Clinical response to CB-154 and the pituitary response to thyrotropin-releasing hormone-gonadotropin-releasing hormone in patients with galactorrhea-amenorrhea.

Ten patients with galactorrhea and amenorrhea were treated with 2-bromo-alpha-ergocryptine (CB-154). All patients had normal anteroposterior and lateral x-rays of the sella turcica and normal or low gonadotropin levels. Before treatment, serum prolactin (PRL) levels were between 80 and 1575 ng/ml. Prior to initiating therapy, six patients were further evaluated by the intravenous administration of thyrotropin-releasing of a pituitary etiology in all patients. During treatment, PRL levels were measured at monthly intervals. After 1 month, serum PRL concentrations were reduced between 13% and 99%. In eight subjects there was complete cessation of galactorrhea. During treatment, nine patients resumed ovulatory menstrual cycles and three patients conceived. After discontinuing therapy, five of seven subjects had a recurrence of galactorrhea, amenorrhea, and hyperprolactinemia.


The risks and prevention of Neisseria gonorrhoeae transfer in fresh ejaculate donor insemination.

The risk of transferring gonorrhea with donor insemination was investigated in this study. Thirteen semen specimens contaminated by Neisseria gonorrhoeae were placed in containers as used in donor artificial insemination (AID) and cultured serially. Since most fresh ejaculates are used rapidly, a 2-hour peroid was used as the end-point. Ten of the thirteen ejaculates were positive on initial and 2-hour delay cultures. Three were negative by both cultures. The epidemiology of gonorrhea is reviewed, and those cases of gonorrhea reported following AID are discussed. The use of frozen semen as advocated by some is compared with the use of fresh ejaculates. It is shown that fresh ejaculates, which are more practical for many physicians and have better fertilizing capacity, can be used if proper cultures are obtained at the time of insemination. It is suggested that either frozen ejaculates with negative culture or fresh ejaculates screened by smear and cultured at the time of insemination be utilized. The use of fresh semen is possible, since results of appropriate cultures could be available and treatment instituted before clinical disease occurs.


Vasectomy and vasovasostomy in rhesus monkeys: the effect of circulating antisperm antibodies on fertility.

Rhesus monkeys develop agglutinating and complement-dependent antisperm antibodies after vasectomy. In order to study whether these antibodies affect fertility after vasovasostomy, 15 animals were given vasectomies and 6 months later vasovasostomies. Subsequently, each was mated with females of proven fertility. Five controls were given sham operations and similarly treated. During this period, each aimal was bled for serum to monitor the humoral immune response, ejaculated for semen analyses, and palpated for granuloma or fistula development. All control animals had a transient decrease in sperm density after sham vasectomy and vasovasostomy operations. The surgical procedures of vasectomy and subsequent vasovasostomy resulted in more animals having sperm of poor motility and quality. All of vasovasostomies were surgically successful in that sperm were again present in the ejaculate of each animal. The amount of sperm in the ejaculate could not be correlated with the ease of surgical procedure, presence or absence of macrophages in the ejaculum, motility, or forward progression. Only animals that had been vasectomized developed circulating antisperm antibodies. Sustained, elevated levels of antisperm antibodies most commonly occurred in monkeys that had high initial total sperm counts. Six of the experimental animals retained high levels of sperm-immobilizing antibodies after vasovasostomy. Of these, two were found to be infertile and two were classed as subfertile. Of the nine experimental animals without sustained antisperm antibody production, only one was classed as subfertile. This suggests that antisperm antibodies may in some cases impair the restoration of fertility after vasovasostomy.


Lactic acidosis complicating diabetic ketosis in a patient with hyperthyroidism.

The present report describes a patient with insulin-dependent diabetes who developed simultaneously lactic acidosis and ketoacidosis following insulin deprivation. Administration of insulin at low doses rapidly corrected both ketosis and lactic acidosis. There had been neither circulatory collapse, nor phenformin intake, and hepatic function was normal. The development of lactic acidosis in this case was possibly precipitated by hyperthyroidism. A review of the literature indicates that lactic acidosis is a very rare complication of diabetic ketosis per se.


Ethology and man: science or myth?

What is known of human evolution gives us little help in describing the biological nature of man: even our fossil history is obscure, and most statements on the evolution of human behaviour are guesses. The fact that primitive man was a predator on other species does not signify that man is "naturally aggressive" to his own kind. The notion of an inherent drive to aggression has no scientific foundation. Knowledge of the conduct of other species can lead to no valid conclusions about human behaviour. The same limitations apply to interpretations of modern man based on what is known of human hunter-gatherers. Ethology can contribute to human studies (1) by providing methods of observing and analysing behaviour, and (2) by providing hypotheses that can be tested. Zoologically-based hypotheses on the ill effects of crowding have been useful but have proved to be wrong. Others on the effects of stimulation in early life, and on breast-feeding and milk composition, have been more fruitful. Abnormal conduct, such as that of Kanner's syndrome, can be usefully studied by ethological methods. Man is a learner and a teacher, whose knowledge of himself increases slowly with the growth of critical research.


Accumulation of dopamine in mouse pancreatic B-cells following injection of L-DOPA. Localization to secretory granules and inhibition of insulin secretion.

Accumulation and subcellular localization of dopamine (DA) in pancreatic B-cells and its effects on insulin secretion were investigated in mice following a single injection of L-3,4-dihydroxyphenylalanine (L-DOPA). Electron microscopic autoradiography showed that 3H-DA formed from administered 3H-DOPA was present over B-cells as well as over other types of islet cells. Pretreatment of the animals with a decarboxylase inhibitor greatly reduced the number of autoradiographic grains. In the B-cells the 3H-DA-grains were associated with the secretory granules. The location of the label may suggest an incorporation in the periphery of the beta-granule, rather than in the dense core, supposed to contain insulin. Accumulation of DA in the B-cells following L-DOPA administration was found to inhibit partially the insulin secretory response to different insulin secretagogues (glucose, glibenclamide and L-isopropylnoradrenaline (L-IPNA). Treatment with monoamine oxidase inhibitor + L-DOPA induced an almost total suppression of L-IPNA-stimulated insulin secretion, whereas glucose-induced insulin release was still only partially inhibited. Pretreatment with a decarboxylase inhibitor abolished the effects of L-DOPA. It is suggested that intracellularly accumulated DA in the B-cell exerts an inhibitory action on insulin releasing mechanisms induced by different secretagogues and that this action might involve interference with a calcium translocation process at the level of the secretory granule.


Comparative effects of phenformin, metformin and glibenclamide on metabolic rhythms in maturity-onset diabetics.

Twelve hour metabolic rhythms have been performed on six maturity-onset diabetic subjects during successive periods of therapy with phenformin, metformin, and glibenclamide. Moderate control of blood glucose concentration was achieved with phenformin and  metformin, the lowest concentrations being found with glibenclamide. Mean blood lactate concentration was grossly elevated during phenformin therapy, moderately elevated with metformin and normal during glibenclamide treatment. Similar patterns were found for the lactate/pyruvate ratio, alanine, glycerol and ketone bodies. Serum triglyceride concentrations were significantly higher during phenformin treatment than with the other two regimes. Serum insulin concentration was higher on glibenclamide than with either biguanide. Most of these effects of the biguanides could be accounted for by an inhibitory effect on hepatic gluconeogenesis. It is concluded that the use of biguanides as hypoglycaemic agents in diabetes is associated with the production of multiple metabolic abnormalities.


Extrapancreatic effect of glibenclamide: stimulation of duodenal insulin-releasing activity (DIRA) in man.

Experiments were performed in order to study a possible participation of gastrointestinal factors in the insulinotrophic action of glibenclamide in man. Six healthy volunteers received 5 mg glibenclamide in 50 ml saline orally. Biopsies were taken from the duodenal mucosa before and after administration of the drug. The duodenal insulin-releasing activity (DIRA) was assayed in the extracts of the biopsy material by using an in situ pancreas preparation of rat. The corresponding drug, IRI and glucose levels were measured in peripheral blood. The values of IRI correlated with both the prior elevation of DIRA and the increasing levels of the drug in the blood. These data indicate that glibenclamide might stimulate the release of gut factor(s) which, in turn, could possibly sensitize the pancrease response to the drug.


[An evaluation of ventricular function in coronary artery disease patients using the isometric (handgrip) test (author's transl)].

Left ventricular contractile function was evaluated in twenty patients with coronary artery disease before and during handgrip by use of Vpm, VCEIO and Vmax (obtained with a simplified calculation) from high-fidelity pressure measurements. In eight patients the response to handgrip was characterized by an increase of the contractile indices accompanied by small changes of left ventricular end-diastolic pressure (not exceeding + 4 mm Hg) and was considered normal. Twelve patients who showed abnormal or pathologic reaction to isometric exercise repeated handgrip after administration of 0.4 mg of spray nitroglycerin. Thus eight patients normalized their response to exercise, while in four we observed again a pathologic reaction to handgrip. It is concluded that is a useful and simple manoeuvre for a better evaluation of left centricular function in patients with coronary artery disease; the repetition of the test after administration of nitroglycerin can be useful in identifying those patients with greatest contractile reserve.


Drug metabolism in adult rat hepatocytes in primary monolayer culture.

Primary monolayer culture of adult rat hepatocytes represents a novel and potentially useful technique to study many aspects of hepatic physiology for extended periods of time in vitro (J Cell Biol 59:722-734, 1973). In examining the hepatic drug-metabolizing system in these cells, we have discovered that the conditions of cell culture exert rapid, selective, and reproducible changes in microsomal enzymes. In the 24- to 48-hr period immediately following preparation and culture of the isolated parenchymal cells, the level of the drug-binding microsomal hemoprotein, cytochrome P-450, measured in extracts of cell homogenates, declined to less than 20% of its in vivo level, whereas NADPH-cytochrome c reductase activity was only moderately reduced and glucose-6-phosphatase activity remained unchanged. The activity of aminopyrine-N-demethylase and aniline hydroxylase also fell, paralleling the level of cytochrome P-450. By contrast, p-nitroanisole O-demethylase activity was unchanged in the cultured hepatocytes despite evidence (type I binding spectrum, NADPH requirement, inhibition by carbon monoxide or by SKF 525A) that p-nitroanisole O-demethylase is a cytochrome P-450-dependent enzyme. In culture, as in vivo, aromatic polycyclic hydrocarbons stimulated p-nitroanisole O-demethylase and aryl hydrocarbon (benzo [a] pyrene) hydroxylase activities; however, this effect was unaccompanied by a detectable increase in total carbon monoxide-binding hemoprotein. The data indicate that the profile of microsomal oxidase enzymes and their control undergo striking changes as hepatocytes adapt to cell culture.


Treatment of Crohn's disease with home parenteral nutrition.

