eight of the 32 patients with myoclonic astatic epilepsy and there was improvement greater than 50 per cent in a further eight patients. Other anticonvulsants were often withdrawn and always reduced. 21 patients received sodium valproate alone from the start of treatment and all other drugs were withdrawn in another 38. Sodium valproate alone controlled all fits in four children with absences, in 18 with absences with automatisms, 10 with tonic-clonic seizures and 22 with myoclonic epilepsy. Side-effects were rare, mild and often temporary. Potentiation of barbiturates and benzodiazepines occurred, especially clonazepam, which should be avoided. Many patients were more alert. Sodium valproate appears to be the drug of choice for epilepsies associated with generalized spike-and-wave discharges, myoclonic epilepsies or photosensitive epilepsies, and is of especial value in children and mentally retarded patients because it lacks sedating effects and often induces liveliness.


Distribution of carbonic anhydrase within the gastric mucosa.

Bullfrog fundic mucosa was separated into fractions containing predominantly surface epithelial cells, mucous neck cells, and and oxyntic cells, respectively, and the carbonic anhydrase concentration of each fraction and of antral mucosa was measured. There were no significant differences between the enzyme concentrations of these fractions. The mean level was 12.4 units of activity per milligram of protein. The cellular localization of carbonic anhydrase in the gastric fundic and antral mucosa of the bullfrog, rat, rabbit, and dog was studied using a specific histochemical technique. The enzyme was shown to be present in both parietal and nonparietal epithelium of all species and to characterize a specific cell type in the dog antrum. The widespread distribution of high levels of carbonic anhydrase activity in both acid secreting and nonsecreting cells of the gastric mucosa suggests a possible role for the enzyme beyond one solely relating to acid secretion.


Geographic distribution and habitat diversity of the Barbary Macaque sylvanus L.

During a 15-month behavioral study in Morocco and a 3-month survey in Morocco and Algeria, the present distribution of the Barbary macaque was determined. In Algeria, monkeys are found in seven constricted and disjunct localities in the Grande and Petite Kabylie mountain ranges. These localities are severely restricted in space and are located in remote or inaccessible areas which support only small populations. Their habitats include mixed cedar and holm oak forests, humid Portuguese and cork oak mixes and gorges dominated by scrub vegetation. In only two regions (Guerrouch and Agfadou) can population of reasonable size be found; even there they do not approach the abundance found in the central Middle Atlas zone of Morocco. Distribution was more extensive earlier in this century and some areas have become unoccupied within the past 15 years. Today, their absolute numbers and population densities are low in all but two locations. Algeria contains approximately 23% (5,500 maximum) of the total number of surviving Barbary macaques in North Africa. About 77% of the total number of Barbary macaques occur in Morocco. Moroccan habitats include high cedar forests, cedar/holm-oak mixtures and pure holm oak forests. Macaque distribution in the High Atlas is restricted to the Ourika valley where only a small relict population survives. There are between five and eight small, disjunct forest pockets in the Rif which support small groups of monkeys. In the Middle Atlas, monkeys are found in high numbers and in relatively wide stretches of distribution, although there are constricted areas of low densities in this region also. 65% (14,000 maximum) of the animals and their highest densities occur in the high mixed cedar forests of the Central zone, and mixed cedar forest appears to be the preferred habitat for the species. With the exception of the Central zone, their remaining distribution is typically disjunct and constricted, and population densities aer low. As in Algeria, distribution in Morocco was wider earlier this century, and several areas have recently become unoccupied.


Neonatal-maternal weight relationship in macaques: an example of intrageneric scaling.

In the genus Macaca, represented by seven species, neonatal weight scales at a power of maternal weight of a = 0.49. This value is lower than those for subordinal and ordinal scaling of neonatal weight in primates (a = 0.63-0.78), but it is higher than the value for intrageneric scaling of brain weight in Macaca (a = 0.18). A causal interpretation is attempted for differing values of a at different taxonomic levels and between different organ systems.


Differential habitat utilizati of four Cercopithecidae in a Kenyan forest.

The presence of Papio cynocephalus, Cercopithecus mitis, Cercopithecus aethiops and Colobus angolensis in a small coastal forest is attributed to their differential habitat utilization. Data on diet, space-use patterns and polyspecific associations are given. Habitat partitioning is found to be resource-related and is accomplished through differential reliance on resources as well as different means of exploiting overlapping resources. The implications of the data for understanding the relationship between ecology and social structure are discussed.


[The treatment of primary generalized epilepsies with dipropyl acetate (DPA)].

79 patients with primary generalized epilepsies have been treated with DPA in a medium dosage of 51 mg/kg bodyweight/day, range 14 to 125 mg/kg/day, for a medium time of 22 months, range 2 to 49 months. 51 children out of this group had been treated previously and were therapy resistant to other medications. 27 children got DPA for their first medication. 34 patients were treated with DPA as a single drug, 45 were treated in combination with other medications. Therapeutic success was found to be remarkable good in impulsive petit mal (n = 4, all patients without any more seizures), in absences (n = 52, complete success in 84%), and in primary generalized grand mal seizures with spike-waves in the EEG alone or in combination with petit mal (n = 30, 87% success). However, centrencephalic myoclonic-astatic seizures (n = 17, no more seizures in 35%) were influenced significantly less. Side effects were rarely seen, mostly they could be observed in those patients treated with DPA and another medication. Side effects never induced interruption of treatment with DPA.


Comparison of TRF, propranolol-glucagon, insulin and glucose stimulation tests in acromegaly.

In 7 acromegalic patients growth hormone responses were studied following administration of synthetic TRF, propranolol-glucagon, insulin, and glucose p.o. Except for the glucose tolerance test, a good reproducibility of the STH response was observed. In 5 out of the 7 patients, there was a distinct rise in the plasma STH level after TRF. All patients with a positive insulin tolerance test responded to TRF, as did the two late responders to glucagon; the early responder to the latter test did not respond to TRF. It has been suggested (Liuzzi et al. 1974a) that TRF might be used as a screening test for detecting hypothalamic dependency of the acromegaly. This study suggests that further study is required before accepting this hypothesis and that a response to a combination of tests (TRF, glucagon, insulin) might be a better screening method.


Intrahepatic transplantation of pancreatic islets in the rat.

Islets of Langerhans from isogeneic donor rats were transplanted directly into the hepatic parenchyma of recipients which had been made severely diabetic by streptozotocin (glycaemia ranging between 400 and 1090 mg%). Complete control lasting up to 13 months was achieved in 65% of recipients by using 600-800 islets. Following intravenous glucose administration, each rat responded similarly to normal rats with a rapid but reduced release of insulin. Cytoimmunofluorescence and electron microscopic studies demonstrated the presence of both functional insulin and glucagon cells, within the transplanted islets. It is suggested that for various reasons direct intrahepatic transplantation might become the preferred method for islets.


Influence of D-thyroxine on plasma thyroid hormone levels and TSH secretion.

Triiodothyronine (T3), thyroxine (T4), basal TSH and TSH after stimulation with TRH were determined in healthy subjects and patients treated with D-thyroxine (DT4). After a dosage of 6 mg DT4 the D/L T4 plasma concentration rose about 4-fold 4 hours after application and was only moderately elevated 14 hours later. To achieve constantly elevated T4 levels 3 mg DT4 were applied in the further experiment every 12 hours. The D/L T4 plasma concentration rose 2.5-4-fold and there was a small but significant increase of the D/L T3 plasma concentration. 74 hours after onset of treatment basal TSH was below detectable limits and the increase of TSH 30 min after injection of 200 mug TRH (TRH test) was only about 15% compared to zero time. The time course of TSH suppression was investigated after treatment with DT4 and LT4 (single dosage of 3 mg). TRH-tests were performed before, 10, 26, 50 and 74 hours after the first dosage of D or LT4. There was no difference in the time course of basal TSH and TSH stimulated by TRH. In 10 patients on DT4 long-term therapy, basal and stimulated TSH were found to be below the detectable limits of 0.4 mug/ml. Our results show that (1) plasma half-life of DT4 is less than 1 day, (2) TSH suppression after D and LT4 treatment is very similar, and (3) in patients on long-term DT4 treatment, TSH plasma concentration is below detectable limits even after stimulation with TRH.


