[Action mechanism of sodium polyacrylates against stomach ulcer]. Three sodium polyacrylates (PAS) with different viscosities were examined for antiulcerogenic properties. Rats were fed with sugar containing a PAS for 4 hr and subjected to 12 hr reserpinization or stress by water immersion. Lower incidence of ulceration and larger amount of stomach content after the experiment were found in the PAS administered groups. The higher viscous PAS was more effective in prevention of ulceration and in prolongation of gastric emptying. Intragastric administration of the drugs to pylorus ligated rats reduced forestomach ulceration. However, differences in the antiulcerogenic activity of three drugs were not observed. The effects on the gastric secretion of three PAS (50 mg) were also examined, using pylorus ligated (6 hr) rats. In PAS groups, the free acid in the gastric juice was reduced to some extent. There was, however, no relation between the antiacid effect and viscosity. The drugs inhibited the peptic digestion in vitro, but the difference of viscosity was not related to the antipeptic effect. It is concluded that prolongation of gastric emptying may be a major factor in the antiulcerogenic activity of PAS.


The effect of fatty acids and ketone bodies on the biosynthesis of insulin in isolated pancreatic islets of obese hyperglycemic mice. Collagenase-isolated islets from obese hyperglycemic mice were used to study the effects of fatty acids and ketone bodies on insulin biosynthesis. The biosynthetic rate was quantitated as the incorporation of 3H-leucine into proinsulin and insulin, which were separated from the crude islet homogenates with anti-insulin serum coupled to cyanogen bromide activated Sepharose 4B. Palmitate and oleate did not stimulate the incorporation into proinsulin and insulin while 16.7 mM glucose caused a nine-fold increase of 3H-leucine incorporation during a 3 h incubation as compared to the basal buffer alone. A weak stimulatory effect was seen with 5 mM octanoate. The ketone bodies had a weak stimulating effect on islet protein biosynthesis, but no preferential stimulus to the insulin biosynthesis was found.


Evidence for the D-cell of the pancreas secreting somatostatin. By immunofluorescence, somatostatin-, glucagon- and insulin-containing cells are localized in serial sections of the pigeon pancreas. The distribution of the somatostatin immunofluorescent-cells corresponds to that of the D-cells (A1-cells), which are particularly numerous in this animal species. This observation, coupled with the finding of D, A and B-cells at the ultrastructural level, indicates that the D-cell is responsible for the secretion of somatostatin.


A naturally occurring Cu-thionein in Saccharomyces cerevisiae. A naturally occurring monodisperse Cu-thionein was prepared using ammonium sulfate precipitation followed by ion exchange (DEAE 23) and gel chromatography (Sephadex G-75). The chromatographic steps were repeated at least twice, or until the Cu-thionein remained homogeneous when subjected to analytical polyacrylamide disc electrophoresis. The molecular weight of this copper protein was 9500+/-500. Up to 24.3% cysteine residues were determined, indicating the relationship to the metallothioneins. Aromatic amino acids were virtually absent, while there were about three times as many acidic amino acid residues, including aspartate and glutamate, as in metallothioneins. 10 g atoms of Cu were measured per mole of protein. The copper binding strength of thionein was extremely high. Displacement by protons (pH 1.5) and gel chromatography or dialysis employing EDTA were not effective. Dialysis against diethyldithiocarbamate produced a protein essentially free of copper. Both the ultraviolet properties and the circular dichroism measurements proved identical with those properties reported for artificially prepared Cu-thionein (see ref.[1]. The major absorption was in the far ultraviolet region with a weak shoulder at 270 nm attributable to copper charge-transfer transititions. 6 Cotton extrema were seen at 213, 283 and 302 nm (negative) and 245, 328 and 359 nm (positive). The possible role of Cu-thionein as an electron transport system was discussed.


Metabolic transformation of colchicine, III. Inhibition of phosphatases by the metabolite O10-demethylcolchicine (colchiceine) and their reactivation by divalent cations. Colchicine and some of its metabolites and derivatives were investigated regarding their inhibitory action against alkaline phosphatases from E. coli and calf intestine, and against acid phosphatase from potato. Colchicine itself was essentially ineffective, also its metabolites O2-demethyl- and O3-demethylcolchicine. The metabolite O10-demethylcolchicine (= colchiceine) was found to be a powerful, non-competitive inhibitor for all three phosphatases with Ki in the range of 0.45-6.0mM. Amongst the derivatives of colchicine, colchiceyltaurine proved to be a non-competitive inhibitor of acid phosphatase (Ki=3.0mM); desacetylcolchiceine behaved like colchiceine. The inhbitory actions of colchiceine and desacetylcholchiceine could be completely reversed by the addition of Mg2, Ca2, or Zn2, respectively. This interaction with divalent cations demonstrates a novel type of action of the metabolite colchiceine when compared with its mother alkaloid and is possibly of major biological significance.


Interactions of concanavalin A with isolated pancreatic islets. Pancreatic islets of mice were isolated by the collagenase method. After a preincubation period of 20 min they were incubated in Krebs-Ringer-albumin buffer in the presence of glucose. Concanavalin A (2 mg/ml) inhibits the glucose-induced release of insulin; this same property is shown by Concanavalin A bound to nonphagocytosable beads of Sephrose at lower concentrations (0.4 mg/ml). In pancreatic islets stimulated by tolbutamide, glibenclamide or arginine the secretion of insulin is not inhibited in the presence of Concanavalin A. The incorporation of tritiated leucine is not influenced by Concanavalin A. Since desialized pancreatic islets react in the same way the glucose-induced secretion of insulin appears to be a membrane-dependent process.


Thyroglossal duct carcinoma.

Thyroglossal duct carcinoma. Primary carcinoma of the thyroglossal duct is rare. This discussion reports two cases and reviews the 50 previously reported in the literature. The criteria for diagnosis include evidence of a thyroglossal duct remnant and a normal thyroid gland. The differentiation from cystic metastases to lymph nodes is pointed out. The histologic types parallel those of carcinoma of the thyroid gland, papillary carcinoma being the most common and having a generally favorable prognosis. The clinical presentation of these tumors is similar to that with benign cysts and thus is of limited value in the diagnosis.


Recent developments in foetal haemoglobin research. There have been numerous new contributions to the knowledge of foetal haemoglobin over the last few years. It is, therefore, timely to review them together. They throw light on the arrangement on the chromosome of non-alpha chain genes, and on the condition generally known as Hereditary Persistence of Foetal Haemoglobin (HPFH) and have contributed to other aspects of human ontogeny and physiology.


Apparent "in situ" clone of cytogenetically marked ataxia-telangiectasia lymphocytes. Six adjacent metaphases, each with the same cytogenetic aberration of a group D chromosome, most probably a No. 14, were observed in a field of a slide from a 96-hour culture of lymphocytes from an individual with ataxia-telangiectasia (AT). None of the 304 other metaphases examined from this or other simultaneous cultures of this individual showed such an aberration. It seems most likely that an "in situ" marked clone has been observed and this supports interpretation of consistent cytogenetic abnormalities in those with AT as having clonal origin. The method of slide preparation employed which involves placing, rather than dropping, the cell suspension on the slide may facilitate detection of "in situ" clones.


[Possibilities of a quality control in the diagnosis of tuberculosis (author's transl)]. A method is described which allows a quality control for demonstrating mycobacteria in the diangosis of tuberculosis. It does not test the differentiating procedures, but the manner of culturing and concentrating the bacteria by applying a statistical evaluation of serial experiments, thus allowing the statistically proven determination of the bacterial count. For this an evaluation method given by Cavalli-Storza was employed.


[Cellular immune reaction in acute and chronic viral hepatitis (author's transl)]. The course of viral hepatitis is markedly influenced by the immunologic response to the infective agent. The immune defense of viral diseases is specially connected to an intact function of T lymphocytes. Investigation of T cell function during acute and chronic viral hepatitis showed an altered immune response, recognizable from the relative number of T lymphocytes, the affinity and ability of lymphocytes to be stimulated by phytohemagglutinin and the demonstration of in vivo activated lymphocytes. Although the interpretation of some of these findings is still difficult, the study of the various cellular immune reactions permits a better understanding of the pathogenesis and course of viral hepatitis.


The potentiality of antibody-producing cells. I Bispecific cell occurrence in double stimulated cultures of syngeneic or allogeneic spleen cells of the mouse. Cultures of spleen cells from Swiss, C57Bl or DBA/2 mice stimulated with 2,4,6-trinitrophenyl (TNP) conjugated sheep erythrocytes (SRBC) were used for studying the in vitro responses to TNP and native SRBC antigens and the frequency of occurrence of cells responding to both antigens. The response was revealed by plating the cultured cells with both native SRBC and TNP-conjugated pigeon erythrocytes (TNP--PRBC). Specific responses were obtained in all the cultures. Bispecific haemolytic plaque-forming cells (PFC) were detected in almost all cultures of individual Swiss mice cells with a frequency of 2-5--14 PFC/10(6) cells recovered. In DBA/2 cell cultures bispecific PFC were found in half the cultures (2-5--8-3/10(6) cells) and in C57Bl cell cultures in 30 per cent of the cultures (7--21/10(6) cells). When cells from individual Swiss mice immunized in vivo with TNP--SRBC were as an allogeneic culture from the 2nd day after immunization in the presence of TNP--SRBC, the frequency of bispecific PFC increased from 8 to 30/10(6) cells. Mixed allogeneic cultures of normal C57Bl and DBA/2 cells yielded high specific responses with regular occurrence of bispecific PFC only when the numbers of cells cultured together was small. However, when allogeneic cells were mixed 24 hours after starting the cultures, all responses were stimulated and bispecific PFC were found in considerable numbers (4--33/10(6) cells). Cross-reactivity between TNP--PRBC and native SRBC antigens was studied by culturing cells with each of the antigens and plating the cells with both, or by immunizing in vivo with SRBC or PRBC and culturing the cells with both antigens from the 2nd to the 5th day after immunization with both antigens. In no instance did bispecific PFC exceed background levels (0-1--0-6/10(6) cells) in these control experiments.


DNA synthesis in rabbit spleen explants. Its relationship to cell division. The synthesis of DNA, as measured by the incorporation of [3H]-thymidine into the nuclei of cells of cultured explants of rabbit spleen, has been studied and correlated with estimation of DNA content of these nuclei using Feulgen densitometry, with and without colcemid arrest. Observations of the proportion of nuclei labelled with [3H]thymidine and their DNA content indicated that many cells were incorporating this precursor for DNA synthesis without the concomitant increases in DNA expected of cells in division cycle. Colcemid arrest failed to produce the changes in DNA content expected of a cell population as rapidly dividing as the labelling kinetics suggested. The results indicated that [3H]thymidine-labelled material, possibly DNA, was being transferred between cells and this accounted for the increased labelling index seen after pulsing with [3H]thymidine. The suppression of [3H]thymidine uptake by hydroxyurea indicated that this was not repair synthesis of damaged DNA.