Home parenteral nutrition was used to treat 9 patients with severe Crohn's enterocolitis. Seven patients had a short bowel syndrome after multiple resections of bowel. In 2 patients home parenteral nutrition was used as primary therapy. The treatment was well tolerated and proved successful in 8 of 9 patients. Three patients have been able to discontinue parenteral infusions and currently are in remission. The main complications were associated with the access device which was replaced in 3 patients. Five patients currently have abnormal liver function tests without progressive liver disease. It is concluded that home parenteral nutrition is an important new therapeutic modality which can reduce or even eliminate the need for repeated or prolonged hospitalization of patients with short bowel syndrome complicating severe Crohn's disease. In addition, the technique of home parenteral nutrition, because it is relatively simple and safe, lends itself to early intervention in severe fulminant cases of Crohn's disease. This approach can result in healing of fistulae and abscesses and greatly shortening the period of hospitalization. The patient is more rapidly rehabilitated, and his fear of early relapse and recurrent malnutrition is minimized, thus facilitating a prolonged period of bowel rest which can lead to eventual remission. Home parenteral nutrition should be kept in mind as a possible alternative to early surgical intervention in selected cases of severe Crohn's disease.


[Antibacterial and antimycotic activity of the amniotic fluid against selected germs (author's transl)].

60 samples of amniotic fluid from 60 patients were obtained between 14 and 42 weeks gestation by dates and tested for their antibacterial activity against staphlococcus aureus strains and brucella abortus and their antimycotic activity against Candida albicans. The antibacterial and antimycotic activity of the amniotic fluid was confirmed. This activity increases steadily from the 15th week and reaches its maximum around term between 36 and 42 weeks. The amniotic samples obtained at 20 weeks gestation showed a weak antibacterial activity especially against staphlococcus aureus of human origin. The above results are important because of the increasing frequency of early and late diagnostic amniocentesis in pregnancy.


The importance of nutritional support for the geriatric surgical patient.

An appropriately designed program of nutritional support will allow the geriatric  patient to undergo necessary surgical therapy with fewer risks and improved results. The selection of the route of administration and the specific agent(s) will depend on the functional status of the gastrointestinal tract as well as the patient's protein and calorie requirements. Table 6 summarizes our recommendations in the delection process. Our obvious preference is the gastrointestinal tract route if possible. When this optimum method of nutritional management is not available, sequential or concurrent supplemented feedings or intravenous hyperalimentation may be required to achieve adequate protein calorie intake. Regardless of the route chosen, careful monitoring of the geriatric patient's response to nutritional therapy is required. Only through careful planning and execution of nutritional therapy can optimum results be achieved in this group of patients.


Influence of sex on the radiation-induced life span modifications in Drosophila melanogaster.

Drosophila melanogaster is exposed to 0, 25, 50, 75 kr of 60Co gamma-irradiation at 1,000 r/min, on the 4th day of its imaginal life. As a result, the life span of the flies is reduced for both sexes. Females, however, are more radiosensitive than males. In females, the radiation-induced life span shortening does not vary with the doses administered; in males, on the contrary, the life expectancy decreases as the dose is increased.


Polyphenol oxidase as a factor in aging.

Polyphenol oxidase oxidizes several proteins that contain tyrosine. During aging, when synthesis and replacement of catabolized proteins are lagging, the relative proportion of oxidized species might reach high values, thus impairing physiological function. A mathematical analysis verifies this assumption.


Wyatt v. Stickney: assessing the impact in Alabama.

In 1971 U.S. District Judge Frank M. Johnson, Jr., ruled that patients involuntarily committed to Alabama mental institutions have a constitutional right to treatment. The following year he issued a court order containing 35 minimum constitutional standards for adequate treatment of the mentally ill and appointed human rights committees at the institutions to oversee their implementation. Focusing primarily on Bryce Hospital in Tuscaloosa, the author examines the extent to which compliance with the standards has been achieved, problems that have arisen in implementing the standards, and issues that still must be dealt with. He briefly discusses changes in the state's mental health system that preceded the filing of the case, and he emphasizes that those changes may have contributed significantly to the improvements that have occurred in mental health care in Alabama after Wyatt.


Incidence of seizures and EEG abnormalities among offspring of epileptic patients.

The marriage rate of epileptic patients was 62% in males and 78% in females. Compared with the rates in the general population, the male patients had a 15% lower rate, but there was no difference in females. There were 263 patients with at least one offspring selected for the study. There were 234 sons and 272 daughters (506 total, 1.9 per patient). Distribution by types of seizure was awakening grand mal, absence or myoclonic petit mal in 24%, grand mal with no aura in 21%, grand mal during sleep in 23%, diffuse grand mal in 7%, grand mal with aura in 13%, psychomotor seizure in 9%, and focal seizure in 3%. The probands were composed of 79% idiopathic and 21% symptomatic in pathogenetic classification. An epileptic EEG abnormality was demonstrated in 22% of male and 44% of female probands. The incidence of seizures among offspring was 2.4% (4.2% age-corrected) in a narrow sense (epilepsy) and 9.1% in a broad sense including febrile convulsions. The latter morbidity was 11.0% for the idopathic and 3.2% for the symptomatic group; 11.0% for female and 6.9% for male probands; 10.2% for sons and 8.1% for daughters. The figure was higher for the probands with the age range at onset of seizure of 0--4 years (20.6%) and 20--29 years (12.6%) than for those with other age ranges; higher for those with awakening grand mal, absence, myoclonic petit mal, for those with family history of epilepsy than those without it. Possible correlation of types of seizure between probands and offspring was demonstrated. Thirty-seven percent of offspring exhibited epileptic EEG abnormalities, and the ratio of epileptic EEG abnormalities to clinical manifestation is about 4:1. Possible existence of familial aggregation of EGG abnormalities and of two kinds of families with large or small epileptic predisposition was indicated. The importance of the role of hereditary and environmental factors in epileptic pathogenesis is proved, and the results of an investigation of congenital malformation among offspring of epileptic mothers are presented. These results were considered to be useful for genetic counseling of epileptic patients.


beta-Galactosidase deficiency in juvenile and adult patients. Report of six Japanese cases and review of literature.

Six juvenile and adult patients with progressive neurological diseases and beta-galactosidase deficiency were reported. Any diseases known to date were denied. These cases together with ten case reports in the literature were reviewed and were classified into three groups from clinical and biochemical points. Group 1 patients were characterized by progressive ataxia and myoclonus with gargoyl changes and macular cherry-red spots. In this syndrome beta-galactosidase activity seems to be secondarily affected by other biochemical defects. A group 2 patient showed similar neurological manifestations without gargoyle changes or macular cherry-red spots. Patients with these clinical features not associated with beta-galactosidase deficiency have also been described in the literature. Group 3 patients had progressive pyramidal and extrapyramidal disease without gargoyl changes or macular cherry-red spots. These cases may represent juvenile and adult type GM1-gangliosidosis. Accumulation of GM1 has not yet been demonstrated.


Characterization of alpha-galactosidase isoenzymes in normal and Fabry human-Chinese Hamster somatic cell hybrids.

The alpha-galactosidases in normal man-Chinese hamster somatic cell hybrids were investigation with antibodies specific for human alpha-galactosidase A and antibodies specific for Chinese hamster alpha-galactosidase. It was found that an isoenzyme in hybrid cells, which has an electrophoretic mobility between that of human alpha-galactosidase A and Chinese hamster alpha-galactosidase, contains immunologic determinants of both human and Chinese hamster origin, suggesting that it is a heteropolymeric molecule. Moreover, the locus for human alpha-galactosidase, which was found to be X-linked, is the locus coding for alpha-galactosidase A. Hybrids isolated after fusion of Chinese hamster cells with cells of a patient with Fabry's disease did not express human alpha-galactosidase A or the heteropolymeric molecule even in the presence of the active human X chromosome, indicating that the deficiency of alpha-galactosidase A in Fabry's disease is probably due to a mutation in a structural gene resulting in the inability to form immunologically detectable and functionally active molecules of alpha-galactosidase A.


A method for establishing cell lines from Drosophila melanogaster embryos.

A simple method is presented for establishing continuous cell lines from Drosophila melanogaster embryos. Subculturing is performed after the first 8 weeks and at 2-week intervals thereafter. Initial plating densities of 5 x 10(4) to 5 x 10(5) cells per cm2 are required for maintaining the subcultures. Cell lines were established from wild-type embryos, from embryos bearing chromosomal rearrangements and from embryos bearing recessive mutations. Permanent lines have doubling times of 24 to 48 hr and have been maintained for as long as 13 months and 25 subcultures.


[Reservoir for bacteria of urinary tract infections in urologic patients (author's transl)].

Many infections of the urinary tract, especially after surgical treatment are nosocomial infections. The etiological bacteria could only be found in the urine of the infected patients and in the faecal flora of nearly 40% of all stationary patients. The so called "problem bacteria": Klebsiella, Proteus rettgeri, pseudomonas showed increasing drug resistance during the observed period between 1972-1975. The bacteria isolated from the faecal flora also showed multiple drug resistance. Thus we concluded that the antibiotic regime in these patients is very important for the selection and the increase of the problem bacteria both in urine and in the faecal flora.


[Change of germs in several wards of hospital during the last twenty years under the therapy with antibiotics (author's transl)].

Statistical examinations concerning the change of germs and their resistance against antibiotics during the last twenty years were performed. The examinations dealed with the material of a bacteriological institute, the material of a department for nephrology and extracorporal dialysis, the material of a center for reanimation, and the material of an urological clinic.


[Human blood fractionation and the risk of hepatitis].

Surveying the development of plasma fractionation the measures to minimize the hazard of transmitting viral hepatitis through plasma fractions are reviewed.


[Blood group compatibility testing for blood transfusions].

Acute and chronic dangers of blood-transfusion are listed. Three serologic demands in selecting donorblood are identity in the ABO-system compatibility in the Rh-system and a negative cross-match in the mode of a three-step Coombs-test. The often claimed consideration of the Kell-system is shown to be numerical in the background of transfusion-practice. The highest responsibility is taken by the physician in transfusing patients with Coombs-positive auto-immune-diseases in which the cross-match is positive and so present antibodies could be hidden.


Relationship between Habs serotypes and 2-amino sugar composition of Pseudomonas aeruginosa.

Thirteen Habs-serotype strains could be classified into 11 groups depending on the characteristic patterns of 2-amino sugar composition; strains of serotypes 2, 5, and 7 had the same pattern, and each other serotype strain had its own distinctive pattern. In our classification, fucosamine and quinovosamine were of importance. We found that fucosamine from strain P14 (Habs serotype 1) was of the D configuration only, unlike DL-fucosamine from strain N10 (Habs serotype 11).