The Tamang: a case of hybridization.

The Tamang, a Mongoloid and hybrid population of the eastern part of the cis-Himalayan region of West Bengal, have been studied for an estimation of the Tibetan and Nepalese components. The degree of admixture of the two parental stocks is found to be about 59 and 41%, respectively. The deviation between the observed and expected gene frequencies of the hybrid is insignificant.


Utilization of electron microscopy in the prenatal diagnosis of genetic disease.

The use of electron microscopy as a further method of diagnosis of disease in cultured skin fibroblasts and cultured amniotic fluid fibroblasts is presented. It was demonstrated that Tay-Sachs disease, Fabry's disease, and metachromatic leukodystrophy had distinctive abnormalities in both cultured skin fibroblasts and cultured amniotic fluid fibroblasts. It was shown that control of culturing conditions made it possible to distinguish normal cell lines from certain cell lines carrying known genetic diseases.


[Monomeric IgM in lymphoproliferative, autoimmune and infectious diseases in man (author's transl)].

7S-IgM is the first specific antibody during evolution of vertebrates. The detection of monomeric IgM in monoclonal grammopathies, autoimmune and infectious diseases is believed to be an atavistic primary IgM response in cases of elevated humoral immune reactions and disorders of lymphatic tissues.


Human eosinophils, acidic tetrapeptides (ECF-A) and histamine. Interactions in vitro and in vivo.

The ECF-A acidic tetrapeptides Val-Gly-Ser-Glu, Ala-Gly-Ser-Glu and the analogue Val-Gly-Asp-Glu were selectively chemotactic for human eosinophils over a narrow dose range although eosinophils from different individuals varied in their dose-response pattern. Histamine abrogated the chemotactic properties of the individual tetrapeptides. When Val-Gly-Ser-Glu and Ala-Gly-Ser-Glu were combined in various concentrations the resultant chemotaxis was either negligible or no greater than that produced when each peptide was tested separately. Val-Gly-Ser-Glu and Ala-Gly-Ser-Glu both promoted eosinophil accumulation when applied to the abraded skin of man or i.d. to the marmoset. Biopsies of marmoset skin revealed that peptide-induced eosinophilia was not associated with mast-cell degranulation. Histamine, which was chemotactic in vitro, did not lead to appreciable eosinophil accumulation in vivo, and combinations of histamine and the acidic tetrapeptides evoked little or no cutaneous eosinophil infiltration either in man or the marmoset. These studies suggest that there is a complex interaction between histamine and the ECF-A tetrapeptides; however, the tetrapeptides alone can promote the recruitment and localization of eosinophils by a mechanism apparently independent of mast-cell degranulation.


Depression of T-cell function and normality of B-cell response in protein calorie malnutrition.

Antibody response to a T-cell dependent antigen, sheep erythrocytes and to a B-cell mitogen, purified lipopolysaccharide (LPS), has been studied in mice kept on protein deficient (2 and 4 per cent casein) diets. The number of plaque-forming cells (PFC) to SRBC were 20-5 +/- 7-7 per million spleen cells in protein-deficient animals compared to 261-0 +/- 31-1 in parallel controls maintained on a protein rich diet (18 per cent casein). No difference was observed in number of PFC formed in controls and deficient animals to LPS, values were 161-4 +/- 19-7, 158-5 +/- 14-2, & 162-3 +/- 31-9 in control (18 per cent casein) and deficient groups (4 per cent and 2 per cent casein) respectively. The delayed hypersensitivity skin reaction to SRBC measured in foot pads was significantly lower in mice on 4 per cent casein diet compared to controls. These studies suggest that the effect of protein deficiency is primarily on T-cell function and not on the B-cell response;


[Complete parenteral nutrition in childhood].

Complete parenteral nutrition by means of continuous intravenous drip into large veins, over a long period, has already shown itself to be of therapeutic use in the field of pediatrics. While the nutritional-physiological aspect of this therapeutic method is clear -- it depends on an accurate and sufficient supply of water, salts, glucose, amino-acids, triglycerides and essential fatty acids, vitamins and trace elements -- in practice it presents difficulties which though often serious are not insurmountable. Especially are to cite the septic complications, which are nearly imperceptible during their development. They are only to accept in view of the often almost hopeless condition of the child, who would be lost without complete parenteral feeding. Similar problems arise through metabolic complications which are caused non rarely only by insufficient dosages and lack of careful supervision but it is generally possible to correct these at an early stage. Complete parenteral nutrition must be exactly indicated. It is indeed a great therapeutic help, particularly when over a longer period -- at least a week or longer -- oral nutrition is impossible or insufficient. Practicall experiences will be reported. A retrospective assessment of 50 cases has shown the tendency (through taking too much care?) to hold the quantity of liquids and supply of calories too short and to administer too much carbohydrates and too little fat.


[Starvation and surgical intervention].

The metabolic processes under conditions of stress and starvation are not identical. Moreover, they have to be differentiated, as far as carbohydrate-, fat- and amino-acid-metabolism are concerned, because both situations are not compatible. It is therefore required, to avoid any kind of starvation and particularly to substitute for adequate substrates during the pre- and postoperative period.


[Modification of immunoglobulins during combined peroral and parenteral nutrition in colitis, ileitis and rectal carcinoma].

Reaction of immunoglobulins (IgA, IgG, IgM) during a four week period under condition with a special parenteral and peroral alimentation was observed in three groups of patients (rectum neoplasm, ulcerative colitis, Crohn's disease) during a period of two weeks all patients received 150 g aminoacids and 510 g carbohydrates by central venous catheter. In the following 14 days parenteral nutrition was limited and oral alimentation began. All patients showed a remarkable approvment of recreation. Globulins rose in between normal spectrum. The most marked changes with a significant uprising of IgG showed patients with rectum carcinoma treated cryosurgically.


[Specificity of ATP depletion in the liver following intravenous injection of energy yielding substrates].

The decrease of ATP-concentration in tissues after rapid intravenous injection is not specific, neither with regard to the organ nor with regard to the substrate. It can be achieved by any substrate which is channeled into metabolism by phosphorylation or activation using ATP, if the organ has the appropriate enzymatic outfit.


Cell-mediated resistance to infection with Listeria monocytogenes in nude mice.

Congenitally dysthymic nude (nu/nu) NMRI mice showed increased resistance to viable Listeria monocytogenes cells during the initial phase of infection as compared with euthymic control mice. The intravenous mean lethal dose (LD50), as determined for euthymic mice after an observation time of 7 and 14 days, amounted consistently to 6 X 10(4) Listeria. The corresponding values determined in nude mice were found to be increased by either 20-fold (1.2 X 10(6) Listeria after an observation time of 7 days) or 4-fold (2.4 X 10(5) Listeria after an observation time of 14 days). The transfer of spleen cells from immune euthymic donor mice into chronically infected nude mice caused almost complete elimination of Listeria within 1 week. The injection of dextran sulfate 24 h before a secondary infection with L. monocytogenes caused loss of antibacterial resistance in both chronically infected nude mice and Listeria-immune euthymic mice, this being expressed by a rapid increase in the numbers of bacteria in the spleens as well as the occurrence of serious signs of illness.


Long-term studies on rhesus monkeys (Macaca mulatta) immunized against Plasmodium knowlesi.

