Monolayer cultures of gonadotrophs separated by velocity sedimentation: heterogeneity in response to luteinizing hormone-releasing hormone. Isolated pituitary cells from 14-day-old male and female and adult male rats were separated into enriched populations of gonadotrophs by velocity sedimentation at unit gravity. The cells, collected from five gradient fractions, were established in monolayer culture. Hormone content and hormonal secretion in response to LHRH were measured after 3 days in culture. Stored FSH and LH declined during culture, but the relative differences between fractions and between animal groups were maintained. In all fractions, LHRH stimulated the release of both FSH and LH in a dose-response fashion. The secretory potential per gonadotroph changed with fraction and the pattern was highly characteristic for each animal group. In 14-day-old females, both FSH and LH secretion increased 20 times from the fraction with the smallest to the fraction with the largest gonadotrophs. In preparations from 14-day-old males, only the release of FSH consistently rose with gonadotroph cell size. With cells from adult males, FSH as well as LH release increased with cell size, but the pattern was different for each hormone. Secretion did not always correlate with the cellular content of hormone before stimulation. The present findings suggest that gonadotrophs isolated from animals in a different physiological or developmental status may retain in short term culture certain of their previous functional characteristics. There is evidence that the gonadotroph cell population is heterogeneous not only in terms of the magnitude of the response to LHRH but also in terms of the relative proportion of FSH and LH secreted. Changes in the relative proportion of certain variants of gonadotrophs may be a cellular basis for differential regulation of FSH and LH secretion by a single releasing hormone.


Further studies on the role of calcium in glucocorticoid-induced lymphocytolysis. In a previous study comparing the effects of triamcinolone acetonide (TA) and a divalent cation ionophore (A23187) on rat thymocytes, we proposed an important contributory role for Ca2+ in glucocorticoid-induced cytolysis. The plausibility of this hypothesis was tested further in thymic lymphocytes (thymocytes) and lymph node lymphocytes (LN-lymphocytes). Thymocytes incubated in a Ca2+-containing medium responded to TA or A23187 with a concentration-dependent decrease in the number of viable cells. TA-induced cytolysis was reduced in Ca-free medium and was equally supported by Ca2+ and Sr2+, but not by Ba2+. A23187-induced cytolysis was supported by Ca2+ greater than Sr2+, but not by Ba2+. Thymocytes were also lysed by increasing concentrations of Ca2+ even in the absence of TA and A23187. LN-Lymphocytes, however, were less sensitive to the cytotoxic effects of Ca2+ under the same conditions. In the presence of Ca2+, thymocytes were lysed to a greater extent than LN-lymphocytes by TA, whereas the sensitivities to A23187-induced cytolysis were the same in both populations. Omission of Ca2+ from the incubation medium inhibited the cytolytic response to TA only in thymocytes. In contrast, A23187-induced cytolysis was impaired in Ca-free medium in both thymocytes and LN-lymphocytes. These observations confirm the previous findings on Ca2+ dependence of glucocorticoid-induced cytolysis in thymocytes. This pathway, however, may not be involved in glucocorticoid-induced cytolysis of LN-lymphocytes. Thus, a more basic and as yet undefined mechanism probably mediates the lymphocytolytic process.


32P-labeling patterns in rat pancreatic islets: tissue source of the radiophosphate released after glucose stimulation. Immediately after stimulation with glucose in vitro, isolated rat pancreatic islets prelabeled with [32P]orthophosphate release a pulse of [32P]orthophosphate into the media (the "phosphate flush"). Islets have been rapidly frozen before, during, and after this pulse to assess the concurrent changes in the distribution of tissue radioactivity. Under the present experimental conditions, approximately 90% of the islet radioactivity was soluble in perchloric acid (PCA-soluble) immediately before stimulation and slightly more than half of that was present as [32P]orthophosphate. The tissue pool of [32P]orthophosphate declined 55% and 62% after 7 and 14 min of stimulation which, respectively, incorporated the peak and the end of the heightened efflux of radioactivity. The net decrementa in tissue orthophosphate could account for all of the radioactivity which was released during the "phosphate flush." During the 14-min period of stimulation, labeled ATP and GTP (which had accounted for 13% and 4% of total PCA-soluble radioactivity before stimulation) increased 51% and 35%, respectively, and labeled ADP and AMP (which had accounted for 5.4% and 1.5% of PCA-soluble counts) fell 36% and 77%, respectively. Certain other PCA-soluble components, such as phosphorylcholine and phosphorylethanolamine, and total PCA-insoluble radioactivity were not demonstrably altered. The findings indicate that the "phosphate flush" originates from a labile pool of tissue orthophosphate. It remains to be established whether the simultaneous changes in the turnover of selected nucleotides are coupled to the translocation of orthophosphate or are mediated separately.


Modifications of luteinizing hormone biosynthesis and release by gonadotropin-releasing hormone, cycloheximide, and actinomycin D. In the first experiment, the influence of synthetic gonadotropin-releasing hormone (GnRH) on the time course of [3H]glucosamine ([3H]GA) and [14C]alanine ([14C]A) incorporation into LH by quartered rat anterior pituitary glands and the sequence of release of radiolabeled and total immunoreactive LH (IR-LH) was investigated. Radiolabeled LH was measured by immunoprecipitation and total IR-LH was measured by RIA. After lag periods of 5 and 20 min, respectively, incorporation of [14C]A and [3H]GA into tissue LH increased linearly for 4 h. GnRH stimulated incorporation of [3H]GA only. The ratio of [14C]A-LH to [3H]GA-LH (14C:3H ratio) in the tissue decreased significantly with time and with GnRH treatment. In the second experiment, replenishment of GnRH in the medium every 0.5 h elevated the release rate (release during each sequential 0.5 h) of both [3H]GA and [14C]A-labeled LH within 1.5 h. The release rate of radiolabeled LH increased linearly until 3.5 h. The 14C:3H ratio in LH released during each time interval was reduced by GnRH. The release rate of IR-LH increased linearly with time, plateaued by 1.5-3 h, and started to decline. In other experiments, cycloheximide blocked synthesis of [14C]A-LH and greatly reduced the GnRH-induced synthesis and release of [3H]GA-LH, but reduced release of IR-LH by only 25%. Actinomycin D had no effect on GnRH-induced synthesis and release of LH at 2 h, but significantly reduced both at 4 h. These data suggest that 1) the time course for the release of preexisting IR-LH differs from that for newly synthesized LH, 2) the newly synthesized LH released in response to high levels of GnRH has more sugar residues than that released under basal conditions, 3) the GnRH-induced LH release can occur under conditions in which LH synthesis has been blocked, and 4) synthesis of messenger RNA is not required for GnRH-induced LH release or short term LH synthesis but seems to be required for continued synthesis and subsequent release of LH.


Pancreatic beta-cell replication: relation to insulin secretion. The relationship between beta-cell replication and insulin release was investigated utilizing neonatal rat pancreatic monolayer cell cultures. Glucose-induced insulin release was either inhibited using diazoxide or mannoheptulose, or potentiated using theophylline. The corresponding effects on the frequency of beta-cell replication were determined by incubating cultures with [3H]thymidine and estimating the frequency of beta-cell labeling in aldehyde thionine-stained radioautographs. beta-Cell replication and insulin release were shown to be dissociable processes in two ways. First, in the presence of diazoxide (1-100 microgram/ml), insulin release was inhibited by as much as 86%, while the frequency of beta-cell replication was not reduced. Second, in the presence of theophylline (1 mM), insulin release was increased by 23%, while beta-cell replication was inhibited. Finally, the inhibition of both beta-cell replication and insulin release by mannoheptulose (5.5 mM) indicated that glucose utilization may be important for both of these beta-cell processes.


Ovarian steroid modulation of gonadotropin secretion and pituitary responsiveness to luteinizing hormone-releasing hormone in the female hamster. The significance of ovarian estradiol (E2) and progesterone secretion in the regulation of pituitary LH and FSH secretion and pituitary responses to LHRH was investigated in the hamster. Cycling females showed increased LH and FSH responses to LHRH on the morning of proestrus as compared to the responses observed on diestrus day 2. Pituitary responsiveness to LHRH declined on the evening of proestrus, after the preovulatory LH/FSH release. The secondary increase in serum FSH concentration on the morning of estrus was accompanied by a selective increase in the pituitary FHS response to exogenous LHRH. Hamsters ovariectomized (ovx) on diestrus day 2 exhibited daily afternoon LH surges but not FSH surges for at least 10 days after ovx. The magnitude of the LH surges in ovx hamsters was approximately 30-50% of that observed in proestrous females. The pituitary LH response to exogenous LHRH in ovx animals was about 25% as great as in proestrus hamsters. Serum FSH concentrations in ovx females increased by only 30% after LHRH injection, while similar treatment with LHRH resulted in 3- to 4-fold increments in serum FSH in proestrous hamsters. Implantation of E2 capsules in ovx hamsters resulted in increased gonadotropin responses to exogenous LHRH. Serum LH concentrations in the E2-implanted, LHRH-injected animals were as great as those observed after LHRH injection in proestrous females. Administration of LHRH, LH, or progesterone on the morning of proestrus failed to detectably alter the timing or magnitude of the proestrus afternoon FSH surge. The present results suggest that the increasing serum titers of estrogen on diestrus and early proestrus result in increased pituitary sensitivity to LHRH, and this increased sensitivity probably contributes to the magnitude of the preovulatory LH surge. The increases in LH and progesterone which occur during the afternoon do not seem to be responsible for triggering the proestrous FSH surge.


Effect of the bonito insulin on the insulin release from monolayer culture of rat pancreas. The effect of bonito insulin on insulin release was examined in the monolayer culture of rat pancreatic beta-cells. The beta-cells were preincubated for 5 to 20 hr with or without a small dose (100 microunits/ml) of bonito insulin in the medium containing 100 mg% glucose. And then, they were incubated in 300 mg% glucose alone or together with bonito insulin for 5 hr. There was no significant difference between the IRI release from these beta-cells with or without bonito insulin. The concentration of bonito insulin was augmented from 100 microunits/ml to 500, 1,000 and 2,000 microunits/ml. A significant inhibitory effect on the glucose-induced insulin release was observed only after the preincubation for 20 hr with 2,000 microunits/ml of bonito insulin.


The effect of luteinizing hormone releasing hormone (LH-RH) and oestrogen on RNA synthesis in anterior pituitary and different brain regions of rats. Oestradiol-17beta caused a marked reduction of RNA content in the cortex, hippocampus, brain stem, hypothalamus and RNA synthesis in the pituitary. LH-RH had facilitatory effect on the cortical and inhibitory influence on the hypothalamic RNA synthesis in vitro, and suppressed the pituitary RNA synthesis both in vivo and in vitro. The possible regulatory role of the LH-RH in the nucleic acid metabolism in discussed.


