Canadian surgery during the French regime, 1608 to 1763. The history of medicine in Canada, from the establishment of Quebec by Champlain in 1608 to the ceding of Canada to the British by the French at the Treaty of Paris in 1763, represents one of the most colourful periods in the history of Canadian surgery. Physicians were notable by their absence and what medicine was available in La Nouvelle France was provided almost exclusively by surgeons, or apothecaries, or individuals who posed as such. Sketches from the lives of five surgeons (Bonnerme, Giffard, Goupil, Bouchard and Sarrazin), an apothecary (Hébert) and a physician; (Gaultier), are presented to highlight various facets of medical care and the leadership role played by medical practitioners in the development of Canada during that period. In addition the study reveals the conflict between the philosophical approach of the physician and the pragmatic approach of the surgeon in 17th century France.


Adriamycin in advanced urinary tract cancer: experience in 42 patients and review of the literature. Forty-two patients with measureable, advanced, genito-urinary cancer received Adriamycin. Significant clinical responses (CR/PR) occurred in only five (14%) of 35 adequately treated patients. Four additional patients achieved a minimal response anf four others showed stabilization of their disease for 4 to 9 months. Review of the literature suggests an overall response rate of 20%. Patients with a urinary diversion lived significantly longer (P 0.006) than patients who did not have this procedure (42.5 months vs 21.5 months). The need to define various patient characteristics in future chemotherapeutic trials in bladder cancer is discussed.


Spontaneous regression of pulmonary leiomyomas during pregnancy. Multiple pulmonary nodular densities simulating metastastic cancer were discovered in a routine chest roentgenogram of a 30-year-old pregnant woman. Lung biopsy revealed nodules composed of smooth muscle and collagenous tissue containing entrapped glandular elements. The lesions were initially interpreted as multiple pulmonary fibroleiomyomatous hamartomas (MPFLH). During pregnancy and the post-partum period, the pulmonary nodules regressed spontaneously. Critical analysis of the published cases as well as our own case indicates that multiple pulmonary fibroleiomyomatous hamartomas (MPFLH) cannot be distinguished from benign metastasizing leiomyoma (BML) by either clinical, roentgenographic, or pathologic criteria and that all represent pulmonary metastases from a primary uterine neoplasm. The spontaneous regression of the pulmonary nodules in the present case as well as the increased risk for development of progressive pulmonary insufficiency in the pre-menopausal patients indicates an apparent hormonal dependence. Total abdominal hysterectomy and bilateral salpingo-oophorectomy appears to be the treatment of choice.


Biliary cystadenoma and cystadenocarcinoma: report of 14 cases and review of the literature. The clinical and pathologic features and long-term follow-up of eight patients with biliary cystadenoma and six patients with biliary cystadenocarcinoma are reported and the previous literature is reviewed. All the cystadenomas were in middle-aged women, but the six cystadenocarcinomas occurred in both male (4) and female (2) patients. The majority of the patients with cystadenoma and half of those with cystadenocarcinoma presented with an abdominal mass. Four of the patients whose cystadenoma was excised are alive and well for periods of time ranging from 2 1/2 to 13 years. Two of the patients with cystadenocarcinoma have survived for three years and for three years and eight months, respectively, after subtotal hepatic lobectomy. Morphologically the cystadenocarcinomas differ from the cystadenomas in that the former have cellular pleomorphism and anaplasia and infiltration of the underlying fribrous stroma; they can invade adjacent viscera and may occasionally metastasize to distant sites. The presence of benign epithelium in most cystadenocarcinomas supports their origin from cystadenoma.


A phase III study comparing the clinical utility of four regimens of 5-fluorouracil: a preliminary report. A clinical trial involving 462 colon, rectum, and breast cancer patients randomized among four different dosage regimens of 5-FU (an intravenous loading course, a weekly intravenous schedule, a nontoxic schedule, and an oral schedule) has shown a significantly better response among colon-rectum cancer patients for the intravenous loading course. In addition, duration of response and time to progression are also significantly better. Overall survival is approaching significance for the colon rectum group (p value .082). In contrast, breast cancer patients show little difference between treatments. Toxicity is somewhat higher for the loading course.


Idiopathic acquired sideroblastic anemia terminating in acute myelofibrosis: case report and review of leterature. Acute myelofibrosis is a rare but distinct accelerated variant of agnogenic myeloid metaplasia that is characterized by marked anemia, peripheral blood myeloblastosis and normoblastosis, a lack of teardrop poikilocytosis, and prominent myelofibrosis. There is usually no palpable hepatosplenomegaly or lymph node enlargement. The clinical course is remarkable short. We describe a 63-year-old man who presented with idiopathic acquired sideroblastic anemia and subsequently developed acute myelofibrosis. Intensive polychemotherapy with vincristine, cytosine arabinoside, and prednisone and a later trial of oxymetholone therapy were ineffective. He died 134 days after the diagnosis of acute myelofibrosis was established. The 11 previously reported cases of acute myelofibrosis are reviewed, and the relationships of acute myelofibrosis to other myeloproliferative disorders and to idiopathic acquired sideroblastic anemia are discussed.


Comparison of lymphangiograms and gallium scans in the non-Hodgkin's lymphomas. A total of 92 lymphangiograms (L.G.) and gallium scans (G.S.) done in 51 patients with non-Hodgkin's lymphoma were evaluated to assess their usefulness in detecting infradiaphragmatic disease. It was found that the correlation between both procedures was fairly good before treatment (85%) but decreased considerably after therapy. A significant number of patients persistently had positive L.G. after treatment, with a negative G.S. No histologic examination of tissues was performed, however, to confirm the radiographic findings. A higher number of equivocal G.S. were found both before and after treatment. No significant difference in correlation was found between the histiocytic and lymphocytic lymphomas but the former were found to have a striking frequency of negative L.G. and G.S. in the infradiaphragmatic area.


Peter: an infant with a myelomeningocele. Caring for a child with a myelomeningocele is not a task that one person can accomplish on his own. An interdisciplinary team approach, early parental involvement in the baby's care, a thorough knowledge of community resources, and parental understanding of the long range implications will greatly influence and affect Peter's early years. With support from both professionals and relatives and friends, it is hoped that they will develop a positive attitude to this challenge and continue to demonstrate their love and concern for Peter.


Mutagenicity of N-butyl-N-(4-hydroxybutyl)nitrosamine, a bladder carcinogen, and related compounds. N-Butyl-N-(4-hydroxybutyl)nitrosamine (BBN), which specifically induces bladder tumors, was shown to be mutagenic to Salmonella typhimurium strains TA 1535 and TA100 in the presence of an S-9 mix prepared from the liver of rats treated with polychlorinated biphenyl. Reduced nicotinamide adenine dinucleotide was a more effective cofactor than reduced nicotinamide adenine dinucleotide phosphate in the activation of BBN by the rat liver S-9 fraction, N-Butyl-N-(3-carboxypropyl)nitrosamine, reported to be the main urinary metabolite of BBN as well as of N,N-dibutylnitrosamine and to induce urinary bladder tumors specifically, was found to be mutagenic without metabolic activation by the S-9 mix. The mutagenicities of 31 compounds related structurally or metabolically to BBN and N,N-dibutylnitrosamine were tested. Of these compounds, 13 have previously been demonstrated to be carcinogenic, and nine have been shown to be noncarcinogenic. All the carcinogenic compounds were found to be mutagenic to strain TA1535 with or without the S-9 mix. Four of the nine noncarcinogenic compounds were also mutagenic. These "false-positive" compounds were predicted, in fact, to be carcinogenic.


The effects of additional flora on the response of salmonella mutants lodged in the gastrointestinal tract. A histidine auxotroph of Salmonella typhimurium, strain TA1538, will lodge for several months in the gastrointestinal tract of otherwise germ-free rats and of rats additionally associated with bacteria characteristic of the normal flora such as Lactobacillus plantarum and Bacteroides vulgatus. In the presence of the additional flora, the concentration of strain TA1538 is diminished in the stomach but not in the lower gastrointestinal tract or in the feces. Following the ingestion of 2-nitrofluorene, there is an increase in the concentration of revertants in the feces which reflects that observed in the colon and cecum. A dose-response relationship can be demonstrated between the amount of 2-nitrofluorene ingested and the concentration of revertants in the feces. A given dose of 2-nitrofluorene, however, produces fewer revertants in the feces of rats with the additional flora than in the feces of rats associated only with strain TA1538. It is not clear whether the decreased number of revertants in the feces in the presence of the additional flora is a result of metabolic transformations of 2-nitrofluorene by B. vulgatus, which can be demonstrated in vitro, or a result of the displacement of strain TA1538 from the stomach. The rat associated with strain TA1538, or other Ames tester strains, may be useful for detecting carcinogens as mutagens within the gastrointestinal tract and for determining the influence of various constituents of the bacterial flora on the concentration of mutagenic compounds.


Direct toxic effects of immunopotentiators on monocytic, myelomonocytic, and histiocytic or macrophage tumor cells in culture. Four murine monocyte, myelomonocyte, and histiocyte or macrophage tumor cell lines adapted to culture were growth inhibited by one or more of the following macrophage-activating substances: Mycobacterium bovis, Bacillus Calmette-Guérin strain, zymosan, lipopolysaccharide, and dextran sulfate, as well as tuberculin purified protein derivative, but not latex beads. Lipopolysaccharide was effective with one line at 4 ng/ml. All four lines actively phagocytosed zymosan and latex beads. In many cases the growth inhibition was apparently immediate but only cytostatic, and cell proliferation resumed upon removal of the drug. Bacillus Calmette-Guérin, live or boiled, was toxic to some of the tumor lines. Synthesis of lysozyme by all the cell lines in the monocyte series and production of granulocyte colony-stimulating factor by the myelomonocytic leukemia were not inhibited during several days of zero growth conditions in the presence of drugs. Since these agents had no direct effect on other hematopoietic tumor types (myeloma, T-lymphoma, mastocytoma) at the same or up to 10(4) higher concentrations, it is proposed that the sensitive tumors retain specific receptors for immunostimulants, either at the cell surface or within the cell in the case of phagocytosable particles. The binding of these agents to physiological receptors leads to stimulation and mitogenesis in normal macrophages and lymphocytes but leads to growth inhibition without affecting differenetiated functions in the corresponding tumor lines.