Studies carried out on four rhesus monkeys (Macaca mulatta) that had been vaccinated against Plasmodium knowlesi show that the immunized animals were protected against a challenge with a heterologous strain of P. knowlesi. This protection was shown to be present even 4 years after the immunization schedule has been completed. The effect could not be attributed toprevious infections with the parasite, since four control rhesus monkeys that had recovered from one to four challenges with P. knowlesi died when exposed to the heterologous strain. Data obtained from the lymphocyte transformation test and the radioimmunoassay are also presented.


Purification of a monocytosis-producing activity from Listeria monocytogenes.

The monocytosis-producing activity (MPA) from Listeria monocytogenes is shown to be associated with the cell envelope. Both MPA and immunosuppressive activity were readily extracted with aqueous solvents and separated as two independent activities by gel filtration and ion-exchange chromatography. As little as 10 microng of the resulting MPA-containing fraction caused a ninefold elevation in the level of circulating monocytes. The molecular weight of the fraction was approximately 22,000, and it contained phosphorus and carbohydrate.


Evaluation of experimentally induced Fusobacterium necrophorum infections in mice.

Two strains of mice, Swiss Webster and DBA/2Cr, were injected intraperitoneally or intravenously with varying dosages of Fusobacterium necrophorum. The ability to eliminate the infection was assessed by quantitative enumeration of the organisms present in the blood, liver, and spleen, Three- to 4-week-old DBA/2Cr mice were highly resistant to both routes of injection. The intraperitoneal injection of older mice failed to demonstrate a dose-effect relationship whereas an intravenous injection of as few as 10(4) cells of F. necrophorum produced progressively necrotic leg abscesses, apparently involving the lymphonodus ischiadicus which filters the site of injection. Mortality was increased with sensitization by a previous sublethal injection. Also, an ethanol-killed cell vaccine delayed the onset of lethal infection, whereas repeated sublethal live cell injections provided nonspecific protection since mice vaccinated with the growth medium were equally protected. The development of leg abscesses after intravenous injection visibly demonstrated the pathogenicity of F. necrophorum and may provide a suitable model for the evaluation of vaccines and the effectiveness of antibiotics.


In vitro interaction of Neisseria gonorrhoeae type 1 and type 4 with tissue culture cells.

As a basis for studies of gonococcal pathogenicity, tissue culture cells were infected with type 1 or type 4 Neisseria gonorrhoeae to determine intracellular viability. A simple and objective means of measurement was devised, based on the uptake of tritiated protein and deoxyribonucleic acid precursors by cycloheximide-inhibited cells infected with gonococci. Cycloheximide was found to inhibit protein synthesis by over 97% tissue culture cells at a concentration of 100 microng/ml. In contrast, N, gonorrhoeae was found to be highly resistant to this antibiotic, and protein synthesis was unaffected by concentrations up to 1,000 microng/ml. Extracellular gonococci were eliminated by treatment with high concentrations of penicillin during cycloheximide inhibition and prior to the addition of radioisotope. Levels of protein and deoxyribonucleic acid synthesis by N. gonorrhoeae in the cycloheximide-treated cells were significantly higher in T1-infected cells (RE2, HeLa, or HEp-2) than in the corresponding T4-infected cells. No differences were observed in tissue cell susceptibility to gonococcal infection. Intracytoplasmic localization of N. gonorrhoeae was confirmed by electron microscopy.


Relationship of pili to colonial morphology among pathogenic and nonpathogenic species of Neisseria.

Growth in colonies with type 1 morphology and the presence of pili are characteristics that have been associated with virulence of gonococci for humans. To determine whether the presence of pili per se might be responsible for colony type 1 morphology, the relationship of pili to colony type was examined in various species of Neisseria. Short pili (175 to 210 nm in length) were seen only on nonpathogenic neisseria, whereas long pili (up to 4,300 nm) were seen on organisms of both nonpathogenic and pathogenic species. Although long pili, similar to those found on organisms from high-domed, type 1 colonies of gonococci, were observed on organisms from high-domed, type 1 colonies of nonpathogenic Neisseria species, they were also observed on low-convex, type 4 colonies of meningococci and nonpathogenic neisseria. Among meningococci there was no difference in the morphology of colonies consisting of organisms with many long pili and colonies consisting of organisms that completely lacked pili. Thus, there was no consistent relationship of pili to colonial morphology. Unless the pili of N. gonorrhoeae are unique among Neisseria species in their influence on colonial morphology, it is likely that factors other than pili determine colony type 1 morphology of gonococci. Whether these same factors, either alone or in conjunction with pili, are also responsible for gonococcal virulence warrants further investigation.


Pyocin sensitivity of Neisseria gonorrhoeae and its feasibility as an epidemiological tool.

Pyocin inhibition of Neisseria gonorrhoeae and its feasibility as a gonococcal typing scheme were examined. Mitomycin C-induced pyocin lysates of Pseudomonas aeruginosa were able to selectively inhibit the growth of gonococcal strains. The particles associated with the inhibitory activity were non-dialyzable, heat labile, Pronase sensitive, trypsin resistant, and of large molecular weight by membrane and gel filtration techniques. The inhibitory activity was shown to be specific by absorption with sensitive and insensitive strains of N. gonorrhoeae and P. aeruginosa. Partial purification of pyocin lysates by ammonium sulfate precipitation followed by ultracentrifugation revealed phagelike particles consistent with high-molecular-weight R-type pyocines. These particles were associated with increased inhibitory activity and could be seen associated with the gonococcal cell surface. One hundred and six gonococcal strains could be differentiated on the basis of their sensitivity of 23 pyocin extracts. Thirty different patterns of pyocin inhibition were seen. Isolates from different body sites from the same patient could generally be identified as being similar strains. Strains isolated from known consorts had the same patterns. In general, agreement between pyocin typing and available epidemiological information was good.


Production of a capsule of Neisseria gonorrhoeae.

Neisseria gonorrhoeae can be induced to form a large capsule that can be visualized with India ink. Encapsulated gonococci appeared to be resistant to non-antibody-mediated phagocytosis compared with unencapsulated gonococci of the same strain and colony type.


Reagin-like activity of serum in human peridonotal disease.

The serum from patients with periodontitis produced a passive cutaneous anaphylaxis reaction in monkeys when tested against subgingival plaque. This detection of reaginic antibodies shows that patients with periodontitis make immunoglobulin E antibodies to plaque constituents.


Differences in buoyant-density properties of Coxiella burnetii and Rickettsia rickettsii.

The pronounced change in the buoyant density of Coxiella burnetii in CsCl gradients that was caused by treatment with formalin or ultraviolet radiation was not observed with Rickettsia rickettsii.


Genital inoculation of male baboons with Neisseria gonorrhoeae.

Eight male baboons inoculated intraurethrally with Neisseria gonorrhoeae failed to shed gonococci or develop serum antibody. Urethral inoculation, preceded by epididymal inoculation, elicited an anamnestic antibody response.


An immunological enhancement phenomenon in experimental brucella infection of the chicks.

The passive transfer of chicken anti-brucella immune globulins promote the survival of Brucella abortus in the spleen of chickens infected with this bacteria. This enhancing effect was reduced significantly when the immune globulins were absorbed with heat-killed brucella cells to remove all the anti-brucella agglutinins and immune adherence antibodies. Therefore, it is most probable that the enhancing effect is related to the presence of brucella antibodies.


Vulnerability of specific rat chromosomes to in vitro chemically induced damage.