The delayed effect of sodium valproate on the photoconvulsive response in man. The acute effect of a single oral dose of sodium valproate on the photoconvulsive response has been studied. In 7 of 9 patients naive to this drug photosensitivity was abolished or reduced. A similar effect was seen in only 2 of 7 already receiving chronic sodium valproate therapy. The effect appeared 1 to 5 hr (mean, 3 hr) after attainment of peak sodium valproate concentration and lasted up to 5 days.


Effect of anticonvulsants on spiking activity induced by cortical freezing in cats. In an attempt to know whether the existing anticonvulsants act on epileptogenic focus, the effect on SA induced by freezing of the visual cortex was examined in gallamine-immobilized cats. The SA was localized in the neighbor or ipsilateral cortex of the freezing area; little epileptiform activity was produced in the contralateral anterior cortex, and ipsilateral thalamus and hippocampus. Spike frequency and its amplitude were stable over 8 hr. Diazepam suppressed SA and decreased spike frequency and its amplitude. Dipropylacetate and acetazolamide also suppressed SA. On the other hand, phenobarbital, carbamazepine, and a high dose of phenytoin enhanced SA and increased the spike frequency. Low doses of phenytoin and trimethadione were without effect in this aspect. Taurine suppressed SA and changed the spikes to wave-like forms. The EEG arousal response was depressed with phenobarbital, carbamazepine, diazepam, and a high dose of phenytoin, but not with the other drugs examined. From these results, it is suggested that diazepam, dipropylacetate, acetazolamide, and taurine depress the epileptogenic focus activity itself without relation to the activating system.


Valproic acid binding to human serum albumin and determination of free fraction in the presence of anticonvulsants and free fatty acids. The interaction between valproic acid (VPA) and human serum albumin (HSA) was investigated using the equilibrium dialysis technique under various conditions. Solutions of VPA in HSA (2 x 10(-4) M) were dialyzed against isotonic phosphate buffer at 37 degrees C. Protein and buffer compartments  were assayed for VPA by GLC. The free fraction (alpha) of VPA increased from 0.13 at 27 microgram/ml to 0.49 at 103 microgram/ml. Scatchard plots were linear, indicating the existence of one type of binding site. The mean (+/- % SD) number of binding sites per macromolecule was 2.06 +/- 3.7% and the mean (+/- % SD) association constant was 2.69 x 10(4) +/- 15.0% liters/mole. The effects of three anticonvulsants (phenytoin, phenobarbital, and carbamazepine) and four major free fatty acids (FFA) (stearic, palmitic, oleic, and linoleic) on alpha were studied. The free fraction, 0.18, was not affected by phenobarbital (20 and 40 microgram/ml), carbamazepine (10 and 20 microgram/ml) or phenytoin (20 and 40 microgram/ml). Each of the four FFA caused a significant increase in alpha: 19--48% increase at 100 microgram/ml of FFA and 88--118% at 200 microgram/ml.


Protein interactions in the outer membrane of Escherichia coli. Specific protein interactions in Escherichia coli outer membrane were analyzed using chemical cross-linking with truly cleavable reagents and symmetrical two-dimensional sodium dodecyl sulphate/polyacrylamide gel electrophoresis. The major outer membrane proteins were shown to form cross-linked complexes. These include multimers of lambda receptor, protein I, II, III and the free form of lipoprotein. Lipoprotein was also found to be cross-linked to proteins II and III. The identity of many of these complexes was verified using appropriate mutants missing the proteins in question. No new protein interactions were detected in the mutants even when three of the major proteins were missing. Proteins II, III and the free form of lipoprotein could also be cross-linked to the peptidoglycan layer of the cell wall.


Peptidyl transferase of bacterial ribosome: resistance to proteinase K. 70-S ribosomes and 50-S ribosomal subunits from Escherichia coli D10 were treated with proteinase K for increasing periods of time. Peptidyl transferase activity and sparsomycin-induced binding of (U)C-A-C-C-A-[3H]Leu-Ac were tested in the treated particles, the binding of the substrate being more sensitive to the protease than peptide bond formation. Comparison of the amounts of proteins present in the treated particles with the residual activity indicates that only proteins L3 and L14 are released at a similar rate to that at which peptidyl transferase activity is lost. Proteins related to this ribosomal activity by other techniques are lost at a faster rate than the activity itself. In addition, the results indicate that sparsomycin stimulates the binding of the substrate by a different mechanism from that which inhibits peptide bond formation.


Immunofluorescent serum gliadin antibodies in children with coeliac disease and various malabsorptive disorders. I. Technique, clinical evaluation and diagnostic use of a gliadin antibody assay using pyruvic aldehyde-treated human red cells. An immunofluorescent gliadin antibody assay is described using pyruvic aldehyde-stabilized human erythrocytes coated with gliadin. Fifty coeliac children all had high serum IgG-antigliadin titres during a normal diet or a challenge with gluten. On a gluten-free diet (30 children), titres were much lower. In patients followed-up for one year on a gluten-free diet, an initial rise in titres was followed by a slow decline. On challenge, IgG-antigliadin titres showed a slow rise or persistence at the same level in most patients. Fifty-two percent of control children with malabosrptive disorders, but without the typical "flat" mucosal lesion on jejunal biopsy, were shown to have positive titres in their sera, as were 6% of normal children and 4% of adult blood donors. The fluorescent antibody technique was compared with methods commonly used to detect wheat-protein antibodies, and was found to be superior to all of them. The immunofluorescent gliadin antibody assay appears to be useful in following-up children with coeliac disease, and in selecting patients for jejunal biopsy, although it does not replace biopsy.


Torsion dystonia: a case with delayed remission following thalamotomy and phenothiazine therapy. Torsion dystonia in 13-year-old girl progressed to severe opisthotonus and dystonic posturing of the limbs, accompanied by severe psychic changes, cachexia and calcification around joints. Drug therapy was ineffective. Ventrolateral thalamotomy and chlorpromazine brought about a delayed but remarkable amelioration of all symptoms and signs, still present 4 years later.


Renal transplantation: patients with previous saphenous vein-coronary artery bypass grafts. 2 patients underwent saphenous vein-coronary artery bypass grafting before successful renal transplantation. Both patients were able to undergo appropriate therapy for various complications, which further emphasizes the resilience that these patients can have after adequate coronary revascularization.


Insulin secretion and carbohydrate metabolism in the dystrophic mouse. Dystrophic mice were investigated with regard to their regulation of blood glucose and insulin secretion in vivo. The following were also measured: tissue glycogen levels, activity of the glycogenolytic hydrolase, acid amyloglucosidase, and in vitro glucose utilization by liver, muscle and adipose tissue. Basal levels of blood glucose and plasma insulin of dystrophic mice were essentially within the same range as in the clinically unaffected littermate controls. Dystrophic mice had a decreased tolerance to glucose and glibenclamide; the secretion of insulin in response to these secretagogues was moderately reduced. Insulin release following beta-adrenergic stimulation, however, was increased in the dystrophic mice. Glycogen levels and acid amyloglucosidase activity were increased in dystrophic muscles but were normal in liver. Acid amyloglucosidase activity in pancreatic islets was lower in the dystrophic mouse. Glucose utilization in vitro appeared normal in liver tissue from dystrophic mice;  in dystrophic muscle there was a threefold increase in 14CO2-production with no concomitant increase in either glycogen or 14C-incorporation into glycogen. 14CO2 production and 14C-incorporation into lipid and glycogen were increased in dystrophic adipose tissue. We suggest that the decreased glucose tolerance, and the reduced insulin response to glucose in the dystrophic mouse are compensated by an increased glucose utilization in muscle and adipose tissue and an increased beta-adrenergic-mediated secretion of insulin.


Paracetamol versus placebo: effects on post-operative course. Paracetamol was compared with placebo in a double-blind crossover study, in which essentially the same operation was performed on two separate occasions in 24 healthy patients, namely surgical removal of bilateral impacted wisdom teeth. Commencing on the day of surgery, either paracetamol (Panodil; 1.0 g x 4 for 2 days, then 0.5 g x 4 for the next 2 days) or placebo tablets were given, followed by crossover to the alternative treatment at the second operation about 4 weeks later. Several objective and subjective assessments were recorded for paired comparison of post-operative courses. Swelling on the 3rd day after operation when paracetamol was given averaged 71% of that measured when placebo was given (p less than 0.05). After paracetamol, a tendency was noted towards reduced local hyperpyrexia and less post-operative bleeding. The pain and preference scores were clearly in favour of paracetamol. The results provide evidence to suggest that paracetamol may reduce an acute, posttraumatic inflammatory reaction.


Pulmonary and systemic vascular effects of exogenous prostaglandin I2 in fetal lambs. Effects of PGI2 upon pulmonary vascular resistance and systemic arterial pressure of near term fetuses were evaluated in anesthetized, exteriorized, unventilated fetal lambs by means of an open-chest, pump-perfused lung preparation. These effects were compared to those of PGE1 and PGE2 in the same animals. Intrapulmonary arterial infusions of PGI2 produced dose-dependent decreases in pulmonary vascular resistance and systemic arterial pressure. Effects of PGI2 upon pulmonary vascular resistance are intermediate with respect to those of PGE1 (greater) and PGE2 (less). PGI2 is a potent hypotensive compound when administered to fetal lambs, producing dose-dependent decreases in systemic arterial pressure.  When the three prostaglandins are infused into the pulmonary artery, effects on systemic arterial pressure of PGE1 and PGI2 are nearly identical. Effects of PGE2 are considerably less. PGI2 is a potent dilator of fetal lamb vessels and could play a role in circulatory control during perinatal transition.


Murine yolk sac hematopoiesis studied with the diffusion chamber technique. The proliferation and differentiation of visceral and of parietal yolk sac cells from 9-day-old mouse embryos were studied in diffusion chambers (DC). After culture, the visceral yolk sac yielded predominantly macrophages whereas the parietal yolk sac displayed mainly plasmocyte-like cells. It is shown the latter do not synthesize immunoglobulin but rather elaborate Reichert's membrane. Neither visceral nor parietal yolk sac contained detectable pluripotent stem cells (CFUs) prior to or after DC culture, a finding which is discussed in the light of the current literature.


Determination of the numerical density of spherical and ellipsoidal particles in the zona glomerulosa, based on distribution measurements. II. Application of the methods. Stereologic investigations of the adrenal zona glomerulosa of rats were carried out in control, potassium loaded and heparin treated animals. The nuclei of the cells were considered as spheres, and the mitochondria as prolate (= elongated) ellipsoids of revolution. Size and shape distribution, numerical densities and mean volumes of particles were calculated. Control measurements were performed in order to test the acceptability of estimation errors. These stereologic methods proved to be suitable in experimental studies.