Stimulation of thymus- and bone marrow-derived lymphocytes by tumor cells in culture. In vitro lymphocyte stimulation by mitomycin-blocked tumor cells can be used to measure tumor-specific immune responses. In order to determine the responding cell type(s) in this reaction, lymph node and spleen cell populations were specifically depleted of thymus- or bone marrow-derived cells by the use of the appropriate antisera and complement or by immunoadsorption of the Fc receptor-bearing cells to antibody-coated sheep red blood cell monolayers. The compositions of both the original and the modified lymphocyte populations were determined by (a) viability counting following treatment with antisera and complement, (b) direct and indirect immunofluorescence, (c) antibody-coated erythrocyte rosette formation, and (d) response to thymus- and bone marrow-derived cell mitogens. In the lymph node cell populations, only the thymus-derived cells were stimulated by the tumor cells. However, both bone marrow- and thymus-derived cells from tumor-immune spleens underwent stimulation when exposed to tumor cells in culture.


The contribution of metronidazole and two metabolites to the mutagenic activity detected in urine of treated humans and mice. The urine of two patients receiving therapeutic doses of the trichomonacide, metronidazole, was analyzed for mutagenic activity using the histidine auxotroph TA1535 of Salmonella typhimurium. The activity detected in the urine was significantly higher than could be accounted for by the presence of the administered drug. Chromatographic analysis of the urine indicated the presence of the metabolite 1-(2-hydroxyethyl)-2-hydroxymethyl-5-nitroimidazole, which when tested in vitro with TA1535 was found to be ten times more active than metronidazole. An additional urinary metabolite, 1-acetic acid-2-methyl-5-nitroimadazole, was found to be inactive when similarly tested. The in vitro mutagenic activity of metronidazole and the two metabolites was unchanged by the addition of phenobarbital- or Aroclor-induced rat liver homogenate to the test system. In addition, metronidazole and the hydroxymethyl metabolite reverted S. typhimurium TA100 but not TA1537, TA1538, or TA98, and the acetic acid metabolite failed to revert any of the tester strains. In studies with mice, metronidazole was required in excess of the human dose in order for significant amounts of the hydroxymethyl metabolite to be detected in the urine. Urine from mice pretreated with the hepatotoxin, carbon tetrachloride, prior to the administration of metronidazole demonstrated approximately a 50% reduction in mutagenic activity, and the formation of the urinary metabolites was inhibited. These findings indicate the production of metabolites from the parent compound by the liver of the intact animal which could not be determined by use of the standard in vitro liver homogenate system.


Principal cell types in the pancreatic islet of a teleost fish, Xiphorphorus helleri H. By means of correlative light and electron microscopy, five pancreatic islet cell categories are described in the teleost fish, Xiphophorus helleri, each of which has specific light microscopic appearance and fine structure. Different histochemical techniques have been used, including immunofluorescence with antiporcine insulin and glucagon sera. In addition to B- and A1-cells, two categories of A2-cells have been observed both reacting with antiporcine glucagon serum: A2-cells with round granules gave a positive reaction for tryptophan; have been observed, both reacting with antiporcine glucagon serum: A2-cells with crystalline granules gave a negative reaction with the same staining technique on the same section. The "clear cells", the last category, were not specifically stained by any of the staining methods carried out in this investigation. The influence of fixation on staining affinities and on ultrastructure was shown to be considerable.


Multiple arrhythmias detected during nocturnal monitoring in patients with congenital complete heart block. Twenty patients with congenital complete heart block (CHB) were monitored with ECG tape recordings while awake and asleep. Episodes of marked ventricular slowing during sleep (R-R greater than 3000 msec., i.e. less than 20 beats/min) were noted in 35% (7/20). Most of these sudden R-R prolongations were 2:1 or 3:1 exit block of the junctional focus. Atrial and ventricular rate changes were mostly concordant in 2/3 but the remainder demonstrated fixed ventricular rates while atrial rates varied normally, suggesting a "sick" or "lazy" junctional focus. Other arrhythmias were found in 60% (12/20), including several types of rhythms in three patients. One child with previously unsuspected superimposed multiple arrhythmias later had a syncopal episode despite proven supra-Hisian block. The multiple patterns of atrial and ventricular rate changes found indicate complex feedback mechanisms, suggesting that congenital CHB is not a single entity. Since a significant number of patients demonstrated one or another of these unexpected and potentially hazardous findings, we recommend that congenital block patients be carefully studied, including Holter monitoring done on a regular basis.


Automated computer analysis for enzyme-multiplied immunological techniques. The dose/response curve in enzyme-multiplied immunoassays (EMIT) and related techniques may be described by the same "four-parameter logistic function" as has proven useful in radioimmunoassays, immunoradiometric assays (IRMA), and two-site immunoradiometric ("sandwhich") assays. This function provides an empirical description of the dose/response curve by use of an objective least-squares regression analysis, with the advantages of computerization and automation. This facilitates further statistical analyses, quality control, estimation of precision for an unknown, and improved, objective estimation of the sensitivity or minimal detectable dose.


Superiority of dobutamine over dopamine for augmentation of cardiac output in patients with chronic low output cardiac failure. Dobutamine is a newly developed catecholamine reported to have minimal direct vascular effects relative to its inotropic activity and to have less chronotropic and arrhythmogenic properties than other catecholamines used in the treatment of low output states. In this study, the acute hemodynamic effects of dobutamine were compared to those of dopamine in 13 patients with chronic low output cardiac failure. At dosages adjusted to achieve similar increments in cardiac output, dobutamine reduced left ventricular filling pressure (LVEP) from 24 +/- 2 mm Hg (SEM) to 17+/- 2 mm Hg, while dopamine increased LVEP to 30 +/- 3 mm Hg and in six patients caused arterial O2 saturation to fall below 90%. This poor response to dopamine was probably the result of its vasoconstrictive effects and illustrates the potential advantages of using a cardioselective agent such as dobutamine when the desired goal of therapy is to improve ventricular function by direct inotropic stimulation.


Anticonvulsants in serum, determined with a fully mechanized enzyme analyzer. The "EMIT" enzyme immunoassay system (Syva) that involves use of glucose-6-phosphate dehydrogenase (EC 1.1.1.49) as the enzyme label has been adapted to a fully mechanized kinetic enzyme analyzer for analysis of phenobarbital, phenytoin, and primidone. This procedure, compared with the gas-chromatographic procedure of Kupferberg [Clin. Chim. Acta 29, 283 (1970)], gave a weighted regression line--forced through the origin--of y=(1.01+/-0.04) x for phenobarbital, y=(0.95+/-0.04) x for primidone. Within-run coefficients of variation based on single determinations were 9,11, and 22% for primidone, phenobarbital, and phenytoin, respectively. Run-to-run assay CV was 6% for primidone and phenobarbital and 13% for phenytoin, based on the means of triplicate determinations of a sample with a mid-range concentration.


Enzyme immunoassay of serum thyroxine with the "Autochemist" multichannel analyzer. We have modified the homogeneous enzyme immunoassay of thyroxine and have adapted it to the AutoChemist. Both enzyme immunoassay and radioimmunnoassay procedures were performed on 2016 patients' sera on which thyroxine determinations had been ordered. We find the automated enzyme immunoassay to be precise and accurate, and for the first time it permits total automation of an immunoassay on a multichannel biochemical-profiling instrument. More importantly, it represents the first time that a thyroid-function test has been included in a multitest biochemical screening profile performed on a single automated instrument.


Development and use of analytical systems based on mass spectrometry. Contemporary analytical systems based on mass spectrometry include as components a gas chromatograph, a mass spectrometer, and a computer. The form of operation is usually in electron impact ionization mode for identification and structural studies, and in chemical ionization mode for quantitative analyses. Important stages in the development of these systems included the design of "molecule separators" for the concentration of solutes in the gas phase, the use of mass spectrometers as specific ion detectors, the introduction of chemical ionization techniques, and the development of computer-based operation, data acquisition, and data analysis capabilities. A current line of investigation is concerned with the design and use of systems based on atmospheric pressure ionization. Samples are ionized in a small reaction chamber external to the low-pressure region of a quadrupole mass analyzer. The primary source of electrons is a 63Ni foil or a corona discharge. The ionization process leading to positive ions involves a sequence of ion molecule reactions, usually electrons leads to carrier gas ions leads to reagent ions leads to sample component ions. Negative ions may be formed by direct electron attachment, or by ion molecule reactions that include new types of elimination reactions. The source will accept a variety of gases and solvents. The sample may be introduced in the gas phase without solvents, by probe injection, or in the effluent stream from a gas chromatograph. Samples may be introduced in the liquid phase in solvents by injection after the fashion of gas chromatography or in the effluent stream from a high-performance liquid chromatograph. The novel aspects of atmospheric pressure ionization mass spectrometry lie in its versatility and high sensitivity of detection. Few clinical chemistry laboratories now use these systems. Significant future uses are likely to be in analytical work involving therapeutic drug monitoring and studies of drug metabolism, and in analyses for environmental biohazards including pesticides, herbicides, polyhalobiphenyls, dibenzodioxins, and other toxic compounds.


A list of selected reference materials in clinical chemistry. This is a compilation of reference materials suggested for scientists considering a laboratory reference collection in clinical chemistry, or for librarians seeking to develop or add to an up-t-o-date library collection. Nine areas are represented: general clinical chemistry; analytical methods, procedures, and techniques; calculations and statistics; the clinical laboratory; drugs and metabolism; enzymology; instrumentation, interpretation of data; and ready reference.


Clinical relevance of polyamines as biochemical markers of tumor kinetics. The polyamines, spermidine and spermine, and their diamine precursor, putrescine, constitute a unidirectional biosynthetic pathway whose biosynthetic enzymes and accumulation patterns appear to play important roles in the regulation of growth processes. Concentrations of these compounds in physiological fluids are low or undetectable under normal conditions, are elevated in patients with metastatic cancer, and are thought to reflect growth (putrescine concentrations) and cell turnover (spermidine concentrations) of the organism. Cancers are a broad spectrum of diseases in which there are altered growth fractions and cell-turnover fractions, and therefore cancer chemotherapeutic agents have been developed to take tumor kinetics into account. Because changes in polyamines in physiological fluids reflect cell kinetics, this review compiles evidence of their efficacy as biochemical markers of cancer and suggests their possible usefulness to clinicians in rapidly assessing tumor response to chemotherapy or to multimodality therapy.


Serum albumin: recent progress in the understanding of its structure and biosynthesis. Major discoveries have been made in the past few years on the structure and mode of biosynthesis of serum albumin. The complete amino acid sequence of this protein has been determined, and its covalent structure shown to be a single peptide chain grouped into a series of nine disulfide-bonded loops. These loops appear to associate into three similar domains. By study of isolated fragments of the molecule it can be demonstrated that the binding of billirubin and the primary binding of long-chain fatty acids are functions of separate domains. The biosynthesis of albumin has been found to involve a precursor form, termed "proalbumin", in which a basic hexapeptide is attached to the amino end of the chain. Similar precursor forms are now known to have a role in the formation of other secreted proteins, but in the case of albumin the purpose of the additional peptide is not clear. Clinical methodology for albumin assay has advanced but little despite--or perhaps in part because of--the increasing use of automation. Hope for improvement is foreseen in the advent of immunochemical procedures and in a better understanding of the specificity of dye-binding reactions.