The treatment of rat embryo secondary cultures with DMBA or DMBA-3H for 5, 9, or 24 h resulted in chromosome damage consisting mainly of chromatid type aberrations. There was an increase in the percentage of labelled nuclei and metaphases with increasing length of exposure. In terms of incidence of chromatid lesions, the largest telocentric chromosome (No. 2) was the most susceptible of the autosomes. Banding pattern analysis demonstrated that the region associated with negative band 2q24 of the No. 2 chromosome had the highest number of lesions. An increased accumulation of DMBA-3H label occurred in approximately the same chromatid area of a small fraction of cells exposed for either 5 or 9 h prior to mitosis. The complete loss of DMBA-3H chromosomal labelling after DNAse treatment suggests that the visible grains represent carcinogen-bound DNA. After DMBA and BrdUrd, there was an increase in the number of sister chromatid exchanges compared to controls treated with BrdUrd only; the location of the exchange points on chromosome No. 2 was similar in samples treated with either DMBA and BrdUrd or BrdUrd alone. Additional experiments with thymidine-3H showed that the non-random chromatid lesions on chromosome No. 2 may result from endogenous radiation from the incorporated tritium. These studies demonstrate that a specific chromosome may be affected by diverse agents and that chromatid lesions frequently occur at the site of sister chromatid exchanges.


Radioimmunoassay for the major structural protein of Mason-Pfizer monkey virus: Attempts to detect the presence of antigen or antibody in humans.

The 25,000 dalton protein of Mason-Pfizer monkey virus (MPMV) was isolated by gel filtration chromatography. In agreement with results from other laboratories, antisera to type-C and the non-type-C bovine leukemia and equine infectious anemia viruses did not precipitate 125I-labelled MPMV p25. In addition, these viruses did not cross-react in a competition radioimmunoassay for MPMV p25. Twenty-one human tissues (15 breast carcinomas, 2 normal breasts, 3 acute myelogenous leukemias and 1 sarcoma) were fractionated by detergent solubilization, ammonium sulfate precipitation, and DE-52 anion exchange chromatography. These methods were shown to be highly effective for purification of MPMV p25. Under assay conditions which minimized incubation damage to the 125I-MPMV p25, all tissues failed to react in the competition radioimmunoassay (RIAT). Two hundred and two human sera or plasma specimens, including those from patients with breast cancer and 33 age-matched controls, from 50 patients with hematologic malignancies, from 12 patients with amyotrophic lateral sclerosis, and from 14 patients with systemic lupus erythematosis, were examined for antibodies to MPMV p25. With the exception of two multiple myeloma plasma which produced artifactual false positive reactions based on hypergammaglobulinemia, a known complication of salt precipitation radioimmunoassays, the remainder of the specimens were negative for evidence of MPMV p25 antibodies.


Comparative pharmacokinetic investigations with tritium-labeled ergot alkaloids after oral and intravenous administration man.

Pharmacokinetic studies were carried out with nine tritium-labeled ergot alkaloids (dihydroergotmine, dihydroergotoxine, dihydroergostine, dihydroergocornine, dihydroergovaline, dihydroergonine, ergotamine, 1-methyl-ergotamine, and bromocriptine). Each drug was administered to 6 subjects in a randomized cross-over design as single oral and intravenous doses. Plasma levels and urinary excretion of tritium-labeled material were analyzed on a phenomenological basis by non-linear regression as a sum of exponentials. All substances showed the highest plasma concentration about 2 hours after oral administration (range 1.0-2.7 hours). The mean invasion half-life was 0.5 hours (range 0.32-1.12 hours). The mean elimination half-lives ranged from 1.4-6.2 hours for the alpha-phase and from 13 to 50 hours for the beta-phase, the longest values being observed with bromocriptine. From cumulative urinary excretion data after oral and after intravenous administration, a quotient of absorption was calculated. Values between 25 and 30% were found for most dihydrogenated drugs, namely dihydroergotamine, dihydroergotoxine, dihydroergostine, and dihydroergocornine, the only exceptions being dihydroergovaline and dihydroergonine, which were less well absorbed. Ergotamine and 1-methyl-ergotamine had an absorption quotient of about two-thirds and bromocriptine was nearly completely absorbed.


Light and electron microscopy of the anterior chamber angle structures following surgical disinsertion of the ciliary muscle in the cynomolgus monkey.

Light and electron microscopic studies were done on 11 cynomolgus monkey eyes which had undergone total iris removal followed by surgical disinsertion of the ciliary muscle from the scleral spur 4.7 to 14.4 months earlier. Anterior chamber perfusion to measure gross outflow facility had been performed one to nine times postoperatively. Over most of the circumference in most eyes (1) the ciliary muscle had been retrodisplaced from the scleral spur and had reattached to the sclera more posteriorly; (2) ciliary muscle, trabecular meshwork, and Schlemm's canal appeared normal. A cyclodialysis cleft was never seen. Fixation of some eyes in the in vivo and in vitro presence of pilocarpine demonstrated the contractibility of the retrodisplaced muscle. In isolated areas where the ciliary body had been surgically cut, scar tissue of varying thickness connected scleral spur, sclera, ciliary body, zonule, and lens capsule, but did not infiltrate trabecular meshwork or Schlemm's canal. In such sectors, plasma cell-like cells replaced trabecular endothelial cells and were also present in the scar tissue, ciliary muscle, and surrounding vessel walls in the scar and sclera. In sectors of two eyes, a previously existing trabecular operculum extended posteriorly and completely covered the meshwork. The meshwork in these sectors was poorly perfused by aqueous humor, and electron-dense deposits were present beneath the inner wall of Schlemm's canal. Four totally iridectomized and two unoperated eyes from these monkeys were also examined; ciliary muscle, trabecular meshwork, and Schlemm's canal appeared normal in all, despite the numerous anterior chamber perfusions.


Formation and drainage of aqueous humor following total iris removal and ciliary muscle disinsertion in the cynomolgus monkey.

Aqueous humor formation (AHF), uveoscleral flow (U), gross outflow facility (Cg), true facility of outflow from the anterior chamber (Ct1), true facility of outflow from anterior chamber into the general circulation (Ct2), and the pressure sensitivity of AHF (pseudofacility; Cps) were determined in cynomologus monkey eyes which had undergone total iris removal or iris removal followed by ciliary muscle disinsertion. AHF was in the normal range in iridectomized-only eyes. AHF was present but reduced in "disinserted" eyes. U was markedly reduced and similar in both types of eyes. Cps was normal and Cg, Ct1, and Ct2 were low in "disinserted" eyes. Cg, Ct1, and Ct2 were nearly equal in the "disinserted" eyes, indicating that gross facility consisted almost entirely of pressure-dependent flow from anterior chamber into the general circulation, presumably via the conventional drainage routes.


Aldose reductase and sorbitol dehydrogenase distribution in substructures of normal and diabetic rat lens.

Aldose reductase (AR) and sorbitol dehydrogenase (SDH) make up the sorbitol pathway, which has been implicated in the pathogenesis of sugar cataracts. The levels of the two enzymes were determined with quantitative histochemical techniques in the epithelium, cortex, and nucleus of normal and diabetic rat lenses. The ratio of activity of AR to SDH was found to be nearly 50 to 1 in all substructures. This would strongly favor sorbitol accumulation and subsequent cataract formation.


Regeneration of corneal epithelial basement membrane following thermal cauterization.

Thermal cauterization of the corneas of nine rhesus monkeys was performed by multiple thermokeratophore applications at temperatures of 90 degrees or 120 degrees C. Significant clinical observations included the resteepening of corneal curvature, delayed epithelial healing, stromal haze or scarring, and peripheral neovascularization. Reestablishment of tight adhesion of the regenerated epithelium to Bowman's layer required approximately 6 weeks. By transmission and scanning electron microscopy, the original basement membrane seemed relatively intact immediately following cauterization but disappeared within 1 week. The regeneration of new basement membrane complexes began at 1 week with the appearance of short discontinuous segments of basement material with hemidesmosomes and similarly required approximately 6 weeks for complete restoration.


Experimental immunogenic rubeosis iridis.