Neisseria gonorrhoeae and ABO isohemagglutinins.

Depending on the ABO blood group, gonorrhea may affect the titers of isohemagglutinins compared with those of uninfected controls. The isohemagglutinin titers in group O patients were significantly increased (P less than 0.001) against erythrocytes A, B, and AB. In group A patients, only the titer against AB erythrocytes was significantly increased. In group B patients, the titer against AB erythrocytes was significantly lower (P less than 0.001) as compared with that in sera of healthy persons. In six of eight volunteers, an increase in isohemagglutinin titer was observed after an injection of small doses of killed gonococci. However, guinea pigs, rabbit, or small monkeys, i.e., species for which gonococci are not apthogenic, when immunized with gonococci either did not form ABO hemagglutinins or did so with very low titers. In white gonorrhea patients, there was a significantly higher frequency of group B individuals over those with group A, AB, or O. No such correlation was found in black patients. Isohemagglutinins from human sera could be absorbed by cultured gonococci. The implications of these findings in the pathogenesis of gonococcus infection and the problems associated with vaccine development are briefly discussed.


Lack of protection afforded by ribonucleic acid preparations from Mycobacterium tuberculosis against Mycobacterium leprae infections in mice.

Mycobacterial ribonucleic acid preparations from H37Ra, an attenuated strain of Mycobacterium tuberculosis, provide their usual marked protection against M. tuberculosis challenge; however, they provided no protection against Mycobacterium leprae challenge. Suspensions of intact H37Ra were not effective against M. leprae. Suspensions of BCG gave their usual distinct protection against M. leprae challenge.


Partial purification and properties of an antibacterial product of peritoneal exudate cell cultures from BCG-infected guinea pigs.

Peritoneal exudates elicited in BCG-infected guinea pigs with caseinate yield cell cultures that have been shown to produce soluble material capable of sterilizing certain bacteria if the cultures are incubated with the specific antigen purified protein derivative or the lectin phytohemagglutinin. This material is now shown to have the following properties: (i) strongly adsorbable to glass; (ii) strongly adsorbable to cation- and not to anion-exchange resins but not elutable with mineral acid or solutions of high ionic strength; (iii) strongly adsorbable to cellulose nitrate membrane filter materials and quantitatively elutable with dilute HCl, providing a convenient method for partial purification; (iv) relatively stable over a wide range of pH and temperature; (v) antagonized by polyanions and by iron ions; (vi) active against the three gram-positive bacilli tested and not against the other organisms tested: (vii) more active in alkaline than in acidic media; and (viii) inactivated by proteolytic enzymes.


Exotoxin production by clinical isolates of pseudomonas aeruginosa.

Seventy-five consecutive clinical Pseudomonas aeruginosa isolates were tested for in vitro exotoxin production. Exotoxin was demonstrated in culture filtrates biologically, by its ability to produce characteristic dermonecrotic lesions in guinea pigs, and seriologically, by counterimmunoelectrophoresis (CIE) with rabbit antiserum elicited with purified exotoxin. By these two methods, exotoxin was detected in 87 and 89% of P. aeruginosa strains, respectively (r = 0.48, P less than 0.001). Although less sensitive than CIE in detecting exotoxin immunodiffusion demonstrated a reaction of antigenic identity in most cases. Exotoxin was produced by all seven Fisher-Devlin immunotypes and by untypable strains. In contrast, exotoxin was not detected in the culture filtrates of 16 non-aeruginosa pseudomonas isolates and 48 non-pseudomonas organisms. The production of biologically similar antigenically closely related exotoxins is thus a characteristic of the majority of P. aeruginosa strains derived from diverse clinical sources.


Indirect immunofluorescence antibodies in natural and acquired Rickettsia tsutsugamushi infections of Philippine rodents.

Antibodies against Rickettsia tsutsugamushi detected by the indirect fluorescent-antibody test (IFAT) were present in most rats trapped from a human focus of scrub typhus in the Philippines. Rickettsiae were isolated only from rats with positive IFATs. Naturally acquired antibodies persisted for at least 11 months, and antibodies resulting from experimental infections of rats persisted for at least 7 months. A common Philippine rodent, Rattus mindanensis, tolerated experimental infections with both local and standard Karp strains of R. tsutsugamushi, and such infections always produced a positive IFAT.


Ethylenediaminetetraacetic acid-sensitive antiphagocytic activity of Neisseria gonorrhoeae.

Colonial types of Neisseria gonorrhoeae were examined for the presence of pilus-independent antiphagocytic activity. Type 3 and depiliated type 1 gonococci had a shearing- and protease-resistant antiphagocytic activity that was eliminated by treatment with ethylenediaminetetraacetic acid (EDTA) and that was not present on type 4 bacteria. Incubation of EDTA-treated bacteria 37 degrees C for 90 min resulted in fas prevented by antibiotics that block the final assembly of cell wall macromolecules that depend on the C55-isoprenoid carrier for export. These include both lipopolysaccharide and peptidoglycan. Restoration was, however, unaffected by drugs that interfere with the synthesis of peptidoglycan, but not that of lipopolysaccharide, and by inhibitors of protein synthesis. These data suggested that gonococci have an antiphagocytic mechanism in addition to the previously described determinant (presumably pili) that was removed by blending or by treatment with proteases. Of the two antiphagocytic activities, type 1 had both, type 3 had only the EDTA-sensitive component, and type 4 had neither.


Penicillin sensitivity and serum resistance are independent attributes of strains of Neisseria gonorrhoeae causing disseminated gonococcal infection.

We have determined that isolates of Neisseria gonorrhoeae from patients with disseminated gonococcal infection (DGI) are different from randomly collected isolates from patients with uncomplicated (local) disease. Our comparison was based on the six phenotypic properties of: sensitivity to penicillin (PenS), erythromycin, and streptomycin; resistance to the bactericidal effects of pooled human sera; requirements for arginine, hypoxanthine, and uracil (AHU-); and sensitivity to toxic agar. Although the marked association among these traits made analysis difficult, several factors independently related to virulence were defined. The DGI isolates were significnatly more PenS and resistant to serum, even when the other variables were held constant. An apparent correlation between AHU- auxotype and virulence was shown to be due to the PenS property of most AHU- isolates. Thus, certain mutations to antibiotic resistance as well as susceptibility to sera, may result in loss of virulence in the gonococcus, perhaps through alteration of cell envelope structure.


Mitogenic activity of cell wall components from smooth and rough strains of Brucella abortus.

On the basis of [3H]thymidine incorporation by normal mouse spleen cell cultures, cell wall preparations from a smooth (45/0) strain and a rough (45/20) strain of Brucella abortus were strongly mitogenic. On the other hand, none of several subcomponents extracted from the cell wall preparations, including aqueous and phenolic lipopolysaccharides, was active. These results contrast with the marked mitogenic activity of lipopolysaccharides isolated from other gram-negative bacteria such as Salmonella typhimurium.


Antitumor activity of a Brucella abortus preparation.

Mice injected intraperitoneally with sarcoma-180 cells develop ascites and eventually die. Intraperitoneal injection of a nonviable, aqueous... ether-extracted Brucella abortus preparation (BRU-PEL) as early as 7 days before or as late as 7 days after injection of tumor cells significantly inhibited development of ascites and protected against death. BRU-PEL was not effective if injected after ascites was grossly apparent. BRU-PEL was significantly more active than a Corynebacterium parvum preparation administered in the same way.


Cellular immunity to toxoplasma and besnoitia in hamsters: specificity and the effects of cortisol.

The inhibitory effects of cortisol on cellular immunity were studied in vitro by using hamster peritoneal exudate cells. Two obligate, intracellular protozoa--Toxoplasma gondii and Besnoitia jellisoni-- were used to control for specificity of effects. Results indicate that immune lymphocytes specifically confer immunity to (or "arm") macrophages that specifically express immunity. This arming can be inhibited by 5 microng of cortisol per ml. Macrophages that have been armed already will continue to express immunity (by limiting parasite growth specifically) in the presence of 5 microng of cortisol per ml. Cortisol levels of 20 microng/ml are required to inhibit the expression of immunity by armed macrophages. It was also found that lymphocytes, from hamsters given 20 mg of cortisol subcutaneously 2 days before the harvest of cells, did not arm macrophages, whereas macrophages from these same animals could be armed by immune lymphocytes from untreated hamsters. Therefore, it was concluded that in relation to cellular immunity, lymphocytes are more sensitive to cortisol than are macrophages. Since antibody to these parasites is almost always present in vivo, we also tested the effects of cortisol on the disposition of antibody-modified organisms by activated (not armed) macrophages, and found that 50 microng of cortisol per ml was needed to inhibit macrophage effects on antibody-treated organisms.


Ultrastructural investigations on surface structures involved in Coxiella burnetii phase variation.

By using the cytochemical staining procedure with concanavalin A, horseradish peroxidase, and diaminobenzidine, no surface carbohydrates with terminal alpha-glucosyl or sterically closely related residues could be detected on the cell walls of Coxiella burnetii phases I and II. Using a polycationized ferritin derivative as a cytochemical probe, anionic binding sites were visualized in the electron microscope on cell membranes of C. burnetii phase II, but not on phase I organisms. The sites appeared to be masked in phase I particles. Anionic sites could be demonstrated on phase I organisms after treatment with NaIO4 or dimethyl sulfoxide. A number of different biological properties of C. burnetii phases I and II may depend on the presence or absence of a net negative charge on the surface of the cell walls of these organisms.


Additional evidence that a new virulence-associated antigen of Neisseria gonorrhoeae is not of pilus origin.

Triton-X-100-treated pilus suspensions prepared from Neisseria gonorrhoeae produced a single line of precipitate in immunodiffusion tests. This line was distinct from that of the virulence-associated antigen.


[The agar diffusion method round table discussion on the methods and indications for determining blood levels of aminoglycosides (author's transl)].

The present-day technique of determining the concentration of aminoglycosides in blood by agar diffusion is described including the apparatuses, set-up, and reliability tests. The results of comparative investigations showed a higher degree of conformity in Mueller-Hinton agar than in standard agar. Due to its accuracy the agar diffusion method is especially suited for the calculation of dosage recommendations and for the prediction of the maximal and minimal blood levels during treatment with aminoglycosides.


Sensitivity of gram-negative bacteria to six aminoglycoside antibiotics.

An in vitro study of the susceptibility of 201 newly isolated strains of gramnegative bacteria to six aminoglycoside antibiotics (kanamycin, amikacin, gentamicin, tobramycin, sisomicin and netilmicin) was performed by the twofold dilution method in fluid medium. Both the minimal inhibitory concentration and the minimal bacteridical concentration were determined. Overall, tobramycin seemed the most effective of the drugs studied. Netilmicin, the new derivative from sisomicin, compared favourably with the other drugs tested, but may, on theoretical grounds, offer the additional advantage of retained efficacy in the face of developing bacterial resistance. Not unexpectedly, amikacin appeared to be the most promising of the drugs studied in its action against Pseudomonas aeruginosa. Amikacin and netilmicin appeared to be the most effective of this group of antibiotics against Klebsiella species.