Correlations between the neurobiology of colour vision and the psycholinguistics of colour naming. Neurobiological experiments demonstrate that colour sensation is perceived by the brain by processes which, in principle, follow the opponent colour pairs scheme proposed by Hering in 1874. Tests on colour naming in various European, Asian and Central American languages have shown that the opponent scheme is also reflected in psycholinguistics. The linguistic evolution of colour terms proposed by Berlin and Kay (1969) is correlated directly with the ontogenetic development of language in children as elucidated by Jakobson (1941). Colour vision is therefore a suitable field for interdisciplinary investigations of brain processes and linguistics.


Effect of acetylsalicylate on surgical bleeding, postoperative mortality and allograft survival in rats undergoing heart transplantation. 18 rats were treated with L-ASA before heart transplantation and daily thereafter until death or rejection. 22 animals acted as controls. A significantly higher post-operative mortality rate, without any significant modification of the transplant survival time, was found in L-ASA-treated group.


Somatostatin content and release of isolated pancreatic islets from obese-hyperglycemic mice. The somatostatin content in pancreatic islets of obese-hyperglycemic mice was much lower than in the islets of normal mice. Also the release of somatostatin was decreased from the islets obtained from the obese-hyperglycemic mice. Tissue culture for 1 week changed neither the content of, nor the amount of somatostatin released from, the pancreatic islets.


Effect of suppression of endocrine or exocrine pancreatic function in hemorrhagic shock. Somatostatin did not influence the pathologic consequences of hemorrhagic shock, but pancreatic duct ligation prevented the post-oligemic decline of arterial pressure and formation of toxic factors. These results indicate that pancreatic acinar cells release myocardial depressant factor and are important in the pathophysiology of shock.


Reactions of cysteine, its derivatives, glutathione coenzyme A, and dihydrolipoic acid with isothiocyanates. Isothiocyanates react with deprotonated SH groups of investigated compounds to give the esters of N-monosubstituted dithiocarbamic acid. In the presence of the SH and NH2 groups (Cys, GSH), isothiocyanates react primarily with the SH groups. The reactions are dependent on the pKa SH values.


lambda-Prophage induction by furocoumarin photosensitization. Furocoumarin photosensitization induces lambda-prophage from lysogenic Escherichia coli cells; this effect is clearly due to the photoreaction that furocoumarins give with DNA, and it appears connected with the formation of monoadducts rather than of diadducts (cross-links).


Family interaction and communication deviance in disturbed and normal families: a review of research. Recent family interaction studies are reviewed with an emphasis on looking for dimensions along which disturbed and normal families differ. Several areas of consistency in the literature were found, including: family coalition patterns, patterns of conflict, flexibility versus rigidity, family effectiveness and efficiency, and deviant styles of communication. It was concluded that several measures reliably discriminate disturbed from normal families and that one type of measure in particular is a reliable predictor of thought disorder in offspring. Implications for clinical practice and future research are discussed.


[S-Acyl derivatives of thiosalicylamides having antifungal activity. II]. Some S-acyl derivatives of N-alkylthiosalicylamides [Table I: substances (I leads to XXXI)] were prepared and tested for antifungal activity. The substances, most of which had not been previously reported, were prepared by condensation of 2-mercapto-N-alkylbenzamides with suitable acylating agents. The antifungal activity of the compounds was tested in vitro against Candida albicans and Trichophyton mentagrophytes. For some compounds the was tested activity against the above strains fungicidal, Candida tropicalis and Saccharomyces cerevisiae. Many of the compounds proved to have high antifungal activity comparable with that of Clotrimazol. The results extended knowledge on the structure-antifungal activity relationships of this class of compounds. The compounds with the highest antifungal activity were: 2-acetylmercapto-N,n-heptylbenzamide (XXVIII); 2-acetylmercapto-5-Cl-N,n-propylbenzamide (XIV); 2-acetylmercapto-N,n-octylbenzamide (XXXI); 2-acetylmercapto-N,n-pentylbenzamide (XXV); 2-acetylmercapto-N,n-hexylbenzamide (XXVII).


Failed intrauterine device contraception and limb reduction deformities: a case-control study. Three recent case reports have suggested an association between failed intrauterine device (IUD) contraception and limb reduction deformities in the resulting offspring. To clarify further the purported teratogenic role of the IUD in the etiology of these defects, we conducted a case-control study of 96 mothers who had given birth to infants with limb defects. Interview data about IUD exposure at conception among these mothers were compared with interview data from 2 sets of controls: (1) 915 mothers of infants with other major defects and (2) a subset of the first group consisting of 169 mothers with chromosomally defective infants. No significant increase in the incidence of IUD use was found for cases when compared with either control group. Estimates of the relative risk were 1.28 (95% confidence limits [CL]of 0.29 to 5.67) and 1.78 (95% CL of 0.25 to 12.82) when the first and second groups of controls were used, respectively. These risk figures were not significantly different from 1.0. The evidence accumulated thus far militates against a teratogenic role for the IUD.


Pituitary adenoma and oral contraceptives: a case-control study. A search of the centralized data resource available at the Mayo Clinic for all cases of pituitary adenoma diagnosed in the population of Olmstead County, Minnesota, disclosed an increasing incidence of this tumor in women of childbearing age. The sex, age, and temporal relationships suggest that, if this increase is real, oral contraceptives should be considered as one of the possible etiologic factors. A case-control study, however, did not reveal an association of prior use of oral contraceptives with pituitary tumor--relative risk, 0.5; 95% confidence interval, 0.1 to 2.2. No association was found with other possible risk factors, i.e., prior head injury, radiation therapy, seizures, and smoking. Thus, unless other etiologic agents can be identified, it appears that the increasing incidence is due to advances in diagnostic and surgical technology rather than to a specific etiologic factor.


Possible luteolytic effects of luteinizing hormone-releasing hormone in normal women. The administration of five subcutaneous 250-microgram doses of lutienizing hormone (LH)-releasing hormone (LHRH) at 4-hour intervals, the first injection being given at 8 A.M. on 1 or 2 consecutive days between days 1 and 9 following the LH surge in normal women, shortened the luteal phase from 1 to 4 days in 16 of 17 treatment cycles. There was a better efficiency of treatment when LHRH was administered on days 6 to 9 after the LH surge as compared with days 1 to 5. In fact, the luteal phase was shortened from 3.3 +/- 0.2 days versus 1.4 +/- 0.2 days (P less than 0.01) and the serum progesterone level was decreased to 44% +/- 6% versus 71% +/- 6% of control levels (P less than 0.01) when the neurohormone was injected late as compared with early in the luteal phase. The present data raise the possibility of a luteolytic effect of LHRH in normal women and indicate the interest of such a near-physiologic approach for the control of luteal function and time of appearance of menses.


Gonadotropin-like substance in the preimplanted rabbit blastocyst. The presence of a gonadotropin-like substance in preimplanted rabbit blastocyst fluid was determined bu radioreceptorassay of human chorionic gonadotropin (hCG)/luteinizing hormone (LH), using receptors prepared from bovine corpus luteum, rat testis, and rat ovaries. An average of 16.6 microliter of fluid containing 0.83 ng of luteotropic material was recovered from each blastocyst. An intense fluorescence was exhibited by the trophoblastic cells of the blastocysts treated with fluorescein-conjugated gamma-globulin isolated from antiserum raised against human chorionic gonadotropin. Serum concentrations of LH as well as estradiol, progesterone, and 17 alpha-hydroxyprogesterone up to day 6 after mating were determined in pregnant rabbits and compared with those in pseudopregnant and normal rabbits. In pregnant rabbits after mating, a surge of 62 +/- 15 ng of luteinizing hormone and 73 +/- 22 ng of progesterone/ml of serum occurred which returned to basal levels at 6 hours and on day 1, respectively. Secondary increases in serum luteinizing hormone of 26 +/- 12 to 36 +/- 16 ng/ml on days 3 and 5 and in serum progesterone of 16 +/- 4 and 14 +/- 5 ng/ml on days 5 and 6, respectively, were observed in pregnant rabbits.


A controlled comparative trial of a new antidepressant, ciclazindol. The author describes a controlled, double-blind, comparative trial of a new tetracyclic compound, ciclazindol (WY 23409), against amitriptyline in the treatment of thirty-five patients admitted to hospital with depressive illness. Each patient was randomly allocated to three weeks treatment with either 50 mg b.d. ciclazindol or 50 mg b.d. amitriptyline. In the event of a poor response the dose level was raised to 75 mg b.d. Separation of cases of endogenous depression and severity of depression were assessed by the Levine-Pilowsky Depression Questionnaire, a self-rating technique. Severity of depression was also assessed using the Hamilton Rating Scale. No significant difference was noted between the drugs in either the degree or the rate of response nor when the endogenous cases alone were studied. The interesting observation was made that only one-third of ciclazindol patients gained weight compared to almost three-quarters of the amitriptyline group and the mean weight gain of the latter was over double that of the ciclazindol group. The author concludes that ciclazindol offers promise and merits further study using higher dosage levels once its full safety trials have been completed.


Multicentre study of 'Flagyl' in the prevention of post-operative anaerobic infection. In a study carried out with 565 patients undergoing gynaecological and general surgical procedures, metronidazole ('Flagyl') was used pre- and post-operatively and the incidence of post-operative infection, particularly that due to anaerobic organisms, was recorded. This was not a controlled study but, by comparison with other series, it would appear that the use of oral metronidazole had substantially reduced the likelihood of supervention of anaerobic sepsis. It is probable that the use of intravenous metronidazole therapy would have reduced the incidence of anaerobic sepsis still further had this preparation been available at the time.


Aldactone and acebutolol in treatment of hypertension. Twenty-one essential hypertensives were randomly treated with Aldactone, acebutolol or a combination of the two drugs. Statistically significant falls in systolic and diastolic pressures were demonstrated during treatment with Aldactone 100 mg daily, also when acebutolol and Aldactone 200 mg daily were administered. When the combination of Aldactone and acebutolol was given, diastolic pressures were lowered to a greater degree than seen with individual treatments.


A comparison of indoprofen and indomethacin in the supportive treatment of soft-tissue injuries in football players: a pilot study. Twenty-four professional footballers suffering from recent soft-tissue injuries were included in a double-blind pilot trial comparing the efficacy and tolerability of indoprofen 800 mg/day and indomethacin 80 mg/day. Twelve subjects were allocated at random to each of the two drugs. Mean reduction of pain and swelling by the 3rd day of treatment was greater in the indoprofen group, as was the rate of disappearance of these symptoms, calculated in terms of t 1/2. A statistically significant (p less than 0.05) superiority of indoprofen versus indomethacin was, however, reached only for pain elicited by active movement. No adverse reactions were recorded with either drug.