Use of saliva in therapeutic drug monitoring. We measured the concentrations of phenobarbital, phenytoin, primidone, ethosuximide, antipyrine, and caffeine in paired samples of saliva and plasma by gas chromatograph-mass spectrometer-computer (GC/MS/COM) and enzyme immunoassay. Mixed saliva was collected for the antipyrine and caffeine studies, parotid saliva for the phenobarbital, primidone, ethosuximide and phenytoin studies. The saliva/plasma (S/P) ratios (by weight) obtained by GC/MS/COM were: phenobarbital, 0.31-0.37; phenytoin, 0.11; ethosuximide, 1.04; antipyrine, 0.83-0.95; caffeine, 0.55. The S/P ratio obtained by enzyme immunoassay were: phenobarbital, 0.32; phenytoin, 0.12; primidone, 0.85. The concentrations of phenytoin, primidone, ethosuximide and antipyrine in saliva correspond to the free fraction of the drug in plasma. When we analyzed samples containing phenobarbital or phenytoin (plasma or saliva) by both techniques, we found that the enzyme immunoassay values were generally higher than GC/MS/COM values, suggesting that the metabolites as well as the parent drug were measured in the immunoassay.


Urinary excretion of immunologically reactive metabolite(s) after intranasal administration of cocaine, as followed by enzyme immunoassay. Using the enzyme immunoassay technique (EMIT), we determined the time course of urinary excretion of benzoylecgonine in 16 surgical patients and three volunteers who received intranasal cocaine. After doses varying from 13 to 130 mg, the test for benzoylecgonine was positive in 1 to 4 h, peaked at about 10 to 12 h, remained positive for 18 to 27 h, and became negative after about 17 h. This information should be considered by drug dependency treatment programs in which the EMIT procedure is used to screen urines for cocaine use.


Identification of urinary metabolites of sodium dipropylacetate in man; potential sources of interference in organic acid screening procedures. Organic acid screening of urine samples from two children with neurological disease demonstrated the presence of two unknown metabolites. The children were receiving an antiepileptic drug, sodium dipropylacetate. The major abnormal compound has been shown by gas chromatography-mass spectrometry to be 3-oxodipropylacetic acid, a previously unidentified metabolite of dipropylacetate in man, while the minor metabolite was indentified as 2-(n-propyl)-glutaric acid.


Glomerular lesions in patients with asymptomatic persistent andorthostatic proteinuria discovered on routine medical examination. The incidence of asymptomatic proteinuria in young healthy men entering the army service in Singapore was 0.94%; 0.56% had intermittent orthostatic proteinuria and 0.38% with persistent proteinuria. Renal biopsies from 45 cases with persistent proteinuria and 10 cases with orthostatic proteinuria were studied by light microscopy, electron microscopic and immunofluorescent antibody techniques. Three cases with orthostatic proteinuria showed a minimal lesion ('Nil'), and 7 a minimal lesion with increased centrilobular mesangial matrix and mild focal and segmental mesangial hypercellularity. Focal and segmental capillary loop changes were seen in two cases. No immunoglobulin deposits were found in orthostatic proteinuria. A raised anti-streptolysin O titer was found in 3 cases. 13.3% of cases with persistent proteinuria showed a minimal lesion ('Nil'); 66.6% a minimal lesion with increased centrilobular mesangial matrix and mild focal and segmental mesangial hypercellularity: Focal global sclerosis, focal segmental sclerosing glomerulonephritis and diffuse proliferative GN (mesangial hypercellularity) were each found in 6.7% of patients in this group. Focal and segmental changes in capillary loops were found in 30% of cases. Changes in visceral epithelium such as the appearance of cytoplasmic swelling, vesicles and dense aggregates, and areas of focal foot process fusion were common findings. Mesangial deposits of IgA were found in 34.3%, with an IgA-IgG-beta1C globulin combination in 28.6% of cases. IgM-beta1C globulin was present in 5.7% of cases with persistent proteinuria. Early complement components, C1q and C4, and IgA secretory piece were absent. A raised antistreptolysin O titer was found in 25% of cases. The changes inthe glomeruli may represent the end or healing stages of an early bout of glomerulonephritis. Changed hemodynamic responses on assuming an upright position may play a role in the loss of protein across the glomerular basement membrane in patients with orthostatic proteinuria.


Dense deposit disease: long term follow-up of three cases of recurrence after transplantation. The incidence and early recurrence after transplantation prove the specificity of the appearance of an electron dense alteration of kidney basement membrane often called dense intra-membranous deposit disease. Three new cases with dense deposit disease affecting the original kidneys have been followed-up after transplantation for periods ranging from 4 to 8 years and illustrate the natural history of the recurrence. Serial kidney biopsies showed the predominance of dense deposits near the mesangial area and the vascular pole. These deposits were also seen in some tubular basement membranes. Absence of cell proliferation was noted in all biopsies performed. Immunofluorescence studies revealed fixation of C3 alone. Histological signs of recurrence are compatible with the absence of clinical and biological signs. Transient or permanent proteinuria and microhematuria were common findings. Serum complement levels, measured after transplantation, were low in all three cases. Despite recurrence of the original glomerulonephritis, long-term survival of the graft was commonly observed, two cases being followed-up for 7 and 8 years. Patients with dense intra-membranous deposits glomerulonephritis should not be excluded from a transplantation program. One of the three cases reported here illustrates the exceptional association of recurrence of dense intramembranous deposits, de novo membranous glomerulonephritis and chronic rejection.


Nephropathy in cyanotic congenital heart disease. Three children with cyanotic congenital heart disease who developed transient proteinuria and edema are described. One died of an intercurrent illness but the other two are now well. Renal biopsy findings in all three children demonstrated a mesangial proliferative glomerulonephritis on light microscopy. An unusual ultrastructural appearance of localized electron-dense thickening of the basement membrane of the capillary loops was seen in all three and collagen fibers were present in the mesangium of two. There was slight fusion of foot processes in two specimens and marked fusion in the third. Immunofluorescence in two patients demonstrated IgM staining in both and fibrin in one. The cause of the glomerular lesions is unknown but, among the many possible factors involved, anoxia and increased venous pressure may be important.


Antihypertensive effects of tolamolol.

Antihypertensive effects of tolamolol. The antihypertensive and renin-lowering efficacy and side effects of tolamolol, a beta adrenergic blocking drug with cardioselectivity, were examined in 10 patients with mild essential hypertension while on regular diet. Tolamol, at a dose of 300 to 900 mg per day, given over a period of 2 to 4 wk significantly decreased systolic and diastolic blood pressures in both the recumbent and standing positions. Normal blood pressure (140/90 mm Hg or less) was attained in 8 subjects. Mean heart rate and ambulatory midday plasma renin activity (PRA) decreased significantly; however, there was no significant correlation between blood pressure decrease and either the pretreatment PRA or decrease in PRA. Body weight did not change significantly. No adverse side effects were detected and no changes in the liver or renal function or in the blood count were observed. It is concluded that tolamolol is effective in lowering blood pressure and PRA in patients with hypertension.


Analgesic efficacy of an orally administered combination of pentazocine and aspirin. With observations on the use and statistical efficiency of GLOBAL subjective efficacy ratings. The analgesic efficacy of a combination of pentazocine and aspirin (PA) in the ration 1:13 was compared with that of 650 mg of aspirin alone (A650) and with placebo (PBO), in 98 patients with postoperative pain. Two dose levels of the combination were compared: the lower dose (PA-L) consisted of pentazocine 25 mg and aspirin 325 mg, while the higher dose (PA-H) consisted of pentazocine 50 mg and aspirin 650 mg. All active treatments performed significantly better than PBO. PA-L performed as well as A650, while PA-H performed significantly better than A650. In addition to the usual subjective measures of analgesia, we obtained in 74 patients an evaluation of the overall (GLOBAL) performance of the treatment. This was rated on an ordinal scale of 1 ("poor") to 5 ("excellent"). On the GLOBAL scale, PBO had a mean score of 2.4 (fair-good); A650 and PA-L had scores of 3.6 and 3.9 respectively (good-very good): and PA-H had a score of 4.5 (very good-excellent). In five of six comparisons between treatment means, GLOBAL had the best discriminating power of all six measures. In the two comparisons of greatest interest (A650 against PBO and PA-H against A650), GLOBAL was more than twice as efficient as the TOTAL (summed pain score) measure. In comparing the statistical efficiency of different measures of the same analgesic effect, there is a problem in determining what are "clinically equivalent differences" on the various analgesic scales employed. We propose the use of the observed sample differences and the safeguard of repeating the comparisons over several studies to minimize the effect of random-origin bias.


Spironolactone and hydrochlorothiazide in normal-renin and low-renin essential hypertension. The relative blood pressure response and side effects of spironolactone (S), 400 mg/day, and hydrochlorothiazide (H), 100 mg/day, were evaluated in a double-masked crossed comparison. Subjects were 24 essential hypertensives, 13 normal and 11 with low renin activity. Upright renin levels were determined in the AM after 3 days of moderately low sodium diet (less than 100 mEq/day) and at the end of each drug treatment period. The values were not revealed to one investigator until completion of the study. After 4-week placebo control periods, either S or H was given in divided doses for 6 weeks. Blood pressure measurements and side effects were evaluated at biweekly intervals. The fall in blood pressure from control was the same for each drug and was independent of renin activity. Side effects occurred more often in patients treated with S, and H was judged superior by risk/benefit analysis. We conclude that S is no more beneficial than H in hypertensive patients with either low or normal renin activity but induces a higher incidence of adverse effects in the dose used in this study. We found no evidence for a greater effectiveness of S in low-renin essential hypertension.


Sealant retention and effects on occlusal caries after 2 years in a public program. A clinical trial of one application of Nuva-Seal fissure sealant was conducted under School Dental Service conditions in a low-income area of London. The sealant was applied to one of a cariesfree pair of permanent teeth by schools' dentists, the other tooth being left as a control. One examiner, who did not apply the sealant, conducted examinations at baseline, after 6 months, and after 2 years. There were 118 children, initially aged 5-12, who remained in the trial after 2 years. The sealant was fully retained on 27.4 % of the teeth, partially retained on 40.6 %, and missing on 17.9 %. The remaining 14.1 % of teeth had been restored. There was no statistically significant difference between subjects who had less decay in sealed teeth and those who had less decay in control teeth. The sealant was 13.6 % effective (not significant, P less than 0.4) in preventing caries in sealed teeth. These results are less favorable than those in other trials reported. The potential role of fissure sealants in a School Dental Service is discussed.