The antigens of pigeon breeder's disease. I. Studies on unfractionated pigeon dropping extracts.

Approximately 6% of the dry organic dust from pigeon coops was extracted in the form of soluble, nondialyzable components. These pigeon dropping extracts contain a number of stable protein and glycoprotein antigens together with pigments and other components that cause nonspecific precipitation of normal and immune sera. Based on their migration in immunoelectrophoresis, the antigens have been designated PDEB, 1, 2, 3 or 4.


Quantitation of beta 1c/1A globulin (C3) in inbred mice: variation dependent upon strain, age, sex and environment.

The effect of age, sex, genetic backgrounds and environmental factors on mouse C3 level was studied. In general, C3 level increased as a mouse became older. Little difference in C3 level was observed between male and female of all ages. C3 level was found in a great variation from strain to strain, but we could not demonstrate any major role of H-2 in determining serum C3 level. Serum C3 level is not shown to be linked to the serum level of H-2 linked serum protein Ss (C4). On the other hand a close correlation between C3 level and C5 level was demonstrated. Serum C3 markedly increased, along with C5, when mice were injected with turpentine oil, suggesting both components are acute phase reactants. The degree of increase in C3 level in response to turpentine oil injection appeared to be controlled by a single or a very few genes probably not linked to H-2.


Differential airway reactivity to carbachol and physostigmine sulfate in rhesus monkeys with and without reagin-mediated respiratory responses.

Bronchial airway reactivity to carbachol (CAR) and physostigmine (PHY) was individually measured in three rhesus monkeys with and three rhesus monkeys without reagin-mediated respiratory responses (RR). The three monkeys with reagin-mediated RR demonstrated bronchial hyperreactivity to CAR (p less than 0.25) but not PHY when compared with the three control monkeys. No correlation between airway reactivity to CAR and PHY was noted in all six monkeys. We conclude that monkeys with reagin-mediated RR possess airway hyperreactivity to the exogenously administered acetylcholine analogue CAR, but not to the endogenous acetylcholine which is released in vivo by PHY.


Three immunoglobulin classes in the pigeon (Columbia livia).

Three classes of immunoglobulins have been identified in the pigeon. IgG and IgM were purified from pigeon serum whereas IgA was isolated from pigeon hepatic bile. Pigeon IgG and IgM had the same properties and immunohistological distribution as their chicken homologues. Pigeon IgA was identified on the following grounds: (1) it contains the same light chains as pigeon IgM and IgG; (2) it is relatively abundant in exocrine secretions such as bile, egg white, cropmilk and intestinal fluid, whereas it is present only in small amounts in serum; (3) it occurs in the cytoplasm of the majority of the immunocytes from the intestinal mucosa; (4) its electrophoretic mobility and molecular size are similar to those of chicken IgA. Surprisingly, no immunoglobulin-containing cells could be detected in sections of the wall of the cropmilk gland, despite the high IgA content of the cropmilk.


Avarice, inefficiency, and inequality: an international health care tale.

The paper is concerned with impact of a medical profession, physicians, on the delivery of health care. The basic economic motivation of self-interest and avarice has led this profession to produce health care outcomes which are inequitable and inefficient. In the first section of the paper the regional geographical distribution of physicians in four disparate health systems--England, Ireland, France, and West Germany-is analyzed and found to be highly unequal. The next section is concerned with the efficacy of therapies and the cost-effectiveness of health care delivery systems in a variety of countries. The final section discusses how health care can be more equitably and more efficiently delivered. It is argued that both markets and bureaucaracies are likely to be inadequate unless carefully monitored. In particular, there is a great need to investigate the cost-effectiveness of therapies and then persuade physicians, via pecuniary and nonpecuniary incentives, to behave in a manner which leads to  more equitable and efficient health care outcomes.


Rational prescribing of drugs.

A rational prescription is defined as one which proposes the best pharmacological approach to obtain the therapeutic objective. To achieve this goal requires a careful consideration of factors related to: patient, disease, medication, and doctor. Concepts of risk/benefit and cost/benefit are of relevance for the therapeutic decision. The doctor also has to make an effort to assure the compliance of his patients. This requires a good understanding of the personality and of the socio-cultural background of the patient. Social pharmacology helps to appreciate and understand the role of these factors and to use them for improvement of therapeutic results.


External biliary-pancreatic fistulas.

Twenty-seven patients treated for pancreatic and/or biliary-cutaneous fistulas have been reviewed. Four patients died mainly because of cardiopulmonary and septic complications. Spontaneous sealing of the fistula occurred in 81% of the conservatively treated cases (48% of all cases). All the LO fistulas but only 68% of the HO fistulas treated conservatively sealed spontaneously. Eleven patients were treated surgically. There were three deaths and three failures (reappearance of fistula). All the patients who died had been operated on within three months after the appearance of HO fistulas. There was no mortality among the patients with LO fistulas or among patients operated on at a later stage. We have reached the following conclusions: 1. There is a significant difference in prognosis between low output and high output fistulas. 2. In LO fistulas, there is no need for a surgical intervention aimed to close the fistula unless it persists for at least one year. 3. In HO fistulas, if a corrective operation is necessary, it should be withheld for at least three months whenever possible. 4. Roux-en-Y fistulojejunostomy is considered to be the procedure of choice. 5. Infection and premature colsure of the external part of the fistulous tract should be avoided by insertion of drains and repeated surgical drainage, where necessary. 6. High caloric feeding, elemental diet and intravenous hyperalimentation are very important factors that enhance recovery in the surgically and conservatively treated patients.


Experimental macular edema after lens extraction.

Macular edema has been observed frequently in man after cataract extraction, but pathogenic mechanisms remain unclear. Seven eyes of four young adult rhesus monkeys underwent lens extraction. The retinas and maculae of these eyes were examined by ophthalmoscopy fundus photography, fluorescein angiography, light and electron microscopy, and the horseradish peroxidase tracer technique. In the macular region, the blood-retinal barrier at the retinal vasculature was disrupted in three of the seven eyes. All three eyes had had vitreous loss during lens extraction. Horseradish peroxidase was observed both intracellularly and extracellularly in the maculae. In contrast, the blood-retinal barrier at both the retinal pigment epithelium and the retinal vasculature of the peripheral retina in most eyes was intact. We conclude that macular edema secondary to lens extraction is due to disruption of the blood-retinal barrier at the levels of the retinal vasculature and the retinal pigment epithelium.


Aldose reductase in retinal mural cells.

Cultured mural cells (intramural pericytes) from adult rhesus retinal capillaries were examined for the presence of the sorbitol pathway. A radioimmunoassay for human aldose reductase, cross-reactive with rhesus lens aldose reductase, showed the presence of this enzyme in our cultured cells. Mural cells grown in culture media containing normal (10 mM) and high (40 mM) levels of glucose were examined for polyol accumulation by gas-liquid chromatography. Cells incubated in high glucose medium showed a threefold increase in sorbitol concentration over cells grown at low glucose levels. After 30 days in high glucose medium, mural cells formed dense multilayered areas with extensive cellular debris. These findings suggest the presence of the sorbitol pathway in cultured retinal mural cells and cellular degeneration in high glucose medium; this may have possible implications in the pathogenesis of diabetic retinopathy.


Ultrastructure of the hyaloid vasculature in primates.

The histology and ultrastructure of the hyaloid vascular system was studied in 16 monkeys, ages 85 days in utero to 3 years old. The well-developed hyaloid artery is identifiable ultrastructurally as an arteriole consisting of a nonfenestrated intima, a multilayered smooth muscular media, a connective tissue adventitia, and a perivascular sheath. By the fourth week after birth, the vessel walls appear hyalinized and acellular; the lumen is occluded by a thrombus. The time of the complete disappearance of the hyaloid artery varied considerably in individual monkeys; remnants of the hyaloid artery were often present on the disc (Bergmeister's papilla) in adult life. The vasa hyaloidea propria and tunica vasculosa lentis are small branches of the hyaloid artery which fill the primary vitreous. Ultrastructurally, they are Type A-1-alpha capillaries having a nonfenestrated endothelium, incomplete pericyte layer, and basement membranes surrounding each. Fluorescein angiography of several newborn monkeys revealed nonleakage of dye from the hyaloid artery and its branches, thus correlating with the presence of endothelial tight functions.


Experimental myelin intrusion in the nerve head.

On the assumption that myelin intrusion into the papilla and peripapillary region might occur with traumatic lesions of the optic nerve, a study was made of the clinical and histopathologic changes that might be expected. Nine monkey eyes were subjected to hemostat compression of the nerve close to the globe and studied over variable periods up to 28 days. Myelin was demonstrable ophthalmoscopically, followed by variable and increasing amounts of hemorrhage. The myelin was demonstrable histopathologically only during the first 2 weeks after the manipulation and was then masked by the associated hemorrhage and gliosis. The optic nerve showed expected myelinolytic reactions.


Visual acuity and behavior of monocularly deprived monkeys after retinal lesions.

One eyelid of each of three Macaca nemestrina monkeys was sutured shut at 3 weeks of age. One animal had the central 10 degrees of the open eye retina lesioned at 3 weeks of age (EL1), a second at 9 months of age (LL1), and the third had no lesion (MD1). After a reverse suture at 9 months of age, the deprived eye was tested for grating acuity, visual behavior, and visual field. EL1 tested positively on all behavioral tasks by 1 month and showed good visual ability to negotiate a playroom but never performed better than 20/1,250 on grating acuity. MD1 showed little visual behavior in the playroom but tested at 20/400 acuity. LL1 rapidly recovered all aspects of visual behavior and tested at 20/80 visual acuity. Perimetry shows that LL1 and EL1 respond mainly in the central portion of the deprived eye visual field but not to the periphery or the monocular segment. We conclude that removal of the open eye retina after the critical period allows much visual recovery by the deprived eye but that removal of the open eye retina within the critical period does not prevent many of the effects of monocular deprivation.


The effect of Candida albicans on the isolation of N. gonorrhoeae.