We  have developed a primate model of rubeosis iridis in monkeys systemically sensitized to crystalline beef insulin. After intravitreal insulin injection, the dose-related immunogenic inflammation includes cells, flare, fibrin, and blood in the anterior chamber. With more severe inflammation, posterior synechiae, iris bombé, and cataracts occur. Of particular importance, new blood vessels develop within the stroma and on the anterior surface of the iris. Following injection of small amounts of insulin, the anterior surface vessels may regress over time, and the iris regains its normal appearance and coloration. However, the new stromal vessels persist and are cuffed by inflammatory cells including plasma cells. After injection of large amounts of insulin, more extensive structural alterations develop as noted above in conjunction with persistent iris anterior surface and stromal neovascularization. The relationship of rubeosis iridis to clinical inflammatory syndromes and to previous laboratory studies is discussed. Stromal neovascularization was a consistent finding in this experimental model even when anterior surface vessels regressed. On the basis of these experimental data and a review of publications describing human pathology, we believe that a broadening of the classic definition of rubeosis iridis is waranted to include a recognition of the stromal component of the clinical and pathologic findings.


Paraureteral diverticula. Associated renal morphology and embryogenesis.

A study was made of paraureteric diverticula, the associated ureteric orifices, and the accompanying kidneys. It was shown that the kidney was more abnormal as the ureteric orifice position lay more laterally and into the diverticulum. The fascial arrangements of the terminal ureter and the related diverticulum were examined. When the ureter opened totally within the diverticulum, both structures showed a single investment of fascia, suggesting a common embryologic orgin of the two structures. When the ureter opened either separately into the bladder or on the verge of the diverticulum, the urter and the adjacent diverticulum each had separate fascial investment, suggesting a different etiology for the formation of this type of diverticulum. A total of 25 single-system specimens were employed for the above study. In the series there were an additional three duplex systems associated with paraureteric diverticula in shich the kidney morphology followed the above pattern but the fascial arrangements were different. Based on the observations of this investigation, it is suggested that the renal morphology is governed by the position of the ureteric orifice in the diverticulum and that the term vesical hiatal diverticula be used to denote those diverticula occurring in the vicinity of the ureteric orifice. It is further suggested that the diverticulum which engulfs the ureteric orifice be called paraureteral, and those with separate orifices in the bladder be termed periureteral to differentiate the embryologic from the pulsion etiologies.


[Etiology of perioral dermatitis].

The studies of Röckl und Schubert supported the theory that fusobacteria are often found in perioral dermatitis. When we characterize perioral dermatitis as fusobacteriosis, we do not have any doubt that fluorinated corticosteroids represent a very important pathogenetic factor. Corticoids enable opportune fusobacteria to become pathogenic. In our opinion without these bacteria the beginning of the perioral dermatitis seems impossible. Other supporting causes, which are relevant to sex distribution (oral contraceptives, moisturizing creams) are described.


[Histology of skin changes caused by external corticoid preparations].

Skin changes produced in 70 persons by prolonged topical application of corticosteroids were studied with light microscopy. Epidermal atrophy, alterations of the skin adnexa and changes in the dermal tissue are characterizing the so called corticosteroid-skin. It is suggested that the changes are dependent as well on duration and intensity of the corticosteroid treatment, as on individual factors.


Monoclonal cryoglobulinemia with macroglobulinemia in a dog.

A 9-year old female Doberman Pinscher had a 5-month history of epistaxis. On routine testing of the dog's serum, blockage of the intake tubes of the equipment resulted. At this point, cryoglobulinemia was suspected and was later confirmed to be due to a monoclonal cryoglobulin (IgM), by means of immunochemical studies. Clinical recovery was achieved by use of cytotoxic drugs.


Preparation and properties of derivatives of virginiamycin S.

Reduction of virginiamycin S with sodium borohydride produces allo- and normal-dihydro-virginiamycin S. Reduction of the tosylhydrazone of virginiamycin S with sodium cyanoborohydride affords deoxyvirginiamycin S. These compounds are less active than virginiamycin S. Like virginiamycin S they enhance the activity of virginiamycin M1.


Synthesis of cell envelope components by anucleate cells (minicells) of Bacillus subtilis.

Minicells produced by Bacillus subtilis CU403 (divIVB1) are capable of mucopeptide biosynthesis as shown by the incorporation of L-alanine, D-alanine, and N-acetylglucosamine into trichloroacetic acid-precipitable material, which can be degraded to trichloroacetic acid-soluble material by lysozyme digestion. Incorporation of the precursors is sensitive to vancomycin and D-cycloserine and insensitive to chloramphenicol. Penicillin inhibits the incorporation of D- and L-alanine N-acetylglucosamine at concentrations in excess of 10 mug of penicillin per ml; however, minicells are insensitive to penicillin-induced lysis. The material synthesized in minicells from N-acetylglucosamine is not subject to turnover during a subsequent 6-h incubation period. [2-3H]glycerol is converted to a cold trichloroacetic acid-precipitable form by minicells. This synthesis is not inhibited by vancomycin, penicillin, D-cycloserine, or chloramphenicol. Fractionation of the material synthesized from glycerol into hot trichloroacetic acid-soluble material and chloroform/methanol-extractable material indicates that minicells convert glycerol into teichoic acid and lipid.


Selective inhibition of Bacillus subtilis sporulation by acridine orange and promethazine.

Two structurally similar compounds were found to inhibit sporulation in Bacillus subtilis 168. A dye, acridine orange, and an antischizophrenic drug, promethazine, blocked spore formation at concentrations subinhibitory to vegetative growth, while allowing synthesis of serine protease, antibiotic, and certain catabolite-repressed enzymes. The sporulation process was sensitive to promethazine through T2, whereas acridine orange was inhibitory until T4. The drug-treated cells were able to support the replication of phages phie and phi29, although the lytic cycles were altered slightly. The selective inhibition of sporulation by these compounds may be related to the affinity of some sporulation-specific genes to intercalating compounds.


Characterization of a translocation unit encoding resistance to mercuric ions that occurs on a nonconjugative plasmid in Pseudomonas aeruginosa.

The nonconjugative plasmid, pVS1, has a molecular weight of 18.5 X 10(6) and confers resistance to sulfonamides and to mercuric ions. In Pseudomonas aeruginosa PAO, the transfer can be mobilized by a variety of conjugative plasmids, and the process does not require a functional recombination system in the donor. Hybrid plasmids that arise by the relocation of the mer gene onto the mobilizing plasmid can be isolated readily, and, as far as can be determined, these hybrids retain the genome of the conjugative plasmid in toto. The relocation of mer occurs by a Rec-independent process and leads to a constant increase (about 6 X 10(6) daltons) in the size of the recipient plasmid. This suggests that the mer gene in pVS1 is located on a translocation unit, designated Tn501, of a molecular weight of about 6 X 10(6). The translocation of Tn501 into RP1 is not usually associated with the loss of any known plasmid-mediated function, but transfer-defective or tetracycline-sensitive derivatives do occur at frequencies of about 4%, whereas carbenicillin-sensitive or kanamycin-sensitive variants arise with a frequency of about 0.2% each. It seems therefore that the integration of Tn501 can occur at any one of a minimum of five sites in RP1.


Initiation of Bacillus spore germination by hydrostatic pressure: effect of temperature.

