Naltrexone: physiological and psychological effects of single doses. An ascending series of single doses of the narcotic antagonist naltrexone, ranging from 20 to 160 mg, was administered to 8 abstinent former addicts in order to assess agonistic activity and any toxic side effects. There was little alteration of normal body function. Significant, but small, changes in sublingual temperature (0.4 degrees F decrease), and diastolic blood pressure (1.7 mm Hg increase) were induced. Among the battery of tests assessing behavioral or mood-feeling variables, only 2 showed significant between-condition effects: facilitated performance on the Cross-out Test (attention and perception), and a dose-related decrease in Morphine-Benzedrine Group (MBG) scores of the Addiction Research Center Inventory (ARCI) (mild euphoria). On the whole, subjects had few subjective reactions or unpleasant side effects. Naltrexone appears to be a safe, nontoxic medication in the dosage range examined.


Amitriptyline plasma levels and therapeutic response. Eighteen depressed outpatients were treated for 6 wk with amitriptyline. Clinical improvement was monitored using the Hamilton Depression Rating Scale administered by two psychiatrists blind to the tricyclic used for treatment, dosage, and plasma levels. Amitriptyline and its desmethyl metabolite, notriptyline, were assayed twice weekly by gas chromatography-mass fragmentography. For the 17 patients having total tricyclic plasma levels between 0 and 250 ng/ml, there was a negative correlation between the Hamilton score and the mean total tricyclic level (p less than 0.01) and amitriptyline level (p less than 0.005). The mean nortriptyline level did not significantly correlate with the Hamilton score. The 10 patients having mean total tricyclic levels above 95 ng/ml had lower median Hamilton scores at week 3 (p less than 0.025) and at week 6 (p less than 0.0025) than those whose tricyclics were lower. The percentage of recovered patients increases significantly as the plasma levels rise to 250 ng/ml, the maximum plasma level considered in this study.


The interaction between halofenate and propranolol. The effect of halofenate on beta adrenergic blockade by propranolol was studied in 4 subjects during chronic drug administration in a randomized, double-blind study. The plasma propranolol concentration was significantly lower during treatment with halofenate than with placebo. The reduction in propranolol levels correlated with a decrease in beta adrenergic blockade. The mechanism for the decrease in plasma concentration has not been determined.


Pharmacokinetics and hemodialyzability of cefazolin in uremic patients. Cefazolin may be used in azotemic patients at reduced dosage as suggested by others. Although hemodialysis removes some of the drug, therapeutic levels are maintained throughout dialysis after a single intravenous dose of 500 mg. Imperfect transport from blood to dialysate is related to protein binding but varies with the degree of azotemia. Small amounts of cefazolin may be administered to patients during dialysis by addition to dialysate.


Clinical comparison of adriamycin and a combination of methyl-CCNU and imidazole carboxamide in disseminated malignant melanoma. The effects of adriamycin were compared to a combination program of methyl-CCNU and imidozole carboxamide (DTIC) in 44 patients with disseminated malignant melanoma. There were objective clinical responses in 6 of 21 patients with the combination of DTIC and methyl-CCNU who received this program as primary treatment and none in 23 patients receiving adriamycin as primary treatment. Secondary responses were not observed with either treatment regimen. Toxicity with the combination program included leukocyte depression (less than 3,000/cu mm) in 25% and platelet depression (less than 100,000/cu mm) in 40% compared to 52% leukocyte depression and 16% platelet depression after adriamycin. There were no responses after the combination treatment program in the absence of myelosuppression. There was nausea and vomiting in virtually all patients, which was moderately severe in one third of the patients receiving the combination and in only 10% of those receiving adriamycin. Alopecia developed in all who received adriamycin but in only 15% of the combination treatment group. The combination treatment response of 28% was of the same order as most response rates previously reported in this disease. This randomized controlled clinical trail found adriamycin without clinical benefit and not worthy of further trial in patients with disseminated malignant melanoma.


A replica plating method for CHO cells using nylon cloth. A new technique of replica plating for CHO cells has been developed in which the pattern of colonies growing in a tissue culture plate is reproduced on a nylon cloth. The efficiency of replica plating onto the nylon cloth is affected by the colony size and the time of cloth-to-plate contact. Maximum replica plating efficiency (80-90%) occurs using 5- to 6-day-old colonies and a contact time of 4 h. The method easily allows 10(4) colonies to be examined for mutant phenotypes. The effectiveness of the technique is demonstrated by isolation of a clone deficient in activity of glucose-6-phosphate dehydrogenase.


Methacycline compared with ampicillin in acute bacterial exacerbations of chronic bronchitis. A double-blind crossover study. In order to objectively document the accepted clinical efficacy of ampicillin in treating bacterial exacerbations of chronic bronchitis, as well as to evaluate the efficacy of methacycline, a double-blind crossover study was designed. Twenty patients with chronic bronchial disease were treated for two separate acute bacterial exacerbations, once with 2 gm of ampicillin daily, and once with 600 mg of methacycline daily, for 14 days. There were a few significant differences when comparing the efficacy of the antimicrobials. For example, the daily volume of sputum significantly went from 35.6 ml initially to 20.5 ml at the end of treatment with methacycline, and from 37.4 to 18.0 ml with ampicillin. Sputum neutrophils excreted per day went from 446 to 147 million with methacycline and from 433 to 94 million with ampicillin. Gram-positive diplococci and cocci on gram stains of sputum significantly decreased form 10.6 to 3.3 with methacycline and from 16.8 to 2.1 with ampicillin. This investigation objectively documents with accepted clinical efficacy of ampicillin and proves methacycline to be an equally effective agent.