Effects of mitogens for mouse B lymphocytes on chicken lymphoid cells. Lipopolysaccharide from E. coli (LPS) and purified protein derivative from tuberculin (PPD) increased the [3H]thymidine incorporation of chicken spleen cells in culture. No such stimulation was observed with dextran sulphate. Thymus and bursa lymphocytes were not stimulated by any of these compounds. Spleen cells from chickens chemically bursectomized with cyclophosphamide treatment showed decreased responses to LPS and PPD, but responded normally to the T-cell mitogen concanavalin A. None of the tested mitogens induced polyclonal antibody synthesis or directly enhanced the primary antibody response to sheep erythrocytes (SRBC) in spleen cell cultures. LPS-coated SRBC, however, enhanced the in vitro antibody response to SRBC. The results suggest a moderate proliferative response of chicken lymphocytes to LPS and PPD, possibly involving B cells, but not further effects comparable to those on mouse B lymphocytes.


Non-specific and DNP--erythrocyte rosette formation after contact sensitization with DNCB. DNA--guinea-pig erythrocyte rosette-forming cells (RFC), as found after contact sensitization of guinea-pigs with DNCB, were shown to be highly sensitive to pretreatment with cyclophosphamide or X-irradiation, which suggests that these cells are B cells. The finding was confirmed by rosette-blocking experiments using an anti-immunoglobulin serum. It has already been shown that they are not directly related to antibody production. Possibly they represent precursors of antibody-producing cells. An increase in rosette formation, using non-antigenically related erythrocytes, was shown to parallel development of cell-mediated immunity (CMI) in the lymph nodes. These non-specifically induced RFC were also found to be B cells. Data were obtained which make it improbable that either proliferation or influx of B cells during development of CMI plays a major role in this increase. An alternative mechanism is suggested, while a possible role for these non-specifically induced RC is proposed.


Dosage adjustments of cefazolin according to the pharmacokinetics of this new cephalosporin. Serum and urine levels of cefazolin were studied: 1. following a single 500mg intra-muscular dose of the drug in normal subjects (n = 10), in patients with renal impairment (n = 12) and in patients undergoing maintenance hemodialysis (n = 10); 2. following a single 500 mg intra-venous dose of the drug in normal subjects (n = 10). In normal subjects, serum half-lives averaged 1.75 hour after intravenous injection (Ke = 0.395) and 2.22 hours after intra-muscular injection (Ke = 0.312); urinary recovery of cefazolin over a six hours period amounted to 61% after intra-venous injection and to 47.5% after intra-muscular injection. The renal clearance of the drug approximated 40 ml/mn. A linear correlation (Ke = 0.022 + 0.0028 CrCl) was established between the overall elimination rate-constants and the creatinine clearances in the subjects under investigation. The various pharmacokinetic constants thus obtained can be used to calculate the maintenance doses, loading doses and dosage intervals adjusted according to creatinine clearances. These data actually yield dosage regimes adapted to each individual case according to the degree of renal function.


[Antibody detection in infections with Serratia marcescens (author's transl)]. A report is given on the detection by agglutination and precipitation of antibodies against homologous strains in infections with Serratia marcescens. The results of the investigation indicate that antibody formation is likely if invasive forms are present. The agglutination titers were between 1:40 and 1:640. It can be presumed that, in addition to the patient's antibodies against the O antigen, antibodies are also formed against certain antigens with the character of ferments or toxins.


Properties of lymphocytes in chronic HBAg-positive hepatitis. An approach to pathogenetic problems. Lymphocytes from 22 children affected by chronic hepatitis were studied and a comparison was made with a control group of 32 healthy children in the same age range with respect to PHA dose-response curves. E-rosette formation and membrane immunoglobulin (Ig). From the results it appeared that PHA dose-response curves were displaced to the left (low PHA dose), DNA synthesis was significantly higher for all PHA dosages tested, and that the plasma of chronic hepatitis patients acted neither as inhibitor nor stimulator. The percentage of E-rosette-forming cells decreased, but the total number was normal. The mean percentage of Ig-bearing cells was normal but the single values were spaced over a wide range. It can be assumed that, at least in the chronic stage, there is an enhanced response to PHA and an increased percentage of cells without markers either for T or for B cells.


[Studies on the in vitro effects of cephalothin and gentamicin, alone and in combination, on proteus mirabilis and enterococci (author's transl)]. A combined antibiotic therapy is only useful in a few precisely defined clinical pictures where testing of the chemotherapeutics administered is required to determine their characteristics of action (antagonism, indifference, synergism) on the isolated organism. For the initial therapy of critical acute infections, simultaneous administration of cephalothin and gentamicin proved to be valuable. In the present study, the efficiency of these chemotherapeutics alone and in combination was investigated in a quantitative serial dilution test and with the membrane-filtration method. Thirty strains of Proteus mirabilis and enterococci showed only low sensitivity to the antibiotic alone. In combination, whereby gentamicin was at a constant level comparable to in vivo serum levels, an increase of bacteriostatic and bactericidal action could be demonstrated, especially for Proteus mirabilis. The antibacterial spectrum, the molecular-biological mode of action, clinical experience and possible side-effects of the cephalotin-gentamicin combination are discussed.


The need for orthodontic treatment: a periodontist's point of view. Data at hand suggests that occlusal traumatism is not causally related to gingival inflammation. Nor does occlusal traumatism affect the severity of existing inflammation. While an effect of occlusal traumatism on the spread of inflammation has been postulated, most observations seem to question such an interaction. Malposition of teeth has not been proven to be an etiologic factor in inflammatory periodontal disease nor could the degree of periodontal disease severity be consistently associated with improper tooth position. While malposed teeth may make plaque removal more difficult, specific oral hygiene measures should overcome this handicap. Orthodontic appliances may act as local gingival irritants, either directly or as plaque accumulants, and should be designed to minimize such irritations. Finally, orthodontic treatment, by properly aligning tilted or drifted teeth which also exhibit infraosseous lesions, may improve periodontal therapy at such sites.


A social perspective on need and demand for orthodontic treatment. In addition to needs fostered by actual physical disabilities resulting from malocclusion, need and demand for orthodontic treatment may develop entirely apart from any consideration of health needs. In fact, improvement of physical function is not the primary motivation of many persons who receive orthodontic care. A variety of social factors are related to perceived needs for acceptable dentofacial appearance and to public demand for orthodontic services. Since early times human teeth have been the object of mutilations to satisfy sociocultural expectations. The mutilation, and even removal of teeth, to achieve dental cultural norms in, so-called, primitive societies may appear barbaric to an ethnocentric society. However, a significant sociocultural need can be met equally by the African or Brazilian who has had teeth filed to a point and by the Western adolescent who having had four teeth surgically removed to provide space in the arch then undergoes two years of orthodontic therapy to reposition and straighten the remaining teeth. From a sociological point of view, the need and desire of members of a society to achieve a culturally acceptable body image constitutes neither frivolity nor luxury. If children's teeth do not naturally meet the norm for esthetic dental appearance, the response of families to sociocultural expectations and pressures produces a culturally valid need for orthodontic intervention. Other social factors affecting perceived need and demand for orthodontic care are socioeconomic, desire for upward social mobility, and social change with regard to public attitudes toward the availability of dental health services and the increasing removal of financial barriers. Potential changes in present US orthodontic practice may be required if care is to be delivered to significantly larger segments of the population.


Organization of the Navy Dental Corps.

Organization of the Navy Dental Corps. The US Navy Dental Corps was established by an act of Congress in 1912. Its membership consists of dental officers of the Regular Navy and Naval Reserve. The primary mission of the Corps is to prevent or remedy dental conditions that may interfere with the performance of duty by members of the active naval forces. The organizational structure within which the Dental Corps functions is discussed at some length. The Chief of the Dental Corps, under the Surgeon General, is responsible for the planning and direction of programs providing dental care to Navy and Marine Corps personnel and others authorized by law. Until recently, the majority of dental personnel received only technical assistance control from the Bureau of Medicine and Surgery; but in consonance with the goals of improving management effectiveness and achieving efficient use of personnel and material resources, the Navy Dental Corps has undertaken a program of regionalization of its facilities. The establishment of regional dental centers under one command permits immediate response to the needs of populations and the activities they serve, while at the same time reducing layering of administrative and fiscal burdens of existing dental departments at various commands, which allows more dental officers to devote their time to dental health care delivery.


Leukocyte migration inhibition in nickel dermatitis. Leukocyte migration inhibitory factor assay was employed as an in vitro diagnostic aid in nickel dermatitis, the second most common contact dermatitis in North America. 15 patch test-positive and 5 patch test-negative patients, all giving a past history suggestive of nickel dermatitis, were investigated. Significant inhibition of leukocyte migration in both groups of patients was obtained only with nickel sulfate-albumin conjugate and not with unconjugated nickel sulfate. Specificity of this system was tested by utilizing an unrelated metallic albumin complex, and no inhibition was found. When patch testing is equivocal or contraindicated, this in vitro technique may be a practical alternative.


Production of homocytotropic antibody against acid extracts isolated from group A streptococcal cell walls in intact and adrenalectomized rats. Intact and adrenalectomized rats of male Sprague-Dawley were injected with acid-extracted materials isolated from group A streptococcal cell walls using aluminum gel as an adjuvant. We have clearly demonstrated the production of homocytotropic antibodies against this antigens in rats. However, the homocytotropic antibody titers in adrenalectomized rats were much higher than those in intact rats.


Antiserum specific for reticulin of the bursa of fabricius. Antiserum produced by immunizing rabbits with embryonic chicken spleen cells showed, after appropriate absorption with cell suspensions, specificity for fetal spleen cells in membrane immunofluorescence tests. However, when the original antiserum was absorbed with tissue homogenates and tested on cryostat tissue sections, it reacted specifically with the reticular framework which separates the cortical and medullary lymphocytes in the follicles of the adult bursa of Fabricius. It is suggested that the reticulin may influence bursal lymphoid differentiation.