A multicentre double-blind comparison of oxaprozin aspirin therapy on rheumatoid arthritis. Preliminary clinical studies showed that oxaprozin (4,5 Diphenyl-2-oxazolepropionic acid) has anti-inflammatory and analgesic properties with a plasma half-life of about 40 hours. Consequently, a multicentre, double-blind parallel trial was conducted for 12 weeks at thirteen investigator sites, utilizing 212 patients with classic rheumatoid arthritis and comparing oxaprozin 600 mg/day, oxaprozin 1200 mg/day and aspirin 3900 mg/day. Both the oxaprozin and aspirin-treated patients had statistically significant improvement from baseline periods, in most key categories evaluated. Oxaprozin administered twice a day (b.i.d.) was as effective as aspirin administered four times a day (q.i.d.) and caused significantly less tinnitus (p less than 0.001). Fewer patients receiving high dose oxaprozin (2%) dropped out of the study because of unsatisfactory response than did those receiving aspirin (10%). There were no clinically significant laboratory abnormalities in the gastro-intestinal, renal, hepatic or haematological parameters monitored. This study suggests that oxaprozin is effective and well tolerated in the treatment of rheumatoid arthritis.


Comparison of efficacy of amoxapine and imipramine in a multi-clinic double-blind study using the WHO schedule for a standard assessment of patients with depressive disorders. A multi-clinic double-blind controlled study on amoxapine in comparison with imipramine, using the WHO Schedule for a Standard Assessment of Patients with Depressive Disorders, was performed and the data were analyzed with 111 patients. The assessment of severity of illness and overall improvement indicated clearly the superiority of the antidepressive effect of amoxapine to that of imipramine. The onset of antidepressive effect of amoxapine was clearly more rapid than that of imipramine, and in more than half of the patients in the amoxapine group the improvement was seen within four days following the drug administration. Amoxapine was superior to imipramine in terms of safety and usefulness. The side-effects due to amoxapine appeared less frequently and were less serious than with imipramine. The difference between amoxapine and imipramine was especially remarkable for hypotensive effect. The antidepressive effect of amoxaphine was superior to that of imipramine for almost all symptoms and signs. Amoxapine displayed an especially remarkable effect on psychomotor retardation, depressive feeling, anxiety and tension, somatic complaints and sleep disturbance.


Kinetics of native insulin in diabetic, obese, and aged men. The kinetics of unlabeled porcine insulin were studied in 69 nondiabetic male subjects aged 18-83 yr with obesity indexes of 0.93 - 1.51 and in 12 maturity-onset diabetics age 46-78 yr with obesity indexes of 0.95-1.56 by using the euglycemic clamp technique. Analysis of the insulin kinetic data by using a mathematical model permitted the determination, for each individual, of steady state distribution masses and degradation rate constants. The individuals were grouped to allow comparison of the results on the basis of age, obesity index, or diabetes. The responses over a period of 120 min to an infusion and wash out of insulin show some transient as well as steady state differences with age, obesity, or diabetes. Analysis of these data by use of compartmental models leads to the conclusion that in the steady state the ratio of insulin in extravascular spaces to that in plasma (T/P) is decreased in the moderately obese group (26%) and in the diabetic group (17%) but increased in the older group (13%) when each is compared with the appropriate control. Since extravascular insulin includes both insulin bound to receptors and insulin in the interstitial fluid, the observed changes in the extravascular to plasma mass ratio most likely reflect changes in in vivo binding to receptors, although the magnitude of the change would be modified somewhat by changes in the size of the interstitial spaces relative to plasma. In addition, the rate of entry of new insulin into plasma (BSDR) was increased in the diabetic population (45%; P less than 0.02) as well as in the moderately obese group (27%) but was decreased somewhat in the older group (11%). The following general conclusions can be drawn from the results: The pattern of parameter changes seen with obesity is similar to that seen with maturity-onset diabetes. The decrease in T/P seen with obesity and with maturity-onset diabetes cannot be accounted for solely by changes in fasting plasma insulin levels in these populations. The pattern of changes seen in the older subjects is opposite that seen in the maturity-onset diabetics, which suggests that diabetes is a perturbation distinct from the normal aging process. Finally, the changes in the metabolism of insulin are not large, making it unlikely that they are the sole cause of the major alterations in glucose tolerance seen with aging, obesity, or diabetes.


Syngeneic transplantation of fetal rat pancreas. III. Effect of insulin treatment on the growth and differentiation of the pancreatic implants after reversal of diabetes. Eight 18-316 fetal pancreases were transplanted to syngeneic alloxan diabetic male rats. Some of the recipients were treated with insulin for a 7-day period immediately after transplant. By previously published clinical criteria, three groups of recipients could be identified after reversal of diabetes by the transplanted tissue: insulin-treated rapid reversal; insulin-treated slow reversal; and control (not treated with insulin). Five animals in each group were sacrificed after glucose tolerance testing for morphologic and hormonal analysis of the transplanted tissue. The insulin-,glucagon-, and somatostatin-positive islet cell masses of the fetal pancreatic implants were quantitated. There was a correlation between the beta cell mass of the implants and the glucose tolerance exhibited by the host animals. The rapid response insulin-treated recipients had significantly greater implant beta cell mass and insulin content compared with the other groups. There was no difference in implant alpha cell mass among the groups, but the insulin-treated implants had a significantly greater glucagon content. The delta cell mass of insulin-treated rapid response was less than that of the other two groups. The results are discussed in relation to previously reported morphometric analysis 15 days after transplantation. The relationships of transplanted beta cell mass, beta cell differentiation, transplant site, and cell-to-cell interactions within the transplanted islet to the control of glucose homeostasis are also discussed.


Dental infection in a cirrhotic patient. Source of recurrent sepsis. A patient with alcoholic cirrhosis had multiple episodes of sepsis with Klebsiella pneumonia. Repeated searches for the source of infection finally revealed the organism in the root of a tooth. Evidence indicated that this site was the primary source of infection. The importance of dental infections in alcoholics and the difficulty in diagnosing those infections are emphasized by this case.


[Stereotaxic: measurements of the skull cap and skull base in merino sheep (Ovis aries L.)]. On the 14 skulls of 6-month old merino labms, stereotaxical measurements were carried out. Not only individual variations were found, but they also dependent on sex. A small assymetry was found on the diagrams of frontal measurements of the skull cap and base. The point bregma is more variable than the lambda. The position of the cranial third of the skull base in 6 month old lambs is not yet constant and until adult age it will shift about 30 degrees to 35 degrees dorsalward and forward. Since the skull cap and base do not grow parallely, and their cranial part has not constant position, we do not recommended this age group for chronic experiments with implanted electrodes. Owing to smaller variations of the skull cap and base and nearer position of the sutura sagittalis toward the medial line, male lambs would be more suitable for short-term experiments and for the stereotactic atlas of deep brain structures.


The effects of crowding stress on the pancreatic islets morphology in golden hamsters. Behavior and the pancreatic islets morphology of golden hamsters exposed for different periods (24 h, 15 and 30 d) to crowding stress were investigated. The crowding induced an intensive turmoil and enhanced irritability and aggressiveness, particularly among the female specimens. Within a week, a rank hierarchy seemed to establish and therefore, later the fighting incidence was more reduced. Marked morphological alterations were recorded in the endocrine pancreas, affecting in various degrees all cell types, but especially the insulin-producing cells. The B-cells showed increased sizes, higher incidence of mitotic divisions, a drastical reduction of secretory granules amounts, enlargement of GOLGI complexes, mitochondria swelling and extension of the ergastoplasm. On the other hand, after 30 d, part of these cells displayed pycnotic nuclei, large cytoplasmic vacuoles and increased number of lasosomes and lipid inclusions. Due to the B-cell hyperplasia, the relative number of both A1- and A2-cells per islet occurred diminished and their typical localization modified.  The islets of 15 and 30 d crowded specimens showed enlarged sinusoids and clear peri-insular spaces. The above morphological modifications, which suggest as a whole the global stimulation of gland secretory activity, are presented in relation with other authors findings. The presumably involvements of adrenal-cortex and -medulla hormones in the mediation of stress-induced glycemic and pancreatic alterations are discussed.


Insulin administration in the frog Rana tigrina: studies on the blood glucose and the histology of the endocrine pancreas. The effect of various doses of insulin (25-, 50-, and 150 U/kg body weight) on the blood glucose level and islet-cytology of the frog Rana tigrina was studied until 96 h. Following the hormonal administration the frogs exhibited hypoglycemia, abnormal neuromuscular activity and degranulation of both the insulin secreting beta- and glucagon secreting alpha-(alpha2-) cells of the pancreatic islets. The action of insulin was dose and temperature dependent; the higher the dose and temperature, the greater the hypoglycemia and atrophy of islet tissue. The insulin-induced convulsive activity appears to be due to the direct action of this hormone on the nervous system; the shocks are not influenced by thermal variation. The great sensitivity of Rana tigrina to exogenous insulin seems to be related to only a few alpha2-cells in the endocrine pancreas and consequently, a smaller amount of circulating glucagon in this animal.


[Are Hegar's rods omissible? (author's transl)]. The widening effect of Sulproston 500 microgram IM on the cervix uteri was investigated in a double-blind study conducted in 50 non-pregnant volunteers. In women in the postmenopausal stage who had been given Sulproston, resistance-free widening of the cervix with Hegar's rods was H 8.33 +/- 0.87. Easy passage of the cervix uteri and of the internal os uteri was achieved with Hegar 8.22 +/- 0.66 in the first half of the cycle and with Hegar 8.35 +/- 3.06 in the second half of the cycle. In all three groups, the size of the rods was significantly smaller following intramuscular application of physiological saline solution. The widening effect of PGE2 or PGE2 derivatives on the cervix, which has been known for quite some time in patients with pregnant uterus, applies in equal measure to the non-pregnant cyclic uterus and to the postmenopausal uterus.


Isolation, genetic mapping and some characterization of a mutation in Escherichia coli that affects the processing of ribonuleic acid. Temperature-sensitive mutants were isolated from an rnc (RNase III-) strain of Escherichia coli, and their rRNA metabolism was analyzed on 3% polyacrylamide gels. One of these mutants was unable to produce 23S and 5S rRNAs at the nonpermissive temperature. When an rnc+ allele was introduced to this strain, it remained temperature sensitive. At the nonpermissive temperature, this strain could then produce 23S rRNA but was unable to make normal levels of 5S rRNA. In matings and transduction experiments, the defect in rRNA metabolism and temperature sensitivity behaved as a syndrome caused by a single point mutation, which was mapped at min 23.5 on the E. coli chromosome. This mutation probably affects an enzyme, ribonuclease E (RNase E), which introduces a cut in the nascent rRNA transcript between the 23S and the 5S rRNA cistrons. The mutation rne is recessive with respect to temperature sensitivity and the pattern of rRNA. Revertants able to grow at 43 degrees and with normal metabolism of rRNA were isolated; genetic analysis showed that they do not contain the original rne mutation, suggesting that they were true revertants. By combining the rne mutation with an rnc mutation, double rnc rne strains were synthesized, which behaved very similarly to the original rnc strain from which the rne mutation was isolated. Such strains have RNA metabolism that is similar to that of rnc strains at permissive temperatures, but at the nonpermissive temperature they fail to synthesize p23, m23, and 5S rRNAs. Thus, the experiments reported here, together with previous studies, suggest the existence of a new processing ribonuclease activity in Escherichia coli, which is called ribonuclease E.