Effect of high-concentration ammonium and sodium fluoride rinses on dental caries in schoolchildren. A double-blind clinical trial was conducted in a non-fluoridated community to determine the effect on enamel fluoride and caries experience of daily rinsing in school with 1,000 parts/10(6) solutions of ammonium fluoride or sodium fluoride at pH 4.4. Subjects were 10- to 12-year-old children (n approximately equal to 200/group at baseline), about one-half of whom reported the usage of fluoride supplements. Dental caries (DFS index) and enamel fluoride (in vivo biopsy) were evaluated at baseline, 12 months, and 24 months. Supplement users had higher enamel fluoride levels and less caries experience initially, as well as generally lower caries increments over the study. In year 1, the overall caries reductions (supplement users and non-users combined) were 23 % (ammonium fluoride) and 33 % (sodium fluoride), P less than 0.01. For year 2, treatment effects were significantly greater: 54 % (ammonium fluoride) and 47 % (sodium fluoride). In newly erupted teeth, the effects of the ammonium fluoride (70 % DFS reduction) was significantly greater (P = 0.013) than that of the sodium fluoride (48 % DFS reduction). Enamel fluoride levels at the end of 2 years were 3,124 parts/10(6) (ammonium fluoride), 2,771 parts/10(6) (sodium fluoride), and 2,603 parts/10(6) (placebo), P = 0.025.


A comparative study of subcutaneously administered terbutaline and epinephrine in the treatment of acute bronchial asthma. Terbutaline, a new bronchodilator drug reported to have selective affinity for beta 2-adrenergic receptors, was compared with epinephrine in the treatment of 49 adult subjects with acute bronchial asthma. Under double-blind conditions, 24 subjects received 1.0 mg of terbutaline sulfate, and 25 subjects received 0.5 mg of epinephrine hydrochloride subcutaneously. Spirometric measurements, heart rate, and blood pressure, as well as subjective responses, were recorded prior to, and then at 5, 15, 30, 60, and 120 minutes after administration of the drug. The results indicate that terbutaline is an effective bronchodilator drug in subjects with acute asthma; however, the heart rate rose significantly after administration of terbutaline, with a maximal increase of 25 percent above control. Review of the literature reveals that tachycardia is a consistent finding when subcutaneous doses of terbutaline in excess of 0.25 mg are administered. Stimulation of beta 1-adrenergic receptors in the heart appears to be the most important factor involved in this response. A lesser cardioaccelerator effect was observed after administering epinephrine in a dose producing an equivalent degree of bronchodilatation.


Response profiles: A method of evaluating the activity of beta-lactam antibiotics against Enterobacteria. A method is described which may be used to compare the relative activity of beta-lactam antibiotics against gram-negative bacilli. The method offers several important advantages over conventional methods of evaluation of the antibacterial activity of beta-lactam agents: the bacterial inoculum is simply and accurately standardized; the response of the culture is continuously monitored and information is obtained on both the intrinsic activity of the agent and its susceptibility to enterobacterial beta-lactamases. In this way, differences in the antibacterial activity of compounds which appear similar on the basis of more conventional tests may be revealed.


Effect of fosfomycin on the fecal microflora of man. Use of broad-spectrum antibiotics is known to cause changes in the intestinal microflora of man. The present study was initiated to determine the effect of oral fosfomycin calcium on several groups of microorganisms indigenous to the gut. Data from this double-blind study indicate that 2 g of fosfomycin given daily in four divided doses for 28 days increased the incidence of soft stools. No significant changes in numbers of staphylococci or Candida were observed during treatment. Although there was no significant change in total coliforms, the numbers of E. coli decreased markedly from 10(5) to 10(2) organisms/g feces. Concurrent with this decrease, the Klebsiella-Enterobacter counts increased from 10(3) to 10(6) organisms/g feces. Two weeks posttreatment the E. coli and the Klebsiella-Enterobacter counts approached pretreatment levels. These observations are similar to those reported for ampicillin. During the course of treatment when the E. coli dropped precipitously the few surviving E. coli still were suceptible to fosfomycin as determined by an agar diffusion disc test.


Genetic localization of the resistance to fosfomycin. The possible existence of a transferable resistance to fosfomycin was studied in 50 strains of enterobacteria that were isolated in our hospital and which showed a resistance to fosfomycin of more than 256 mug/ml. E. coli K12 E711 was used as a receptor strain in the conjugation -- the times of conjugation were 20 min and 18 h. We found a transferable resistance in 28% of the strains. In all but one of the cases, only the resistance to fosfomycin was transferred. The recombinant which received the resistance to fosfomycin together with three other resistances was used as the donor for the transduction with the phage P1Kc to the receptor strain which was E. coli UTH 1038. A study was carried out on the influence of the radiation of the transducing phages with ultraviolet light on the frequency of transduction. The findings demonstrated a linear decrease in the frequency in proportion to the increase in the time of radiation.


Fosfomycin and plasmidic resistance.

Fosfomycin and plasmidic resistance. 60 fosfomycin-resistant strains of gram-negative bacilli are submitted to conjugation experiments using as recipient cell E. coli K12 which is nalidixic acid resistant. After mating, the number of fosfomycin-resistant E. coli K2 colonies growing on selection plates containing nalidixic acid and fosfomycin never surpassed the normal rate of mutation for fosfomycin-resistance of the recipient strain. In 65% of the experiments, plasmidic resistance to other antimicrobials was transferred to E. coli K12, but was never accompanied by demonstrable fosfomycin resistance. High rate of normal mutation of recipient strain is signaled as the main problem for detecting the plasmidic nature of fosfomycin resistance. With our criteria regarding this fact we have been unable to confirm the plasmidic nature of fosfomycin resistance.


Bactericidal activity of tinidazole. An in vitro comparison of the effects of tinidazole and metronidazole against Bacteroides fragilis and other Anaerobic bacteria. The new antiprotozoal agent, tinidazole, was found to be bactericidal against all 52 isolates of obligate anaerobic bacteria tested, 42 Bacteroides fragilis, 4 clostridia and 6 peptostreptococci. The minimum bactericidal concentrations of tinidazole for B. fragilis ranged from 0.25 to 4 mug/ml, and those of metronidazole from 0.25 to 8 mug/ml, i.e. several times lower than the serum concentrations achievable after oral administration. In most cases the MIC was identical with MBC or half of it. On the average, tinidazole was slightly more effective against B. fragilis than metronidazole. Although essentially the activities of the two drugs were positively correlated, there was a fourfold difference in their MBC for 10 of the 42 B. fragilis.


Enteropathogenic E. coli gastroenterocolitis in neonates treated with fosfomycin. The effect of calcium salt of fosfomycin in the treatment of 43 neonates suffering from acute gastroenterocolitis produced by enteropathogenic E. coli is evaluated. The minimal inhibitory concentration of these E. coli was, generally, lower than 128 mug/ml. Dosages of 150-200 mg/kg body weight/day were administered orally every 8 h. This treatment lasted for 4 days only. Clinical evolution was favorable in 38 (88%) babies and bacteriological evolution in 30 (70%). In eight cases a different flora to the initial was selected during the treatment with fosfomycin. None of the cases treated showed any toxic alteration attributed to the antibiotic.


Neurosurgical infection treated with fosfomycin and 6-methylprednisolone. A left parietal parasagittal meningioma was removed in a 67-year-old female patient. In the postoperative period she had a neurosurgical cerebral suppurating infection: subdural, epidural and epicranial, connected to the exterior by several fistulas. In the operative revision and after a culture, the germ causing the infection, E. coli, was isolated. Treatment was begun with 16 g of intravenous fosfomycin and 120 mg daily of 6-methylprednisolone intramuscularly, and this treatment cured the patient.


Antibiotherapy of Serratia marcescens septicemia in children. The clinical and bacteriological response of 38 treatments performed on 24 children (11 of them neonates) carrying out separate treatments with carbenicillin (2 treatments), gentamicin [4], fosfomycin [6], and associated treatments with gentamicin plus carbenicillin [6], fosfomycin plus gentamicin [18] and fosfomycin plus carbenicillin [2] are considered. The clinical cure was obtained in 21 children (87.5%). The most effective treatment was fosfomycin plus gentamicin; both antibiotics showed synergism in vitro on isolated Serratia strains. A dosage of 75 mg/kg fosfomycin enables serum levels of about 32 mug/ml during 4-5 h, being this level higher to the MIC of all isolated strains of S. marcescens.


Amoxycillin and co-trimoxazole in acute purulent exacerbations of chronic bronchitis. 100 hospital patients suffered from acute exacerbations of chronic bronchitis. 50 were treated with amoxycillin in a dose of 500 mg, three times a day for 10 days and the results compared with 50 patients treated with co-trimoxazole in a dose up to 480 mg trimethoprim and 2,400 mg of sulphamethoxazole daily in males, and two thirds of this dose in females. The trial was single-blind. During the acute phase of infection, both treatments were equally effective in clinical improvement, conversion of the sputum from purulent to mucoid, diminution of quantity and elimination of pathogenic bacteria. Amoxycillin was quicker in sputum conversion and gave less side effects, but the differences were not significant. During the 2-4 weeks following treatment, only a third of the patients who had received co-trimoxazole remained well and free from purulent relapse, as opposed to 72% who had received amoxycillin, a difference significant at the 2% level.


Combined action of fosfomycin with beta-lactam and aminoglycoside antibiotics. The combined action of fosfomycin with penicillin G, ampicillin and streptomycin was investigated. A total of 11 S. aureus and 10 E. coli were included in these studies. It has been shown that a combination of fosfomycin with penicillin G and streptomycin resulted in a synergistic effect. When S. aureus strains were tested, the combination of fosfomycin with penicillin G was more frequently effective than the combination of fosfomycin with streptomycin. The combinations of fosfomycin with ampicillin and streptomycin showed a similar synergistic effect on 7 of 10 E. coli strains. The antagonistic effect never resulted when the above-mentioned combination of antibiotics were used. The authors postulate that a combination of fosfomycin with beta-lactam or aminoglycoside antibiotics may be used in clinical practice and such a procedure should prevent an emergence of fosfomycin-resistant strains.