Ontogenetic aspects of sexual dimorphism and the primary immune response to sheep erythrocytes in hamsters from prepuberty through senescence. Ontogeny of the primary response to sheep erythrocytes of age-matched groups of male and female hamsters was studied at various chronological ages ranging from prepuberty to senescence. Selected organs were likewise weighed upon sacrifice to obtain developmental patterns. Adrenal weights were higher in the male, and pituitary weights were higher in the female; for both organs typical dimorphism was demonstrable by 36 days. Spleen weight and index favored the female by 46 days. Immunological sex dimorphism first appeared in groups injected at 53 days and autopsied at 58 days and persisted through senescence. Sexual dimorphism of antibody-mediated immunity, previously shown to favor the female in both the primary and secondary immune responses, thus follows the dimorphism of total amount of splenic lymphoid tissue and occurs shortly after realization of sexual maturity in the male. These findings support our previous suggestion of the suppressive effect of androgens on the antibody-mediated immune responsiveness of the male hamster.


Epoxides metabolically produced from some known carcinogens and from some clinically used drugs. I. Differences in mutagenicity. The epoxide metabolites of two clinically used drugs and an experimental psychotropic agent, carbamazepine 10,11-oxide, cyproheptadine 10,11-oxide and cyclobenzaprine 10,11-oxide, were fully devoid of any mutagenic activity under conditions where K-region-epoxide metabolites of some known carcinogens, such as benzo(a)pyrene, proved to be potent frameshift mutational agents for Salmonella typhimurium TA 1537 and TA 1538. All epoxides tested were non-mutagenic for TA 1535, designed to detect substitution mutations. The 10,11-epoxides of the three drugs, carbamazepine, cyproheptadine and cyclobenzaprine, were not cytotoxic to any of the bacterial tester strains used, precluding that mutagenicity might have been overshadowed by cytotoxicity. When the mutagen, precursor, benzo(a)pyrene, was incubated together with TA 1537 and a mammalian microsomal preparation in the presence of a system generating the co-factor necessary for mono-oxygenase activity, activation to mutagenic species was observed which was dramatically increased in the presence of a potent epoxide hydratase inhibitor, 1,1,1-trichloropropene 2,3-oxide, suggesting epoxide(s) as the (or one of the) mutagenically active species metabolically produced in situ. None of these effects was observed with the three medical drugs. Moreover, the observation that the alkene oxide 4-phenylstyrene 7,8-oxide is mutagenic to the two strains TA 1537 and TA 1538 but the K-region arene oxide derived from 7,12-dimethylbenz(a)anthracene is inactive for the latter strain indicates that epoxidation of an aromatic double bond of a polycyclic hydrocarbon is neither a necessary nor a satisfying condition for frameshift mutagenesis to occur.


The development of the nursing labor force in the United States: a basic analysis. In this article, we draw together some basic material on the objectives character of the nursing labor force in the United States. First, we present a statistical and descriptive picture of the nursing labor force as it exists today. Then we analyze the historical development of that labor force up to about 1950, stressing its general integration into a hierarchical and concentrated health industry. Finally, we look at the development of the nursing labor force over the past 25 years in terms of factors such as sex, race, age educational credentials, labor force participation, place of work, and the hierarchical division of labor, discussing the general implications of the changes in these factors for worker organization in the health industry. Our main conclusion is that the nursing labor force is in a kiey position to lead a movement for the restructing of the health industry to serve the health needs of the American people rather than the profit and power requirements of the health employers. The primary, and difficult, task ahead is to build labor organization which will have the vision and unity to work this goal.


The women's health movement.

The women's health movement. This article provides a brief overview of the history, current status, and future directions of the Women's Health Movement. The links between feminism and women's health issues are drawn and reasons for their simultaneous occurrence are suggested. It is shown that health issues are seen by the Movement has suspended in the larger social context from which they arise.


The politics of birth control, 1920-1940: the impact of professionals. Before the 1920s, a birth control movement arose in the United States out of socialist, feminist, and other radical groups concerned with women's rights and sexual freedom. After 1920 the birth control movement became gradually transformed into a respectable, nonradical reform cause, the recipient of large grants from big business, with women's rights secondary to an overriding concern with medical health and population control. This transformation was achieved through the professionalization of the birth control movement-that is, its takeover by professional experts, almost all male, in place of the radical amateur women, fighting for their own interests, who initiated it. The article examines two groups of professionals who were particularly influential in this transformation: doctors and academic eugenists. The former made birth control a medical issue, held back the development of popular sex education, and stifled a previously developing feminist approach to women's birth control needs. The later contributed racism to the birth control movement, helping to transform it into a population control movement with racist and anti-feminist overtones. Both groups, while they made contributions to the technology of contraception, simultaneously held back the spread of birth control by transforming the campaign for it from a popular, participatory cause to a professional staff lobbying operation.


Sexual surgery in late-nineteenth-century America. This article describes how the tendency of post-Civil War medicine accorded with general social tendencies. Doctors' attitudes toward women reflected their anxiety about female emancipation and changing sex roles. The specialty of gynecology emerged in significant part as a reaction to female emancipation. The strong surgical emphasis in American gynecology practice has continued. The operations, clitoridectomy and female castration, were intended to reimpose the traditional sexual order (the term "castration" and "female circumcision" implied women had become men). The paper goes on to describe the beginnings of psychosomatic medicine, the circularity of gynecologists' treatment of women, and the effects of this treatment. The operations were in part the expression of a symbiotic relation between doctor and patient. Because they expressed widespread and deep-seated beliefs, they were difficult to challenge.


Denaturation of thermophilic and mesophilic 6-phosphogluconate dehydrogenase by 8M urea. The denaturation reaction of 6-phosphogluconate dehydrogenase (6-phospho-D-gluconate: NADP oxidoreductase (decarboxylating EC 1.1.1.44) from B. Stearothermophilus and E. coli in 8 M urea has been compared. The rates of denaturation were evaluated from the fluorescence quenching that occurs at 334 nm. The thermophilic enzyme has been found about ten times more resistant to the action of the denaturating agent, and showed a biphasic denaturation process, whereas the E. coli enzyme followed a single first-order kinetics.


Treatment of prostatic cancer with Estracyt (estramustine phosphate). The experience gained with Estracyt, kindly supplied by AB LEO, Sweden, is reported. On the basis of former data in the literature we only used the drug in estrogen resistant and advanced cases. Estracyt (estramustine phosphate) is a nitrogen mustard derivative of the urethan type, attached to oestradiol-17-phosphate. In histologically verified cases, it was administered in daily doses of 300 mg intravenously for three weeks, followed by maintenance doses of 300 mg a week in tablets for three months. During treatment, liver and bone marrow function was checked systematically. The changes in morphological picture were studied by means of biopsies during and at the end of treatment. In agreement with the data in the literature a favourable effect was observed in estrogen resistant patients, with no toxic effect whatever on the bone marrow. At the same time GOT and GPT and BSP retention examinations demonstrated a hepatotoxic side effect. The pathological values returned to normal after withdrawal of the drug. Histological examinations showed that the tumour cells had changed but failed to disappear after treatment.


Ara-A and IDU therapy of human superficial herpetic keratitis. Ara-A ointment was compared to IDU ointment in patients with dendritic herpes simplex virus infection of the corneal epithelium. Twenty-eight patients were treated with Ara-A ointment and twenty-four with IDU ointment. The lesions healed in 5.1 days with Ara-A and in 6.9 days with IDU. This drug was given in a double-controlled manner, so that neither the patient, nor the investigator knew which drug the patient was receiving. The patient groups were comparable as to length of the dendritic lesion and duration of symptoms. The adverse reactions to each of these drugs were comparable and in no case was there any permanent ocular change from drug use.


Use of a platelet-fibrinogen-thrombin mixture as a corneal adhesive: experiments with sutureless lamellar keratoplasty in the rabbit. A platelet-fibrinogen-thrombin mixture utilizing autologous platelets was studied for its potential as a corneal adhesive. In the rabbit it demonstrated sufficient adhesive properties to allow 50 per cent of lamellar keratoplasties (autotransplants) to remain in place without the use of sutures. The mixture retains significant adhesive properties for four to six days. It is simple to prepare and apply. It also appears nonantigenic and nontoxic to the cornea; it does not incite inflammation, nor interfere with corneal clarity or the regrowth of corneal epithelium.


Studies on the effectiveness of liver and bacterial beta-glucuronidase enzymes in the hydrolysis of urinary beta-glucosiduronides of some tryptophan metabolites. The effectiveness of Escherichia coli and bovine liver beta-glucuronidases in the hydrolysis of the urinary beta-glucosiduronides of tryptophan metabolites was studied. Moreover, the effect of the prolonged contact of these conjugates to the urinary enzyme was investigated in the first and second voiding urine samples. It is found that both enzymes have no important role in releasing the free carcinogens from their glucosiduronides. The presence of free carcinogens could be attributed to the spontaneous hydrolysis of some labile conjugates. However, the prolonged contact of the freely active substances during the sleeping hours with the epithelium of the bladder may enhance the process of bladder carcinogenicity. The increased accumulation of these metabolites in the first voiding urine could be interpreted in terms of their rate of excretion rather than by the enzymatic hydrolysis of their conjugates.


Effect of microbial substances of different origins on the growth of Mycobacterium leprae. Mycobacterium leprae multiplied in media enriched with substances originating from other mycobacteria, from non-acid-fast Actinomycetales or from gram-positive or gram-negative Eubacteriales. Most of the M. leprae strains did not grow on a synthetic medium containing the amino acids present in M. smegmatis, but the growth-promoting effect of sonic extracts of this organism indicated that substances of bacterial origin, other than amino acids, do act as growth factors. The identification of the growing, acid-fast microorganisms with M. leprae was verified by their ability to oxidize D-3,4-dihydroxyphenylalanine (D-dopa test). M. leprae did not multiply on Nakamura medium unless at least 40% of medium NM7 enriched by bacterial substances was present. Adequate aeration was essential for multiplication of M. leprae in enriched NM7 and No. 3 media.


Diuresis and improved renal hemodynamics produced by prostaglandin E1 in the dog with norepinephrine-induced acute renal failure. Unilateral acute renal failure was induced in dogs by renal artery infusion of norepinephrine. Renal hemodynamics and function were studied before and 5 days after norepinephrine infusion, and during subsequent renal artery infusion of prostaglandin E1. After norepinephrine, the infused kidney exhibited decreased urine volume, sodium excretion, and creatinine clearance. Cortical blood flow, determined with radioactive microspheres, also decreased. No redistribution of intracortical blood flow occurred. Arteriographically, proximal vessels were constricted and the cortical nephrogram density diminished. During prostaglandin E1 infusion, urine volume, sodium excretion, and cortical blood flow increased. Creatinine clearance did not change significantly. Blood flow was redistributed from the outer third to the inner third of the cortex. Proximal vessels were dilated and the cortical nephrogram density was enhanced. The apparent dissociation of cortical blood flow from creatinine clearance suggests that prostaglandin E1-induced diuresis and natriuresis result from a mechanism other than increased glomerular filtration.