Continuous positive airway pressure without tracheal intubation in spontaneously breathing patients. Indications for the use of therapy with continuous positive airway pressure (CPAP) in spontaneously breathing patients are increasing in number. The value of this technique without tracheal intubation was investigated in 14 patients with acute respiratory distress. In most patients (eight patients, or 57 percent) the technique was successful, as evidenced by avoidance of the necessity for tracheal intubation and improvements in clinical appearance, arterial oxygen pressure, and chest x-ray films. Complications were observed in three patients, but these necessitated discontinuation of therapy in only one. The use of this technique allows avoidance of endotracheal intubation and mechanical ventilation, with their attendant risks.


Metabolic studies with 5-fluorocytosine-6-14C in mouse, rat, rabbit, dog and man. Metabolism of 5-fluorocytosine-6-14C (5-FC) was studied in mice, rats, rabbits and dogs after oral and subcutaneous, single and repeated administration. In the urines of all species, intact 5-FC accounted for more than 90% of the total radioactivity at any time of the various treatment schedules. The average proportion of the urinary metabolites was around 5% in dogs, 3% in rabbits, 2.5% in rats, and 2% in mice of the total radioactivity. At repeated dosage, there was an increase of metabolites in mice but a decrease in rats treated subcutaneously. Neither increase nor decrease was observed in rabbits (treated orally) and dogs. Two metabolites were identified, alpha-fluoro-beta-ureido-propionic acid (FUPA) and alpha-fluoro-beta-alanine, the latter occurring mainly after oral treatment. These compounds represent probably that part of 5-FC which was deaminated to 5-fluorouracil (5-FU) or directly to 5-fluorodihydrouracil. FUPA was the only metabolite found in the urines collected from 4 out of 5 human volunteers during the first 12 h after single oral administration of 3.5 g of the radiolabelled drug. Its maximum proportion was 1.1% of the total radioactivity. No metabolites were detected in the urine neither of the 5th volunteer nor in those of 3 mycosis patients who were given the radioactive dose after they had received regular chemotherapy with unlabelled 5-FC (150 mg/kg/day) for at least 2 weeks. The sensitivity threshold of the method was 0.1-0.4% of the total radioactivity. One of the patients had developed thrombocytopenia which was probably due to 5-FC chemotherapy. The symptoms of 5-FC intolerance were in most of the examined species similar to those observed with 5-FU [9]. However, no quantitative correlation between proportion of metabolites and 5-FC toxicity is apparent except that man is the species in which both metabolism and toxicity are the lowest. It has not been proved yet that 5-FC intolerance occurring in a small percentage of patients receiving 5-FC chemotherapy (mainly leukopenia, thrombocytopenia) results in fact from conversion to 5-FU.


Cyclacillin: microbiological and pharmacological properties and use in chemotherapy of infection - a critical appraisal. The pharmacological and  microbiological properties of cyclacillin were discussed and its usefulness in the management of a variety of infections was reviewed. The antibacterial spectrum and clinical utility of this agent generally parallels that of ampicillin, but there are minor differences in specificity with regard to individual microorganisms. The in vitro activity of cyclacillin against gram-negative pathogens varies greatly, the results fluctuating in accordance with the composition of the assay medium of method. The drug has demonstrated excellent therapeutic effectiveness against experimental infections produced by both gram-positive and gram-negative pathogens, thus making questionable a prediction of therapeutic effectiveness from in vitro susceptibility tests alone. Cyclacillin is more resistant to beta-lactamase hydrolysis than ampicillin, is much better absorbed when given by mouth and, as a result, the levels reached in the blood and in the urine are considerably higher than those obtained with the same dose of ampicillin. Cyclacillin produces fewer and milder side-effects than ampicillin.


Amide Penicillin Wy-12,556: therapeutic activity in animals subjected to repeated bacterial infection. Amide penicillin Wy-12,556 and DBED penicillin were studied for their antibacterial effect in CD-1 mice repeatedly infected with Streptococcus pyogenes C203 and/or Staphylococcus aureus Smith. Animals infected with S. aureus Smith failed to show a difference between the therapeutic activity of DBED penicillin and Wy-12,556. Wy-12,556 was significantly more effective than DBED penicillin in protecting the animals challenged repeatedly with S. pyogenes C203. Wy-12,556 provoked a positive transient chemotactic response in uninfected mice. 24 h after intraperitoneal challenge the exudate showed a high neutrophilic count.


Tumors of the optic nerve.