Suspensions of Bacillus cereus T, B. subtilis, and B. pumilus spores in water or potassium phosphate buffer were germinated by hydrostatic pressures of between 325 and 975 atm. Kinetics of germination at temperatures within the range of 25 to 44 degrees C were determined, and thermodynamic parameters were calculated. The optimum temperature for germination was dependent on pressure, species, suspending medium, and storage time after heat activation. Germination rates increased significantly with small increments of pressure, as indicated by high negative deltaV values of -230 +/- 5 cm3/mol for buffered B. subtilis (500 to 700 atm) and B. pumilus (500 atm) spores and -254 +/- 18 cm3/mol for aqueous B. subtilis (400 to 550 atm) spores at 40 degrees C and -612 +/- 41 cm3/mol for B. cereus (500 to 700 atm) spores at 25 degrees C. The ranges of thermodynamic constants calculated at 40 degrees C for buffered B. pumilus and B. subtilis spores at 500 and 600 atm and for aqueous B. subtilis spores at 500 atm were: Ea = 181,000 to 267,000 J/mol; deltaH = 178,000 to 264,000 J/mol; deltaG = 94,000 to 98,300 J/mol; deltaS = 264 to 544 J/mol per degree K. These values are consistent with the concept that the transformation of a dormant to a germinating spore induced by hydrostatic pressure involves either hydration or a reduction in the visocosity of the spore core and a conformational change of an enzyme.


Transitory germinative excision repair in Bacillus subtilis.

Bacillus subtilis strains UVSSP-42-1 (hcr42 ssp1) and UVSSP-1-1 (hcr1 ssp1) are ultraviolet (UV) radiation sensitive both as dormant spores and as vegetative cells. These strains are unable to excise cyclobutane-type dimers from the deoxyribonucleic acid (DNA) of irradiated vegetative cells and fail to remove spore photoproduct from the DNA of irradiated spores either by excision (controlled by gene hcr) or by spore repair (controlled by gene ssp1). When irradiated soon after spore germination, these strains excise dimers, but not spore photoproduct, from their DNA. This process, termed germinative excision repair, functions only transiently in the germination phase and is responsible for the high UV resistance of germinated spores and for their temporary capacity to host cell reactivate irradiated phages infecting them. The recA1 mutation confers higher UV sensitivity to the germinated spores, but does not interfere with dimer removal by germinative excision repair.


Characterization of a lysine-specific active transport system in Rickettsia prowazeki.

Rickettsia prowazeki possesses an active transport system for lysine with a Kt of influx of 1 muM. Extraction and chromatographic analysis of the accumulated labeled material show the material to be lysine rather than a derivative. This intracellular lysine pool can be exchanged with external unlabeled substrates for at least 10 min; The lysine analogues L-aminoethyl cysteine, N-methyl lysine, hydroxylysine, and D-lysine competitively inhibit uptake of L-lysine, but cadaverine, diaminopimelate, arginine, ornithine, and epsilon-aminocaproate do not. Accumulation of lysine can be inhibited by the energy poisons potassium cyanide, triphenylmethyl phosphonium bromide, and 2,4-dinitrophenol. The effect of potassium cyanide, but not 2,4-dinitrophenol or triphenylmethyl phosphonium bromide, can be overcome by adenosine 5'-triphosphate. Both energy-dependent influx and energy-independent efflux are inhibited by the sulfhydryl reagents N-ethyl maleimide and p-chloromercuriphenyl sulfonic acid.


Sodium effect of growth on aspartate and genetic analysis of a Bacillus subtilis mutant with high aspartase activity.

Most strains of Bacillus subtilis, dervied from the 168 (Marburg) strain, grow slowly on aspartate as sole carbon source. We isolated a mutant (aspH) that grows rapidly on aspartate because it produces aspartase constitutively. Thus, aspartase is needed for rapid growth on aspartate, whereas aspartate-alpha-ketoglutarate aminotransferase is not needed, as was demonstrated by a mutant lacking that enzyme activity. By two--and three-factor crosses using PBSl transduction, the aspH mutation was located between the aroD and the lys markers of the genetic map. Although sodium ions do not affect growth on glucose or L-malate, they specifically stimulate growth on aspartate in both the parent and the aspH mutant strains. Enzyme activities of crude aspartase and fumarase and of purified aspartase do not increase in the presence of sodium. These results show that stimulation by sodium involves some reaction other than the enzymes catabolizing aspartate. The ease of purification from the aspH strain and the stability of aspartase suggest that the B. subtilis enzyme is particularly useful for aspartate determinations.


Immunochemical comparison of phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase among the Enterobacteriaceae.

The bifunctional enzyme of the tryptophan operon, phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase (PRAI-InGPS;EC 4.1.1.48), was characterized by an immunochemical study of six representative members of the Enterobacteriaceae: Escherichia coli, Salmonella typhimurium, Enterobacter aerogenes, Serratia marcescens, Erwinia carotovora, and Proteus vulgaris. PRAI-InGPS was purified from E. coli, and antisera were prepared in rabbits. These antisera were utilized in quantitative microcomplement fixation allowing for a comparison of the overall antigenic surface structure of the various homologous enzymes. These data showed E. coli PRAI-InGPS and S. marcescens and E. carotovora PRAI-InGPS (taken as a group) to have an index of dissimilarity of approximately 10, whereas the other organisms had values intermediate. In addition, antiserum to E. coli tryptophan synthetase beta2 subunit was used in microcomplement fixation to extend the previous comparison of this subunit (Rocha, Crawford, and Mills, 1972) to E. carotovora and P. vulgaris. Indexes of dissimilarity for E. coli compared to P. vulgaris of E. carotovora were 1.0 and 1.7, respectively. Agar immunodiffusion using PRAI-Ingps antisera showed significant cross-reaction among E. coli, E. aerogenes, S. typhimurium, and P. vulgaris whereas the enzymes from S. marcescens and E. carotovora cross-reacted to a lesser extent, with the latter reaction being quite weak. Comparative enzyme neutralization using E. coli PRAI-InGPS antisera showed significant cross-reactions among the enzymes in that all were neutralized at least 25%. The data taken together indicate that the trpC gene products in the Enterobacteriaceae are a homologous group of proteins, that the genetic divergene of the trpC gene is basically the same as the trpA gene, and that both are less conserved than the trpB gene. Furthermore, the PRAI-InGPS, enzyme active site appears to represent a more evolutionarily conserved region of the protein. These findings indicate that, with respect to PRAI-InGPS, similarity to E. coli among the organisms examined is in the following order: (E. aerogenes, S. typhimurium, P. vulgaris) greater than (S. marcescens, E. carotovora).


Isolation and characterization of four plasmids from Bacillus subtilis.

Nineteen Bacillus subtilis isolates obtained from type culture collections were examined for the presence of covalently closed circular duplex deoxyribonucleic acid molecules by the technique of cesium chloride-ethidium bromide density gradient centrifugation. Four of the 19 strains tested carried covalently closed circular molecules. Two of these strains (IFO3022, IFO3215) harbored a similar plasmid with a molecular weight of 5.4 X 10(6). The other two strains (IAM1232, IAM1261) carried 4.9 C 10(6)-and 5.3 X 10(6)-dalton plasmids, respectively. These plasmid-harboring strains did not show phenotypic traits such as antibiotic resistance orbacteriocin production. The plasmid deoxyribonucleic acids were digested by three restriction endonucleases, EcoRI, HindIII, and BamNI, and were classified into three different types from their electrophoretic patterns in agarose gels.


Isolation and preliminary characterization of auxotrophs of a filamentous Cyanobacterium.

Auxotrophic mutants of the filamentous cyanobacterium Anabaena variabilis were isolated by a method in which, after mutagenesis and before penicllin enrichment, mutant and wild-type cells were separated by cavitation. Auxotrophs were identified by their inability to grow on minimal medium, and they were partially characterized by replica plating to media supplemented with single nutrients or specific groups of nutrients. Of the 83 auxotrophs isolated, 65 required an inorganic source of nitrogen for growth. In addition, auxotrophs were isolated that required methionine (six), uracil (two), adenine (one), biotin (two), and nicotinic acid (two). (The number of isolates of each type is indicated in parentheses.) The nutrient requirements of five auxotrophs appeared complex and were not determined. A large proportion of the mutants requiring inorgainic fixed nitrogen was altered in the differentiation of heterocysts. The following morphological aberrancies were observed: abnormally high and abnormally low frequencies of heterocysts; thick, uneven heterocyst envelopes; incompletely developed pore regions; very distinct pore regions; and protoplasts separated from the envelope of the heterocyst. Spontaneously occurring, N2-fixing, prototrophic revertants of mutants with aberrant heterocysts have been isolated at a frequency of 2 X 10(-8) to 4 X 10(-8) of the cells plated. That most such revertants produced morphologically normal heterocysts is consisten with the idea that heterocysts play an essential role in aerobic N2 fixation.