LH and FSH response to synthetic LH-RH in children and adolescents with Turner's and Klinefelter's syndrome. Stimulation with LH-RH (luteinizing hormone-releasing hormone) was performed in 29 children and adolescents with Turner's syndrome (15 less than 13 years old), and in 11 patients with Klinefelter's syndrome (7 before puberty). Synthetic LH-RH was injected intravenously in a dose of 25 mug/m2; plasma LH and FSH were determined radioimmunologically. The results show that patients with Turner's syndrome had pathologically elevated LH and FSH values at all ages. Normal LH and FSH results before and after LH-RH were found in 3 patients with X0/XX karyotype who had spontaneous menstruation. The 2 adult patients with Klinefelter's syndrome showed an exaggerated LH response and excessively high FSH levels before and after LH-RH, consistent with hypergonadotropic hypogonadism. Six out of 7 prepubertal boys with Klinefelter's syndrome, had a normal LH and FSH response to LH-RH. Only 1 prepubertal boy with cryptorchidism and genital maliformation in addition to the Klinefelter's syndrome had pathological values. The normal gonadotropin results in prepubertal patients with Klinefelter's syndrome show that their testes apparently loose the negative feedback activity on gonadotropin secretion only during puberty, at a time when tubular hyalinization appears. This is in contrast to Turner patients with gonadal dysgenesis who have hypergondadotropic hypogonadism already early in childhood.


Plasma gonadotropin (LH and FSH) concentrations in repubertal and pubertal children upon stimulation by LH-releasing hormone. The simultaneous determination of LH and FSH were carried out in a single plasma sample by double antibody radioimmunoassays in a total of 26 prepubertal and pubertal children. The blood samples were taken before and after i.v. application of 25 mug synthetic LH-RH. Plasma LH and FSH concentrations found in prepubertal boys and girls at beginning of the test and at the time of maximum response were compared with those measured in pubertal boys and girls. The mean basal levels of LH and FSH are in group I (8 prepubertal girls): LH 0.6+/-0.7 (SD) mIU/ml, FSH 1.5+/-0.8 mIU/ml; in group II (6 prepubertal boys): LH 1.8+/-1.0 mIU/ml, FSH 2.0+/-0.8 mIU/ml; in group III (6 pubertal girls): LH 4.1+/-2.0 mIU/ml, FSH 4.7+/-2.8 mIU/ml; in group IV (6 pubertal boys): LH 3.5+/-2.3 mIU/ml, FSH2.0+/-1.2 mIU/ml. The mean maximum response of LH and FSH after stimulation are in group I: LH 4.1+/-2.3 mIU/ml, FSH 11.7+/-3.7 mIU/ml; in group II: LH 4.2+/-1.0 mIU/ml, FSH 6.0+/-1.1 mIU/ml; in group III: LH 19.7+/-10.6 mIU/ml, FSH 12.8+/-5.7 mIU/ml; in group IV: LH 13.4+/-5.7 mIU/ml, FSH 3.8+/-0.9 mIU/ml. It is suggested that without stimulation of the pituitary with LH-RH, plasma LH and FSH levels are not reliable parameters to test the pituitary gonadotropin reserve.


Immunofluorescence demonstration of avidin in the immature chick oviduct epithelium after progesterone. Immature chicks were used for the experiments 1-2 days after hatching. A group of chicks were injected with 5 mg of progesterone and sacrificed 1 or 24 hr later. An other group of chicks were injected daily for 9 days with 5 mg of diethylstilbestrol and thereafter with single injection of progesterone. Cryostat sections were incubated with rabbit anti-avidin serum and with fluorescein labelled antirabbit globulin for fluorescence histochemistry. In the control animals no epithelial cells of the oviduct were fluorescence positive independently whether or not the animals were pretreated with diethylstilbestrol. One hour after administration of progesterone epithelial cells showed occasionally a slight synthesis of avidin. 24 hours after the injection of progesterone most, however never all, of the epithelial cells showed avidin in the apical part of the cell. The fluorescence reaction was clearly more intense if the animals were estrogen-primed. Diethylstilbestrol caused a differentiation of oviductal glands which were, however, only occasionally avidin positive after progesterone. These results suggest that primitive oviductal cells can produce avidin without preceding differentiation whereas estrogen causes a differentiation of new line of cells which have regularly lost their capacity of avidin synthesis after progesterone administration.


Fluorescamine as a histochemical reagent: demonstration of polypeptide hormone-secreting cells. Fluorescamine is a useful fluorescence microscopic reagent for the demonstration of certain peptide hormone-secreting cells in formaldehyde-fixed tissues. Among the cells demonstrated are the pituitary GH cells, the gastrin cells, the insulin cells and the thyroid C cells. In the latter cell system degranulation brings about a marked decrease in the intensity of the fluorescamine-induced fluorescence suggesting that the fluorogenic component is associated with the secretory granules. In models neither of the hormones tested--GH, gastrin, insulin--gave fluorescamine-induced fluorescence after pretreatment with formaldehyde. It is suggested that the fluorescamine-induced fluorescence of the endocrine cells demonstrates granular components distinct from the hormones.


Distribution and properties of glucagon immunoreactivity in the digestive tract of various mammals: an immunohistochemical and immunochemical study. Numerous epithelial cells in the oxyntic gland area of the stomach of cat and dog appear to store pancreatic-type glucagon, as evidenced by immunohistochemistry and--in the case of the dog--also by radioimmunoanalysis. No glucagon immunoreactive cells are found in the antropyloric mucosa. Disseminated cells in the intestinal mucosa store gut-type glucagon. In all species studied, these cells predominate in ileum and colon.


Canine salmonellosis: prevalence, epizootiology, signs, and public health significance. Of 53 salmonella serotypes isolated from dogs, S typhimurium and S anatum have been the most commonly isolated. Surveys have disclosed that over 20% of the dogs in a population may be infected. Simultaneous, multiple infections with 2 or more serotypes are not unusual. Nonclinical salmonellosis occurs in most cases. The severe form of the disease is manifested by diarrhea, vomiting, fever, depression, abortion, and death. Dogs may remain carriers and fecal shedders and thus serve as sources of salmonellosis for man and other animals. A number of documented transmissions from dogs to human beings have been recorded. Such infections in man have been severe.


The cellular changes during osteogenesis in bone and bone marrow composite autografts. Osteogenesis in and around autografts has been studied in the hope of identifying the osteoblast precursor. Fresh autografts of cortical bone with its constituent bone marrow were implanted into intramuscular sites in rats. The grafts were removed at intervals over a four week period and examined by light and electron microscopy. A chronological sequence of degenerative, reparative and differentiative events was found and described. The relative contributions of the host and graft components to post-grafting osteogenesis are discussed and the problem of the origin of the osteoblast cells is examined, taking into account present views about the interrelationships between skeletal connective tissue cells and their precursors. It is concluded that, in the investigation described, the presence of host granulation tissue was necessary for osteogenesis and it is postulated that osteoblast progenitor cells can arise from this granulation tissue, their differentiation along the osteogenic line being stimulated in some way by the presence of the graft bone.


The ultrastructure of the anterior pituitary gland of the vole, Microtus agrestis, in normal and experimentally manipulated animals. The ultrastructural appearance of the various types of cell present in the anterior pituitary of the vole has been described. There was a great measure of similarity between the cytological picture in this species and in the rat. Prolactotrophs contained the largest secretory granules, which were of variable shape; the granules of somatotrophs, whilst only slightly smaller than those of prolactotrophs, were invariably round, and of more uniform size; corticotrophs were represented by cells which were extremely angular, and whose secretory granules, besides being smaller than those of somatotrophs, were arrayed around the periphery of the cell below the plasma membrane; gonadotrophs contained granules of a similar size to those found in cortiocotrophs, but were found throughout the cytoplasm of the cells, whic were round to ovoid in shape; thyrotrophs contained the smallest granules of all, the shape of the cell itself bein angular...


The fate of heterotopic isografts and allografts of mouse yolk sac epithelium. As a contribution to the problem of the immunological privilege of the fetus it was decided to investigate the fate of yolk sac grafts in mice. Ten-day post-coitum mouse visceral yolk sac was transplanted to a dermal tunnel in the flank of isogeneic animals. In this site the yolk sac was capable of surviving for at least 15 days past the normal duration of pregnancy. Yolk sac was transplanted to a similar site in allogeneic animals and was found to be destroyed by an immunological reaction within 13 days. Orthotopic skin grafts between the same strains of mice used in the yolk sac transplantation were rejected in first-set fashion within 13 days. A limited series of host animals, previously sensitized by visceral yolk sac, rejected orthotopic skin grafts in second-set fashion. These findings indicate that the cells of the visceral yolk sac in the mouse contain transplantation antigens. Possible reasons why such antigens do not lead to fetal rejection during pregnancy are considered.


Rapamycin (AY-22,989), a new antifungal antibiotic. I. Taxonomy of the producing streptomycete and isolation of the active principle. A streptomycete was isolated from an Easter Island soil sample and found to inhibit Candida albicans, Microsporum gypseum and Trichophyton granulosum. The antibiotic-producing microorganism was characterized and identified as Streptomyces hygroscopicus. The antifungal principle was extracted with organic solvent from the mycelium, isolated in crystalline form and named rapamycin. Rapamycin is mainly active against Candida albicans; minimum inhibitory concentration against ten strains ranged from 0.02 to 0.2 mug/ml. Its apparent activity against Microsporum gypseum and Trichophyton granulosum is lower because of its instability in culture media on prolonged incubation required by these fungi. No activity was observed against gram-positive and gram-negative bacteria. Acute toxicity in mice is low.


Rapamycin (AY-22,989), a new antifungal antibiotic. II. Fermentation, isolation and characterization. Rapamycin is a new antifungal antibiotic produced by Streptomyces hygroscopicus NRRL 5491. It was isolated from the mycelium by solvent extraction, purified by silica gel column chromatography and crystallized as a colorless solid which melts at 183 approximately to 185 degrees C and has the empirical formula C56H89NO14. From its characteristic ultraviolet absorption spectrum rapamycin can be classified as a triene. It is highly active against various Candida species, especially Candida albicans. Its activity is compared with that of amphotericin B, candicidin and nystatin.