[Specificity of introducing the transposon that determines chloramphenicol resistance (Tn9) into the chromosome of Escherichia coli K-12]. Translocation of Tn9 coding for chloramphenicol-resistance from lambdaatt80 genome into bacterial chromosome was studied. Three preferential sites of Tn9 integration, attTn9A, ATTTn9B and attTn9C, were found on the chromosome of Escherichia coli K-12; attTn9A was mapped between purD and rpoB loci, attTn9B was cotransducible with argG, attTn9C was located between 61 and 66 minutes of the standard E. coli K-12 genetic map. The integration of Tn9 in these att sites occurs with almost equal probability. Tn9 integrated in attTn9A shown significantly higher frequency of excision and P1-transduction frequency than Tn9 integrated in either of two other attTn9 sites.


[Regulation of the activity of Escherichia coli deo-operon structural genes: the mutation mapped within the operon boundaries and affecting drm and pup gene activity]. The mutant AIR38 is isolated from Escherichia coli K-12 strain deficient in thymidilate synthetase and deoxyriboaldolase (HfrH, thy, dra)--by selection for low thymine requirement on the medium containing inosine as the carbon source. Under the conditions mentioned the mutant AIR38 (thy, dra) grows at low thymine concentration (2 mkg/ml), and is uncapable to grow in the presence of thymidine (40 mkg/ml). Dra+ derivatives of the AIR38 do no catabolize inozine in the presence of thymidine as well. The mutation AIR38 is mapped within the deo-operon between drm and pup mutation markers. The levels of phosphodeoxyribomutase and purine nucleoside phosphorylase in cell extracts of AIR38 are 2.5-6-fold decreased. In transductional experiments with phage P1 and the mutant AIR38 as recipient the delayed haploidization of merozygotes dra+, AIR+/dra, AIR38, thy and the dominant expression of the sensitivity to thymidine in the presence of inosine as the carbon source are observed. It is supposed that the mutation AIR38 affects the structural gene of purine nucleoside phosphorilase by altering the mode of interaction of this enzyme with the membrane under the conditions of thymine starvation.


[Mutagenic action of nitroso compounds on Escherichia coli cells]. An attempt to induce some forward and back mutations in two Escherichia coli strains (his- and HfrH requiring thiamine) under the action of the carcinogenic nitrosamines--dimethylnitrosamine (DMN) and diethylnitrosamine (DEN)--is described. For this purpose the cells of E. coli were treated with 5% DMN or 1% DEN for 1 hour at 37 degrees C in 0.14 M NaCl. It was shown that the sensitivity of both strains to both nitrose compounds was not the same. DEN was 5-fold as toxic as DMN for the E. coli cells. DMN and DEN induced neither mutations of resistance to 10(-3) M valine, nor reversions in histidine-dependent strain. These mutations were obtained after the cells were treated with 0.1 M NaNO2. Lethal effects of DMN increased more than in 5 times and the toxicity of DEN did not change in hydroxylating mixture, in which nitrosamines derived to active compounds. Under these conditions both carcinogenes showed a mutagenic activity. DEN proved to be about twice as strong mutagenically as DMN. Thus, in our experiments we could see that DMN and DEN could induce both forward and back mutations in E. coli.


Isolation, cloning and characterization of argF gene DNA from Escherichia coli K-12. A 1650 base pair (BP) fragment carrying the entire argF structural gene with its associated control regions was isolated from an EcoRI/BamHI digest of phi80argFilambda cI857 DNA. This segment was cloned using the EcoRI and BamHI cleavage sites in the plasmid pBR322. A preliminary restriction map of the argF region was prepared. RNA polymerase binding studies indicated that the argF promoter is located approx. 30 base pairs from the EcoRI terminus of the cloned DNA segment.


Cloning of the phr gene and amplification of photolyase in Escherichia coli. A new technique is developed for physically enriching recombinant DNA molecules in an in vitro recombination mixture. UV-irradiation of the donor DNA before recombination enables photoreactivating enzyme (PRE) (deoxyribodipyrimidine photolyase, EC 4.1.99.3) to attach to the donor segments in recombinant molecules. This attached protein causes retention of the recombinant molecules on a nitrocellulose filter, while molecules not containing donor DNA pass through. The bound DNA  is repaired of its UV damage and released for insertion into cells by exposure to photoreactivating light in situ, yielding approx. 350-fold enrichment. Although applicable to any gene, this procedure has been used in cloning the Escherichia coli phr gene itself, permitting 100-fold amplification of the gene product in vivo.


Influence of inflammatory bowel disease on intestinal microflora. The microflora of the jejunum, ileum, and colon has been studied from operative samples in Crohn's disease (n = 30), ulcerative colitis (n = 15), and controls (n = 40). There was no significant difference in the flora of patients with ulcerative colitis compared with controls. In Crohn's disease there was a significant increase in E. coli (P less than 0.001) and B. fragilis (P less than 0.001) in the ileum and of E. coli (P less than 0.001) and lactobacilli (P less than 0.01) in the colon. The abnormal ileal flora in Crohn's disease was unrelated to serological evidence of disease activity (indices: ESR, serum albumin, serum seromucoids), diameter of the ileum, or excision of the ileocaecal valve. The abnormal colonic flora in Crohn's disease was not related to presence of macroscopic colitis.


A new host, Simulium argyreatum Meig., for the cytoplasmic polyhedrosis virus of blackflies in Czechoslovakia. The cytoplasmic polyhedrosis virus (CPV) from the midgut of blackfly larvae is recorded from a population of Simulium argyreatum in Czechoslovakia. A 7.7% infection of 2766 sampled larvae was manifested by white spots in their body. 85% of the white-spotted larvae were infected with CPV when sampled in June. In August 9% of 1377 larvae had white spots but only 10.6% of them were with CPV. Out of 823 collected pupae, only 3 imperfect pupae had the virus in apparent form. Other infections producing white spots in larvae were due to the microsporidia Plistophora multispora, P. simulii, P. debaisieuxi, Thelohania fibrata, the fungus Coelomycidium simulii and the mermithids.


[Effect of alpha-mercaptopropionylglycine (alpha-MPG) and sodium dipropylacetate (DPA) on antibody formation (III) (author's transl)]. Mechanisms of the immunostimulative activity. In the present paper, an investigation was carried out to clarify the mechanisms regarding immunostimulative activity of alpha-MPG and DPA. Phagocytosis of peritoneal macrophage was enhanced by high concentration of alpha-MPG, but was unaffected with DPA in vitro. Mice were immunized with an i.v. injection of sheep erythrocytes and 3 days later spleen cells were isolated and incubated with alpha-MPG and DPA FOR 24 HR. Although neither drug affected the number of recovered and living cells, hemolytic plaque forming cells were increased with alpha-MPG and there was a tendency to increase with DPA. Phytohemagglutinin-P and/or lipopolysacchride E. coli-induced blast formation was not reinforced by either drug in spleen cells of mice, and no mitogenic activity was found in the spleen cells. In the early stage of immuno-response to polyvinylpyrrolidone as a T cell independent antigen, the titre of antibody showed no increase after either drug. alpha MPG and DPA apparently act as potentiators in antigen modifications of macrophages.


[Ozone-oxygen therapy for  gynecologic carcinomas. The effect of parenteral-ozone oxygen mixture administration on free fatty acids and triglycerides in patients with gynecologic carcinomas]. As some authors suspect that ozone influences the metabolic process of fat, we tried to analyse the influence of an ozone-oxygen gas which was applied parenterally. 40 patients with gynecological cancer received 10 ml ozone-oxygen gas with a content of 450 gamma ozone and venous blood was removed before and 10 minutes after application. The serum was lyophilized and the level of fatty acids and triglycerids was determined by the method of Randerath (1965). a statistically significant decrease of the concentrations was observed after application of ozone. Different theories as to the cause of this action are discussed.


[In vivo study of the effects of i.v. immunoglobulin therapy. Tissue distribution of native and enzyme-treated human immunoglobulin fractions given i.v. in man]. The tissue distribution of intravenously injected native and enzyme-treated human gammaglobulin has been investigated in human rectum mucosa. FITC-labelled antibodies to human gammaglobulin were used to locate the injected antibody preparation. Three hours after application of these different gammaglobulin-preparations no enrichment in the mucosa was found. Neither the native gammaglobulin nor the enzyme-treated gammaglobulin showed any concentration in the lamina propria of the large bowel mucosa or an intracellular enrichment in the epithel. This study shows that the quicker elimination of enzyme-treated gamma-globulin from the circulation cannot be explained by an intracellular increase.


[Immunological problems in radiotherapy and adjuvant cytostatic chemotherapy of breast cancer]. The possible importance of the immune system in early development and progression of breast cancer is being discussed. The different laboratory methods in controlling the specific and non-specific immune reactivity are summarized. The modificaiton of the immunological parameters by radio- resp. chemotherapy is critically presented on the basis of published data and on own results. An analysis of the data obtained by various immunological methods in respect to their consequences in diagnosis and prognosis is done. An immunological monitoring in the control of radio-, chemo- and immunotherapy in breast cancer patients is proposed.


[Questioning of women in puerperium concerning their experiences at delivery]. 200 patients of the Universitäts-Frauenklinik Würzburg were asked in a quenstionnaire about their attitude towards antenatal care and child birth as performed in a big gynecological hospital. Safety was their main point for choosing a big hospital. Patients' reactions about intensive perinatal care, analgesia, husbands' presence and rooming in are summarized as well as their criticism of organization and treatment during the ante-, peri- and postnatal period. 5 conclusions are drawn to make child birth in a big hospital more humane.


[Development and current possibilities of cerebrospinal fluid diagnosis in the routine laboratory. 2. Studies on protein composition]. In the 1st part of this paper (Schulz et al. 1979) a survey was given of the cytological and the chemical procedures at present available for cerebrospinal fluid diagnostics. The 2nd part describes different laboratory methods to separate the protein mixture of cerebrospinal fluid into protein fractions or single proteins. These surveys intend both an improvement of the quality and a unification of laboratory methods regarding diagnostic examinations of cerebrospinal fluid.