Tumors of the optic nerve. This is a review of primary and secondary tumors of the optic nerve. The emphasis is an optic nerve gliomas and meningiomas. Optic nerve gliomas are slowly growing astrocytic neoplasms of the anterior visual pathways, the majority of which occur within the first two decades of life, with equal sex incidence in about 1 of 200,000 patients presenting with eye complaints. The incidence is greater in neurofibromatosis. The typical presentation is visual impairment in a verbal pre-school child with optic canal enlargement and optic atrophy. An intraorbital location of the tumor leads to axial, irreducible, non-pulsatile proptosis. An intracranial location may disturb hypothalamic and pituitary function and produce hydrocephalus. Ocular findings may also include limited motility on a mechanical-restrictive basis, a pupillary relative afferent defect, nystagmus, and variable, non-specific visual field defects. Roentgenographic studies may show concentric unilateral enlargement of the optic canal with preservation of a well corticated margin, a fossa under the anterior clinoid process in continuity with the optic canal ('J'-shaped sella), and findings of increased intracranial pressure. On pathologic examination the tumor is a smooth, fusiform, intradural enlargement of the optic nerve. Histologically there is proliferation of elongated (pilocytic) astrocytes in reticulated patterns with intervening microcystic spaces containing mucosubstance and surrounding reactive hyperplasia of the arachnoid. Mitoses are not found. The diagnosis is clinical X-ray studies and brain scan should be performed. The differential diagnosis is that of unilateral proptosis in a child and includes acute ethmoiditis, hyperthyrobidism, craniostenosis, other neoplasms, Hand-Schuller-Christian disease, and orbital hemorrhage due to trauma. Surgical resection is performed in cases with unilateral optic nerve involvement, the surgical approach being determined by tumor location. Bilateral or chiasmal cases are treated with radiotherapy when progression occurs. Malignant optic nerve gliomas and optic nerve hyperplasia are also discussed. Optic nerve meningiomas arise from the nerve sheath and are to be distinguished from orbital meningiomas arising from ectopic arachnoidal cells or those secondarily involving the orbit by extension from adjacent sites. Up to 80% of orbital meningiomas occur in females, in two age peaks, 25% in the first decade, and the rest in the 5th decade. Meningiomas present with visual loss and may produce proptosis, papilledema and/or optic atrophy, retinal striae, opticociliary shunts, limitation of extra-ocular movements, and lid edema, Signs of von Recklinghausen's disease should be sought. X-rays are the mainstay of diagnosis. Orbital meningiomas are composed of cells in sheets or in whorls with some spindle shaped cells. Calcifications are typical. Usually the dura is penetrated and the orbit invaded. Primary orbital meningiomas are locally infiltrating but do not metastasize. Complete local excision en bloc is recommended...


[Influence of glucose infusions on proinsulin and insulin syntheses in the islands of langerhans in the rat]. Experiments on RNA synthesis and the observation of an increased secretion of proinsulin during stimulated synthesis of insulin in islets of Langerhans in vitro led to the supposition that insulin synthesis in vivo may be increased to a higher extent than synthesis in vitro. This investigation evaluates the effect of long time infusions of glucose on the synthesis of proinsulin and insulin in islets of Langerhans from the rat. The following results were obtained: 1. the rate of synthesis of proinsulin and insulin increased due to glucose infusions by a factor of 2 after 6 hours and by a factor of 4 after 24 hours; 2. prolonged infusion led to a decrease of insulin synthesis and serum insulin concentrations; 3. the turnover of proinsulin into insulin remained unaffected by the glucose infusion; 4. a specific stimulatory effect on the synthesis of one of the two insulins of the rat was not observed. The conclusion was made, that glucose in vivo stimulates enzyme induction, followed by increased RNA synthesis and mitotic activity. These combined factors lead to an increased insulin synthesis.


The verification of neurones in the pancreas of spiny mice (Acomys cahirinus). The statement that up to now no neural elements could be identified in the islets of Langerhans with the aid of electron microscopy led us to reinvestigate this question of innervation of the pancreas in spiny mice using light microscopy. Single nerve cells as well as nerve fibre bundles could be found in the interlobular connective tissue of the pancreas. By constrast, we could observe neither nerve cells nor nerve fibres in the islets of Langerhans. Extrainsular neuroinsular complexes could not be found either.


Transport of pyrimidine nucleosides in cells of Escherichia coli K 12. 1. The transport of pyrimidine mucleosides into cells of Escherichis coli has been investigated in mutant strains which cannot metabolize these nucleosides.  Such cells transport and concentrate purimidine mucleosides several hindredfold. 2. The transport is inhibited by energy poisons and by sulfhydryl reagents. 3. Pyrimidine mucleosides compete mutually for transport. Adenosine is also a strong competitor while guanosine and inosine are weak competitors. 4. The rate of pyrimidine mucleoside transport is shown to be under control of the cytR and deoR gene products, which are also known to regulate the synthesis of nucleoside-catabolizing enzymes. The transport system is repressed by growth on glucose, as is the synthesis of the enzymes.


Autologous marrow recovery following allogeneic marrow transplantation in a patient with severe aplastic anemia. A 45-year-old woman with severe idiopathic marrow failure was prepared for marrow transplantation by administration of cyclophosphamide (cy) 50 mg/kg on each of four successive days. She then received an intravenous infusion of 20 X 10(9) nucleated marrow cells from an HL-A matched and mixed lymphocyte culture (MLC) non-reactive sister. There was evidence for minimal marrow recovery in 1-2 months and a second marrow infusion was carried out 69 days after the first without additional immunosuppression. There was a continued slow recovery of peripheral blood counts with complete reconstitution of erythropoiesis, return of the white blood cell count to between 3 and 4000/mm3, with 50% granulocytes, and platelets to 60--70,000/mm3, 11 months after the initial grafting attempt. Red cell antigens and gamma globulin allotypes were of recipient type. The MLC and the indirect cell mediated lympholysis (CML) test became positive possibly indicating cellular sensitization to non HL-A antigens. This report of a patient with severe marrow failure documents autologous recovery of marrow function after receiving a large dose of cy and allogeneic marrow. The implications of this are discussed.


Modification of hemopoietic stem cells of BALB/c mice by Rauscher leukemia virus. Bone marrow and spleen from Rauscher leukemia virus (RLV) infected BALB/c mice were tested for proliferative activity at various time intervals after infection using the spleen colony assay. Both bone marrow and spleen derived colonies showed a modified maturation behavior. The erythroid colonies were classified into three consecutive stages of maturation according to morphologic criteria. Both bone marrow and spleen derived erythroid colonies exhibited a retardation of maturation upon RLV infection of the donor mice; the spleen derived erythroid colonies showed the most severe changes. RLV itself could not mimic this phenomenon; moreover it was shown by immunofluorescence that only low amounts of RLV were present in the spleen colonies. It is therefore suggested that RLV modifies part of the stem cell compartment leading to a prolonged cycle time of proerythroblasts and hence to a retardation of colony growth.