Treatment of cefazolin in 45 cases of urinary tract infections. We have treated 45 cases of urinary tract infection with cefazolin (i.m.) and cure was achieved in 35 (77.77%). 26 of the 28 patients (92.85%) with acute cystitis who received 250 mg every 12 h for 6-10 days were cured. 17 patients with pyelonephritis received 500 mg every 12 h during 10-14 days and therapeutical success was achieved in 9 of them (52.94%). Laboratory control was carried out at the initiation of therapy, at the end, and 1 month after therapy. Tolerance to injections of cefazolin was considered good and only in one case appeared a mild eosinophia.


Comparative studies on the evaluation of the effect of new anthelminthics on various intestinal helminthiasis in Iran. Effects of anthelminthics on intestinal helminthiasis. The effect of pyrantel pamoate, levamisole, mebendazole, thiabendazole and bephenium hydroxynaphthoate on various intestinal helminths were evaluated among the inhabitants of four villages in the Dezful area southwest of Iran. A total number of 328 persons, all infected simultaneously with Ascaris and hookworm (Ancylostoma duodenale) and 49.2% with Trichostrongylus spp., were randomly divided into six groups. One group was kept as the control and the other five were each treated with one compound. Follow-up examinations showed that all of the drugs used were highly effective on Ascaris, and the differences in the cure rate were not statistically significant except for bephenium hydroxynaphthoate which showed a lower cure rate. For hookworm, cure rates of 100, 90, and 85% were observed with levamisole, pyrantel pamoate and bephenium hydroxynaphthoate, respectively. Cure rates observed with mebendazole and thiabendazole were 35 and 51%, respectively. For Trichostrongylus, the highest cure rate was achieved with levamisole, followed by thiabendazole and mebendazole. While the percentage of people showing side-effects was rather low for all drugs, thiabendazole and bephenium hydroxynaphthoate produced a higher, and levamisole a lower, percentage of side effects.


Evaluation of two dose levels of loxapine succinate in chronic schizophrenia. In a double-blind, placebo controlled design in chronic schizophrenic inpatients using 50 mg. and 100 mg. LOX, an attempt was made to replicate the findings of a previous study and to establish the dose-level and duration of treatment optimal for this kind of patient. Multiple regression analysis adjusting for dose, age, duration of illness, and baseline values indicated that 100 mg. LOX was an effective dose as previously shown. In contrast to the previous study, significant response effects were found by the 4th week. In addition, the dose was linearly related to response on nearly all the variables. The principal side effects were drowsiness and extrapyramidal signs.


[Transferral of Salmonella from mother to newborn infant (author's transl)]. Two newborn infants were infected with Salmonella originating from their mothers. After infection of a newborn infant with Salmonella typhi from a mother, known for a long time to be a chronic carrier, no pathological findings were observed but the child continued to secrete typhoid bacilli for three years. The infection of the second child with S. panama, from a mother recognized as a carrier post-partum, caused a fulminating diarrhoea accompanied by septicaemia which had a fetal outcome.


Malignant melanoma of the rectal ampulla: report of a case and review of the literature. Documentation of the seventeenth case of melanoma of the rectum is presented. The world literature is reviewed. The effectiveness of immunotherapy with BCG or BCG in combination with CCNU and DTIC remains to be established.


[Immunoblastic sarcoma (reticulum-cell sarcoma) of the gastro-intestinal tract in a case of dermatitis herpetiformis Duhring (author's transl)]. In a 43-year-old man who for 20 years was known to have dermatitis herpetiformis there co-existed, since 1974, a "dermatogenic enteropathy" with gastro-intestional symptoms. Several small-intestine biopsies demonstrated severe partial to subtotal villous atrophy with crypt hyperplasia, as seen in gluten-sensitive enteropathy. At post-mortem there was an immunoblastic sarcoma - so-called reticulum-cell sarcoma - in the upper jejunum and abdominal lymph-nodes, which was presumably the immediate complication of the sprue-like lesions of the small intestines.


Heparin Therapy: regimens and management. Heparin remains the most effective antithrombotic drug. It acts by combining with plasma antithrombin, thereby accelerating the neurtalisation of thrombin and other acitvated coagulation factors. Full-dose intravenous heparin is indicated in all cases of pulmonary embolism and established deep venous thrombosis, unless there exist compelling contraindications. Continuous intravenous infusion of heparin appears to be safer than intermittent injection. Low-dose subcutaneous heparin is effective in preventing the initial occurrence of thigh vein thrombi and in reducing the incidence of fatal pulmonary embolism in general surgical patients over the age of 40. The efficacy of low-dose heparin in preventing pulmonary emboli following hip surgery has not been established. The incidence of severe heparin-induced thrombocytopenia appears to be rising. Platelet counts should be performed in all patients receiving heparin by any mode of administration.


Dantrolene sodium: a review of its pharmacological properties and therapeutic efficacy in spasticity. Dantrolene sodium or dantrolene1 is 1([5-(nitrophenyl)furfurylidend] amino) hydantoin sodium hydrate. It is indicated for use in chronic disorders characterised by skeletal muscle spasticity, such as spinal cord injury, stroke, cerebral palsy and multiple sclerosis. Dantrolene is believed to act directly on the contractile mechanism of skeletal muscle to decrease the force of contraction in the absence of any demonstrated effects on neural pathways, on the neuromuscular junction, or on the excitable properties of the muscle fibre membranes. Controlled trials have demonstrated that dantrolene is superior to placebo in adults or children with spasticity from various causes, as evidenced by clinical assessments of disability and daily activities, and by muscle and reflex responses to mechanical and electrical stimulation. It is somewhat less effective in patients with multiple sclerosis than in those with spasticity from other causes. There has been a general clinical impression in controlled trials that dantrolene caused less sedation than would have been expected from therapeutically comparable doses of diazepam. In 2 controlled trials, there was no significant difference between dantrolene and diazepam in terms of reductions in spasticity, clonus, and hyperreflexia, but side-effects such as drowsiness and inco-ordination occurred significantly more frequently on diazepam. Long-term studies have indicated continuing benefit for patients taking dantrolene, though the incidence of side-effects has often been high and there has been a suggestion of exacerbation of seizures in children with cerebral palsy. Dantrolene may be of value in the medical treatment of spasm of the external urethral sphincter due to neurological and non-neurological disease, and animal studies suggest a potential use in the management of malignant hyperpyrexia. Chemical evidence of liver dysfunction may occur in 0.7 to 1% of patients on long-term treatment with dantrolene, with symptomatic hepatitis in 0.35 to 0.5% and fatal hepatitis in 0.1 to 0.2%. The drug commonly causes transient drowsiness, dizziness, weakness, general malaise, fatigue and diarrhoea at the start of therapy. Muscle weakness may be the principal limiting side-effect in ambulant patients, particularly in those with multiple sclerosis, and therapy could be hazardous in patients with pre-existing bulbar or respiratory weakness. The dosage of dantrolene has been fixed in most controlled trials, though long-term studies have indicated the need for individualisation of dosage. The initial dose is usually 25mg once daily, increasing to 25mg two, three or four times daily, and then by increments of 25mg up to as high as 100mg two, three or four times daily. The lowest dose compatible with optimal response is recommended.


[Hormonal contraceptives and periodontium]. The clinical changes of the gingiva reported in the literature (gingival hyperplasia and new growths like the pregnancy epulis) in connection with oral hormonal contraception as well as the causal connection between sexual hormone activity and the changes of gingiva and oral mucosa are discussed. Our own experiments show that the increase in the sulcus fluid flow rate under administration of combination preparations is largely independent of an increase in the degree of gingival inflammation.


Relationship between luteinizing hormone releasing hormone concentration in hypophysial portal blood and luteinizing hormone release in intact, castrated, and electrochemically-stimulated rats. The concentration of luteinizing hormone releasing hormone (LHRH) in hypophysial portal plasma was determined in pentobarbital anesthetized,intact and castrated rats of both sexes, including proestrous rats following electrochemical stimulation of the medial preoptic area (MPOA). Mean LHRH levels in portal plasma obtained between 1400--1700 h from estrous and diestrous rats and from rats ovariectomized for 8 weeks were similar and ranged from 50--55 pg/ml, but the LHRH levels in proestrous rats were less than 12 pg/ml. In addition, hypophysial portal plasma collected during 1100 to 1400 h from animals orchidectomized for 8 weeks and from intact male rats contained mean LHRH concentrations that ranged from 50--65 pg/ml and 30--35 pg/ml, respectively. Electrochemical stimulation of the MPOA in the female rat on the afternoon of proestrus resulted in a marked increase in the concentration of LHRH in portal plasma. LHRH levels in portal plasma during the 0 to 30, 30 to 60, 60 to 90, 90 to 120, and 120 to 150-min periods after electrochemical stimulation of the MPOA were 105 +/- 24.2, 61 +/- 10.8, 51 +/- 8.2, 36 +/- 5.3, and 32 +/- 4.1 pg/ml, respectively. LHRH levels in portal plasma from the unstimulated rats were not detectable (less than 12 pg/ml) in most of the animals. In another group of proestrous rats, the effect of rabbit anti-LHRH serum or normal rabbit serum (NRS) on the release of LH after electrochemical stimulation of MPOA was examined. Pretreatment of proestrous rats with anti-LHRH serum blocked the release of LH due to MPOA stimulation, whereas pretreatment with NRS did not inhibit LH release. On the basis of these findings, it is concluded that electro-chemical stimulation of the MPOA in proestrous rats increases LHRH levels in portal blood and that the enhanced secretion of LHRH stimulates the release of LH from the pituitary gland.


Effects of testosterone implants and hypothalamic lesions on luteinizing hormone regulation in the castrated male rat. The effects of intrahypothalamic and subcutaneous implants of testosterone (T) and those of hypothalamic lesions on resting levels of circulating LH and pituitary responsiveness to exogenous LHRH were studied in castrated male rats to elucidate hypothalamic and pituitary regulation of LH secretion. Two hundred mug implants of testosterone propionate (TP) in the median eminence region suppressed plasma LH titers before evidence of direct inhibition of pituitary function (as indicated by testing with LHRH) was found. Such implants release appreciable amounts of T into the peripheral circulation in the immediate post-operative period, and SC Silastic (constant release) capsules containing T have similar effects. The findings suggest that, regardless of the site of implant, the initial negative feedback inhibition of LH by T is not dependent on direct action at the pituitary levels but rather appears to be a hypothalamic effect. In the days following exposure to hypothalamic or peripheral implantation of T, however, a progressively developing decline in the response to exogenous LHRH was observed. In order to determine whether this effect results from suppression of endogenous LHRH release, the median eminence-arcuate region was destroyed to remove the source of LHRH. In these animals, the suppression of plasma LH was evident on the first day after the lesion, but pituitary responsiveness to LHRH was unaffected until after one week. When Sialastic capsules were implanted SC into lesioned animals, a more rapid (less than 1 week) inhibition of pituitary responsivity ensued. Suprachiasmatic lesions did not affect basal LH secretion or pituitary responses to LHRH. The data provide evidence for a dual feedback action of T on LH in castrated male rats: an initial inhibitory effect presumably due to hypothalamic inhibition (commencing at around 6h after hypothalamic of SC implantation of T), and a subsequent suppression of pituitary responisveness (after one day) presumably due to direct action of T on the pituitary. In addition to these phenomena, findings in rats bearing median eminence-arcurate lesions suggest that the removal of endogenous LHRH by itself leads to an eventual decline in pituitary responsiveness (greater than one week postoperatively).