Comparison of endocrine secretion by monolayer cultures derived by different procedures from neonatal hamster and rat pancreas. Monolayer cultures derived from neonatal hamster or rat pancreas by two different epithelioid cell-enriching gravity sedimentation procedures varied in ability to maintain uniform levels of insulin secretion with increased culture age. Rat pancreatic cultures were superior in this respect to identically derived hamster preparations, depending on the preparative procedure employed. Quantitative differences in the temporal pattern of insulin secretion by different rat pancreatic culture preparations were ascribed to plating cell density and consequent terminal cell density as a function of preparative procedure such that reduced densities favored sustained secretory levels. These findings suggest the importance of tissue species and preparative procedure in deriving pancreatic monolayer cultures capable of sustained levels of insulin secretion with age.


Formulating therapeutic objectives as a prelude to program evaluation. The author emphasizes the need for more precise formulation of goals for mental health treatment programs. He describes the Veterans Administration's five-year effort at goal formulation using ideas borrowed from operations research and education. With goals from various VA programs as examples, he illustrates how goal statements can be written with more specificity. He also describes how matching rating-scale items can be developed to measure the degree to which each goal is being met.


More rigid costing formulas required under proposed federal regulations. Many hospital laboratories are at a distinct disadvantage in competition with commercial laboratories, but a more reasoned approach to costs and utilization can erase the deficit.


SHUR: a shift of power?

SHUR: a shift of power? HEW's Health Care Financing Administration links uniform reporting and Medicare reimbursement under the provisions of the proposed System for Hospital Uniform Reporting. This summary of a position paper published by the public accounting firm Ernst & Ernst covers two questions: What will SHUR mean to hospitals? and, What can be done about it?


A Catholic health care system: who's responsible? If Catholic hospitals cannot develop cooperative efforts within and among themselves, it will be difficult for them to do so with other health care systems or the government. Their desire to establish a united, comprehensive Catholic health care system is not enough; they must be willing to risk even a partial loss of independence in order to achieve this goal and preserve the Catholic healing ministry.


Calcitonin secreting struma-carcinoid tumor of the ovary. The ultrastructure of strumal carcinoid tumors at times may reveal a far more complex structure than can be ascertained from light microscopy. It may at times be indistinguishable from medullary carcinoma of the thyroid gland. Both tumors appear to be capable of producing 5-hydroxyindolacetic acid and calcitonin. The common shared biologic and ultrastructural features suggest a common origin, from neuroectodermal cells.


Cryptosporidial enteritis in a patient with congenital hypogammaglobulinemia. Cryptosporidium is a potentially pathogenic coccidial protoxoan known to inhabit the striated border of interestinal epithelium in several animal species. Recently two cases of human intestinal cryptosporidiosis have been described. The purpose of this report is to document a third case of chronic debilitating cryptosporidial enteritis in a young boy with congenital hypogammaglobulinemia. The endogenous protozoan life cycle and morphologic features are examined as studied by light and electron microscopy.


Influence of intermittent hyperglycemic glucose levels on the phagocytosis of microorganisms by human granulocytes in vitro. Repeated brief exposures to glucose in concentrations of 0.8 g/100 ml significantly depresses in vitro phagocytic ingestion of opsonized Gram-positive and Gram-negative bacteria by human peripheral polymorphonuclear granulocytes (PMN's), while the same concentration of L-aminoacid mixtures (Freamine) under the same conditions has no influence on it.


Cellular immunity in patients with the Wiskott-Aldrich syndrome before and after administration of transfer factor: a follow-up study. Four patients with Wiskott-Aldrich syndrome were treated with transfer factor (TF), in an attempt to improve their clinical condition. Before and during treatment, cellular reactivity was followed in vivo (delayed type skin reactivity) and in vitro (lymphocyte transformation). In all patients positive skin reactions were occasionally observed during TF treatment. The lymphocyte reactivity in vitro to phytohaemagglutinin (PHA) was slightly diminished, the responding capacity of the mixed lymphocyte culture (MLC) severely impaired and the response to bacterial, viral and fungal antigens absent before and during TF treatment. No clinical improvement was observed after large doses of TF. No correlation between skin test reversal and TF administration was found, and lymphocyte transformation in vitro did not improve. Subsequently, a double blind trial, in which we compared the effect of TF and placebo, was carried out in these patients, which revealed no effect of TF therapy. It is concluded that the results of treatment with TF in these patients are highly controversial.


Isolation of IgE from normal mouse serum. Mouse IgE was isolated from reaginic ascitic fluid and an antiserum to it was produced in rabbits. An immunoadsorbent, prepared with this antiserum,  was used to isolate IgE from normal mouse serum. Mouse serum proteins, isolated by (NH4)2SO4 (40--60%) precipitation, were applied to the anti-IgE immunoadsorbent and eluted with 3 M KI or 3.5 M NaSCN. In addition to containing IgE, the eluate contained other immunoglobulins which were subsequently removed with specific immunoadsorbents to IgG1, IgG2a, IgG2b, IgA and IgM. Further purification was achieved by gel filtration on Bio Gel A 0.5 m, ion exchange chromatography on DEAE cellulose, and Pevikon block electrophoresis. Since non-immunoglobulin components were demonstrated by immunoelectrophoresis, IgE was isolated from this pool with an anti-Fab immunoadsorbent and eluted with 3.5 M NaSCN. Based on recovery of 987 micrograms IgE from 855 ml NMS, IgE concentration was estimated at 1.15 microgram/ml. Analysis of purified IgE by immunoelectrophoresis and immunoprecipitation in gel detected a single component, partially identical to isoelectric focused rat IgE myelomas IR162 and IR331 Antiserum to it could completely abolish 48 h but not 2 h passive cutaneous anaphylaxis (PCA) reactions. Normal IgE, when used as a diluent, could inhibit 48 h PCA reaction of reaginic antibody suggesting that it binds to mouse skin in a way similar to reagin. This demonstrates that IgE can be isolated from normal mouse serum. The availability of this source of mouse IgE should facilitate further use of this animal model in studies of allergic reactions.


Immunosuppressive activity of Fusarium toxins. Effects on antibody synthesis and skin grafts of crude extracts, T2-toxin and diacetoxyscirpenol. The inhibitory effect of crude Fusarium poae extracts, T2-toxin and diacetoxyscirpenol on synthesis of antibody to sheep red blood cells was investigated, as well as the effect of T2-toxin on skin graft rejection. Fusarium crude extracts, T2-toxin and diacetoxyscirpenol cause a significant reduction in thymus weight and inhibit responsiveness to sheep red blood cells. The effect on antibody synthesis was critically dependent on the drug doses used. The effect of an immunosuppressive dose of T2-toxin disappeared within 6 days after the end of the treatment. The administration of T2-toxin subsequent to immunization stops antibody synthesis. T2-toxin significantly prolongs the period required for skin graft rejection.


Suppression of secondary immune response by antilymphocyte serum: time relationship between immunization and administration of antilymphocyte serum. The effect of antilymphocyte serum (ALS) on the secondary humoral immune response to sheep erythrocytes (SRBC) in rats was studied by the Jerne plaque assay technique. Its effect was also studied on the delayed hypersensitivity (DH) response to SRBC by the foot pad swelling test. ALS(N), which was prepared against lymphocytes from normal rats, had no effect on the secondary humoral and cellular response or on the primary cellular response, when administered postantigenically. ALS(I), which was raised against lymph node cells from SRBC immunized rats produced significant immunosuppression of the secondary response to SRBC when administered either before or after the antigenic injections. In the case of DH, ALS(I) behaved just like ALS(N) having no effect on the secondary response and suppressing the primary only when administered prior to the antigen.


Influence of extracellular Ca2+ and Mg2+ on chemotactic factor-induced neutrophil aggregation. The influences of extracellular Ca2+ and Mg2+ on chemotactic factor-induced rabbit polymorphonuclear neutrophil (PMN) aggregation was studied using a recently described Coulter counter assay technique. Compared with those of other PMN functional assays (adhesion, chemotaxis, degranulation, and phagocytosis), the cation requirements for cell aggregation appear unique. Chemotactic factor-induced aggregation did not occur in the absence of either cation. When the concentration of both cations was increased equivalently, aggregation increased. The effect plateaued at 2.8 mM of Ca2+ and Mg2+. When the concentration of one cation alone was increased, aggregation peaked. Further increases inhibited maximal aggregation. By systematic variation of the cation concentrations, optimal aggregation was found at Ca2+ and Mg2+ concentrations of 2.8 mM. At these concentrations, significant aggregation was induced with chemotactic doses of bacterial factor, the chemotactic fragment of human C5, and the synthetic chemotactic tripeptide, formyl-met-leu-phe. Thus, under these conditions, chemotactic factor-induced aggregation of neutrophils may be a useful indicator of interactions of chemotactic factors with neutrophils.


Biologically active derivatives of fatty acids: prostaglandins, thromboxanes, and endoperoxides. The causal role assigned to the E and F prostaglandins in inflammatory processes, implied by the antiinflammatory action of prostaglandin synthetase inhibitors, is not consistent with the findings reported here that a compound (MK-447) capable of increasing levels of these prostaglandins is antiinflammatory in classical animal models of acute inflammation. That both MK-447 and prostaglandin synthetase inhibitors depress the enzymatic formation of PGG2 from arachidonic acid suggests that this endoperoxide plays a pivotal role in acute inflammation. However, in view of the intermediate nature of PGG2, it seems likely that such a pivotal role for this substance is a function of its ability to be converted to other inflammatory mediators. Possible candidates for a causal role are thromboxane A2 (TXA2) prostacyclin (PGI2), both of which derive from PGG2. However, direct evidence is presented to show that an oxygen equivalent released in the enzymatic conversion of PGG2 to PGH2 is a prime factor in inflammation.


Steroids, aspirin, and inflammation.

Steroids, aspirin, and inflammation. The ability of adrenal corticosteroids to both both suppress inflammation and compromise host defenses has been well documented. Recently, a series of in vitro and in vivo experiments, based on our new knowledge of the cell biology of inflammation and the biochemistry of the phagocytic cell itself, has provided new insights into the mechanism of steroid action in the inflammatory process. Evidence is presented that pharmacologic doses of steroids are capable of inhibiting each of the steps in phagocytic-micro-organism interaction: chemotaxis, recognition and opsonization, phagocytosis, membrane fusion, and degranulation. In addition, steroid alteration of the postphagocytic superoxide production, hydrogen peroxide generation, and prostaglandin and thromboxane synthesis is described. The antiinflammatory effects of aspirin and indomethacin can be explained almost entirely by virtue of their ability to inhibit cyclooxygenase, this preventing the transformation of arachidonic acid to both prostaglandins and thromboxanes. The cortisol-induced inhibition of endoperoxides, prostaglandins, and thromboxanes (at a site proximal to the release of arachidonic acid) may well explain those antiinflammatory actions that cortisone shares with aspirin. However, patients treated with nonsteroidal antiinflammatory agents effectively combat infections. In contrast, corticosteroids have more profound effects, as can be seen by the inhibition of superoxide production, with the subsequent decrease in hydrogen peroxide generation and the diminution in release of the antibacterial lysosomal hydrolases within the phagocytic vacuole. Thus, corticosteroids interfere with the killing of microorganisms. This new understanding of the pharmacologic action of cortisol on phagocytic cells explains, we believe, how glucocorticoids alleviate inflammation while, at the same time, they permit multiplication of the offending microorganism within the phagocyte.