Recently, several studies have reported that strains of Candida albicans and perhaps other noraml bacterial flora are capable of inhibiting the growth of Neisseria gonorrhoeae in vitro, and it is possible that this may occur in vivo as well. This possible antagonism between N. gonorrhoeae and normal flora microorganisms poses many questions regarding the reliable laboratory diagnosis of gonorrhea. Mixed cultures of C. albicans and N. gonorrhoeae were grown together on GC base agar plates with varying concentrations of nystatin. Recovery of N. gonorrhoeae was improved as nystatin concentration was increased to 25 IU/ml, while growth of C. albicans was effectively inhibited. At concentrations of 50 and 100 IU/ml nystatin also began to inhibit N. gonorrhoeae. There was a noticeable decrease of nystatin bioactivity against C. albicans when the GC plates were stored at 4 C, thus making it more difficult to isolate N. gonorrhoeae. Growth curves of C. albicans and N. gonorrhoeae grown together in liquid medium did not significantly differ from growth curves of the individual organisms. Statistical analysis of our findings does not support the theory that strains of C. albicans produce a soluble diffusible factor that is inhibitory to N. gonorrhoeae.


Uridine diphosphate galactose 4'-epimerase deficiency. IV. Report of eight cases in three families.

Eight persons who had no activity of uridine diphosphate galactose 4'-epimerase in their circulating blood cells are known today. They were healthy members of three different families. Propositi were newborns discovered in a mass screening test for blood galactose which registered high levels of erythrocyte galactose-1-phosphate. Epimerase deficiency was restricted to circulating blood cells, but in liver biopsy specimens, in cultured skin fibroblasts and in activated lymphocytes epimerase was found to be normally active. Heterozygotes had intermediate red cell epimerase activity. There were no symptoms of galactose intolerance and no pathology related to the enzyme defect. All 8 epimerase deficient persons had a ccdee Rhesus genotype. Attempts at demonstrating genetic linkage between the epimerase and Rhesus loci were unsuccessful because of the difficulty encountered in ascertaining epimerase heterozygosity. Individuals with hereditary UDP-galactose 4'-epimerase deficiency appear to produce an unstable mutant enzyme requiring higher NAD concentration for maximum activity.


Isolation and characterization of lipopolysaccharides from cell walls of blue-green algae of the genus Phormidium.

Lipopolysaccharide (LPS) fractions were isolated from three species of blue-green algae of the genus Phormidium, namely, P. africanum, P. laminosum, and P. uncinatum, by using a phenol-water procedure followed by exhaustive extraction with ammonium oxalate. The materials obtained were shown to be closely related biochemically. Nearly 60% of the LPS consisted of the polysaccharides galactose, glucose, mannose, xylose, arabinose, and rhamnose and an unidentified, fast-moving sugar residue. In addition, glucosamine, galactosamine, and 2-keto-3-deoxyoctonate were detected. Oleic, stearic, and palmitic acids were found in the hydrolysate of the lipid component, which averaged 1.5% of the LPS. Concomitantly, the protein component (7 to 20%) was shown to contain the following amino acids: aspartic acid, threonine, serine glutamic acid, proline, glycine, alanine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine, and arginine. Whole cells, as well as the LPS, of Phormidium possessed antigenic properties.


Specificity of siderophore receptors in membrane vesicles of Bacillus megaterium.

Membrane vesicles of Bacillus megaterium strains SK11 and Ard1 bound the ferrischizokinen and ferriferrioxamine B siderhores (iron transport cofactors). An approximately equimolar uptake of both labels of [3H, 59Fe]ferrischizokinen indicated binding of the intact chelate. Binding reached equilibrium in 2 to 5 min, was temperature independent, and was unaltered by the addition of several energy sources. A 91% dissociation of bound [Fe]ferrischizokinen was achieved in 60 s by the addition of excess ferrischizokinen. Ferriaerobactin, a siderophore which is structurally related to ferrischizokinen, caused no detectable release of bound [59Fe]ferrischizokinen. Of several other ferrigydroxamates tested, only ferriferrichrome A achieved the release (11%) of [Fe]ferrischizokinen. Rapid dissociation (92%) of bound [59Fe]ferriferrioxamine B by the addition of ferriferrioxamine B was observed, and a 67% release of [59Fe]ferriferrioxamine B was caused by ferriA2265, its structural relative. Ferrischizokinen, ferriferrichrome A, and ferrirhodotorulic acid produced a 6, 25, and 29% dissociation, respectively, of [59Fe]ferriferrioxamine B; ferriaerobactin caused no dissociation. [59Fe]ferriaerobactin was bound by the membranes, but its dissociation was not effected by unlabeled ferriaerobactin, suggesting no specific receptors for this chelate. The respective binding affinity constants and maximal binding capacities of membrane vesicles of strain SK11 were 2 x 10(7) M-1 and 280 pmol per mg of protein for ferrischizokinen and 7 x 10(7) M-1 and 37 pmol per mg of protein for ferriferrioxamine B. These values in strain Ard1 were, respectively, 1.4 x 10(7) M-1 and 186 pmol per mg of protein for ferrischizokinen and 11 x 10(7) M-1 and 23 pmol per mg of protein for ferriferrioxamine B. Separate, specific binding sites (receptors) for ferrischizokinen and ferriferrioxamine B exist on the vesicles. The ferrischizokinen receptors have a lower affinity but a higher binding capacity (eightfold) than that shown by the ferriferrioxamine B receptor. These receptors may be components of independent transport systems.


Classification of Bacillus subtilis flagellins.

Purified flagellins derived from 16 strains of Bacillus subtilis were classified into at least five distinct groups on the basis of their reaction with antiflagellar filament antibody and antiflagellin antibody. This classification was in good accord with that derived independently on the basis of amino acid analyses of the flagellins. Flagellar antigenicity appears to provide a useful typological character in classifying B. subtilis strains.


Role of glutamate in the sporogenesis of Bacillus cereus.

Bacillus cereus T, sporulating in a chemically defined medium under optimum conditions, requires substrate quantities of glutamate during the first 4 h of sporogenesis. Seventy percent of the glutamate utilized was catabolized to CO2 during this period, with the remaining glutamate carbon assimilated into various spore constituents, principally protein and nucleic acid. The importance of glutamate as the primary source of reducing potential and energy for early stages of spore formation was investigated. Although the relative efficiency at which tricarboxylic acid cycle intermediates substituted for glutamate was suggestive of oxidation via the tricarboxylic acid cycle, only partial inhibition of glutamate oxidation by fluoroacetate was observed.


Inorganic salts resistance associated with a lactose-fermenting plasmid in Streptococcus lactis.

Present evidence indicates that lactose metabolism in group N streptococci is linked to plasmid deoxyribonucleic acid. Lactose-positive (Lac+) Streptococcus lactis and lactose-negative (Lac-) derivatives were examined for their resistance to various inorganic ions. Lac+ S. lactis strains ML3, M18, and C2 were found more resistant to arsenate (7.5- to 60.2-fold), arsenite (2.25- to 3.0-fold), and chromate (6.6- to 9.4-fold), but more sensitive to copper (10.0- to 13.3-fold) than their Lac- derivatives. These results suggested that genetic information for resistance and/or sensitivity to these ions resides on the "lactose plasmid." Kinetics of ultraviolet irradiation inactivation of transducing ability for lactose metabolism and arsenate resistance confirmed the plasmid location of the two markers. Lac+ transductants from S. lactis C2 received genetic determinants for resistance to arsenate, arsenite, and chromate but not for copper sensitivity. In this case, resistance markers were lost when the transductants became Lac- but the derivatives remained copper resistant. The resistant markers for arsenate and arsenite could not be identified as separate genetic loci, but chromate resistance and copper sensitivity markers were found to be independent genetic loci. The "lactose plasmid" from S. lactis C10 possessed the genetic loci for arsenate and arsenite resistance but not for chromate resistance or copper sensitivity.


Initiation and termination of chromosome replication at 45 degree C in a temperature-sensitive deoxyribonucleic acid initiation mutant of Bacillus subtilis 168, TsB134.

Deoxyribose nucleic acid transfer experiments showed that upon shifting Bacillus subtilis TsB134 to 45 degree C, initiation of new rounds of replication was effectively blocked and the majority of existing rounds terminated.


Hydrolysis of lithocholate sulfate by Pseudomonas aeruginosa.

Pseudomonas aeruginosa was found to be able to hydrolyze bile sulfate. This property was observed when lithocholate sulfate was substituted for the sulfur source in the culture medium. The addition of MgSO4 to the medium inhibited the hydrolysis of the bile sulfate.


Chemical analysis of the phenol-water-extractable materials from Anabaena flos-aquae.

The high molecular-weight carbohydrate substances extracted in the aqueous and phenol phases by phenol-water extraction of Anabaena flos-aquae A-37 were found to be polysaccharides without lipid attached.


Effect of temperature on the growth of Myxococcus xanthus.

The cardinal growth characteristics of Myxococcus xanthus were examined from 14 to 40 degree C, and the examinations indicated that the organism is mesophilic in character. The maximum growth rate (0,3 doublings per h) was between 34 and 36 degree C and the temperature characteristic (micron) is 17,000 cal/mol (71,162 J/mol).


Metabolic role of free mycolic acids in Mycobacterium tuberculosis.

Small amounts of free mycolic acids and trehalose dimycolate that are rapidly formed by Mycobacterium tuberculosis H37Ra are probably derived from mycolyl acetyl trehalose and transferred to the cell wall. However, the transfer of mycolic acids from mycolyl acetyl trehalose to the cell wall still appears to be the more prominent route.


Acetylated methylmannose polysaccharide of Streptomyces.

A polysaccharide composed of 3-O-methyl-D-mannose and D-mannose in a molar ratio of approximately 10:1 and containing 3 to 4 esterified acetyl residues has been isolated from Streptomyces griseus. This acetylated methylmannose polysaccharide (AMMP) is similar to the methylmannose polysaccharide (MMP) of Mycobacterium smegmatis (Gray, G. R., and Ballou, C. E. (1971) J. Biol. Chem. 246, 6835-6842) in its size and composition, the absence of acidic or basic groups, and the lack of a reducing end. It is different, however, in its content of esterified acetyl residues, and it is slightly different in its structure and in its gel filtration properties. The structure of AMMP has been established by proton magnetic resonance spectroscopy, and by combinations of methylation analysis and Smith degradation utilizing non-radioactively labeled polysaccharide and [3H]methyl-labeled polysaccharide obtained from cells grown in the presence of L-[methyl-3H]methionine. It is concluded that AMMP is a linear, nonreducing, neutral polysaccharide composed of a terminal D-mannose residue linked alpha(1 leads to 4) to a chain of 10 consecutive alpha(1 leads to 4)-linked 3-O-methyl-D-mannose residues. The reducing terminal 3-O-methyl-D-mannose residue exists, at least in part, as its alpha-methyl glycoside. The positions of attachment of the ester residues have not been established.


Enzymatic transfer of mannose from mannosyl-phosphoryl-polyprenol to lipid-linked oligosaccharides by pig aorta.