The reversal of glucose repressed prodigiosin production in Serratia marcescens by the cyclic 3'5'-adenosine monophosphate inhibitor theophylline. Glucose was found to cause severe repression of prodigiosin production in Serratia marcescens and a dose related partial reversal was demonstrated by theophylline. It is suggested that this reversal is due to the inhibition of cAMP phosphodiesterase and the concomitant increase in cellular cAMP concentration.


Inhibition of prodigiosin formation in Serratia marcescens by adenosine triphosphate. ATP, inorganic phosphate and ribose inhibited prodigiosin formation in Serratia marcescens, but adenine did not. ATP was not hydrolyzed by the organism during the experiment.


Antibody-induced formation of caps in Toxoplasma gondii. Trophozoites of Toxoplasma gondii from mouse peritoneal exudate move their surface membrane antigens towards one pole of the cell when incubated with antibodies. The phenomenon may be induced in up to 50% of incubated parasites. It is prevented by some metabolic inhibitors and low temperatures (0-4 degrees C). These properties do not change in parasites subpassaged after repeated incubation with antibodies.


Differences in susceptibility of tissues to revascularization studied in ectopic implants. The survival rate of implanted marrow is reduced when the tissue is transferred from one site to another within the first 24 hours but not more than 48 hours after the initial implantation. Splenic implants can be transferred at any time after implantation without affecting the survival rate. The observation suggests a difference in the sensitivity of the two tissues to the avascular period that they experience prior to initiation of angiogenesis.


Induction of immunological memory in mice by RNA extract. RNA extract isolated from spleens of mice immunized with lipopolysaccharide from E. coli induced an immunological memory in normal mice. Application of small amounts of corresponding antigen provoked a specific secondary immune response in RNA primed mice.


Evidence for a selective cytotoxic effect of carrageenan on cells of the immune system in vivo and in vitro. Carrageenan suppressed antibody production to sheep erythrocytes in the mouse only when injected together with or 24 h prior to antigen. Pretreatment of sensitized spleen cell cultures with carrageenan reduced the degree of stimulation obtained using antigen but not PHA.


Stimulation by ATP of protein initiation in a prokaryotic organism, B. stearothermophilus. In contrast to E. coli ribosomes, with B. stearothermophilus ribosomes initiation complex formation is stimulated by ATP as well as GTP, but maximum stimulation occurs when both the nucleotides are present; and their terminal phosphate must be hydrolysable. In the presence of ATP and GTP, B. stearothermophilus ribosomes synthesize a highly phosphorylated guanine derivative, ppGpp, and the role of ATP in initiation might be related to this synthesis. We discarded the role of ATP as being trivial and corresponding solely to the well-known effect on eukaryotic systems.


Tissue pressures and fluid dynamics of the lungs. A pressure more negative than pleural surface pressure has to be applied to water on the pleural surface to keep it from being absorbed; yet, there is a thin layer of liquid in the pleural space under normal physiological conditions. How is the pressure of the liquid related to the pressure on the surface of the pleural membranes? There is evidence that the liquid pressure might be less than the surface pressure, and the balance is achieved by points of contact between the pleural membranes with tissue deformation forces. If it is possible for liquid and surface pressure to be different in the intrapleural space, the same mechanisms might be active within the lung tissue. Information about the tissue pressures can be estimated by determining the pressures acting on the outer surfaces of intrapulmonary vessels. There is evidence that some vessels of the lungs have a surface pressure close to alveolar pressure (alveolar vessels) and others have a surface pressure that falls relative to pleural pressure as alveolar pressure is increased relative to pleural pressure (extra-alveolar vessels). There appears to be a complete dissociation between the surface pressures and the liquid pressure surrounding the pulmonary blood vessels. These observations provide strong support for the concept that the pressure of liquid within tissue need not be the same as the tissue pressure.


Cellular immunity to lymphocyte antigens in human infertility. The leukocyte migration test was used to detect cellular immunity to HL-A antigens present on the husband's lymphocytes and spermatozoa in a group of fertile and infertile couples. Approximately 50% of the married female population appear to be sensitized. There was no difference between the fertile and infertile groups in the frequency of positive results, but a greater number of migration indices were below 0.7 and 0.6 in the infertile group. It is suggested that sperm cells are the most likely cause of immunization.


Effects of luteinizing hormone-releasing hormone on fetal survival in pregnant rabbits. The effect of synthetic luteinizing hormone-releasing hormone (LH-RH) on ovulation, implantation, and fetal survival rate was tested in pregnant rabbits. Single doses ranging from 2 to 900 mug, injected intravenously on the 3rd, 5th, or 6th postmating day, did not prevent implantation. Following the intravenous infusion of 20 to 200 mug of LH-RH on day 10 of pregnancy, the fetal survival rate was comparable to that of control rabbits laparotomized at the same stage of pregnancy. However, after repeated doses of 100 mug of LH-RH injected subcutaneously in a delaying vehicle either once or twice daily between the 8th and 13th days of gestation, multiple ovulations and severe luteolysis of original corpora lutea occurred, and fetal survival to term was highly variable. The results suggest that, depending on the dosage administered, LH-RH may either interrupt pregnancy by luteolysis or support fetal survival by stimulating luteotropic activity.