Developmental changes in pituitary responsiveness to luteinizing hormone-releasing hormone (LHRH) in the female rat: ovarian-adrenal influence during the infantile period. Pituitary LH and FSH repsonses to synthetic LHRH as estimated by increases in plasma FSH and LH 15 and 45 min following its iv injection were enhanced during the first 2 weeks of life, reaching a maximum around day 10-15 and declining thereafter. No AM.-PM. variations in pituitary responsiveness were observed at any age studied. The increased pituitary response found in infantile rats did not appear to be caused by a slower rate of disappearance of LHRH in blood of the younger animals. Ovariectomy-adrenalectomy. (Ovx-Adrx) or Ovx at day 10, but not Adrx alone, resulted in elevated LH and FSH levels 5 days later and almost complete obliteration of the FSH response to LHRH. The LH response was not altered. Treatment with 5alha-dihydrotestosterone (DHT) but not with estradiol benzoate (EB) or testosterone propionate (TP) suppressed the post-Ovx-Adrx rise in plasma LH and FSH. Progesterone (P) potentiated the effect of DHT. Restoration of basal plasma LH and FSH levels (by DHT and/or P) restored FSH responsiveness to exogenous LHRH. EB and TP were ineffective. The LH response was slightly depressed by EB + DHT. It is concluded that the elevated plasma FSH levels in the infantile female rat may be due at least in part to a high degree of pituitary responsiveness to LHRH and/or FSH-RF brought about by steroidal signal of ovarian origin. DHT and P appear to be the steroids responsible for such a stimulatory action.


Assymetry of the myo-inositol transport system in Klebsiella aerogenes. Energy is necessary to create the assymetry of the transport system. 1. In Klebsiella aerogenes the influx of myo-inositol proceeds during the steady state at a rate equal to that of efflux. 2. The kinetic parameters of influx during the steady state are similar to those observed at the initial time of uptake. 3. Efflux and influx processes share the same transport system. 4. The ability of other cyclitols to chase the accumulated scyllo-inositol is dependent on their affinity for the transport system. 5. A counter transport can be observed in preloaded cells only in poisoned cells and under anaerobiosis. 6. The efflux process is not temperature-dependent. 7. The KT of influx in poisoned cells is larger than that in normal cells. 8. Although no saturation kinetics of efflux could be observed, it can be inferred that the KT of efflux is at least 50-fold larger than the KT of influx; 9. The results suggest that in the transport of myo-inositol by K. aerogenes energy coupling enhances the affinity of the carrier for the substrate at the outer side of the cytolplasmic membrane by lowering the KT.


Interactions of yeast tRNAPhe with ribosomes from yeast and Escherichia coli. A fluorescence spectroscopic study. The interaction of ethidium-labeled tRNAPhe from yeast with ribosomes from yeast and Escherichia coli was studied by stead-state measurements of fluorescence intensity and polarization. The ethidium label was covalently inserted into either the anticodon or the dihydrouridine loop of the tRNA. The codon-independent formation of a tRNA-ribosome complex led to only a moderate increase of the observed fluorescence polarization indicating a considerable internal mobility of the labeled parts of the tRNA molecule in the ribosome complex. When the ribosome complex was formed in the presence of poly(U), the probes both in the dihydrouridine loop and in the anticodon loop were strongly immobilized, the latter exhibiting a substantial increase in fluorescence intensity. A smaller intensity change was observed when E. coli ribosomes were used, although the extent of immobilization was found to be similar in this case. Competition experiments with non-labeled tRNAPhe showed that the labeled tRNAPheEtd was readily released from the complex with yeast ribosomes when poly(U) was absent, whereas in the presence of poly(U) it was bound practically irreversibly. The finding that the mobility of a probe in the dihydrouridine loop is affected by the codon-anticodon interaction on the ribosome suggests a conformational change of the ribosome-bound tRNA which may involve opening of the tertiary structure interactions between the dihydrouridine and the TpsiC loop.


Formate hydrogenlyase system in Salmonella typhimurium LT2. Isolation from Salmonella typhimurium of mutants unable to reduce benzyl viologen under anaerobic conditions has allowed the study of the factors involved in the multienzymic formate hydrogenylase system. 1. Depending on the affected activities, different classes of mutants were found: FHL-A mutants have lost formate dehydrogenase 1 and formate dehydrogenase 2 activities; mutations in fdhA (117 min) or fdhB (33 min) lead to such a phenotype. FHL-B and FHL-C mutants have lost formate dehydrogenase 2 activity and part or all of hydrogenase activity, respectively; both types correspond to mutations in the hyd gene (approximately 90 min). FHL-D mutants have lost only formate dehydrogenase 2 activity; fhlD gene maps at 120 min. 2. In some cases, mixtures of extracts from two mutants display formate dehydrogenase 2 and formate hydrogenylase activities. Restoration studies suggest the existence of one factor sensitive to growth conditions and inactivated by oxygen or heating. This factor which is present and active in FHL-C mutants, is probably the one missing in FHL-D mutants. 3. A new scheme for the formate hydrogenylase system is proposed, in which hydrogenase transfers electrons directly to benzyl viologen.


Phosphorylation in vitro and in vivo of ribosomal proteins from Saccharomyces cerevisia. Crude ribosomes from Saccharomyces cerevisiae cultures were phosphorylated in vitro when incubated in the presence of [gamma-32P]ATP. Analysis of the ribosomal proteins with two-dimensional electrophoresis revealed that of the 29 proteins identified in the small subunit, only protein S6 was phosphorylated. Of the 37 proteins identified in the large subunit, one was highly phosphorylated (L3) and two only slightly phosphorylated (L11 and L14). The protein kinase activity associated with the ribosomes was extracted with 1 M KCl and was not dependent on adenosine 3':5'-monophosphate; it preferentially phosphorylated casein and phosvitin, but was less active on histones. Structural ribosomal proteins were also phosphorylated in vivo when the yeast cultures were incubated with [32P]orthophosphate; the radioactivity resistant to hydrolysis by hot perchloric acid was incorporated into the proteins of the two subunits. Radioactive phosphoserine was found by subjecting hydrolysates of ribosomal proteins to high-voltage electrophoresis. After two-dimensional electrophoresis, one poorly phosphorylated protein (S10) was identified in the small subunit. In the large subunit, one protein (L3) was highly labelled, and two proteins (L11 and L24) only slightly labelled.


Rhodanese-Mediated sulfur transfer to succinate dehydrogenase. The interaction of the sulfurtransferase rhodanese (EC 2.8.1.1) with succinate dehydrogenase (EC 1.3.99.1), yeast alcohol dehydrogenase (EC 1.1.1.1) and bovine serum albumin was studied. Succinate dehydrogenase incorporates the sulfane sulfur of [35S]rhodanese and, in the presence of unlabelled rhodanese, also incorporates that of [35S]thiosulfate. Rhodanese releases most of its transferable sulfur and is re-loaded in the presence of thiosulfate. Rhodanese undergoes similar modifications with yeast alcohol dehydrogenase but this latter does not bind 35S in amounts comparable to those incorporated in succinate dehydrogenase: nearly all the 35S released by [35S]rhodanese is with low-molecular-weight compounds. Bovine serum albumin also binds very little sulfur and [35S]rhodanese present in the reaction mixture does not discharge its radioactive sulfur nor does it take up sulfur from thiosulfate. Sulfur release from rhodanese appears to depend on the presence of - SH groups in the acceptor protein. Sulfur incorporated into succinate dehydrogenase was analytically determined as sulfide. A comparison of the optical spectra of succinate dehydrogenase preparations incubated with or without rhodanese indicates that there is an effect of the sulfurtransferase on the iron-sulfur absorption of the flavorprotein. The interaction of rhodanese with succinate dehydrogenase greatly decreases the catalytic activity of rhodanese with respect to thiocyanate formation. This is attributed to modifications in rhodanese associated with the reduction of sulfane sulfur to sulfide. Thiosulfate in part protects from this deactivation. The reconstitutive capacity of succinate dehydrogenase increased in parallel with sulfur incorporated in that enzyme following its interaction with rhodanese.


The stereochemical basis of template function. The behavior of nucelotides with thioketo-substituted pyrimidine bases (4-thiouracil, 2-thiouracil and 2-thiocytosine) or amino-analogue purine bases (2-aminopurine and 2,6-diaminopurine) in transcription and translation was investigated. The experimental results obtained led to the following conclusions. 1. The stereochemical basis of substrate selection in transcription is the geometry of Watson-Crick base pairs A-U (or A-T) and G-C between substrate and template bases. 2. The topology of the active site of Escherichia coli RNA polymerase is precisely adopted to the geometry of Watson-Crick base pairs. 3. The enzyme active site discriminates between A-U (A-T) and G-C base pairs. An essential feature in this discrimination is the 6-NH2 group of the A-U (A-T) base pair and the 2-keto group of cytosine in the G-C base pair. 4. The codon properties of a nucleic acid base in messenger RNA can be predicted on the basis of its specificity in polynucleotide interactions. There seems to be no evidence for the participation of protein topological sites in the control of the specificity of codon-anticodon interactions in translation.


Ribitol-containing lipopolysaccharides from Proteus mirabilis and their serological relationship. Ribitol phosphate was recently identified as a constituent of lipopolysaccharides obtained from 'proteus mirabilis strain D52 giving 1:4-anhydroribitol during acid hydrolysis (Gmeiner, 1975). Two other Proteus mirabilis strains belonging to serogroups O16 and O33 were shown previously to contain an unknown compoound X as lipopolysaccharide constituent (Kotelko et al., 1975). In this report the identification of compound X as 1:4-anhydroribotol by gas-liquid chromatography, mass spectrometry and mass fragmentography is described. Serological investigations using passive hemagglutination, hemagglutination inhbition and semi-quantitative precipitin reactions indicate strongly that ribitol plays a role in the serological specificity of the respective lipopolysaccharides.