A glyoxalase I inhibitor of a new structural type produced by Streptomyces. Many streptomyces strains produced an inhibitor of crude glyoxalase prepared from rat liver which did not inhibit glyoxalase I prepared from yeast. Another inhibitor, C11H14O6, which inhibited glyoxalases prepared from both rat liver and yeast was obtained from a cultured broth of Streptomyces griseosproeus and crystallized. Preincubation of this inhibitor with reuduced glutathione increased its inhibitory activity, which suggested its reaction with reduced glutathione. It showed a strong inhibition of growth of HeLa cells and inhibition of Ehrlich ascites carcinoma by daily injection. It also showed weak inhibition of the solid type of Ehrlich carcinoma and prolonged the survival period of mice inoculated with L-1210 cells.


Characteristics of two broad spectrum antibiotics produced by Mycoplasma sp. RP III. Two extracellular antibiotics produced by Mycoplasma sp. RP III growing in serum-containing media have been purified by solvent extraction and ion-exchange chromatography. Factor I inhibits Gram-positive and Gram-negative bacteria in vitro and a crude preparation protected mice from Pseudomonas and Straphylococcus infections; it is of low cytotoxicity. Factor II, a lipid, inhibits Gram-positive and Gram-negative bacteria, Candida species, and is as cytotoxic as the actinomycins.


Platomycins A and B I Taxonomy of the producing strain and production, isolation and biological properties of platomycins. Two new antibiotics, platomycins A and B, belonging to the phleomycin-bleomycin family, were isolated from the culture filtrate of Streptosporangium sp. MK-78. This strain has been identified as a new variety of Streptosporangium violaceochromogenes. Both platomycins A and B are active against a variety of Gram-positive and Gram-negative bacteria, and also inhibit solid Sarcoma 180 and Ehrlich ascites carcinoma.


Transduction of drug resistance to tetracycline, chloramphenicol, macrolides, lincomycin and clindamycin with phages induced from Streptococcus pyogenes. Strains of Streptococcus pyogenes isolated from pediatric patients with acute infections which were resistant to one or more of the antibiotics, tetracycline (TC), chloramphenicol (CP), macrolide antibiotics (erythromycin, kitasamycin, oleandomycin, josamycin), lincomycin (LCM) and clindamycin (CLM), were used for transduction of drug resistance. These drug-resistant strains were treated with mitomycin C to induce phages and transduction of drug resistance was attempted by means of phages so induced. It was found that transduction of resistance to the above antibiotics was possible. The transductants obtained on TC-containing selective agar plate were resistant to TC alone while those produced on CP- or erythromycin (EM)-containing selective agar plate were resistant to CP, macrolide antibiotics (Mac), LCM and CLM. From this finding, it was inferred that transduction of resistance to TC, CP, Mac, LCM and CLM via phages occurred in two different patterns, i.e., transfer of resistance to TC alone and that of resistance to CP, Mac, LCM and CLM. All of the transductants obtained were found to belong to group A. In T-typing, they were of the same T-12 type as the donor and recipient strains in a majority of cases though some were not typable.


Distribution volumes of [131I]albumin, [14C]sucrose, and 36Cl in sheep lung. We measured the steady-state volumes of distribution for radioactive chloride, sucrose, and albumin in the lung of six anesthetized, spen-thorax sheep. We allowed 2 days for [131I]albumin to equilibrate throughout the body, 2 h for the 36Cl, and a 40-min constant infusion for [14C]sucrose before killing the animal. Five minutes before death, we gave [125I]albumin to tag lung plasma volume. We killed the animals by clamping both lung hila; we then removed the lungs and homogenized them. We measured residual red cell and plasma volume, total extravascular lung water, and the extravascular content of the three tracers. The distribution volumes expressed as fractions of blood-free lung weight were: 36Cl equals 0.44, sucrose equals 0.28, and albumin equals 0.07. If the sucrose distribution volume is taken as the best estimate of the lung's extravascular extracellular space, then chloride clearly overestimates the interstitial fluid volume, being either bound or partially intracellular. On the other hand, albumin clearly underestimates the interstitial fluid volume.


Rapid estimation of pulmonary extravascular water with an instream catheter. An arterial catheter-bearing external conductivity electrodes and a thermistor was used for measurement of lung thermal volume (LTV) by the double-indicator method. Ten milliliters of 3% saline at room temperature were injected, dilution curves measured, and LTV calculated as mean transit time difference, less thermistor time constant, times cardiac output (CO). Comparisons were made, in dogs, between LTV, pulmonary extravascular lung water with Evans blue and tritiated water (PEVWtho), and weighed lung water (WLW). Pulmonary edema was induced with dextran and epinephrine. CO was measured by thermodilution in both the pulmonary artery (PA) and aorta (AO) and dye dilution in the AO. CO from dye dilution was compared with thermodilution (aortic detection) to detect irreversible loss of thermal indicator. Comparisons showed good correspondence of dye and thermal curves (Y = 0.91X - 0.16 1/min; r = 0.93). LTV is about 120% of WLW in near normal lungs, 90% of WLW in extreme edema. PEVWtho was 60-70% WLW.


Mycoplasma hominis: growth, reproduction, and isolation of small viable cells. Improved methods for studying the growth of Mycoplasma hominis (ATCC 14027) have been developed, involving modified growth conditions and preparation of the organisms under minimally distorting conditions. Cells so prepared from batch cultures show relatively uniform exponential growth and appear to be dividing by binary fission; but pleomorphic forms appear upon further incubation. Similar behavior was demonstrated by another laboratory-adapted strain and by three clinical isolates, and therefore seems characteristic of the species. The pleomorphic populations contain small forms having diameters within the 100- to 250-nm size range reported for "elementary bodies." Such forms were isolated from this strain of M. hominis by sequential filtration using gravity alone, after cell aggregates were dispersed by Pronase treatment. Of the small bodies which traversed membranes of 220-nm pore size, a negligible number grew in liquid or on solid media, suggesting that these were not essential reproductive units in a life cycle, but involution forms due to growth in an altered environment.


Yeast sterol esters and their relationship to the growth of yeast. Variation in the percentage of sterols esterified to long-chain fatty acids during cellular growth has been examined. Under all conditions, a constant percentage of sterol esters was maintained during exponential growth. This maintenance level was found to vary with different growth conditions. A sharp increase in the rate of esterification was observed upon entry of the culture into the stationary growth phase. The minor cellular sterol components were found to accumulate after this period of rapid sterol ester synthesis, with a relative decrease in the size of the ergosterol pool. Evidence is presented that sterol esters of ergosterol precursors are unable to be metabolized to ergosterol. Once esterified, the fatty acids do not appear to be scavenged during starvation conditions.


Inhibition of cell division in Escherichia coli K-12 by the R-factor R1 and copy mutants of R1. The effect of the copy number of plasmid R1drd-19 on cell division of Escherichia coli K-12 was studied in populations growing as steady-state cultures at different growth rates, the growth rate being varied by use of different carbon sources. The plasmid copy number was also varied by using copy mutants of the R-factor. The mean cell size was larger in populations carrying an R-factor than in R-factorless populations, an effect that was more pronounced at low growth rates and in populations carrying R-factor copy mutants. The increased cell size was due to formation of elongated cells in a fraction of the population and to an increase in the diameter of all cells. The majority of the cells divided at a normal cell length, but the presence of an R-factor caused some cells to elongate, probably by the uncoupling of chromosome replication and cell division. This can be explained as a competition between the chromosome and plasmid replicons for some replication factor(s), presumably acting on both initiation and elongation of replication. The formation of elongated cells was a reversible process, but occasionally some of the elongated cells reached lengths 20 times that of newborn cells. If cell division did not occur at the normal cell size, the septum was not formed until the cell size was four times that of a newborn cell. When an elongated cell divided, it usually formed a polar septum, thus producing a newborn cell of normal cell length. The ability of plasmid-containing cells to omit one cell division but to retain the capacity of dividing one mass doubling later is compatible with a mechanical model for septum formation and cell division.


Plasmid incompatibility and control of replication: copy mutants of the R-factor R1 in Escherichia coli K-12. Plasmid incompatibility was studied in Escherichia coli K-12. By double-antibiotic selection, clones were constructed that carried the two R-factors R1 and R100, both belonging to the compatibility group FII. After release of the selection pressure, each of the two plasmids was lost at the same rate (8% per generation). Mutants of R-factor R1 showing an increased number of copies per chromosome (copy mutants) were tested for their incompatibility towards R-factor R100. The results indicate that plasmid incompatibility is quantitative and not just a qualitative property. All copy mutants studied affected incompatibility, and there were two classes of mutants: one increasing and one decreasing the incompatiblity exerted towards the test plasmid R100. Evidence is presented that incompatibility is related to the mechanisms that control replication. The implications of such a relation on proposed models for control of replication are discussed. The data do not support the hypothesis that plasmid incompatibility is due to competition for a replicational or segregational site.


Further characterization of a non-essential mutator gene in Escherichia coli K-12. The properties of mutR, a mutator closely linked to thyA, have been further characterized. We have found that the mutator gene is carried on a specialized transducing phage (lambdapcI857 thyA) generated by the excision of lambdacI857 integrated at a secondary attachment site between lysA and thyA. We present three lines of evidence indicating that mutR is a nonessential gene. (i) Deletions of the mutator can be found amoung survivors of heat induction of lambdacI857 when the phage is integrated between lysA and thyA. (ii) Mutations in mutR can be induced with the frameshift mutagen ICR-191. (iii) An amber mutant in mutR has been found. Viable strains could be made by combining the mutator with polB, polA polR, ligts7, and uvrA mutations. The mutator was still able to increase the spontaneous mutation frequency in these genetic backgrounds. When the reversion patterns of a series of well-characterized trpA mutations were analyzed, the results suggested that mutR is more efficient at causing transitions than transversion mutations.


Chromosome-plasmid interaction in Escherichia coli K-12 carrying a thermosensitive plasmid, Rts1, in autonomous and in integrated states. An Hfr strain of Escherichia coli K-12 was obtained by integrative suppression with a thermosensitive plasmid, Rts1. The R plasmid was integrated into the chromosome between rif and thr, and transfer of the chromosome occurred counterclockwise. The thermosensitivity of host cell growth due to the dnaA mutation was markedly but not completely reduced in this integratively suppressed Hfr strain. When the dnaA mutation was removed by transducing the dnaA+ genome to this Hfr, the thermosensitivity of cell growth due to existence of Rts1 was suppressed in contrast to strains carrying it autonomously. Thermosensitivity of cell growth appeared again when the plasmid was detached from the chromosome to exist autonomously. Contrary to the effect on cell growth, the transfer of the chromosome and the plasmid itself and the ability to "restrict" T-even phages were still thermosensitive in all of these strains carrying Rts1, irrespective of its state of existence. The detached plasmid as well as the original Rts1 were segregated upon growth at 42 C. These data are discussed in relation to chromosome-plasmid interaction. One of the most important conculusions is that some plasmid genes, related to their replication, are phenotypically suppressed by the chromosome when it is integrated.