A particulate enzyme preparation prepared from the intimal layer of pig aorta catalyzed the transfer of mannose from mannosyl-phosphoryl-polyprenol (MPP) into a series of oligosaccharides that were linked to lipid. The reaction required detergent with Triton X-100 and NP-40 being best at a concentration of 0.5%. Several other detergents were inactive or only slightly active. The pH optima for this activity was about 7 to 7.5 in Tris buffer and the apparent Km for MPP was about 2 x 10(-7) M. The reaction was not stimulated by the addition of divalent cation and, in fact, was inhibited by the high concentrations of cation. The addition of EDTA did not inhibit the transfer of mannose from MPP and was somewhat stimulatory. The transferase(s) activity was "solubilized" from the particles by treatment with Triton X-100. This solubilized enzyme still formed a series of lipid-linked oligosaccharides from either MPP or GDP-mannose. The oligosaccharides were released from the lipid by mild acid hydrolysis and were separated by paper chromatography. Some five or six radioactive oligosaccharides were formed from either MPP or from GDP-mannose and these oligosaccharides had similar mobilities upon paper chromatography. However, MPP was a better donor for the larger oligosaccharides (i.e. those containing 8, 9, or 10 sugar residues), whereas GDP-mannose was better for formation of the oligosaccharide containing 7 sugar residues. In the presence of EDTA and detergent no MPP was formed from GDP-mannose, but radioactivity was still incorporated into the lipid-linked oligosaccharides. Under these conditions essentially all of the radioactivity was in the oligosaccharide containing 7 sugar residues. Since much of this activity could be released as mannose by acetolysis, GDP-mannose may be the direct mannosyl donor for formation of 1 leads to 6 branches. Oligosaccharides 7, 8, 9, and 10 were isolated and partially characterized in terms of their molecular weights, sugar composition, susceptibility to alpha-mannosidase, and 14C products formed by acetolysis and periodate oxidation. The molecular weights ranged from 1310 for oligosaccharide 7 to 1750 for oligosaccharide 10. Hydrolysis of each oligosaccharide and reduction with NaB3H4 gave the expected ratio of [3H]hexitol to [3H]hexosaminitol based on the molecular weight of the oligosaccharide. However, the hexitol fraction contained [3H]mannitol and [3H]glucitol. Since the amount of radioactivity in glucitol was 2 to 4 times that in mannitol and since only glucosaminitol was found in the amino sugar peak, it seems likely that each 14C-oligosaccharide was contaminated with an unlabeled oligosaccharide of equal molecular weight containing glucose and GlcNAc. Acetolysis of the 14C-oligosaccharides gave rise to 14C peaks of mannose, mannobiose, and mannotriose. In the larger oligosaccharides, most of the radioactivity was in mannobiose whereas in oligosaccharide 7 most of the radioactivity was in mannose...


Bisulfite-induced C changed to U transitions in yeast alanine tRNA.

The reaction of yeast tRNAAla1ab with NaHSO3 at 25 degrees and pH 5.8 has been studied. Five reactive residues have been located. Four of these (C-17 in Loop I, C-36 in the anticodon, C-74 and C-75 near the acceptor end) react to the same extent (42%) under the conditions of the experiment. The other (C-72 in the first base pair of the acceptor stem) reacts much more slowly (8%). No other changes were detected, but kinetic data suggest two or more additional residues may react very slowly. The C changed to U change in the anticodon (igc changed to igu) is a missense change (Ala changed to Thr). Both mechanistic considerations and experimental data from the literature show that HSO3--induced deamination of cytosine residues occurs only at unstacked residues. The quantitative changes for tRNAAla indicate that the stacking lifetimes of C-17, C-36, C-74, and C-75 are about equal. All other cytidine residues are much more tightly stacked. These results are consistent with the folded cloverleaf models that have been proposed from x-ray diffraction studies of yeast tRNAPhe. Residues 48 and 56, which are in single-stranded regions in the unfolded cloverleaf structure, do not react suggesting that they are tightly stacked in solution under the conditions of this experiment. The data also indicate that the anticodon loop is flexible in solution.


Bisulfite-induced C changed to U transitions in yeast valine tRNA.

The reaction of yeast tRNAVallab with NaHSO3 at 25 degrees and pH 5.8 has been studied. Six reactive residues have been located. C-17 in loop I is the most reactive (51% conversion) and C-73 in the first base pair of the acceptor stem the least reactive (8%). Three of the remaining reactive residues (C-39 in loop II, C-75 and C-76 near the acceptor stem) react to the same extent (36 to 38%) under the conditions of the experiment. C-37 in the anticodon reacted to a lesser extent (28%) than C-39 (36%), located just 2 residues away in the anticodon loop. No other changes were detected, but kinetic data suggest one or more additional residues may react very slowly. The C changed to U change in the anticodon (iac changed to iau) is a missense change (Val changed to Ile). Both mechanistic considerations and experimental data from the literature show that HSO3--induced deamination of cytosine residues occurs only at unstacked residues. We interpret the quantitative changes in tRNAVal to indicate that C-17 spends a large portion of its lifetime in an unstacked conformation. The stacking lifetimes of C-37, C-39, C-75, and C-76 seem to be similar but not identical. All other cytidine residues are much more tightly stacked. These results are consistent with the folded cloverleaf models that have been proposed from x-ray diffraction studies of yeast tRNAPhe. Residues C-46, C-49, C-57, and C-61, which are present in the single-stranded regions of the unfolded cloverleaf structure, do not react, suggesting that they are tightly stacked in solution under the conditions of this experiment. The data also suggest that anticodon-loop conformations other than the extremes with five bases stacked on either the 3' or 5' portion of the anticodon stem exist in solution and that the anticodon loop is flexible.


Initiation and translation in vitro of mRNA for MOPC 315 immunoglobulin heavy chain and characterization of translation product.

An initiation study of mineral oil-induced plasmacytoma (MOPC) 315 heavy chain immunoglobulin (H315) in vitro has been conducted using formyl-[35S]methionyl-tRNAfMet and a highly purified 18 S message from MOPC 315 solid tumor in a crude rabbit reticulocyte lysate system. The product was specifically precipitated by antibodies directed against MOPC 315 immunoglobulin and H315. The in vitro H315 products terminally labeled with formyl-[35S]methionine or internally labeled with [3H]leucine were electrophoretically identical with in vivo H315 on sodium dodecyl sulfate-polyacrylamide gels. All of the [35S]-methionine was incorporated at the NH2 terminus, not internally, since there is a near complete recovery of [35S]methionine following one cycle of Edman degradation. The NH2-terminal cyanogen bromide peptide, CN2, of in vivo and in vitro H315 co-migrated exactly on gel electrophoresis under conditions which completely resolved two proteins differing in size by only 14 amino acids. These data strongly suggest that there is no NH2-terminal precursor of H315 in this system. Cyanogen bromide peptide profiles of in vivo and in vitro H315 were chromatographically indistinguishable. Three peptides, CN1, CN2, and CN4, which represent approximately 85% of the total amino acids of H315 were isolated and further characterized by electrophoresis and paper chromatography. All were very similar to the corresponding peptides of authentic H315. We conclude that the fidelity of H315 translation is preserved in vitro.


Sindbis envelope proteins as endogenous acceptors in reactions of guanosine diphosphate-[14C]Mannose with preparations of infected chicken embryo fibroblasts.

Preparations of Sindbis-infected chicken embryo fibroblasts incubated with GDP-[14C]mannose and UDP-N-acetylglucosamine catalyze the glycosylation of endogenous phospholipids and membrane-associated proteins. The proteins are identified as the viral envelope proteins by precipitation with anti-Sindbis antiserum, by comparison with authentic virion glycoproteins on sodium dodecyl sulfate-poly-acrylamide gel electrophoresis, and by comparison of the glycopeptides of the membrane-associated glycoproteins with the glycopeptides from Sindbis virions on gel filtration chromatography. Our results indicate that glycophospholipid participates in the mannosylation of the viral proteins since an inhibitor of oligosaccharide-lipid synthesis also inhibits the labeling of the glycoproteins.


Resolution of two forms of cytochrome P-450 from liver microsomes of rabbit treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Two forms of cytochrome P-450 with different substrate specificities were isolated from liver microsomes of rabbits treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). A specific antibody was produced toward the major form of the cytochrome. The antibody inhibits microsomal acetanilide hydroxylation (80%). It does not cross-react with the minor fraction of the cytochrome or inhibit the hydroxylation of 3,4-benzpyrene or coumarin, the N-demethylation of aminopyrine or the O-deethylation of 7-ethoxycoumarin catalyzed by rabbit liver microsomes. The major form has an estimated Mr = 54,000 and displays an n-octylamine difference spectrum with an absorption maximum at 426 nm and a minimum at 391 nm. When reconstituted, this cytochrome catalyzes acetanilide hydroxylation at a higher rate than microsomes or the minor fraction. The n-octylamine difference spectrum of the minor fraction displays an absorption maximum at 431 nm and a minimum at 410 nm. When reconstituted, this fraction catalyzes the hydroxylation of 3,4-benzpyrene and the O-deethylation of 7-ethoxycoumarin. The two cytochromes appear to be distinct entities and function in different catalytic pathways.


Kinetic evidence for an acyl-enzyme intermediate in D-alanine carboxypeptidases of Bacillus subtilis and Bacillus stearothermophilus.

The kinetics of hydrolysis and transpeptidation of the synthetic substrate diacetyl-L-lysyl-D-alanyl-D-alanine and of the natural substrate UDP-acetylmuramyl pentapeptide and related compounds catalyzed by the D-alanine carboxypeptidases of Bacillus subtilis and Bacillus stearothermophilus in the presence of the nucleophiles hydroxylamine or glycine have been examined. These kinetic data suggest that an acyl-enzyme intermediate is formed in the first step of the reaction and that the transpeptidation is the consequence of the partitioning of this intermediate between water and the nucleophile in the second step.


Analysis of bioglass fixation of hip prostheses.

The analysis of the bonding interface between hip prostheses and bone after functional use in animals was carried out. Scanning electron microscopy (SEM) with energy dispersive analysis (EDX), and Auger electron spectroscopy were used to evaluate the bonding interface. Various methods of postsacrifice sample preparation were used to evaluate the effect of such different methods on the analysis of the bonding interface. Comparison of the results with several rat tibia implant experiments is also presented.


Reversibly contractile nuclear matrix. Its isolation, structure, and composition.