Peripheral blood lymphocyte distribution in scleroderma. Determination in peripheral blood T and B lymphocytes performed in progressive systemic scleroderma (8 patients); dermatomyositis (5 patients); and inflammatory or mesenchymal scleroderma ('mixed' connective tissue disease) (5 patients) demonstrated a marked increase in peripheral B cells with T cell depression in patients with mesenchymal scleroderma. Patients with progressive systemic scleroderma and dermatomyositis demonstrated also a peripheral T depression, but no consistent changes in B cell population were found. The clear relationship between a B lymphocyte increase and elevated immunoglobulins, antinuclear and antiribonucleoprotein antibody and positive direct immunofluorescence of skin (basal membrane and blood vessels) defines inflammatory or mesenchymal scleroderma in which a disturbance in humoral immunity is involved. B lymphocyte determination can help to measure these changes in immunoreactivity.


Glucose tolerance and insulin release, a mathematical approach I. Assay of the beta-cell response after oral glucose loading. From the dose-response relations between glucose and insulin after oral glucose loading, a reproducible parameter for beta-cell response was deduced. The main advantage of this parameter -corrected insulin response, defined as CIR = I- 100/G(G-70)- lies in it independence from the initial or reached glucose level.


Glucose tolerance and insulin release, a mathematical approach. II. Approximation of the peripheral insulin resistance after oral glucose loading. A parameter of peripheral insulin activity (A = 10(4)/IpGp) can be obtained after oral glucose loading by simple calcuation using insulin and glucose levels at the glucose peak. In combination with a glucose-independent parameter of beta-cell function (CIR = 100. I/G(G-70) a parameter of glucose tolerance (GT = A-CIR) is defined. The parameters allow one to separate the contributions of beta-cell function and peripheral insulin resistance to the glucose tolerance observed after glucose loading. Examples, based on the literature and our own work, illustrate the increase of A and GT by cortisone acetate premedication as well as long-term oral contraceptive medication.


Vascular reactivity to angiotensin II and to norepinephrine in diabetic subjects. Vascular responsiveness to infused angiotensin II and to norepinephrine was determined in 14 normal subjects and two groups of diabetic subjects, 16 with no clinically detectable diabetic complications and 14 with diabetic retinopathy but no clinical evidence of nephropathy. All were maintained on a 100-mEq. -Na- 100-mEq. -K diet. Serum electrolytes, 24-hour urinary sodium, creatinine clearance, and plasma renin activity did not differ significantly among the groups. Group mean baseline diastolic pressure in those with retinopathy was higher than in normal subjects but no significantly different from that of uncomplicated diabetics. The pressor dose of angiotensin II (ng./kg./min. to increase diastolic blood pressure 20 mm. Hg) for each group respectively was 11.5 +/-0.9, 12.9+/- 1.3, and 8.3 +/- 1.3, and the slope of the dose-response curve (mm. Hg rise in blood pressure resulting from the infusion of 1 ng./kg./min. following the initial increment in blood pressure) was 2.0 +/-0.2, 1.6+/-0.2, 3.3+/- 0.6. For norepinephrine, the pressor doses were 163 +/- 24, 212+/-21, and123 +/- 11 and slopes were 0.17 +/- 0.03, 0.13 +/- 0.02, and 0.20 +/-0.02. Neither diabetic group differed significantly from normal subjects. Diabetics with retinopathy were more sensitive to angiotensin II, pressor dose (P less than 0.059) and slope (P less than 0.02), and to norepinephrine, pressor dose (P less than 0.006) and slope (P =0.05) than those without complications. These data suggest that vascular reactivity is enhanced in diabetics with retinopathy.


Further studies on the metabolism of D-glucose anomers in pancreatic islets. The alpha and beta anomers of commercially available D-(5-3H) glucose were separated by miniaturized Hudson-Dale procedures based on precipitation with acetic acid. Reflectometric measurements of the reactivity with matrix-bound glucose oxidase showed that the preparations were about 90 per cent pure with respect to anomeric composition. Nonradioactive anomers separated by the same procedures were analyzed by optic polarimetry and gas chromatography. The preparations were about 90 per cent pure with respect to anomeric composition and produced no peaks but D-glucose on trimethylsilylation and chromatography. Microdissected pancreatic islets of noninbred ob/ob-mice exhibited a linear production of 3H2O for three to nine minutes when incubated with 6 mM alpha-D-(5-3H) glucose, beta-D-(5-3H) glucose, or D-(5-3H) glucose in anomeric equilibrium; the three glucose preparations did not differ in their rate of conversion to 3H2O. The rate of 3H2O production increased with glucose concentration (3-21 mM) during incubations for three minutes and, again, there was no evidence for the metabolic activity's being dependent on the anomeric composition of the labeled sugar. When microdissected islets were perifused without glucose and suddenly exposed to 5-6 mM alpha-D-glucose or beta-D-glucose, the concentration of glucose-6-phosphate rose within five minutes and did not differ significantly between experiments with alpha-D-glucose and beta-D-glucose. In the same perifusion experiments, only alpha-D-glucose caused a pronounced stimulation of insulin secretion, the difference from beta-D-glucose being significant. The results indicate that the recognition of glucose as an insulin secretagogue does not only involve metabolism by glucose-6-phosphate. The possible roles of the sorbitol pathway and of hypothetical regulatory sites for the glucose molecule ("receptors") are briefly discussed.


The kinetics of beta-granule formation: a morphometric study. The rate of formation of beta-granules in mouse islet cells has been studied by morphometry using experimental conditions in which insulin secretion is known to be blocked. The number of beta-granules in a cell is increased by 120%, 45 minutes after a continued glucose stimulus of 16.7 mM. The implications of the results are discussed with respect to the construction of kinetic models for the secretion of insulin by exocytosis.