Incorporation of pantothenate into citrate lyase by a pantothenateless mutant of Klebsiella pneumoniae. A pantothenate-requiring mutant of Klebsiella pneumoniae was isolated. The mutant showed an absolute dependence on pantothenate for growth. When grown in the presence of [14C]pantothenate, the mutant incorporated [14C]pantothenate into citrate lyase (3.4 mol/mol of enzyme). Analysis of a double-labeled enzyme ([14C]pantothenate and [3H]acetate) by gel electrophoresis in sodium dodecyl sulfate showed that both 3H and 14C were associated solely with the smallest subunit, the acyl carrier protein of citrate lyase.


Factors affecting the palmitoyl-coenzyme A desaturase of Saccharomyces cerevisiae. The activity and stability of the palmitoyl-coenzyme A (CoA) desaturase complex of Saccharomyces cerevisiae was influenced by several factors. Cells, grown nonaerobically and then incubated with glucose, either in air or under N2, showed a marked increase in desaturase activity. Cycloheximide, added during such incubations, prevented the increase in activity, suggesting de novo synthesis. The stability of the desaturase from cells grown nonaerobically was affected by subsequent treatment of the cells; enzyme from freshly harvested cells, or from cells that were then shaken under nitrogen, readily lost activity upon washing or during density gradient analysis, whereas aerated cells, in the presence or absence of glucose, yielded stable enzyme preparations. The loss of activity in nonaerobic preparations could be reversed by adding soluble supernatant from these homogenates and could be prevented by growing the cells in the presence of palmitoleic acid and ergosterol, but not with several other lipids tested.


Pyrimidine biosynthetic enzymes of Salmonella typhimurium, repressed specifically by growth in the presence of cytidine. The repressive effects of exogenous cytidine on growing cells was examined in a specially constructed strain in which the pool sizes of endogenous uridine 5'-diphosphate and uridine 5'-triphosphate cannot be varied by the addition of uracil and/or uridine to the medium. Five enzymes of the pyrimidine biosynthetic pathway and one enzyme of the arginine biosynthetic pathway were assayed from cells grown under a variety of conditions. Cytidine repressed the synthesis of dihydroorotase (encoded by pyrC), dihydroorotate dehydrogenase (encoded by pyrD), and ornithine transcarbamylase (encoded by argI). Moreover, aspartate transcarbamylase (encoded by pyrB) became further derepressed upon cytidine addition, whereas no change occurred in the levels of the last two enzymes (encoded by pyrE and pyrF) of the pyrimidine pathway. Quantitative nucleotide pool determinations have provided evidence that any individual ribo- or deoxyribonucleoside mono-, di-, or triphosphate of cytosine or uracil is not a repressing metabolite for the pyrimidine biosynthetic enzymes. Other nucleotide derivatives or ratios must be considered.


Isolation, characterization, and mapping of Escherichia coli mutants blocked in the synthesis of ornithine decarboxylase. Several Escherichia coli K-12 mutants blocked in the synthesis of ornithine decarboxylase (OD) were isolated after transduction for serA+ in a strain (MA197) blocked in agmatine ureohydrolase (AUH) with a mutagenized phage lysate of P1. The new double-polyamine mutants were characterized by an unconditional polyamine dependence; either putrescine or spermidine was required for normal growth. The mutational block was varified by the demonstration of a virtual absence of OD activity in cellular extracts. The mutation, designated speC, was mapped by P1 transduction in several strains and was shown to have a cotransduction frequency of 17.2% with serA. Map order was established as serA speB speC metK. A derivative of one of the OD mutants having wild-type levels of AUH and blocked in OD was utilized along with an OD AUH mutant and an OD+ AUH strain to explore the phenomenon of "pathway selection" using growth rate as a parameter. Polyamine pool studies were carried out simultaneously. The results presented here support the hypothesis of pathway selection, implying a preferential utilization of exogenous arginine rather than endogenously produced arginine in polyamine biosynthesis.


Tryptophan synthetase alpha(5.7-S): novel molecular species formed within Escherichia coli. A novel molecular species contributes about 5% of the total tryptophan synthetase of Escherichia coli derepressed for the trp operon enzymes. The new species is identified under conditions in which the dissociation of the two nonidentical subunits of the tryptophan synthetase complex is favored. The new species sediments at 5.7S, catalyzes the conversion of indole-3-glycerol phosphate to indole, and has been designated alpha(5.7-S). Although alpha(5.7-S) is not observed in extracts of trpA or trpB mutant strains deficient in the ability to form tryptophan synthetase alpha or beta2 subunits, respectively, a mixture of the two extracts allows the formation of alpha(5.7-S). Similar results are obtained when a homogeneous alpha protein is mixed with an extract of a trpA mutant strain, suggesting that the interaction of alpha and beta2 proteins is obligatory for alpha(5.7-S) formation. One can obtain a beta2 protein preparation that when mixed with a pure alpha protein gives no alpha(5.7-S). Therefore, the interaction of alpha and beta2 proteins alone is not sufficient for the formation of alpha(5.7-S). When a mixture of alpha and beta2 proteins devoid of alpha(5.7-S) is added to extracts of trp deletion mutants, the novel species can be reconstituted in vitro only when deletions are used that carry at least the operator-proximal part of the trpB gene. Therefore, it is concluded that the alpha(5.7-S) species of tryptophan synthetase results from the interaction of the alpha protein, the beta2 protein, and a third component, beta', specified by the deoxyribonucleic acid defined by the end points of two trp deletion mutants.


Effect of temperature on the size of Escherichia coli cells. The distributions of cell volumes of steady-state Escherichia coli ML30G cultures at various temperatures were measured. For cultures in a minimal medium, the distributions were indistinguishable at several temperatures between 15 and 30 C; at higher temperatures the cells were slightly smaller, and at lower temperatures they were slightly larger. For cultures in a complex medium, the cells were slightly larger at both high and low temperatures of growth. An abrupt change of temperature within the middle range led to a transient change in the distribution of cell volume, suggesting that the size of dividing cells is well regulated. No synchrony of division was induced by a change in temperature.


Proteinase activities of Saccharomyces cerevisiae during sporulation. Sporulation in Saccharomyces cerevisiae occurs in the absence of a exogenous nitrogen source. Thus, the internal amino acid pool and the supply of nitrogen compounds from protein and nucleic acid turnover must be sufficient for new protein synthesis. Since sporulation involves an increased rate of protein turnover, an investigation was conducted of the changes in the specific activity of various proteinases. A minimum of 30% of the vegetative proteins was turned over during the course of sporulation. There was a 10- to 25-fold increase in specific activity of various proteinases, with a maximum activity around 20 h after transfer into the sporulation medium. The increase in activities was due to de novo synthesis since inhibition of protein synthesis by cycloheximide blocks both an increase in proteinase activities and sporulation. There was no increase observed in proteinase activities of nonsporogenic cultures (a and alpha/alpha strains) inoculated into the sporulation medium, suggesting that the increase in proteinase activities is "sporulation specific" and not a consequence of step-down conditions. The elution patterns through diethylaminoethyl-Sephadex chromatography of various proteinases extracted from T0 and T18 cells were similar, and no new species was observed.


Polypeptide nature of growth requirement in yeast extract for Thermoplasma acidophilum. The active component(s) in yeast extract required by Thermoplasma acidophilum for growth is polypeptide in nature. A fraction from yeast extract was isolated and partially characterized as one or more peptides of molecular weight about 1,000 containing 8 to 10 amino acids. Although it was composed largely of basic and dicarboxylic amino acids, only one amino group per molecule was free. The polypeptide(s) appeared to bind avidly to cations. No other organic compounds except glucose were needed by Thermoplasma. Among several hundred known compounds tested, only glutathione plus Fe2+ or Fe3+, clostridial ferredoxin, and spinach ferredoxin elicited any growth response.


Enzymatic lesions in methionine mutants of Aspergillus nidulans: role and regulation of an alternative pathway for cysteine and methionine synthesis. In Aspergillus nidulans the pathway involving cystathionine formation is the main one for homocysteine synthesis. Mutants lacking cystathionine gamma-synthase or beta-cystathionase are auxotrophs suppressible by: (i) mutations in the main pathway of cysteine synthesis (cysA1, cysB1, and cysC1), (ii) mutations causing stimulation of cysteine catabolism (su101), and (iii) mutations in a presumed regulatory gene (suAmeth). A relative shortage of cysteine in the first group of suppressors causes a derepression of homocysteine synthase, the enzyme involved in the alternative pathway of homocysteine synthesis. A similar derepression is observed in the suAmeth strain. Homocysteine synthesized by this pathway serves as precursor for cysteine and methionine synthesis. A mutant with altered homocysteine synthase is a prototroph, indicating that this enzyme is not essential for the fungus.


Comparison of the cell envelope structure of a lipopolysaccharide-defective (heptose-deficient) strain and a smooth strain of Salmonella typhimurium. The cell envelope structure of Salmonella typhimurium LT2, which has a heptose-deficient lipopolysaccharide (LPS), is significantly different from that of an isogenic strain with a normal LPS. The rough strain, when examined by freeze-etching, lacks most surface structures that are routinely present in the smooth strain (surface particles and flagella) and has few transmemberane studs in the cytoplasmic membrane (those present are generally found in aggregates), and the outer membrane cleavage is substantially stronger than that of the smooth strain. These envelope differences were independent of both growth temperature and culture age. Examination of ultrathin sections indicated that the rough strain has an outer membrane which forms a much more defined double-track artifact than the smooth strain. The addition of MgCl2 to the growth medium of the rough strain decreased the extent of outer membrane cleavage, and flagella became evident in freeze-etched preparations. The presence of supplemental MgCl2 in the growth medium, which resulted in these morphological changes in the rough strain, also produced growth at a previously restrictive temperature and a decrease in the leakage of periplasmic enzymes. The smooth strain was unaltered morphologically or physiologically by MgCl2 under identical conditions. It is suggested that the outer membrane of the rough strain is more planar.