From Tetrahymena macronuclei we have isolated a reversibly contractile nucleo-skeleton, i.e., an "expanded" nuclear matrix which reversibly contracts when the total concentration of the bivalent cations, Ca and Mg (3:2), is decreased to 5 mM or increased to 125 mM. During contraction the average diameter of the expanded matrix becomes reduced by about 24%; this corresponds to a volume contraction of about 55%. The reversible contraction of the nuclear matrix does not depend on ATP and cannot be inhibited by salygran. The expanded matrix is obtained by removing carefully from the macronuclei 89.7% of the phospholipid, 99.6% of the DNA, 98.5% of the RNA, and 74.8% of the protein by treatment with Triton X-100 and digestion with DNase and RNase followed by an extraction with 2 M NaCl. Electron microscopy reveals, within the expanded matrix, residual equivalents to the structures characteristic for macronuclei: (a) a residual nuclear envelope with nuclear pore complexes; (b) residual nucleoli at the periphery; (c) a fibrillar internal network. The expanded matrix is essentially composed of proteins (96.2%) and traces of DNA (0.8%), RNA (0.5%), phospholipid (1.6%), and carbohydrates (0.9%). The last, which have been determined by gas chromatography, contain glucose, mannose, and an unidentified sugar in the ratio 1:5.4:5.7. The ratio of acidic to basic amino acids of the expanded matrix is 1.55. Sodium dodecyl sulfate (SDS) gel electrophoresis reveals a predominant protein with a mol wt of 18,000 which is apparently involved in the reversible contractile process. The mechanism of this reversible contraction of the expanded matrix remains to be elucidated, but it differs both from actin-myosin contraction systems and from the contractile spasmoneme system in vorticellids.


Intramolecular heterogeneity of mitochondrial DNA of Drosophila melanogaster.

DNA from purified mitochondria of Drosophila melanogaster can be isolated as supercoiled molecules which when nicked have a contour length of 5.9 micron. Partial denaturation mapping shows regional heterogeneity of base composition with one early denaturing region, with a calculated GC content close to zero, extending over 20% of the genome. DNA isolated from unfertilized eggs shows nuclear and mitochondrial DNA in equal proportions; we found no evidence of other cytoplasmic species.


Sulphonated polystyrene as an optimal substratum for the adhesion and spreading of mesenchymal cells in monovalent and divalent saline solutions.

Cell adhesion and spreading were studied on sulphonated polystyrene dishes in serum-free saline (Mn, Na, Cl, buffer) i.e., without an intervening protein layer. Spreading as a function of surface charge density, SCD, peaked around 2-10 negative charges per square nanometer, corresponding to a monomolecular layer of sulphonate ions. At optimal SCD, macrophages, BHK-C13 and whole mouse embryo secondary cells all showed considerable spreading, even in monovalent saline-more so than on a conventional tissue-culture surface. But outside this narrow range of SCD, or on protein-coated surfaces, the divalent cation was indispensable. The biphasic effect of sulphonation on cell adhesion is consistent with the theory that a substratum need not be biochemically specific, provided it is physiochemically polar, rigid and dense. According to this theory, polystyrene of sub-optimal SCD would not be sufficiently polar, while supra-optimal sulphonation would produce a hydrogel surface, lacking in local rigidity and density, due to osmotic swelling. The principle of polymer exclusion, by a surface hydrogel layer, is also consistent with observations on the inhibitory effects of adsorbed proteins-viz., albumin, collagen, serum and cellular exudate, respectively-contrasted with the ready attachment of cells to a bare, optimally charged substratum, in this minimal in vitro system.


Variations in prolactin and growth hormone production during cellular growth in clonal strains of rat pituitary cells.

A permanent, clonal strain of rat pituitary tumor cells (GH3-cells) spontaneously synthesizes and secretes prolactin (rPRL) and growth hormone (rGH) into the culture medium. The rates of hormone production (microng extracellular hormone/mg cell protein/24 hours) and synthesis (vida infra) as well as the rate of [3H]thymidine incorporation into DNA (DNA synthesis) have been studied. During logarithmic growth rPRL and rGH production increased to 160 and 250% of the value at day 2 after plating, while during the plateau phase of cell growth hormone production decreased to initial values. The fluctuations in rPRL production could be fully explained by variations in the rate of rPRL synthesis: [3H]eucine incorporated into rPRL as measured with immunoprecipitation and polyacryl-amide gel electrophoresis. Also the rates of synthesis and production of rGH showed parallel changes during exponential and plateau phase of growth, but this hormone was probably degraded intracellularly. The relative reduction in the rate of synthesis of rPRL and rGH during the plateau of growth corresponded closely to the fall in the rate of DNA synthesis. The reduction in rPRL synthesis could not be explained through an inhibition by extra-cellular rPRL accumulation or by cell to cell interaction occurring in dense cultures. The intracellular concentrations of both hormones were unaltered during logarithmic growth, but rose to 500% for rPRL and 200% for rGH during the plateau phase. In spite of the marked variations in basal rPRL and rGH production the GH3 cultures of different ages were equally able to increase rPRL and decrease rGH production in response to thyrotropin releasing hormone (3 X 10(-7) M) and 17beta-estradiol (10(-8)M).


Cell density-dependent secretion of plasminogen activator by 3T3 cells.

The expression of extracellular fibrinolytic activity in untransformed 3T3 cell cultures depends on the growth state of the cells. Actively growing 3T3 cultures exhibit a relatively high level of fibrinolysis, which decreases progressively as the cells become confluent and density-inhibited. The low level of fibrinolytic activity in confluent 3T3 cultures is due to a diminution in secretion of plasminogen activator since the intracellular level of plasminogen activator remains high. The amount of plasminogen activator observed in growing 3T3 cultures varies depending upon whether the cells are passaged with trypsin/EDTA solution, or with Ca++ selective chelating agent, ethylene-bis (oxyethylenenitrilo) tetraacetic acid (EGTA). However, in cells passaged using either agent, the amount of plasminogen activator secreted is always greatest when the cells are actively growing and decreases thereafter. In contrast to confluent 3T3 cultures, dense cultures of SV40-virus transformed 3T3 cells continued to secrete relatively large amounts of plasminogen activator. The ability to decrease secretion of plasminogen activator as cells become dense may be an important characteristic of cells which demonstrate density-dependent inhibition of cell multiplication in vitro.


Hormonal control of hyaluronic acid production in fibroblasts and its relation to nucleic acid and protein synthesis.

Whole serum and elevated pH previously had been found to stimulate both cell multiplication and hyaluronic acid production by chick embryo fibroblasts in culture. In a study to determine whether cell multiplication and hyaluronic acid production both respond to a single well-defined substance, insulin was found to stimulate, and cortisol to inhibit both processes coordinately. It appears, therefore, that multiplication and differentiated function in fibroblasts respond to a common underlying regulatory signal. Inhibition of ribosomal RNA synthesis by actinomycin D does not prevent serum stimulation of hyaluronic acid production, but inhibition of total RNA synthesis does. If total RNA synthesis is inhibited only after the hyaluronic acid production has reached a new high level, it continues at that level for the next five hours. The stimulatory treatment causes an increase in the activity of the enzyme hyaluronate synthetase. Inhibition of protein synthesis prevents any increase in hyaluronic acid production, and reduces the basal level of production. Reduction of the availability of Mg2+ in the medium coordinately inhibits DNA synthesis and hyaluronic acid production. The results are discussed in the light of a model for coordinate control growth and metabolism based on the availability of Mg2+.


Pituitary autonomy in hyperprolactinemic secondary amenorrhea: results of hypothalamic-pituitary testing.

Twenty-seven women with secondary amenorrhea of greater than six months duration were subjected to multiple testing of hypothalamo-pituitary function. They were divided into normo-prolactinemic (Group 1 mean serum prolactin (PRL) 9.8 ng/ml; range 6.8 to 13.0 ng/ml; n=9) and hyperprolactinemic (Group 2 mean 37.5 ng/ml; range 19.2 to 93.7 ng/ml; n=18) groups on the basis of 4 weekly baseline determinations. Group 2 had significantly (P less than .05) lower serum LH and urinary pregnanediol levels than did Group 1; there was no statistical difference between the groups in serum FSH, T4, T3 or urinary estrogen measurements. Two women in Group 2 were found to have a pituitary chromophobe adenoma. Group 2 women showed no significant rises in serum PRL following stimulation tests with thyrotropin releasing hormone (TRH, 200 microng iv) and metoclopramide (10 mg orally), which caused significant responses in Group 1. The TSH response to TRH was, however, preserved in Group 2, while it was subnormal in Group 1 subjects. Both groups showed similar FSH and LH responses to luteinizing hormone-releasing hormone (LHRH, 25 microng iv). No significant suppression of serum PRL was seen in Group 2 patients given L-Dopa (500 mg orally),, which produced a significant response (P less than 0.05) in Group 1 subjects, while all patient showed marked reduction in serum PRL values following 2-bromo-alpha-ergocryptine (CB-154, 2.5 mg orally). When compared with other Group 2 members, the 2 cases with proven pituitary adenomata gave similar responses to the stimulation-inhibition tests and were not clearly distinguished on this basis. We conclude: 1. The pattern of PRL responses to dynamic tests, although of pathophysiological interest an autonomous pituitary lesions in patients with hyperprolactinemic secondary amenorrhea. 2. Such dynamic tests, although a pathophysiological interest, provide no clinical information additional to that provided by the mean basal serum PRL value. 3. In clinical practice, such dynamic tests should be confined to patients with mean serum PRL levels at around the upper limit of the normal range.


Response to thyrotropin releasing hormone: an objective criterion for the adequacy of thyrotropin suppression therapy.

Most serum thyrotropin (TSH) assays do not adequately discriminate between normal values and absent TSH. We therefore evaluated the TSH response to thyrotropin releasing hormone (TRH) as a criterion for the adequacy of TSH suppression therapy. Twenty-six outpatients with various thyroid disorders (cancer, 10; nodules, 9; miscellaneous, 4; hypothyroidism after 131I therapy for Graves' disease, 3) were studied. Using the frequent sampling technique (samples every 20 min) in two normal volunteers and one untreated patient who was TRH-responsive, we first confirmed the observation that TSH secretion occurred episodically throughout the 24-h period. In contrast, serum TSH was undetectable (less than 0.6 micronU/ml) throughout the 24-h period in 5 patients on TSH suppression therapy who were TRH-unresponsive and one who had a minimal response to TRH. Thus, TRH-unresponsive patients did not secrete measurable amounts of TSH throughout the 24-h period. To suppress TSH secretion, all patients were treated with L-thyroxine (T4) at doses which resulted in undetectable TSH values in random plasma samples. TRH tests were carried out only when random TSH concentrations were less than 0.6 micronU/ml. Seven of the twenty-six patients (27%) including two with thyroid cancer were TRH-responsive indicating a potential for TSH secretion. In these seven, the T4 dose was adjusted until they were TRH-unresponsive. The mean change in T4 dose of these 7 patients was 20+/-10 (SD) microng/day and this resulted in a mean increase of 1.5+/-1.1 microng/dl for T4 and 20+/-20 ng/dl for T3. For all patients, the mean  T4 dose required for TSH suppression was 172+/-53 microng/day or 2.6+/-0.8 microng per day per kg body weight. Twenty-three of 26 patients required between 100-200 microng/day and the remaining 3, 250-300 microng/day. The T4 dose required to suppress TSH resulted in normal serum concentrations of T4. 9.1+/-2.0 MICRONG/DL, AND T3, 136.7+/-33.6 NG/DL. These T4 doses did not produce a rapid heart rate, either awake or asleep, arrhythmias, or electrocardiographic abnormalties as assessed by 24-h Holter monitor tracings in 11 patients. Our results thus show that the T4 dose which results in an unresponsive TRH test ensures that serum TSH will remain undetectable (less than 0.6 micronU/ml) throughout the 24-h period. An unresponsive TRH test, therefore, appears to be a very useful and reliable index of TSH suppression.