Isolation and long term preservation of pancreatic islets from mouse, rat and guinea pig. This paper reports techniques for the isolation and long term preservation of pancreatic islets from the mouse, rat and guinea pig. Islets have been isolated using a modification of a free hand microdisection procedure described by Hellerström in 1964 [1]. Isolated islets have been subjected to three preservation systems and their viability following storage assessed by light microscopy of sections stained with Gomor's aldehyde fuchsin [2] and by measuring the insulin release from islets in vitro in response to a glucose stimulus. The systems were: a) Simple cold storage in Hank's balanced salt solution at 4 degrees C. Following 15 h cold storage, histological and functional survival was 100%. This dropped to 10% at 48 h. There were no survivors following 72 h storage. b) Sub zero cell storage. In Group I (freezing rate 1 degrees C/min) histological survival was 35% and functional survival 20%. In Group II (freezing rate 5 degreees C/min with 24 h culture period after rewarming) histological survival was approximately 87% and functional survival 75%. c) Organ Culture. Islets from the guinea pig, rat and mouse showed minimal morphologic damage when cultured for 21 days in a simple organ culture system. At 28 days, histological survial was approximately 30%. Following organ culture we were unable to correlate histological and functional survival.


High concentration of somatostatin immunoreactivity in chicken pancreas. Extracts of discrete lobes of chicken pancreas were assayed for somatostatin (SRIF), insulin and glucagon immunoreactivity. The distribution of hormone concentration was correlated appropriately with the known distribution of A, B, and D-cells. Concentrations of all three hormones were highest in  the splenic lobe. The glucagon content of the ventral and dorsal lobes was low. Evidence is presented that the SRIF-like material in the pancreatic extracts is very similar to synthetic cyclic SRIF; parallel immunoassay displacement curves and similar chromatographic elution profiles were obtained. The SRIF concentration in chicken pancreas is 21 times higher than that found in the rat. Chicken pancreas may provide a useful model for studies of somatostatin physiology.


Polyploidy in the pancreas of the normal and diabetic mutant mouse. The DNA content of endocrine and exocrine pancreatic nuclei of normal C57BL/KsJ and diabetic mutant C57BL/Ks-db/db mice was measured by Feulgen microdensitometry. The exocrine and endocrine pancreatic nuclei of the 4-week-old normal, 12-week-old normal, and 4-week-old (prehyperglycaemic) diabetic mutant mice contained diploid and tetraploid cells, while the 12-week-old (established hyperglycaemic) mutant contained diploid, tetraploid, and octaploid nuclei. The polyploidy in the endocrine pancreas of all these mice was confined to the B-cells, while the A-cells were always diploid.


The ultrastructure of polyploid B-cells in the islets of normal mice. Light and electron microscopic studies of diploid, tetraploid and octaploid B-cells in the islets of normal C57BL/KsJ mice revealed that polyploid cells were characterized by a wider range of granulated states than diploid B-cells. The maximum granule densities were similar for polyploid and diploid cells; however, some polyploid cells were almost devoid of granules, while the least granulated diploid cells contained intermediate granule densities. The tetraploid cell also appeared to be characterized by an increased mitochondrial stage which suggests compensation for the greater degree of degranulation. These observations were confirmed by morphometric analysis. Two interpretations of the apparent polyploidy are discussed; that polyploid B-cells may be more responsive to insulin releasing stimuli than diploid B-cells and that tetraploid cells may only be diploid cells in the G2 phase of the mitotic cycle.


Carcinogenic effect of 1-butyl-1-nitrosourea on female Sprague-Dawley rats. Carcinogenic effect of a single oral administration of 300 or 200 mg/kg body weight of 1-butyl-1-nitrosourea (BNU) and continuous oral administration of 400 ppm solution of BNU in the drinking water for 5, 10, 15, and 20 weeks to female SD rats was studied. In addition, the number of spleen cells capable of forming plaques (PFC) against primary immunization with sheep red blood cells was investigated in various stages of the animal experiments. With a single oral administration of BNU, tumors developed in 31/50 (62%) rats between the 25th and 75th week. They were most frequently seen in the mammary gland (40%), followed by the stomach (10%), kidneys (8%), and ovary (8%). Leukemia was found in 8%. No dose-effect relationship was observed in these 2 experimental groups. On the other hand, tumors developed in 67/77 (88%) of the rats that received BNU in their drinking water. The incidence of tumors was highest in leukemia (61%), followed by mammary tumors (26%), intestinal tumors (12%), and ear duct tumors (8%). There was a dose-effect relationship among the 4 groups in the latent period and target organs for tumor development. Although the PFC count of the rats receiving BNU for 5 weeks recovered gradually to about 50% of the control level at the end of the 25th experimental week, it remained less than 10% of the control level for the whole experimental period in those receiving BNU longer than 10 weeks. Therefore, it was apparent that the tumors developed, proliferated, and finally killed the host rats in highly immunosuppressive state.


Inhibition of tumor growth in vivo and in vitro by macrophages from rats treated with a streptococcal preparation, OK-432. The role of macrophages from rats treated with a streptococcal preparation (OK-432) for inhibition of syngeneic tumor growth was examined both in vivo and in vitro. Treatment of rats with OK-432 intraperitoneally resulted in a transient inhibition of ascites tumor growth and concomitant increase in survival time. Peritoneal exudate cells from these rats completely suppressed tumor growth when admixed with tumor cells and transferred subcutaneously to syngeneic recipient rats. No such effect was observed with tumor cells from saline-treated control rats. The macrophages were responsible for the neutralization of tumor growth, since the removal of adherent cells from peritoneal exudate cells of rats treated with OK-432 resulted in the abrogation of their antitumor activity. To further investigate the antitumor activity of macrophages, an in vitro cytotoxicity assay was carried out on several tumor cell lines. A complete target cell destruction was brought about by macrophages from the rats treated with OK-432 but not from the control animals. The tumor cytotoxicity mediated by macrophages activated with OK-432 was regarded as non-specific because they could damage all the target cell lines tested.