Altered outer membrane protein in different colonial types of Neisseria gonorrhoeae.

Dark-colored colony types of Neisseria gonorrhoeae (T3 and dark variants of T1 and T2) had markedly increased amounts of an approximately 28,000-dalton outer membrane protein, as compared with light-colored colony types (T4 and light variants of T1 and T2). The presence of this protein appeared to be unrelated to piliation. The apparent molecular weight of this protein on sodium dodecyl sulfate-polyacrylamide gels varied, depending on methods used to solubilize envelope proteins. In view of the location of this protein on the outer membrane, this protein could be important to the pathogenicity or antigenicity of the organism as well as to colonial characteristics in vitro.


Inhibition of iron uptake and deoxyribonucleic acid synthesis by Desferal in a mutant strain of Bacillus subtilis.

In the Bacillus subtilis mutant 1D-4, the hydroxamate Desferal inhibited growth, iron uptake, and deoxyribonucleic acid synthesis but did not quantitatively affect synthesis of ribonucleic acid and protein.


Properties of derivatives of the Pseudomonas plasmid pVS1 that have inherited carbenicillin resistance from RP1.

A procedure is described for the isolation, in Pseudomonas aeruginosa PAO, of bacteria carrying derivatives of pVS1 that inherited the carbenicillin-resistance determinant from RP1 either alone or together with that for aeruginocin resistance. Such bacteria occur among the transconjugant progeny from both recombination-proficient or -deficient pVS1+ RP1+ donors, suggesting that the formation of these plasmids is due to the translocation of TnA from RP1 into pVS1. It is possible, therefore, that the aeruginocin-resistance determinant is part of TnA or is closely linked to it. Unexpectedly, none of these plasmids showed the 3 x 10(6)- to 4 x 10(6)-dalton increase in size predicted for TnA+ derivatives of PVS1. It is suggested that an interaction between TnA and the Tn501 translocation unit in pVS1 could account for this result.


A fluorescent probe response to the interaction of pyocin R1 with sensitive cells.

Additon of pyocin R1, a bacteriocin of Pseudomonas aeruginosa, to sensitive cells caused a fluorescence increase of 8-anilino-1-naphthalenesulfonate (ANS) in the cell suspension. The reaction was rapid, starting with a short time lag after adsorption of pyocin onto the cells and finishing within several minutes. The fluorescence response was attributed to the interaction of the cell body and ANS, not to that of the medium outside the cells and ANS. The maximal amplitude of fluorescence after pyocin addition was dependent on temperature, and the relation appeared to be biphasic. Similarly, Arrhenius plots of the initial rate of fluorescence change were biphasic. The transition of slopes in both cases occurred in the temperature range between 18 and 19 degrees. These results suggest that ANS interacts with lipids in the cell envelope and that pyocin causes a structural change of the cell envelope leading to increased fluorescence of ANS.


[Studies on the facial muscles in the aged].

The dissection of the facial musculature in the elderly made on 2 bodies, confirmed the mode of evolution noted earlier in the Primates. The reduction in the m. platysma myoides was confirmed; this muscle showed only two parts: a rudimentary pars buccalis and a relatively developed pars mentalis. This reduction was accompanied by fundamental transformations since the m. quadratus of the chin appeared at the anterior part of the pars mentalis of the platysma myoides. The derivatives of the deep m. sphincter coli showed an arrangement very similar to that of the Simians. However some modifications already observed became stronger, particularly the verticilisation of the m. caninus and the separation of the two m. zygomaticus. Curiously, the extrinsic ear musculature persisted and remained well developed despite its absence of activity.


Dependence of ionophore- and caffeine-induced calcium release from sarcoplasmic reticulum vesicles on external and internal calcium ion concentrations.

The effects of the ionophore, X537A, and caffeine on ATP-dependent calcium transport by fragmented sarcoplasmic reticulum were studied in the absence (calcium storage) or presence (calcium uptake) of calcium-precipitating anions. The ionophore caused rapid calcium release after calcium storage, the final level of calcium storage being the same whether a given concentration of X537A was added prior to initiation of the reaction or after calcium storage had reached a steady state. Although 10 to 12 muM X537A caused approximately 90% inhibition of oxalate-supported calcium uptake when added prior to the start of the reaction, this ionophore concentration caused only a small calcium release when added after a calcium oxalate precipitate had formed within the vesicles, and only slight inhibition of calcium uptake velocity when added during the calcium uptake reaction. When low initial calcium loads limited calcium uptake to 0.4 mumol of calcium/mg of protein, subsequent calcium additions in the absence of the ionophore led to renewed calcium uptake. Uptake of the subsequent calcium additions was not significantly inhibited by 10 to 12 muM X537A. These phenomena are most readily understood in terms of constraints imposed by fixed Cai (calcium ion concentration inside the vesicles) on the pump-leak situation in sarcoplasmic reticulum vesicles containing a large amount of an insoluble calcium precipitate, where most of the calcium is within the vesicles and Cai is maintained at a relatively low level. These constraints restrict calcium loss after calcium permeability is increased because calcium release can end when the calcium pump is stimulated by the increased Cao (calcium concentration outside the vesicles) so as to compensate for the increased efflux rate. In contrast, an increased permeability in vesicles that have stored calcium in the absence of a calcium-precipitating ion causes a much larger portion of the internal calcium store to be released. Under these conditions calcium storage capacity is low so that release of stored calcium is less able to raise Cao to levels where the calcium pump can compensate for the increased efflux rate. The constraints imposed by anion-supported calcium uptake explain the finding that more calcium is released by X537A or caffeine when these agents are added at higher levels of Cao, and that more calcium leaves the vesicles in response to a given increase in calcium permeability at higher Cai. Although such calcium release is amplified by increased Cao, the amplification is attributable to the constraints described above and does not represent a "calcium-triggered calcium release."


Dependence of calcium permeability of sarcoplasmic reticulum vesicles on external and internal calcium ion concentrations.

The ability of sarcoplasmic reticulum vesicles to retain calcium following ATP-supported calcium uptake in the presence of the calcium-precipitating anions oxalate and phosphate depends on Cao (calcium ion concentration outside the vesicles) and Cai (calcium ion concentration within the vesicles). Calcium efflux rates at any level of Cai are accelerated when Cao is increased. Higher Cao at the time that calcium uptake reactions reach steady state is associated with a spontaneous calcium release that reflects this effect of increased Cao. Increasing Cai at any level of Cao causes little or no acceleration of calcium efflux rate so that calcium permeability coefficients, estimated by dividing calcium efflux rates by Cai, the "driving force", are inversely proportional to Cai. Calcium permability coefficients thus correlate, as a first approximation, with the ratio Cai/Cao, decreasing 1000-fold as this ratio increases over a 3000-fold range (Cao = 0.1 to 3.3 muM, Cai =4 to 750 muM). Oscillations in both the calcium content of the vesicles and Cao are seen as calcium uptake reactions approach steady state, suggesting that calcium permeability undergoes time-dependent variations. Sudden reduction of Cao to levels that markedly inhibit calcium influx via the calcium pump unmasks a calcium efflux that decreases slowly over 60 to 90 s. The maximal calcium permeability observed in the present study would allow the calcium efflux rate from the sarcoplasmic reticulum at a Cai of 100 muM to be approximately 10(-10) mol/cm2/s, which is about 1 order of magnitude less than that estimated for the sarcoplasmic reticulum of activated skeletal muscle in vivo. The release of most of the stored calcium in some experiments indicates that the observed permeability changes can occur over a large portion of the surface of the sarcoplasmic reticulum.