Hormonal mechanism of milk secretion in the newborn.

The mechanism inducing milk secretion in 10 puerperal women and in their children (Witch's milk) was studied by means of sequential hormone measurements starting at delivery and continued during the first postpartum week. Determinations included prolactin (PRL), growth hormone (GH), estradiol 17beta (E217beta), and progesterone (PG). Hormonal levels in the newborns closely paralleled those of the mothers. In both groups, the onset of milk secretion coincided with the disappearance of sex steroids from plasma in the presence of elevated PRL concentrations. Response to TRH (8 microng/kg) revealed adequate PRL reserve, and failure of this peptide to modify basal GH in the neonates was suggestive of appropriate hypothalamic control. The role of estrogens in the induction of pituitary hyperplasia during pregnancy is discussed.


Changes in the prolactin response to thyrotropin-releasing hormone (TRH) during the menstrual cycle of normal women.

Serum prolactin (PRL) and thyrotropin (TSH) levels were measured after iv administration of 200 microng of synthetic thyrotropin-releasing hormone (TRH) in 20 normal women ages 18 to 34. Ten women received TRH on days 7 to 8 of the menstrual cycle and 10 women received TRH on days 21-22. Although there was no difference in the dose of TRH relative to body weight in the two groups of women, the peak PRL level after TRH stimulation was greater in the women studied on day 21-22 (48.5+/-5.7 ng/ml, mean+/-SE) than on day 7-8 (35.2+/-4.2 ng/ml) of the cycle (P less than 0.05). In contrast, TSH rose to a greater degree in the preovulatory phase (13.8+/-1.8 micronU/ml) than the luteal phase (7.7+/-0.7 micronU/ml of the cycle (P less than .01). Studies of the PRL and TSH response after TRH administration should take the  phase of the menstrual cycle into account.


Evaluation of the rapid hippurate hydrolysis test with enterococcal group D streptococci.

The rapid hippurate hydrolysis test was evaluated with the conventional test, using 17 group A streptococci, 9 non-enterococcal group D streptococci, 108 enterococcal group D streptococci, and 2 strains of Listeria monocytogenes. There was complete correlation between the rapid and conventional tests with all organisms except enterococcal group D. The rapid hippurate hydrolysis method was more sensitive with the enterococci; 95.4% were positive with the rapid method, and 9.3% were positive with the conventional method. Thin-layer chromatography (TLC) was performed on all isolates to determine if the end product of hydrolysis, glycine, was indeed present. The TLC results were in agreement with the rapid and conventional methods for group A streptococci, nonenterococcal group D streptococci, and L. monocytogenes. TLC results were in total agreement with the rapid hippurate hydrolysis test for the enterococcal group D isolates, thus verifying the accuracy of this more sensitive test. Trace amounts of glycine were found in the substrate, indicating the need for including an uninoculated substrate control as well as stock strains of group A and B beta-hemolytic streptococci (negative and positive controls, respectively) each time the rapid hippurate hydrolysis test is performed.


Smear results in the diagnosis of mycobacterioses using blue light fluorescence microscopy.

Examination of 6,880 sputum specimens from untreated patients disclosed that 3.1% were positive for mycobacteria by fluorescence microscopy, and 92.5% of these has positive cultures. There was a positive correlation between the number of organisms seen on smear and growth on culture. All specimens with positive smears and negative cultures contained rare or few acid-fast bacilli on the smear. Eighty-two percent of the specimens with positive cultures and negative smears yielded less than 25 colonies, whereas there were greater than 25 colonies from 93% of the specimens with positive smears and cultures. In a low prevalence, general hospital population, the primary acid-fast smear continued to be a reliable diagnostic tool.


New tests for characterization of mumps virus antibodies: hemolysis inhibition, single radial immunodiffusion with immobilized virions, and mixed hemadsorption.

Hemolysis inhibition (HLI), single radial immunodiffusion (SRID) with immobilized virions, and mixed hemadsorption tests were used for measuring antibodies against mumps virus. Rabbit hyperimmune sera against mumps and early and late human convalescent sera were analyzed. All three tests identified antibodies against both hemagglutinin and the second major envelope component, hemolysin (fusion factor). The sensitivity of the HLI test corresponded to that of the hemagglutination inhibition (HI) test, but in some sera HLI antibodies occurred in greater quantity than HI antibodies. The SRID test readily identified rises in antibody titers in connection with acute infection. Due to its simplicity and lack of sensitivity to nonspecific inhibitors, it is recommended for use in this context. The mixed hemadsorption test showed a high sensitivity for specific identification of mumps antibodies. It therefore may be suitable for use in screening for immunity to mumps.


Isolation and characterization of a rough colony type of Neisseria gonorrhoeae.

A new colony type of Neisseria gonorrhoeae was detected in the primary cultures from 8 of 180 men with gonococcal urethritis. This colony type contrasts with those previously described by having a rough and irregular surface. In six of the eight cases, the rough form predominated. The distinctive morphology of the rough colony variant could be maintained indefinitely by selective subculture. By electron microscopy, organisms taken from rough colonies of each of the eight isolates were piliated. Antimicrobial susceptibilities of type 1 and rough clones derived from the same patients were identical for ampicillin, penicillin, tetracycline, and spectinomycin. After inoculation of rough colonies into subcutaneous chambers in mice and guinea pigs, type 1 colonies predominated in cultures of material obtained from the chambers. This new piliated colony type of N. gonorrhoeae may provide an opportunity to investigate factors other than pili that contribute to gonococcal virulence.


Simplified media for isolating Neisseria gonorrhoeae.

Two simplified media for isolating Neisseria gonorrhoeae are described. They differ from the modified Thayer-Martin medium in the enrichment and antibiotics used for preparation. The enrichment for one, enrichment 4 medium, contains only four ingredients, and amphotericin B is used instead of nystatin. This medium is comparable to the modified Thayer-Martin medium for isolating N.gonorrhoeae. It is convenient to prepare and is only one-third the cost of Thayer Martin medium. It is a suitable alternative to the modified Thayer-Martin. The enrichment 5 medium is a hemoglobin-free version of enrichment 4 medium. It is somewhat more selective for contaminants but is also more inhibitory for N. gonorrhoeae.


Cellular immunity in chronic Chagas' disease.

The cellular immune response was assessed in 20 patients with chronic Chagas' disease (American trypanosomiasis). Thymus-derived lymphocyte function was determined in vivo by cutaneous reactivity to several antigens including a soluble preparation derived from Trypanosoma cruzi and sensitization to 2,4-dinitrochlorobenzene. The in vitro T-cell reactivity was investigated by the proliferative response to phytohemagglutinin and to T. cruzi antigen and by inhibition of leukocyte migration with the specific antigen. In addition, the proportion and absolute numbers of peripheral blood T and B-lymphocytes were determined by rosette formation. This research indicates that the general and specific cellular immune response, evaluated by the tests herein mentioned, is well preserved in patients, with Chagas' disease. We conclude that chronic Chagas' disease is not associated with deficiency in cellular immunity, nor does it lead to it. Conceivably, the active participation of delayed hypersensitivity may play an important role in the expression of the human chagasic lesions.


Effects of Gaseous CO2 and bicarbonate on the growth of Neisseria gonorrhoeae.

The quantitative and qualitative growth response of Neisseria gonorrhoeae strains was assessed under the following conditions: incubation in a candle jar (approximately 2.5% ambient CO2) on medium without bicarbonate, incubation in air on medium without bicarbonate, incubation in air on medium with bicarbonate, and incubation in air in a sealed jar on medium with bicarbonate. Incubation in the presence of ambient CO2 (candle jar) resulted in the highest plating efficiencies for the three laboratory strains 9, 62, and 2686. The addition of NaHCO3 to the medium enhanced the growth response in air of all three strains, particularly if incubation was carried out in a closed environment (sealed jar). The qualitative growth response of 34 clinical isolates and laboratory strains was assessed under the same conditions of incubation after the plating of an inoculum containing approximately 2 X 10(6) bacteria. The strains were divided into different classes based on their growth responses. About 40% of the strains grew as well on bicarbonate-containing medium incubated in air in sealed jars as they did on medium without bicarbonate incubated in a candle jar. Ten percent of the strains showed only slight growth on bicarbonate containing medium incubated in sealed jars and appeared to have an almost obligate requirement for ambient CO2. Twenty percent of the strains apparently had partially lost their requirement for gaseous C02 and showed slight growth in air on medium without bicarbonate and slight to moderate growth in air on medium containing NaHCO3. The remaining 30% seemed to have completely lost their requirement for gaseous C02 and/or the bicarbonate anion and grew almost as well in air on medium without bicarbonate as they did in either ambient C02 (candle jar) or on medium containing bicarbonate incubated in a sealed jar. These results suggest that N. gonorrhoeae strains may vary widely in their requirements for CO2 and/or the HCO3-anion. Incubation in the presence of ambient CO2 tends to maximize the growth response on solid medium of those strains, which require it for growth. The presence of ambient CO2 is particularly important if growth is to be obtained after the plating of small inocula. Medium conatining 0.1% NaHCO3, if incubated in a closed environment (sealed jar), apperas to be equivalent to medium without bicarbonate incubated in ambient CO2 in supporting the growth of some but not all strains of N. gonorrhoeae.


Screening for cross-reacting capsular polysaccharide K antigens of Escherichia coli using antiserum agar.

Agar plates containing antiserum against group B meningococcus or Haemophilus influenzae type b were used to determine the prevalence of cross-reacting K1 and K100 capsular polysaccharide antigens in 265 isolates of disease-causing Escherichia coli. K1 antigen was found in 22% of isolates from various sites. K100 antigen was found in only three isolates. This technique is a convenient method to detect specific E. coli K antigens for evaluation as possible factors important in the virulence of the organism.