Effect of protein-bound polysaccharide preparation, PS-K, on the immune response of mice to sheep red blood cells. The protein-bound polysaccharide preparation, PS-K, was found to potentiate the production of hemolytic plaque-forming cells of spleen and serum hemagglutinin in C57BL/6 mice, when they were immunized with a low dose of sheep red blood cells. PS-K was administered intraperitoneally at a dose of 150 mg/kg daily for 5 days. Ten days later, the mice were immunized with either 4 X 10(8) or 1 X 10(7) sheep red blood cells. Both the number of plaque-forming cells in their spleen and the amount of serum hemagglutinin were significantly higher in the PS-K treated mice when 1 X10(7) sheep red blood cells were used as an immunizing dose.


Loperamide: a new antidiarrheal agent in the treatment of chronic diarrhea. Twenty-seven patients with chronic diarrhea due to Crohn's disease, ulcerative colitis, short bowel syndrome, and idiopathic (functional) causes participated in a multiphase study (open, double blind, and long term open) of loperamide HCl, a new single entity, oral antidiarrheal agent. In the open phase of the study, loperamide effectively relieved symptoms of diarrhea in 21 of 27 patients. The average number of stools dropped from eight in the initial relapse period to two stools after 1 month of treatment (P = 0.0001). Efficacy was confirmed in the double blind and long term open phases of the study. Four patients who were not relieved while on therapy had discontinued the drug because of abdominal cramping. No other side effects attributable to the drug were observed. Loperamide has been found to be a safe and effective agent for the treatment of chronic diarrhea.


A double blind crossover comparison of loperamide with diphenoxylate in the symptomatic treatment of chronic diarrhea. Loperamide, a novel antidiarrheal agent, was compared with diphenoxylate in a double blind crossover study of 23 patients with chronic diarrhea of various etiologies. Both agents were found to be capable of controlling or greatly reducing chronic diarrhea. Loperamide was superior to diphenoxylate in its abiltiy to decrease the frequency and improve the consistency of the stools, even at a 2.5-fold lower dose level.


Contamination of the small intestine by enterotoxigenic coliform bacteria among the rural population of Haiti. Coliform bacteria were isolated by either aerobic or anaerobic culture techniques from aspirates of the proximal small intestine of 4 of 5 Haitians with tropical sprue, but not from any of 10 well nourished Haitians who had milder gastrointestinal complaints and abnormalities. Klebsiella (Klebsiella pneumoniae and Klebsiella ozaenae) was cultured from the jejunal aspirates of 2 sprue patients and Escherichiae coli from the other 2. Fifteen colonies of coliform bacteria cultured from each aspirate were specifically identified by their biotype. In three instances, every colony in each aspirate was the same; In three instances, every colony in each aspirate was the same; in the fourth aspirate, two biotypes of E. coli were present, one of which grew under both aerobic and anaerobic culture conditions and another which grew only under anaerobic conditions on initial isolation. A randomly selected strain of each of the five coliform biotypes isolated was examined for enterotoxigenicity by determining the effect of variously prepared cell-free preparations on water transport in the rat jejunum using standard marker perfusion techniques. Every strain tested was toxigenic; one produced only a heat-stable toxin, one produced a heat-labile toxin only, and three elaborated both forms of enterotoxin. These observations indicate that most Haitians with tropical sprue have colonization of the proximal small intestine by a specific strain of enterotoxigenic coliform bacteria, but such is not the case among Haitians who have milder intestinal abnormalities.


Overwhelming watery diarrhea associated with a cryptosporidium in an immunosuppressed patient. A 39-year-old man with severe bullous pemphigoid developed overwhelming diarrhea after 5 weeks' treatment with 150 mg of cyclophosphamide and 60 mg of prednisolone daily. Jejunal and ileal biopsies showed severe mucosal injury and tiny 2- to 4-mu organisms on the epithelial surfaces. Similar organisms were seen in smears of jejunal fluid. Electron microscopic examination of jejunal biopsies showed these spherical bodies to be trophozoites, schizonts, microgametocytes, and macrogametocytes typical of the genus Cryptosporidium. Diarrhea resolved 2 weeks after discontinuation of cyclophosphamide and coincided with disappearance of Cryptosporidia from the jejunal biopsies. Immunosuppression may have predisposed this patient to cryptosporidial diarrhea. Cryptosporidiosis is another infection which can be diagnosed by small bowel biopsy. When immunosuppressed patients develop severe diarrhea, opportunistic infection with this and other organisms should be considered as the possible cause.


[Balantidium coli infection of the female genitals (author's transl)]. Two cases of therapy-resistant fluorine are reported in which Balantidium coli from the class of the cilia (Ciliophora, Ciliata, Infusoria) was identified. After a personal experiment, the Balantidia must be considered extremely pathogenic. In international literature, no further indication of a ciliata infection of the female genitals was found.