Tonofilament protein from newborn rat epidermis. Isolation, localization, and biosynthesis of marker of epidermal differentiation.

A novel extraction procedure, previously used on the cell walls of dermatophytes, has been applied to the epidermis of newborn rat. A leucine-rich fraction was isolated which contained over 60% of the total epidermal radioactivity from [3H]leucine in 15 to 20% of the total protein. This fraction was further purified by chromatography in DEAE-cellulose and Sephadex G-200. The protein with the highest specific activity from [3H]leucine was isolated and gave a single band in sodium dodecyl sulfate polyacrylamide gels of molecular weight = 58,000. Antibody to this protein gave a single precipitin band by immunodiffusion and immunoelectrophoresis in agar with the purified protein. This antibody ultrastructurally immunolocalized specifically over tonofilaments in all layers of the epidermis, but showed no reaction in the dermis. The synthesis of this protein in vitro was inhibited by puromycin but not by actinomycin D, suggesting ribosomal synthesis involving a relatively long lived messenger.


Glycoprotein biosynthesis in myeloma cells. Characterization on nonglycosylated immunoglobulin light chain secreted in presence of 2-deoxy-D-glucose.

The biosynthesis and secretion of a glycosylated, K-type immunoglobulin light chain (K-46) was studied in a mouse myeloma tumor, mineral oil plasmacytoma-46B. Viable single cell suspensions were prepared from excised tumors and optimal conditions were established for incorporation of amino acid and carbohydrate precursors into the protein synthesized and secreted by the cells. The glucose analog, 2-deoxy-D-glucose, was utilized as an inhibitor of glycosylation to determine the role of glycosylation in the biosynthesis, intracellular transport, and export of the protein from the cell. It was determined that 6 mM 2-deoxyglucose prevents the incorporation of glucosamine, mannose, and galactose into secreted protein, but permits the incorporation of leucine at approximately 40% of control values. The nonglycosylated protein, secreted in the presence of 2-deoxyglucose, was characterized as a nonglycosylated form of K-46 light chain by the following criteria: (a) electrophoresis in polyacrylamide gels containing sodium dodecyl sulfate, (b) reactivity of the nonglycosylated protein with antisera prepared against native, fully glycosylated, K-46 light chain, (c) analysis of the protein by gel filtration techniques, (d) behavior of the protein on lectin-derivatized Sepharose, and (e) analysis of tryptic peptides derived from the protein. We have concluded that 2-deoxyglucose-inhibited cells synthesize and secrete the normal polypeptide chain of K-46 devoid of its carbohydrate side chain indicating that glycosylation is not an essential step in the biosynthesis, intracellular transport, or export of this protein that is normally synthesized and secreted in a glycosylated form. Under conditions of 2-deoxyglucose inhibition, the nonglycosylated form of K-46 light chain constitutes a significantly greater proportion of accumulated intracellular protein, suggesting that the biosynthesis of the polypeptide chain of K-46 light chain proceeds at a nearly normal rate, but that the absence of the carbohydrate side chain of the protein retards, but does not prevent, the intracellular transport of the protein and its export from the tumor cell.


Fatigue characteristics of posterior elements of vertebrae.

The fatigue strength of vertebral posterior elements was determined by applying repetitive forces to the inferior joints between the articular processes of human rhesus-monkey vertebrae. The results indicate that in vivo stresses resulting from unusual activities for sustained periods can induce fatigue fracture of the lumbar vertebrae. Comparison of these data with those of other investigators suggests the use of ultimate strength as a normalizing factor to correlate fatigue-strength data obtained under different test conditions.


The key role of the lateral malleolus in displaced fractures of the ankle.

The reason why late degenerative arthritis developed in some patients who had sustained displaced bimalleolar fractures of the ankle was investigated. The roentgenograms indicated that incomplete reduction of the lateral malleolus and a residual talar tilt were present. When bimalleolar fractures were created in cadavera the talus could be anatomically repositioned only when the lateral malleolus was accurately reduced. Fifty-three patients with bimalleolar fractures were treated by anatomically fixing the lateral malleolus with a four-hole plate. There was an anatomical reduction of the talus and medial malleolus in each instance and there were no late cases of degenerative arthritis when these patients were followed for from six months to nine years. We concluded that the lateral malleolus is the key to the anatomical reduction of bimalleolar fractures, because the displacement of the talus faithfully followed that of the lateral malleolus.


Dysplasia of the odontoid process in Morquio's syndrome causing quadriparesis.

Eleven patients with documented Morquio's syndrome who had dysplasia of the odontoid process and resulting atlanto-axial instability were reviewed. They were found to be at risk for acute traumatic quadriparesis, chronic myelopathy of a variable and often rapid rate of progression, and sudden death by respiratory arrest. The evidence suggests that early prophylactic posterior cervical fusion is indicated for patients with this malformation and that once quadriparesis is established, recovery of function is limited.


Ectopic ossification after total hip arthroplasty. Predisposing factors, frequency, and effect on results.

In a statistical analysis of 398 consecutive patients with 507 total hip arthroplasties which was designed to identify factors predisposing to ectopic ossification and to determine the frequency of ossification and its effect on the results, it was found that male patients with considerable bilateral osteophytic osteoarthritis were statistically most likely to have ectopic ossification, especially if ossification had existed before arthroplasty consequent to previous surgery. Ectopic ossification which was first noted six weeks after total hip arthroplasty in 96 per cent of the cases did not change in amount thereafter, though the bone did mature. Both the range of motion of the hip and the function of the patient were affected by the ectopic bone during the first postoperative year, but after that only the range of hip motion was influenced.


Increase in plasma growth hormone levels following thyrotropin-releasing hormone injection in children with primary hypothyroidism.

Thyrotropin-releasing hormone (TRH) induced a significant increase in plasma growth hormone (GH) levels in 4 of 8 children with primary hypothyroidism, while a slight decrease was observed in 8 control children. Base-line plasma prolactin (PRL) levels and peak responses to TRH were higher in hypothyroid children than in controls. These data may indicate the existence of dysfunction of central nervous system mechanisms of control of GH and PRL secretion in subjects with primary hypothyroidism.


Complement-fixing antibody response to rotavirus infection.

A human rotavirus complement-fixing (CF) antigen, prepared by purification of large volumes of fluid feces collected from children with winter diarrhea, was used to study the development and persistence of antibody in children with diarrhea and the prevalence of rotavirus antibody in Melbourne. In children with diarrhea, antibody rises were detectable within 4 to 6 weeks of the onset of illness, and the titers usually remained elevated for the next 1 to 2 years. CF antibody did not develop in two children with proven rotavirus infection aged less than 6 months, an age at which poor CF responses to other viruses have also been observed. A study of CF antibody levels in the general community showed that in Melbourne, most children have been infected with human rotavirus by the age of 3 years.


Field evaluation of New York City medium in the biological environment-CO2 chamber in recovery of Neisseria gonorrhoeae and urogenital mycoplasmas.

The biological environment-CO2 chamber containing New York City (NYC) medium was evaluated in a clinical field trial in comparison with Transgrow and NYC medium in plates for the recovery of Neisseria gonorrhoeae and with NYC medium for urogenital mycoplasmas. The results of the study indicated that the biological environment chamber-CO2 culturing system with NYC medium is an effective method for the handling, transport, and culture of N. gonorrhoeae, large-colony mycoplasmas, and T-mycoplasmas if the delay in transport and incubation does not exceed 24 h.