Prostaglandins, thromboxanes, and polymorphonuclear leukocytes: mediation and modulation of inflammation. When appropriately stimulated (even in the absence of phagocytosis), human polymorphonuclear leukocytes release and/or generate proinflammatory materials and substances capable of provoking tissue injury. These include hydrolases and nonenzymatic substances ordinarily contained within lysosomes, as well as oxygen-derived free radicals. It is now possible to add prostaglandins and thromboxanes to this list. Whereas prostaglandins are capable of eliciting many phenomena associated with inflammation, their effects on cyclic nucleotide metabolism may render these compounds antiinflammatory. Thus, the very cells that release mediators of inflammation provide a mechanism for modulating the inflammatory response.


Significance of an in vitro phenomenon in which murine erythrocytes are lysed by autologous spleen cells and spherules of Coccidioides immitis. Spleen cells from mice immunized with a variety of antigens and incubated in vitro with killed spherules of Coccidioides immitis lyse six to eight times more autologous murine erythrocytes than normal spleen cells and spherules. Cellular and biochemical events in this phenomenon were investigated to ascertain its significance. Kinetic studies suggested that hemolysis results from the activation of some immune cells by spherules. The capacity of spherules to activate these cells is rather unusual because, of the inert particles tested, only zymosan A and crude chitin demonstrated comparable activity. Furthermore, although the hemolytic phenomenon occurred in serum-free medium, more lysis was effected by immune cells and opsonized spherules or zymosan A than by immune cells and untreated fungal particles. Sheep, chicken, and human erythrocytes were not lysed in the hemolytic phenomenon; however, hemoglobin in chicken and sheep erythrocytes was oxidized. Both the murine erythrocyte lysis and oxidation of ovine hemoglobin correlated with the reduction of Nitro Blue Tetrazolium by immune cells adherent to spherules, and both phenomena appeared to be mediated by H2O2 released into the medium by activated cells. Spleen cells reactive with spherules could not be depleted by treatment with iron carbonyl, antiimmunoglobulin plus complement, or anti-brain-associated theta plus complement, but they were partially or completely depleted after rosette formation with erythrocytes coated with antibody or murine complement. Using light and electron microscopy, we noted that immune spleens contained more neutrophils than normal spleens, that these neutrophils reduced Nitro Blue Tetrazolium after stimulation with phorbol myristate acetate, and that they were the most prevalent cell type adherent to spherules after incubation in vitro.


Adhesion of commensal bacteria to the large intestine wall in humans. Biopsies taken during colonoscopic examination of the human large bowel were used to examine the relationship of the commensal bacterial to the mucosal epithelial cell surface. Bacteria were seen adhering to the exposed epithelial cell surface and also to the mucus sheet. Isolation of aerobic organisms showed that Escherichia coli are closely associated with the gut wall throughout the large intestine. One strain of E. coli predominated in each biopsy, and this strain was present along the whole length of bowel. Adhesion of bacteria to the gut wall does occur in vivo and may be one of the factors involved in the ability of an organism to colonize and persist.


Importance of the intestinal inflammatory reaction in salmonella-mediated intestinal secretion. The ability of Salmonella typhimurium to invade the intestinal epithelium is essential to the pathogenesis of salmonella-induced intestinal secretion. This invasion is accompanied by an intense acute inflammatory reaction. The present study tests the hypothesis that the acute inflammatory reaction may have a role in the pathogenesis of salmonella-induced secretion. Two groups of rabbits infected with S. typhimurium were studied: normal animals and animals pretreated with nitrogen mustard. Nitrogen mustard depletes the polymorphonuclear leukocyte pool and thereby prevents the formation of an acute inflammatory reaction. In vivo ligated ileal loops were constructed and infected 72 h after nitrogen mustard administration when polymorphonuclear leukocytes were undetectable. Nitrogen mustard treatment markedly inhibited salmonella-induced  secretion. Ileal histology in normal animals infected with S. typhimurium revealed an intense acute inflammatory reaction, while in animals pretreated with nitrogen mustard only a rare polymorphonuclear leukocyte was seen. The antisecretory effect of nitrogen mustard was not merely a nonspecific effect since nitrogen mustard treatment did not inhibit cholera toxin-induced secretion and did not alter either ileal morphology nor the activities of various intestinal enzymes in normal animals. Nitrogen mustard also did not alter the virulence of the inoculated S. typhimurium. These data suggest that the mucosal inflammatory reaction induced by salmonella invasion may be important to the pathogenesis of the salmonella secretory process. The mechanism by which the inflammatory reaction stimulates secretion is not known.


Cultivation of Ureaplasma urealyticum in continuous culture. Continuous culture of Ureaplasma urealyticum is reported with a steady-state cell biomass of greater than 10(6) cells per ml. Thus, large cell numbers can be easily obtained; in addition, the system provides a powerful means for exploring what nutrients(s) limits the growth yield of this organism. Urea is shown not to be the growth-limiting nutrient in conventional media, although when provided in excess it appears to be completely hydrolyzed.


Mitomycin C stimulates production of a toxin in Shigella species that causes morphological changes in Chinese hamster ovary cells. Shigella dysenteriae produced a toxin that caused morphological changes in Chinese hamster ovary cells which was different from the neurotoxin (Y. Takeda, K. Okamoto, and T. Miwatani, Infect. Immun. 18:546--548, 1977). It was found that the toxin was produced not only by S. dysenteriae but also by S. flexneri, S. boydii, and S. sonnei. The production of the toxin was stimulated by the addition of 1 microgram of mitomycin C per ml to the culture in the early logarithmic growth phase.


Lymphoid cells in endotoxin-induced production of the amyloid-related serum amyloid A protein. Endotoxin-treated mice exhibit a rapid rise in the level of the serum amyloid A (SAA) protein, but this effect is not observed in endotoxin-resistant C3H/HeJ mice. To evaluate the role of lymphoid cells in the production of SAA protein, C3H/HeJ mice were adoptively transfused with endotoxin-sensitive (C3H/HeN) bone marrow cells. After such adoptive transfer, endotoxin treatment of C3H/HeJ mice resulted in high serum levels of SAA protein. The ability of chimeric mice to make SAA protein correlated with the presence of endotoxin-sensitive B lymphocytes and macrophages. These findings suggest that lymphocytes and/or macrophages play an important role in initiating SAA protein synthesis after endotoxin treatment and suggest a possible mechanism by which chronic infection or inflammation leads to amyloidosis.


Dissociation of leukocyte alkaline phosphatase from the bactericidal activity of neutrophils. We have identified an apparently healthy individual with no history of repeated infections whose neutrophils were virtually devoid of alkaline phosphatase activity by either spectrophotometric or histochemical assay. His cells showed normal bactericidal activity toward both Staphylococcus aureus and Escherichia coli when tested in vitro. We have also demonstrated that L-p-bromolevamisole, a potent inhibitor of alkaline phosphatase in many tissues, is likewise an effective inhibitor of the neutrophilic enzyme. Concentrations of this compound which caused nearly complete inhibition of the neutrophilic enzyme did not impair the ability of intact cells to kill bacteria. These results suggest that leukocyte alkaline phosphatase is not required for the normal bactericidal activity of neutrophils.


Transfer by bone marrow cells of increased natural resistance to Klebsiella pneumoniae induced by lipopolysaccharide in genetically deficient C3H/He mice. In a previous study we demonstrated that lipopolysaccharide failed to elicit nonspecific resistance in C3H/He lipopolysaccharide low-responder mice against Klebsiella infection in contrast to its activity in a closely related histocompatible high-responder subline, C3HeB/Fe. Complete restoration of lipopolysaccharide-induced protection against 10(5) Klebsiella was obtained in the present study by transferring bone marrow from high-responder mice to the highly deficient C3H/He mice. The ability of C3H/He mice to clear and destroy bacteria in 5 h was also transferred by C3HeB/Fe marrow cells. In contrast, when high-responder C3HeB/Fe mice were reconstituted with low-responder bone marrow, the clearance and destruction of K. pneumoniae were similar to what is observed in the high-responder strain, but survival was only temporary. Collectively, our data show that the failure of C3H/He mice to respond to lipopolysaccharide with nonspecific immunity is due to a defect in two types of bone-marrow-derived cells--radioresistant and radiosensitive.


Relationship between enterotoxin production and serotype in enterotoxigenic Escherichia coli. We examined the relationship between serotype and enterotoxin production in 109 enterotoxigenic Escherichia coli strains isolated from 109 patients with severe cholera-like diarrhea in Dacca, Bangladesh. Of 69 strains producing both heat-labile and heat-stable toxins, 59 (86%) belonged to the one of four O serogroups, and 56 (81%) of these strains belonged to one of six O:K:H serotypes. In contrast, 34 strains producing only heat-stable toxin were distributed among 15 O serogroups, and six strains producing only heat-labile toxin were distributed among six O serogroups. Twelve strains producing heat-labile and heat-stable toxins and five strains producing heat-stable toxin were found which had the same serotype (O78:K-:H12) and biotype. It appears that at least in one geographic setting E. coli strains producing both heat-labile and heat-stable toxins are more restricted in their O groups and O:K:H serotypes than E. coli that produce only heat-stable toxin and that certain serobiotypes may commonly include strains which produce both toxin types.


Hemagglutination patterns of enterotoxigenic and enteropathogenic Escherichia coli determined with human, bovine, chicken, and guinea pig erythrocytes in the presence and absence of mannose. A hemagglutination (HA)-typing system has been developed for the presumptive identification of enterotoxigenic Escherichia coli (ETEC) possessing the colonization factor antigens (CFA) CFA/I or CFA/II. E. coli isolates are grown on CFA agar and tested for mannose-sensitive (MS) or mannose-resistant (MR) HA of human, bovine, chicken, and guinea pig erythrocytes. CFA/I-positive ETEC exhibit MRHA with human, bovine, and chicken erythrocytes, but no HA with guinea pig erythrocytes. CFA/II-positive ETEC produce HA (MRHA) only with bovine and chicken erythrocytes. Common pili appear to be the primary MS-hemagglutinin of E. coli because the prototype strain K-12 exhibits HA (MSHA) with all but bovine erythrocytes. However, only 6.6% (23 of 351) of E. coli belonging to the classical enteropathogenic E. coli serogroups (EPEC) possessed the same HA pattern as strain K-12; 42% of the EPEC cultures (146 of 351) were similar to K-12 in producing MSHA with chicken and guinea pig erythrocytes and no HA with bovine erythrocytes, but different in that these produced either no HA or MRHA with human erythrocytes. These EPEC-associated HA patterns were assigned to a separate category, termed HA type III. Non-EPEC serogroups associated with sporadic diarrhea (i.e., the facultatively enteropathogenic E. coli, or FEEC) and 41% (19 of 46) of available Salmonella isolates also produced HA type III patterns. This observation is of considerable interest because many FEEC possess somatic antigens cross-reactive with Salmonella. Although the biochemical basis for this result has not been established, the data reported herein suggest a relationship between the HA type III phenotype and virulence (enteropathogenicity) in both the EPEC and FEEC serogroups. We propose that HA typing be used in conjunction with serotyping of E. coli to determine the degree of association of HA type III E. coli with sporadic diarrhea in infants and young children.