The effect of initiation factor IF-1 on the dissociation of 70-S ribosomes of Escherichia coli. By means of exchange studies, in which 3H-labelled 50-S subunits and unlabelled 70-S ribosomes from Escherichia coli MRE 600 were used, it has been demonstrated that the 30-S subunit is the only target for IF-3 in the dissociation of 70-S ribosomes. The interference of IF-3 with the dynamic equilibrium of 70-S in equilibrium 50-S + 30-S occurs by binding of the factor to the 30-S subunit. The 30-S-IF-3 complex in impaired in the association reaction, which implies that IF-3 is acting as an anti-association factor. The action of IF-1 is two-fold. Firstly IF-1 increases the rate of exhcange of the ribosomal subparticles in the 70-S ribosome without changing the position of the equilibrium. Thus the spontaneous equilibrium is attained more rapidly in the presence of IF-1. This kinetic effect of IF-1 is also demonstrated in the IF-3-mediated dissociation of 70-S ribosomes. Secondly IF-1 is able to increase the IF-3-mediated dissociation. It seems likely that the explanation for the latter phenomenon must be sought in the binding of IF-1 to 70-S ribosomes, resulting in a loosening of the ribosomes structure, as well as to 30-S. IF-3 complex, thaereby slowing down the association reactions of the subunits.


Calorimetric studies on melting of tRNA Phe (yeast). The heat effects involved in thermal unfolding of tRNAPhe from yeast have been determined in various buffer systems by direct differential scanning calorimetry. Perfect reversibility of the melting process has been demonstrated for measurements in the absence of Mg2+ ions. The overall molar transition enthalpy, delta Ht = 298 +/- 15 kcal mol-1 (1247 +/- 63 kJ mol-1), has been shown to be independent of the NaCl concentration and the nature of the buffers used in this study. Delta Ht is identical in the presence and in the absence of Mg2+ ions within the margin of experimental error. This experimental result implies a vanishing or very small heat capacity change to be associated with melting. Decomposition of the calorimetrically determined complex transition curves, on the assumption that the experimental melting profile represents the sum of independent two-state transitions, results in five transitions which have been assigned to melting of different structural domains of the tRNA.


Myo-Inositol transport in Klebsiella aerogenes. scyllo-Inositol, a non-metabolizable substrate for the study of the myo-inositol transport system. 1. Scyllo-Inositol is transported against a concentration gradient by Klebsiella aerogenes, but is not metabolized. 2. The apparent KT of scyllo-inositol is 0.05 X 10(-3) M while that of myo-inositol is 0.23 X 10(-3) M. The V values are respectively 32 and 12 nmol min(-1) mg cells(-1) (dry weight). 3. Both cyclitols bind to the same carrier, since typical competitive inhibition kinetics are observed. 4. Neither phosphoenolpyruvate phosphotransferase nor a periplasmic binding protein seems to be involved in the scyllo-inositol transport system. 5. The induction of the transport system is not repressed by malate, furmarate, citrate, succinate or pyruvate; however, acetate represses the induction. The activity of the transport system is increased when the growth medium is supplemented with adenosine 3':5'-monophosphate.


Progeria: a cell culture study and clinical report of familial incidence. This report relates the case histories of two sisters who demonstrated the typical symptoms of progeria at birth. One of these children had died previous to this study. The familial occurrence underlines the thesis that progeria is an autosomal-recessive disorder. The examination of the cultured skin fibroblasts from the younger child showed a clear decrease in cell growth. On the other hand, the immunfluorescent examination of skin biopsies and cultured skin fibroblasts revealed no atypical distribution of collagen types.


Effect in bronchial asthma of a new beta-adrenergic blocking drug atenolol (ICI 66, 082). The bronchial effect of intravenous atenolol (ICI 66.082) has been studied in a double-blind cross over trial in 10 patients with pronounced, labile bronchial asthama. A single i.v. dose of atenolol 3 mg. sufficient to cause a fall in heart rate and systolic blood pressure at rest, induced only a slight and clincially almost negligible impairment of ventilatory function. An ordinary therapeutic dose of salbutamol by inhalation far outweighed the bronchial effect of atenolol. The drug appears promising with regard to its cardio-selective properties.


Effect of an inhibitor of pepsin proteolytic activity in the Exalto-Mann-Williamson ulcer. The role of pepsin proteolytic activity in the Exalto-Mann-Williamson ulcer has previously not been established. Our studies show that a potent inhibitor of pepsin activity, when administered orally to E-M-W dogs, reduced both the incidence and severity of jejunal ulceration. These results indicate that pepsin may play a major role in the development of ulceration in this model.


Rosette forming cells in the spleens of mice with lymphoid and myeloid leukemia. The kinetics of the appearance of direct (E) or induced (EA and EAC) rosettes in the spleens of mice inoculated with lymphoid and myeloid leukemias were followed under the assumption that leukemic infiltration of the spleens would result in progressive increase of rosette-forming cells if the malignant cells possessed appropriate receptors.


Recovery of hematologic competence without engraftment following attempted bone marrow transplantation for aplastic anemia: Report of a case with diffusion chamber studies. An 18-year-old male with severe aplastic anemia associated with hepatitis was prepared for bone marrow transplantation with cyclophosphamide (50 mg/kg/day) for 4 consecutive days. He then received 3.47 X 10(8) nucleated bone marrow cells per kilogram of body weight from his HL-A identical sister by intravenous infusion. Although subsequent karyotyping showed that engraftment was not achieved, prompt hematologic recovery ensued by the third week post-transplant. Diffusion chamber culture of the patient's bone marrow stroma, obtained prior to cyclophosphamide therapy, was carried out. A prompt increase in the number of cells in the chambers was observed. A differential cell count of the contents was similar to that seen when normal bone marrow is cultured in a similar manner.


Predominance of granulocytopoiesis in bone marrow grafts in the omenta of mice treated with erythropoietin. The effects of erythropoietin on the differentiation of murine bone marrow injected into the omenta of x-irradiated mice were investigated. Experimental hosts were injected with 2.5 units of erythropoietin on days 0-7 and sacrificed on day 10. Control hosts were injected with saline or sheep serum. After 10 days the grafts were greater than 95% granulocytic regardless of host treatment. Since these grafts contain multipotent hematopoietic stem cells and the experimental hosts were exposed to large doses of erythropoietin, the results of this experiment indicate that a specialized microenvironment is required for murine erythropoiesis in vivo.


Changes in the immunocompetence and surface markers of lymphocytes in stored blood. Changes in lymphocyte responses to mitogens and other lymphocyte markers were studied during a 14-day period of storage of blood at 4 degrees C and 22 degrees C in plastic bags. The following tests were done: (a) absolute lymphocyte and neutrophil counts, (b) blastogenic response to PHA, PWM, Con-A, and LPS and (c) determination of T and B cells using E-rosette and EAC-rosette techniques. The neutrophils disappeared rapidly at 22 degrees C. The absolute lymphocyte count doubled in a week. The responses to PHA, PWM, and Con-A declined rapidly and were absent by the 6th day. The response to LPS disappeared on day 14. The ability to form E-rosettes and EAC-rosettes was lost by days 4 and 6, respectively. The results obtained at 4 degrees C were similar except that these changes were delayed varying from 1-7 days.


Immuno-electronmicroscopic study of human EA rosettes. The immuno-electronmicroscopic study of human EA rosettes, using ferritin-labelled anti-human IgG antiserum, showed clusters of ferritin granules at the points of contact between erythrocytes and lymphoid cells, indicating that the links between Fc fragment of IgG on the surface of erythrocytes and specific receptor on the surface of lymphoid cell correspond to the sites of morphological interaction between the 2 cell types.


On the morphology of live spermatozoa of human semen. Live and dead spermatozoa of human semen were distinguished and evaluated separately with respect to their morphology after supravital staining with buffered eosin. This evaluation was shown to be almost as reliable as that of the commonly stained smear. Different samples from the same individual were found to be similar with respect to both the total population and the live population. Different types of abnormalities were not equally common in the live and dead populations. Tapered heads were more common in the live population and abnormal midpieces more common in the dead, whereas abnormal tails and cytoplasmic droplets occurred with the same frequency in both populations. However, neither the frequency of all spermatozoal abnormalities nor that of abnormal heads or midpieces studied separately showed strong correlations in the two populations, in contrast to the frequencies of tapered heads, cytoplasmic droplets, and abnormal tails. No constant relationship was found between the live population and the entire sperm population with respect to numerical differences or quotients between the figures for abnormal cell frequency.Therefore, study of the morphology of the live sperm population gives supplementary information about the qualities of semen not attained by previous methods of study.


Isolation of a spermatozoal immobilization factor from Escherichia coli filtrates. A low-molecular weight spermatozoal immobilization factor (SIF) has been isolated from Escherichia coli cultures. This factor is stable to heating, freezing, and lyophilization, and immobilizes but does not kill spermatozoa. The concentration of spermatozoa influenced the effectiveness of the SIF in immobilizing spermatozoa, indicating that bacterial infections could have a greater influence on the oligospermic ejaculate than on a normal specimen. SIF is a stable factor and binds to spermatozoa via a reversible mechanism.


In vitro studies of intestinal endotoxin absorption. I. Kinetics of absorption in the isolated everted gut sac. Previous studies have shown in a qualitative manner that endotoxin can cross gut epithelium, but precise quantitation has not been possible. The present studies were undertaken to measure quantitatively the mucosal to serosal unidirectional flux of endotoxin with the use of an in vitro rat gut sac preparation. 51Cr-Labeled endotoxin was placed in the mucosal bath in concentrations ranging from 0.05 to 2.0 mg per ml. Over a 2-hr period of time, a small amount of endotoxin was transported transmurally, which was shown chromatographically to be similar to the starting material and which retained its toxic and immunogenic properties. It was first shown that the presence of 2.0 mg per ml of endotoxin in the mucosal bath did not significantly alter the tissue's histology or permeability to 3-O-methyl-D-glucose. When unidirectional fluxes were measured, it was found that the flux was not proportional to the endotoxin concentration as would be expected with a passively permeable solute, but rather its transport system became "saturated," displaying a maximum transport rate of 4.72 (mug per cm) per 2 hr and a Km of 0.425 mg per ml. The isolated gut sac provides an excellent model for the precise study of factors involved in endotoxin absorption.


Immunohistochemical localization of human pancreatic polypeptide (HPP) in the human endocrine pancreas. In order to clearly identify the cell type responsible for production of human pancreatic polypeptide (HPP), a newly postulated hormone, we have conducted an immunohistochemical detection of this peptide in the human endocrine pancreas at both light and electron microscopy levels. It was shown that HPP containing cells are located both in endocrine and exocrine pancreatic tissue. In the islets, the positive cells are very few in number and are characterized by an elongated shape and the presence of small secretory granules of about 150 nm in diameter. The presence of HPP in an endocrine cell type reinforce the hypothesis that this peptide is a hormone.