Outer membrane of Salmonella typhimurium: chemical analysis and freeze-fracture studies with lipopolysaccharide mutants. The outer membrane layer of the cell wall was isolated from wild-type Salmonella typhimurium LT2 as well as from its mutants producing lipopolysaccharides with shorter saccharide chains. Chemical analysis of these preparations indicated the following. (i) The number of lipopolysaccharide molecules per unit area was constant, regardless of the length of the saccharide side chain in lipopolysaccharide. (ii) In contrast, in "deep rough" (Rd or Re) mutants producing the lipopolysaccharides with very short saccharide chains, the amount of outer membrane protein per unit surface area decreased to about 60% of the value in the wild type. (iii) In the wild type, the amount of phospholipids is slightly less than what is needed to cover one side of the membrane as a monolayer. In comparison with the wild type, the outer membrane of Rd and Re mutants contains about 70% more phospholipids, which therefore must be distributed in both the outer and inner leaflets of the membrane. Freeze-fracture studies showed that the outer membrane of Re mutants were easily fractured, but fracture became increasingly difficult in strains producing lipopolysaccharides with longer side chains. The convex fracture face was always nearly smooth, but the concave fracture face or the outer half of the membrane was densely covered with particles 8 to 10 nm in diameter. The density of particles was decreased in Re mutants to the same extent as the reduction in proteins, suggesting the largely proteinaceous nature of particles. A model for the supramolecular structure of the outer membrane is presented on the basis of these and other results.


On the processive mechanism of Escherichia coli DNA polymerase I. A procedure has been developed to assess whether polymerization of nucleotides by DNA polymerases is processive, that is, whether a succession of polymerization steps occurs without release of the enzyme from the template. The method involves measurement of the ratio of deoxyguanylate to deocycytidylate incorporated in the course of replicating a segment of the right hand cohesive end of phage lambaDNA with the sequence 5'-G-G-G-C-G-G-C-G-3'. In the case of the Escherichia coli DNA polymerase I, each enzyme molecule completes synthesis of the sequence before dissociation occurs. Furthermore, at both 6 and 22 degrees, the polymerase remains bound to the lambdaDNA template after synthesis has completed. Template challenge experiments, in which the polymerase is allowed to begin synthesis in the presence of a molar excess of lambdaDNA before addition of a very large excess of calf thymus DNA, show that under the conditions used, productive binding of polymerase to lambdaDNA is a slow process requiring 1 to 2 hours. After synthesis has been completed, polymerase remains bound to the lambdaDNA in spite of the availability of new primer termini. The association, polymerization, and dissociation rates measured in these experiments suggest that the polymerization reaction catalyzed by DNA polymerase I is processive, and that hundreds of nucleotides may be polymerized between each association and dissociation.


Kinetic analysis of Escherichia coli deoxyribonucleic acid polymerase I. The kinetic properties of Escherichia coli DNA polymerase I were simplified to those of a 1 deoxynucleotide substrate reaction by the use of polynucleotide templates. With poly(dA)-oligo(dT) as the template-primer complex, Mg2+ decreases the Km of the substrate dTTP but has little or no effect on the Km of the substrate Mg-dTTP, suggesting that multiple pathways involving the binding of Mg2+, dTTP, and Mg-dTTP are operative in forming the active complex. The Km of free Mg2+, extrapolated to zero concentration of substrate (830 = 62 muM), agrees within a factor of 2 with the dissociation constant of magnesium from 4 +/- 1 sites on the enzyme determined previously by binding studies (Slater, J.P., Tamir, I., Loeb, L.A., and Mildvan, A.S. (1972) J. Biol. Chem. 247, 6784-6794). The maximal turnover number with poly(dA) as template is 5.7 +/- 0.7 s-1. Changing the nature of the base in the polydeoxynucleotide template alters the maximal rate of polydeoxynucleotide synthesis by an overall factor of 31 with poly(dC) is greater than poly(dT) is greater than poly(dA) is greater than poly(dG), indicating that pyrimidine templates are copied faster than purine templates. Changing the sugar structure from poly(dA) to poly(rA) causes a 3-fold increase in the rate of template copying. A study of the kinetic effects of all noncomplementary deoxynucleotides with all deoxynucleotide templates, as well as with poly(rA)-oligo(dT), yields complex patterns of activation and inhibition requiring from 1 to 2 additional binding sites for the noncomplementary nucleotides. The kinetically determined affinities of the active site of the enzyme-template-primer complex for the complementary free nucleotide (as measured by Km) generally exceed those for the noncomplementary neuclotides (as measured by KI slope) by 1 or more than 3 orders of magnitude.


Inhibition of growth of microbial mutants by trans-octadecenoates. A series of positional isomers of trans-octadecenoic acid were tested for their ability to support growth of microbial mutants that could not synthesize unsaturated fatty acids. The bacterial strain used (a variant of Escherichia coli 30E) grew with supplements of the trans isomers only at high temperatures (38 degrees) and with acids containing the trans-ethylenic bond between carbon atoms 8 through 13. The yeast mutant (Saccharomyces cerevisiae KD46) grew only with the 9-trans-octadecenoate giving cell yields about one-fifth those obtained with oleate. Although the trans isomers had little effect on the growth of the bacteria in the presence of oleate, they inhibited the growth of yeast with oleate. Inhibition was strongest for the 4, 6, 7, 11, and 12 isomers, almost negligible for the 8 isomer and of differing intermediate degrees for the others. The inhibitory effects had no correlation with the melting points of the acids and appeared to reflect selective action(s) on the metabolism of the cell. When the net yield of the yeast cultures with oleate was lowered by the effect of added trans acids, there was a marked accumulation of triglycerides and nonesterified acids in the cells. The marked increase in triglyceride content while phospholipid per cell remained relatively constant suggest that trans acids, in addition to forming inadequate membrane lipids, may also interfere with a basic control point in lipid metabolism.


Relaxation complexes of plasmid DNA and protein. I. Strand-specific association of protein and DNA in the relaxed complexes of plasmids ColE1 and ColE2. The ColE1 and ColE2 relaxation complexes of supercoiled DNA and protein were purified from Escherichia coli cells. Protein remains associated with the open circular DNA of these complexes after induction of relaxation with sodium dodecyl sulfate. The protein is associated specifically with the strand that possesses a site-specific break in the Co1E1 and ColE2 relaxed complexes. This protein remains associated with the DNA after centrifugation of the relaxed complex in a neutral or alkaline (pH 12.5) cesium chloride gradient or treatment with 8 M urea, 2 M NaSCN, 2M LiCl, 0.2 M sodium acetate, pH 4.6, and 70% formamide at 60 degrees.


Relaxation complexes of plasmid DNA and protein. II. Characterization of the proteins associated with the unrelaxed and relaxed complexes of plasmid ColE1. The proteins of the DNA-protein relaxation complex of plasmid ColE1 were labeled with [3H]leucine by growth of ColE1 containing Escherichia coli cells in the presence of this radioactive labeled amino acid. Three major [3H]leucine-labeled proteins are found associated with the supercoiled DNA in the ColE1 relaxation complex. The molecular weights of these proteins, determined by sodium dodecyl sulfate-acrylamide gel electrophoresis, are 60,000, 16,000, and 11,000, respectively. Induction of relaxation of the supercoiled DNA by treatment of the complex with sodium dodecyl sulfate results in a dissociation of the two smaller proteins from the DNA. The 60,000 protein, however, remains associated specifically with the nicked strand of the open circular DNA. The strand-specific association of this protein with the relaxed DNA resists heat denaturation of the DNA, sedimentation through an alkaline (pH 12.5) sucrose gradient, and centrifugation to equilibrium in an alkaline (pH 12.5) CsCl gradient.


Relaxation complexes of poasmid DNA and protein. III. Association of protein with the 5' terminus of the broken DNA strand in the relaxed complex of plasmid ColE1. The location of the protein in the open circular DNA form of the ColE1 DNA-protein relaxation complex, induced by treatment with sodium dodecyl sulfate, has been studied using several enzymes of DNA metabolism. Escherichia coli exonucleases I and III are able to degrade extensively the nicked strand of the relaxed complex from the 3' end. DNA polymerase I can initiate synthesis using the relaxed complex as template-primer and specifically extends the 3' end of the nicked strand. The 5' end of the sodium dodecyl sulfate-relaxed complex, however, is blocked to the 5'-3' hydrolitic action T7 exonuclease. This block remains after trypsin treatment of the sodium dodecyl sulfate-relaxed complex but is removed by Pronase treatment. T4 DNA ligase is unable to seal either the sodium dodecyl sulfate-relaxed complex or the Pronase-treated relaxed complex even after pretreatment of the relaxed complex with T4 DNA polymerase and polynucleotide kinase. However, pretreatment with DNA polymerase I and the four deoxyribonucleoside triphosphates facilitates ligase closure of the Pronase-treated relaxed complex but not the sodium dodecyl sulfate-relaxed complex. These studies indicate that the protein in the relaxed ColE1 complex is located at or near the 5' end of the nicked strand.


The development of a computer-assisted search for anomalous compounds (CASAC). An automated system for the recognition of anomalies in multi-compound mixtures is described. The mixtures to be investigated are injected into a combined gas chromatograph-mass spectrometer and low resolution spectra are acquired by repetitive scanning. Using an on-line computer, all of these spectra are compared with a pre-recorded file of spectra obtained by identical analyses of a "normal" mixture. The matching procedure has been designed to allow for differences in retention times. The CASAC program calculates and plots the degree of coincidence and in this way determines whether the sample spectrum contains fewer or more fragments than the corresponding library spectrum. The system has been applied to studies on pathological urine samples.


The effects of estrogen and progesterone on the functional capacity of the gonadotrophs. The functional capacity of the gonadotrophs under the influence of exogenous estrogen and progesterone was assessed by repeated stimulation with submaximal doses of LH-releasing factor (LRF) (10 mug at 2-h intervals) of subjects during the early follicular phase of the cycle and of hypogonadal women. The initial increment of peak serum LH and FSH concentrations after the first administration of LRF, was used to describe the pituitary sensitivity, and the integrated release during the 10 hours of LRF-stimulated pulses was utilized to approximate the pituitary gonadotropin reserve. During the early follicular phase, response to LRF stimulations was relatively stable with corresponding release of LH and FSH. An augmentation of sensitivity, as well as the reserve, for both LH and FSH was elicited by an incremental change in circulating estradiol levels, a change imposed by daily administration of estradiol benzoate for 4 days during the early follicular phase. Under the same conditions, the addition of progesterone (10 mg, im) at the end of the estradiol benzoate treatment induced a marked amplification of the estrogen-augmented pituitary gonadotropin sensitivity and reserve. The pituitary sensitivity, relatively higher than the reserve in hypogonadal subjects, was reversed by the administration of ethinyl estradiol (20-50 mug/day) for 7 days. These data indicate that the functional capacity of the gondotrophs is profoundly modulated by estrogen through relative changes in pituitary sensitivity and reserve, and that progesterone in low doses exhibited an amplifying effect on estrogen-primed gonadotrophs in both the pituitary sensitivity and the reserve.