Adjuvants in the induction of suppressor cells. The effect of different mycobacterial adjuvants on the parameters of delayed hypersensitivity was investigated in strain 13 guinea pigs. The composition of the tubercle bacilli and the type of vehicle in which the antigen was presented determined the presence and extent of suppressor cell activity. When antigen was introduced in complete Freund adjuvant, both adherent and nonadherent cells had suppressive properties, with the suppressive effect demonstrable in vitro from 1 to 5 weeks after sensitization. Suppressor activity was indicated in vivo by a reduction of delayed footpad hypersensitivity in animals presensitized with complete Freund adjuvant.


Capacity of virulent Treponema pallidum (Nichols) for deoxyribonucleic acid synthesis. Treponema pallidum (Nichols) was extracted from infected rabbit tissue, and cell lysates were prepared for monitoring thymidine kinase and deoxyribonucleic acid polymerase activities. No thymidine kinase could be demonstrated in preparations of T. pallidum or the cultivable T. phagedenis biotype Reiter. Significant levels of deoxyribonucleic acid polymerase were detected in both treponemal samples. Interestingly, comparisons of polymerase activity among a spectrum of bacterial genera revealed a direct correlation between enzyme concentrations and estimated generation time. Incorporation of [3H]uridine and [3H]thymidine into macromolecules by intact T. pallidum and the Reiter treponeme was examined. Selective ribonuclease-deoxyribonuclease digestion and cesium chloride gradient banding demonstrated that T. pallidum, independent of the host, and T. phagedenis were capable of synthesizing deoxyribonucleic acid only from the [3H]-uridine precursor.


Strain specificity of opsonins for group B streptococci types II and III. Strains of types II and III group B streptococci do not appear to be uniformly susceptible to opsonization by antibody-containing human sera, as studied using both a chemiluminescence and a radiolabeled bacterial uptake technique. We could not demonstrate a correlation of serum-sensitive or resistant strains with capsular antigen quantities, although serum absorption studies with whole organisms and HCl, trichloroacetic acid, and saline extracts indicated that the antibody to type-specific capsular polysaccharide is important in opsonizing both serum-resistant and serum-sensitive strains. Since trypsin treatment produced significantly enhanced opsonization of serum-resistant and serum-sensitive strains, proteins present on some group B streptococci may be important antiphagocytic factors.


Characteristics of lipid A-protein complex from endotoxin of Shigella dysenteriae type 1 (S and R strains). Mild acetic acid hydrolysis of endotoxin (lipopolysaccharide-protein complex) of Shigella dysenteriae type 1 (S and R forms) yielded a lipid A-protein complex that consisted of amino acids, fatty acids, and sugar and, in terms of chemical composition, displayed no marked differences between the S and R forms. Its protein portion (53 to 56%) consisted of at least 16 amino acids. In the fatty acid portion (14 to 18%), myristic, 3-hydroxymyristic, palmitic, and stearic acids accounted for 50%. The sugar portion (10 to 12%) consisted solely of glucosamine. The remainder was unidentified substances, most of which contained phosphorus. Lipid A-protein complexes derived from both S and R forms were not toxic for mice in doses up to 1,000 microgram/mouse, but their Linulus test activity had increased considerably as compared with the starting lipopolysaccharide-protein complex material: from 10(-6) to 10(-10--10(-12) mg/ml. The lipid A-protein complexes were readily soluble in a water solution of triethylamine, in dimethyl sulfoxide, and in pyridine.


Comparison of sulfamethoxazole alone and combined with trimethoprim in urinary tract infections. In a double blind, randomized study, sulfamethoxazole was compared alone and in combination with trimethoprim as commonly used in therapeutic regimes for the treatment of uncomplicated acute urinary tract infections in out-patients. The cure of sulfamethoxazole alone was 92.2%, and for sulfamethoxazole plus trimethoprim 97.6%. The rate of side-effects for the former was 5%, for the latter 21.8%. If the failure rate plus the rate of occurrence of rash, which necessitated discontinuing the drug, are combined, it appears that 8.8% of the patients were at a disadvantage receiving sulfamethoxazole compared to 9.7% for the combination of sulfamethoxazole plus trimethoprim. When considering the cure rate and rate of side-effects together, therefore, the position of sulfamethoxazole alone as a suitable drug in this type of infection is defended.


Antibacterial efficacy of cefazolin and cephradine in neutropenic mice. The activity of cefazolin (CEZ) and cephradine (CED) was studied in experimentally infected neutropenic mice. Neutropenia was induced by 600 rad whole-body irradiation; an infection was induced by the injection of 5 X 10(6) Escherichia coli into the thigh on Day 5 after irradiation. Antibiotics were administered 1 h later, and antibacterial activity was estimated from bacteria counts made in the homogenized individual thighs 3 h after infection. The effect of a low dose of each of the cephalosporins on the infection was significantly lower in the absence of granulocytes than in animals with intact host defence; at higher dosages the effect of both antibiotics on the infection was the same in neutropenic and unirradiated mice. In the neutropenic mice, CEZ was 2.95 times more active than CED against E. coli in vivo, this difference in activity being similar to that found earlier in normal mice.


Influence of carbonyl iron treatment on lymphocyte subpopulations. The influence of carbonyl iron treatment on different human mononuclear cell subsets was investigated. There is evidence that, in addition to removal of phagocytic cells, lymphocytes are depleted, B cells being more affected than other subpopulations.


Effect of the MER tubercle bacillus fraction on the responsiveness of mice to T-independent antigens. The effect of treatment with the methanol extraction residue (MER) mycobacterial fraction on the immunological responsiveness of BALB/c mice to the T-independent antigens pneumococcal polysaccharide type III (SIIII) and trinitrophenyl-lipopolysaccharide conjugate (TNP-LPS) was ascertained. Pretreatment with MER prevented the establishment of immunological paralysis by threshold doses (10 or 15 microgram) of SIII and by a paralyzing dose of 100 microgram TNP-LPS. The induction of immunological paralysis by SIII was unaffected by treatment with the bacterial adjuvant Corynebacterium parvum and with the B cell mitogens PPD, LPS (Escherichia coli lipopolysaccharide), and dextran sulfate.


Characterization of pig lymphocyte subpopulations by adherence to nylon wool. The property of adherence to nylon wool was used to separate and characterize pig blood lymphocyte subpopulations. Thymus-dependent null cells proved the least adherent lymphocytes, and B cells, bearing surface Ig or Fc receptor, the most adherent. sIg+ lymphocytes detected by immunofluorescence and by 51Cr release cytotoxicity showed similar frequency and adherence properties. Lymphocytes rosetting with SRBC in PBS or in dextran had similar adherence patterns and showed both non-adherent and adherent components. Lymphocytes bearing major histocompatibility complex (MHC) Ia-like antigens shown by eosin exclusion or 51Cr release cytotoxicity were enriched in the adherent fraction, but also occurred in significant numbers in the non-adherent fraction. Non-adherent Ia+ cells were predominantly sIg- and probably T cells, whereas the adherent Ia+ group included the sIg+ cells and some sIg- cells.


The mechanism of action of soluble lymphocyte mediators. VI. Effect of migration inhibitory factor (MIF) on macrophage microtubules. The effect of migration inhibitory factor (MIF) on macrophage microtubules was examined by functional, biochemical and morphological methods. It was found that: (1) the microtubule-stabilizing agent deuterium oxide (D2O) inhibits spontaneous macrophage migration from capillaries and enhances the migration blocking effect of MIF; (2) MIF does not modify the amount of total tubulin in macrophages, as determined by an 3H-colchicine binding assay, but increases significantly the proportion of tubulin present in polymeric form; (3) macrophages exposed to MIF and examined by immunofluorescence with specific antitubulin antibody demonstrate a striking increase in the percentage of cells with a well-organized microtubular network, characterized by a high density of thick fibrils and prominent paranuclear microtubule organizing centers. It is concluded that MIF inhibits cellular motility by inducing the formation of numerous microtubules, probably by enhancing tubulin polymerization.


Regulation of lymphocyte activation by serum factors. The effect of autologous normal mouse serum on activating properties of the polyclonal B cell activators lipopolysaccharide and purified protein derivative was studied. 'Low' concentrations (0.1-1%) were found to be slightly stimulatory, whereas 'high' concentrations (greater than 10%) were clearly inhibitory both for the induction of plaque-forming cells and for the DNA synthetic response. Similar effects were recorded in unstimulated cultures. Absorption of normal mouse serum with the erythrocytes used as target cells in the plaque assay resulted in less suppression or sometimes even in enhanced responses.


Non-specific potentiation of IgE by parasitic infections in man. Infection with the helminthic parasites Ascaris lumbricoides and/or Necator americanus (hookworm) induces the production in man of high levels of serum IgE. The specificity of this IgE antibody when measured by RAST to a wide range of allergens was restricted in general to the helminthic antigens. Absorption of the sera with immunosorbents produced by coupling extracts of A. lumbricoides to CNBr activated Sepharose 4B established that Ascaris antigen specific IgE antibodies contributed a minor fraction of the total serum IgE. These observations suggest that parasitic infections in man as in laboratory animals potentiate the production of high levels of IgE with specificity unrelated to that of the parasite antigens. While the specificity of this potentiated IgE was not established, it is not directed towards inhalant allergens.


Study of pregnancy-associated alpha2-glycoprotein in relation to populations of human blood leucocytes. Pregnancy-associated alpha2-glycoprotein (alpha2-PAG) was identified by immunofluorescence on the surface membrane of mononuclear cells from normal individuals with an extensive range of plasma concentrations of the glycoprotein. The incidence of positive cells did not correlate with plasma levels but was significantly raised in chronic lymphocytic leukaemia. Anti-alpha2-PAG antibody interfered with E and Fc rosette formation but not with production of C3 rosettes. A large proportion of C3 receptor-bearing cells (B lymphocytes) but only a tiny fraction of E-rosette-forming cells (T lymphocytes) were positive for the protein. Antibody directed against human Ia antigen markedly reduced staining for alpha2-PAG.