[Two-phase respiration of apnoeic newborn, basic research and clinical value (author's transl)]. Studies on fetal human and calf lungs proved an 1: 2,5 increase rate of the transpulmonary blood-flow after inflation of the alveolar spaces. The extent of the inflation depends on the function pressure X time. Based on the rupture rate of the pulmonary tissue, the factor pressure is limited to 20-30 ch H2O. Consequently the inflation only can be obtained by increasing the factor time. Conditional on the flow resistance in the variably calibrated branches of the pulmonary system, the maximal pressure of 30 cm H2O has to act repeated continuously at least 10 seconds over an endotracheal tubus. Only then even the periphere alveoles will fully be inflated (phase I equals monotone inflation phase). An intermittend pressure breathing of maximal plus 30 cm H2O respectively-30 cm H2O will be supporting. After the total inflation short pressure waves of plus 30 cm H2O in physiological frequency (equals 30/minute) will do for further ventilation of the alveoles (phase II equals rhythmical ventilation phase).


[Epidemiology and diagnosis of vaginal discharge. First Report (author's transl)]. In 700 patients, 1336 tests of vaginal discharge were performed by culture, wet mount with the phase contrast microscope and the Papanicolaou stain. The diagnosis of yeast is best done by culture. Yeast was found only in 68% of the culture positive cases by the wet mount method and in 47% of the cases by the Papanicolaou stain. Wet mount and Papanicolaou stain results for yeast are therefore only useful if they are positive. In the diagnosis of trichomonas, culture and wet mount are of the same effeciency. The incidence of yeast and trichomonas was dependant upon the age of the patients. In pregnancy and in cases with diabetes, yeast was found more often but no increase of yeast was found in cases on oral contraception. The diagnosis of bacterial mixed infections poses the most difficult problem and can only be improved by expensive culture methods.


Homologous and heterologous protection of mice with group A streptococcal M protein vaccines. Purified streptococcal M proteins precipitated with alum (APM) were used to immunize mice. A trivalent vaccine of serotypes 1, 3, and 12 protected mice against challenges by homologous live streptococci and also conferred protection against serotypes 6 and 14 but not against a strain of group B streptococci. Monovalent APM vaccines afforded homologous protection and restricted heterologous protection. The extent of heterologous protection was a function of serotype combinations and was also dose dependent. Rabbit antisera exhibiting strong opsonic activities were active in vitro and in passive mouse protection only for homologous serotypes. Mouse antisera did not passively transfer protection and were not bactericidal in vitro. It was concluded that homologous and heterologous active mouse protection was most likely a result of shared antigenic determinants of the various M proteins although protection of mice could not be measured as a function of circulating anti-M antibodies.


Induction of Escherichia coli and Vibrio cholerae enterotoxins by an inhibitor of protein synthesis. Enterotoxigenic Escherichia coli or Vibrio cholerae 569B (Inaba) grown in the presence of the antibiotic lincomycin, an inhibitor of protein synthesis, produced elevated levels of heat-labile enterotoxin or choleragen, respectively, as assayed by both vascular permeability factor and capacity to elicit fluid accumulation in rabbit ileal loops. This induction of enterotoxin did not reflect either a coupling of lincomycin resistance with increased enterotoxigenicity or an effect of lincomycin on cellular release of enterotoxin, since spontaneously isolated lincomycin-resistant mutants of both E. coli and V. cholerae still required lincomycin for induction, and large increases in E. coli permeability factor activity were found intracellularly as well as extracellularly. After the period of exponential growth, E. coli became refractory to induction by lincomycin, although most of the induced enterotoxin activity appeared only after this period. No increase in copy number of the enterotoxin plasmid in E. coli 711 (P307) was found in induced cells by analysis of deoxyribonucleic acid reassociation kinetics. These and other data suggest that synthesis of enterotoxin, or at least its accumulation, is normally limited by cellular factors whose synthesis is preferentially inhibited by lincomycin. A possible connection between this phenomenon and lincomycin-associated diarrhea is considered.


Plasma-dependent chemotaxis of macrophages toward BCG cell walls and the mycobacterial glycolipid P3. BCG cell walls, associated with oil droplets in the form of emulsions in saline, generate macrophage chemotactic activity from fresh guinea pig plasma. Serum and heat-inactivated plasma were inactive, suggesting involvement of complement or fibrinogen-derived chemotactic factors. Suspensions of cell walls and oil droplets each generated chemotactic activity from plasma, and the activity of the cell wall vaccine was due to the additive effects of these two components. A mycobacterial glycolipid (P3), which is a constituent of BCG cell walls, also had plasma-dependent chemotactic activity. The results suggest that macrophage chemotaxis may be an important part of the immunopotentiating activity of these mycobacterial products.


Depressed antibody responses to a thymus-dependent antigen in toxoplasmosis. The immunodepressive effect of Toxoplasma gondii infection in mice was studied, using sheep erythrocytes (SRBC) as the testing antigen and serum hemagglutinins, hemolysins, and both direct and indirect splenic plaque-forming cells (PFC) to SRBC as assays. In the primary antibody response, immunoglobulin M (IgM), hemagglutinins, and hemolysins and both IgM- and IgG-secreting PFC were depressed in animals immunized after infection. Maximum immunodepression occurred during the first 3 weeks of Toxoplasma infection. When the secondary antibody response was studied, results varied. Mice immunized with SRBC after being infected with T. gondii had a depression in both IgM and IgG PFC. Mice immunized with SRBC before being infected with T. gondii and then given a challenge dose of SRBC had a delay, but no an actual depression, in IgG hemagglutinins and hemolysins and IgG-secreting PFC. These studies show that the immunodepression associated with Toxoplasma infection is complicated, and they provide no definitive explanation for the mechanism.


Relationship between antigenic stimulation and increased splenic peroxidase levels during the immune response. Immunization of mice with either soluble or particulate antigens induced a marked increase in peroxidase activity of spleen cell homogenates. Animals immunized with sheep erythrocytes showed maximum splenic peroxidase activity at 2 days. By the 4th day after immunization, when the hemolytic antibody plaque-forming cell response was maximal, peroxidase activity returned to normal levels. Increased splenic peroxidase activity also occurred in mice immunized with rabbit erythrocytes, as well as with rabbit serum or bovine serum albumin. No change in splenic peroxidase activity occurred in mice injected with syngeneic mouse erythrocytes or serum. Both glass-adherent spleen cell populations, morphologically consisting of 90 to 95% macrophages, as well as nonadherent cells, consisting of more than 90% lymphocytes by the same criteria, showed peroxidase activity. Immunization of mice with sheep erythrocytes resulted in an intracellular redistribution of the peroxidase activity among several distinct subcellular fractions prepared by differential centrifugation. Maximum redistribution occurred with granule-associated enzyme activity. A possible relationship between peroxidase activity with functional activity of lymphocytes and macrophages during humor immune responses seemed likely.


Some factors affecting selective adherence of microorganisms in the bovine mammary gland. Some parameters affecting the adherence of microbes to the ductular epithelium of the bovine mammary gland were studied. Adherence increased from teat sinus to lactiferous sinus to the large ducts, and cells from the lactiferous sinus to the large ducts, and cells from the lactiferous sinus were used for all other experiments. There was no difference in adherence to cells from different quarters of the same cow, but there were significant differences between cows. Scanning electron microscopy suggested that the cells of the ductular epithelium undergo dynamic changes that probably result in secretion and/or desquamation. Adherence to cells could be demonstrated only at a late stage of these changes. The adherence of organisms associated with mastitis was studied using an in vitro test. Adherence generally paralleled prevalence as cause of disease, with Staphylococcus aureus and Streptococcus agalactiae adhering best. Strain variation suggested that virulence was related to adherence with S. agalactiae and S. dysgalactiae but not with S. aureus. It is proposed that specific adherence is an important aspect of pathogenesis of mastitis due to S. aureus and S. agalactiae.


Mitogenic stimulation of murine spleen cells: relation to susceptibility to Salmonella infection. The screening of several inbred strains of mice suggested that the capacity of their spleen cells to respond to the mitogenic effect of lipopolysaccharide (LPS) of gram-negative bacteria was correlated with their resistance to intraperitoneal infection with Salmonella typhimurium. An infection of LPS into mice caused changes in the in vitro responsiveness of their spleen cells to the mitogenic effects of LPS and phytohemagglutinin. Pretreatment of mice with whole ultraviolet (UV)-killed bacteria led to a marked rise in the in vitro response of the spleen cells to UV-killed bacteria, but not to LPS or or phytohemagglutinin. This enhanced response to UV-killed bacteria was not specific for the O antigens of the bacteria.


Humoral immunity in vaginal candidiasis. Serum antibody titers to Candida albicans were estimated in 37 women with recurrent vaginal candidiasis and in 148 normal American and Finnish subjects, using the passive-hemagglutination technique. The antibody titers ranged from 0 to 16 in normal individuals and 4 to 256 in vaginal candidiasis patients. Antibodies to C. albicans in the sera of vaginal candidiasis patients were found to be the secretory immunoglobulin A type, as determined by gel filtration and double-diffusion tests. The results were confirmed by the indir-ct fluorescent-antibody technique. Our findings suggest that, in vaginal candidiasis, the antibody response is mainly local, consisting of secretory immunoglobulin A, some of which finds its way into systemic circulation.


Repetitive counterelectrophoresis on agar gel for the immunological identification of esterases produced by strains of Lancefield's group A, B, and C streptococci. A method, repetitive counterelectrophoresis (RCE), was devised for detecting specific antibody to streptococcal esterase (STE). Reference antisera were prepared by immunizing rabbits with STE of streptococcal strains as follows: SS379 (group A, type 40), 69882 (group A, type 49), and H36B (group B, type Ib). Some human sera, derived from patients with scarlet fever, were also used as references. By this method, we have confirmed the immunological specificity of the STE produced by strains SS379 (STE-AI), 69882 (STE-AII), H36B (STE-B), and Austin (STE-C, group C) and have shown that the STE produced by strain 10706 (group C) is immunologically identical with STE-AI. Each STE presented a distinct colored line with the respective homologous antibody upon development of enzyme activity except for STE-AII, which formed a round spot with the homologous antibody. Horse activating factor (Hayano and Tanaka, 1973) formed a round spot with each STE. The factor in serum that reacted specifically with STE seemed to correspond to gamma globulin.