Cell contacts in human islets of Langerhans. The freeze-fracturing technique was applied to fresh human islets of Langerhans. With this technique, the inside of cellular membranes was revealed, and specific membrane differentiations, hitherto unknown, were observed in the plasma membrane of the endocrine cells. The specific membrane differentiations represent two types of intercellular junctions, namely the tight junction, which determines a closure of the extracellular space and the gap junction which allows molecules and ions to diffuse from one cell to another (sharing the gap junction) without leaking in the extracellular space (intercellular coupling). The presence of such junctions may be important for the secretory behavior of the cells within the islet.


A study of streptokinase therapy in acute pulmonary embolism. A study of Streptokinase therapy in major pulmonary emboli is reported and attention is drawn to some of the factors which can influence the results of this as yet controversial therapy.


Serum LH and FSH response in four-hour infusions of luteinizing hormone-releasing hormone in normal men, Sertoli cell only syndrome, and Klinefelter's syndrome. Constant intravenous infusion of 1 mug/min of luteinizing hormone-releasing hormone (LHRH) for 4 hours into normal men produced a biphasic pattern of LH secretion. The initial LH peak occurred 15-45 minutes after the commencement of the infusion, following which LH levels plateaued or declined, subsequently rising as the infusion continued. No biphasic pattern of secretion could be identified for FSH. During the infusion, no consistent testosterone rise could be identified despite significant elevation of LH levels. Similar infusions of LHRH in men with Sertoli cell only syndrome produced exaggerated responses in terms of both LH and FSH. In contrast, a single intravenous bolus of 100 mug/ LHRH produced an exaggerated response in only one of the 3 men studied, suggesting that the prolonged infusion may be a more discerning dynamic test. Four-hour infusions of LHRH in men with untreated Klinefelter's syndrome produced exaggerated responses for both LH and FSH. After 6 weeks of treatment with long-acting testosterone esters administered weekly (250 mg), the exaggerated responses for both FSH and LH were decreased. For LH, the decrease principally affected the initial phase of LH release, and the pattern obtained resembled more closely the biphasic pattern seen in normal men.


Comparison of short and long-term treatment with synthetic LH-releasing hormone and clomiphene citrate in male hypothalamic hypogonadism. The effects of short and long-term treatment with clomiphene citrate and with synthetic LH-releasing hormone (LHRH) on gonadotropin release were studied in 5 male patients with hypothalamic hypogonadism. Neither short-term treatment with clomiphene citrate (50 mg or 200 mg daily for 4 days) nor long-term treatment (25 mg or 100 mg for 30-62 days) was effective in increasing serum LH and FSH in any patients. On the other hand, 5 cases showed slight or no rise in LH after administration of a single dose of 100 mug LH-releasing hormone (LHRH) and higher rises after administration of 400 mug. Four cases who were given 200 mug LHRH daily for 3 weeks showed an increasing response of serum LH and reached maximal LH levels at 2 weeks, followed by no further increase. In two of the four cases serum LH level reached normal adult male ranges. There was no increase of serum testosterone during LHRH treatment. These results suggest that long-term treatment with synthetic LHRH may be effective for gonadotropin restoration in some patients with hypothalamic hypogonadism in which long-term treatment of clomiphene citrate was shown to be ineffective.


Plasma gonadotropins after administration of LH-releasing hormone in patients with renal or hepatic failure. The pituitary gonadotropin response to the administration of synthetic LH-releasing hormone (LH RH) in 10 patients with renal failure and 5 patients with hepatic insufficiency was determined. The basal plasma LH and FSH levels were either normal or slightly elevated in both groups compared with the controls, but after the administration of LHRH, the peak levels (mean +/- SE mIU/ml) of LH (72.0 +/- 16.6) and FSH (18.7 +/- 4.4) in the patients with renal failure, and of LH (61.5 +/- 20.8) and FSH (13.2 +/- 2.8) in patients with hepatic insufficiency, were not significantly different from those in 11 normal controls (LH, 75.6 +/- 12.0; FSH, 12.4 +/- 2.9). However, significant elevations in plasma gonadotropin levels persisted in these patients during the second and third hours after the administration of LHRH. These studies do not resolve the question of whether the prolonged elevation in plasma LH and FSH levels after LHRH administration is due to enhanced secretion, to a diminished rate of metabolic clearance of LHRH, and/or to the gonadotropins, or both. Evidence from this and previous studies suggests that there may be defects in the secretory functions of both the hypothalamus and pituitary gland in some of these patients.


Serum LH and FSH responses to synthetic gonadotropin-releasing hormone in normal men. The effects of dose and of age on the serum LH and FSH responses to synthetic gonadotropin-releasing hormone (GnRH) were determined in normal men. GnRH was given as in iv bolus in doses from 2 to 500 mug to each of nine normal men, 20-39 years old. The mean (+/- SE) maximum delta LH (maximum increment above the basal concentration) in response to each dose of GnRH was 2 mug, 7.2 +/- 1.7 mIU/ml; 10 mug, 15.7 +/- 3.4 mIU/ml; 50 mug, 37.7 +/- 9.6 mIU/ml; 250 mug, 47.8 +/- 11.2 mIU/ml; and 500 mug, 49.3 +/- 11.2 mIU/ml. The mean delta FSH in response to each dose was 2 mug, 0.5 +/- 0.2 mIU/ml; 10 mug, 0.8 +/- 0.2 mIU/ml; 50 mug, 1.2 +/- 0.3 mIU/ml; 250 mug, 2.0 +/- 0.3 mIU/ml; and 500 mug, 2.4 +/- 0.4 mIU/ml. Since both the LH and FSH responses to the 250 mug dose were significantly greater than those to smaller doses, but the responses to the 500 mug dose were not still larger, 250 mug was used as the standard test done for subsequent studies. The effect of age was tested by administering a 250 mug dose of GnRH as an iv bolus to 42 carefully selected normal men, 14 in each of three age groups, 20-39, 40-59, and 60-79 years old. Dialyzable serum testosterone levels decreased with increasing age, from 21.7 +/- 4.6 to 16.0 +/- 4.9 to 14.3 +/- 3.8 ng/dl (mean +/- SD), and the basal serum LH levels increased slightly, from 7.4 +/- 3.4 to 9.6 +/- 3.1 to 10.8 +/- 3.2 mIU/ml, suggesting that a slight degree of primary Leydig cell failure occurs with increasing age. There was no corresponding increase in mean maximum delta LH in response to GnRH with increasing age, despite the elevation in mean basal LH levels. The maximum delta LH/basal LH ratio, in fact, declined from 6.8 to 5.1 to 3.3 in the three age groups. The basal serum FSH levels did not change with increasing age, but the mean delta FSH values decreased from 3.0 +/- 0.5 to 2.9 +/- 1.1 +/- 0.3 mIU/ml. These data demonstrate decreased LH and FSH responses to synthetic GnRH, as well as some primary Leydig cell failure, in the male senescence.


New technique for quantitative bacteriological sampling of wounds by velvet pads: clinical sampling trial. A new surface sampling method, the velvet pad rinse technique, was compared with the velvet pad imprint method, in a randomized bacteriological sampling trial, using surgical wounds to evaluate quantitative results. The efficiency of bacterial recovery was increased 20-fold by the new technique, and the improvement was ascribed in particular to the introduction of a procedure of rinsing the velvet pad after sampling.


Reproducibility of the analytab (API 20E) system. The reproducibility of the Analytab (API 20E) system for identification of Enterobacteriaceae was evaluated with 110 clinical isolates. Each isolate was identified by two technologists at different times. Genus-species identification was 97.3% reproducible; however, only 55.5% of the strains gave identical reactions in all 20 of the API 20E biochemical tests on repeat testing. Of those strains which varied, 56% possessed only one variable biochemical test. The reproducibility for each biochemical test was calculated and ranged from 89 to 100%. A subset of 20 of the most variable strains was tested further under conditions of varying incubation time (15 and 22 h) and inoculum concentration (10(7), 10(5), and 10(3) colony-forming units per ml), and by having four technologists interpret the test results. The reproducibility for each biochemical test for these 20 variable strains ranged from 86 to 99%. Less variation in interpretation by technologists was seen at an incubation time of 22 h and an inoculum concentration of 10(7) colony-forming units per ml. Consideration of the reproducibility for each biochemical test can aid in determining the probability that two isolates suspected of being the same strain, but with API profiles which differ by one or more biochemical test results, are in fact the same strain. Variables such as inoculum size, incubation time, technologist interpretation, and strip variability affect the API test results and should be standardized to minimize their effects.


Identification of the elementary bodies of Chlamydia trachomatis in the electron microscope by an indirect immunoferritin technique. An indirect immunoferritin (IIF) technique is described for recognizing the elementary bodies (EB) of Chlamydia trachomatis in unsectioned preparations. Both the EB of a genital strain of C. trachomatis grown in irradiated McCoy cells and EB in clinical specimens obtained from patients attending a venereal disease clinic were identified by the IIF test in the electron microscope. Cell culture-grown EB were detected by ferritin staining for up to 4 weeks after the organisms had lost their infectivity for tissue cultures. The IIF test was of comparable sensitivity to isolation methods in detecting chlamydiae in clinical specimens. Other possible applications of the IIF technique are discussed.


Indirect fluorescent antibody procedure for the rapid detection and identification of Bacteroides and Fusobacterium in clinical specimens. An indirect fluorescent antibody (IFA) technique was evaluated as a procedure for rapid detection and identification of members of the Bacteroidaceae. Antisera were prepared against 31 members of this family, including species of Bacteroides and Fusobacterium commonly isolated from human infections. The antisera had demonstrated species and/or subspecies specificity. Thirty clinical specimens were studied. Of 13 specimens yielding Bacteroidaceae, for which antisera were available, 23 were presumptively diagnosed by IFA to contain subspecies of B. fragilis and/or Fusobacterium species. Of 17 specimens yielding negative culture results, two were positive by IFA on direct smear. Frequently the in vivo morphology of cells detected in direct smears by this procedure closely mimicked that of cellular debris, tissue cells, and leukocytes. Polyvalent antisera pools facilitated use of the IFA procedure as a practical tool for rapid diagnosis of infections involving the Bacteroidaceae.


Germ tube and chlamydospore formation by Candida albicans on a new medium. A new medium composed of "cream of rice" infusion, oxgall, Tween 80, and agar is described for the sequential development of germ tubes and chlamydospores by Candida albicans. The procedure used (Dalmau's technique) is an improvement over the fluid substrate procedures previously advocated for germ tube formation. That the same preparation is then used for chlamydospore production is of practical importance for the clinical mycology laboratory.