Lambda cin-1, a new mutation which enhances lysogenization by bacteriophage lambda, and the genetic structure of the lambda cy region. Seven lambda cy mutants have been mapped within a small region located approximately halfway between the rightward boundary of the imm434 region and the lambda cII gene. The seven mutants lie at four sites separated by a total distance of about 12 nucleotide pairs, as estimated from recombination frequencies. Six of the seven mutants lie on the right side of the cy fine structure map, spanning a total distance of about 3-5 nucleotide pairs. Lying approximately 11-21 nucleotide pairs to the left of the leftmost cy mutant is a newly described mutation called cin-1, for c independent. The cin-1 mutation allows some lysogenization when coupled with any cy, cII or cIII mutant, but not when coupled with a defective cI gene. The cin-1 mutation, like cy mutants, has a cis-dominant action upon the cI gene in mixed infections. The observation that gammaimm434 cin-1-cy2001 lysogenizes efficiently, but not gammaimm434 cin-1 cy2001 cII68 nor any other gammaimm434 cin-1 cy derivative, is interpreted to mean that all of the cy mutants on the right side of the cy fine structure map inactivate a binding site for cII/cIII function, but that cy2001, the single mutant on the left side of the cy fine structure map, does not inactivate that binding site.


Two chromosomal genes required for killing expression in killer strains of Saccharomyces cerevisiae. The killer character of yeast is determined by a 1.4 X 10(6) molecular weight double-stranded RNA plasmid and at least 12 chromosomal genes. Wild-type strains of yeast that carry this plasmid (killers) secret a toxin which is lethal only to strains not carrying this plasmid (sensitives).--We have isolated 28 independent recessive chromosomal mutants of a killer strain that have lost the ability to secrete an active toxin but remain resistant to the effects of the toxin and continue to carry the complete cytoplasmic killer genome. These mutants define two complementation groups, kex1 and kex2. Kex1 is located on chromosome VII between ade5 and lys5. Kex2 is located on chromosome XIV, but it does not show meiotic linkage to any gene previously located on this chromosome.--When the killer plasmid of kex1 or kex2 strains is eliminated by curing with heat or cycloheximide, the strains become sensitive to killing. The mutant phenotype reappears among the meiotic segregants in a cross with a normal killer. Thus, the kex phenotype does not require an alteration of the killer plasmid.--Kex1 and kex2 strains each contain near-normal levels of the 1.4 x 10(6) molecular weight double-stranded RNA, whose presence is correlated with the presence of the killer genome.


Substrate-specific enzyme variation in natural populations of Drosophila pseudoobscura. By using a number of different alcohols as substrates, eight alcohol dehydrogenase loci were discovered in Drosophila pseudoobscura. Each of these loci can take more than one substrate. Several of these loci differed in their tissue specificities and activity patterns during development. The genic variation in natural populations was studied at four of these loci and three of them were polymorphic. A quantitative study of substrate-specific differences among alleles of the same locus produced negative results. This result appears to be typical of most studies done on this aspect. From this it was concluded that the substrate specificity of enzymes is not an important factor in determining the greater amount of genic variation at Group II loci than at Group I loci, as proposed by KOJIMA, GILLESPIE and TOBARI (1970). There are several observations which suggest a different explanation for the differences in the genic variability at Group I and Group II loci: (1) There are, on an average, more isozyme loci (loci with similar substrate specificity) for enzymes in Group II than in Group I; (2) The null alleles are far more common at Group II loci than at Group I loci; (3) There is significant heterogeneity in the number of alleles and the heterozygosities at loci within each of these two groups of enzymes; (4) Relatively higher levels of genic variation are observed at Group II loci even in populations which appear to be living in homogeneous environments; and (5) Some loci (e.g. esterases) are highly polymorphic in most species investigated by gel electrophoresis techniques. Based on these general observations, it is proposed that (1) the substrate-specific differences are between isozyme loci and not between alleles of a given locus, and (2) neutral alleles are proportionately far more common at loci at Group II than at loci in Group I, because the former is under less selection constraint than the latter.


Mitotic crossing over and nondisjunction in translocation heterozygotes of Aspergillus. To analyze mitotic recombination in translocation heterozygotes of A. nidulans two sets of well-marked diploids were constructed, homo- or heterozygous for the reciprocal translocations T1 (IL;VIIR) or T2 (IL;VIIR) and heterozygous for selective markers on IL. It was found that from all translocation heterozygotes some of the expected mitotic crossover types could be selected. Such crossovers are monosomic for one translocated segment and trisomic for the other and recovery depends on the relative viabilities of these unbalanced types. The obtained segregants show characteristically reduced growth rates and conidiation dependent on sizes and types of mono- and trisomic segments, and all spontaneously produce normal diploid sectors. Such secondary diploid types either arose in one step of compensating crossing over in the other involved arm, or--more conspicously--in two steps of non-disjunction via a trisomic intermediate.--In both of the analyzed translocations the segments translocated to IL were extremely long, while those translocatated from IL were relatively short. The break in I for T1 (I;VII) was located distal to the main selective marker in IL, while that of T2 (1;VIII) had been mapped proximal but closely linked to it. Therefore, as expected, the selected primary crossover from the two diploids with T2 (I;VIII) in coupling or in repulsion to the selective marker, showed the same chromosomal imbalance and poor growth. These could however be distinguished visually because they spontaneously produced different trisomic intermediates in the next step, in accordance with the different arrangement of the aneuploid segments. On the other hand, from diploids heterozygous for T1 (I;VII) mitotic crossovers could only be selected when the selective markers were in coupling with the translocation; these crossovers were relatively well-growing and produced frequent secondary segregants of the expected trisomic, 2n + VII, type. For both translocations it was impossible to recover the reciprocal crossover types (which would be trisomic for the distal segments of I and monosomic for most of groups VII or VIII) presumably because these were too inviable to form conidia.--In addition to the selected segregants of expected types a variety of unexpected ones were isolated. The conditions of selection used favour visual detection of aneuploid types, even if these produce only a few conidial heads and are not at a selective advantage. For T2 (I;VIII) these "non-selected" unbalanced segregants were mainly "reciprocal" crossovers of the same phenotype and imbalance as the selected ones. For T1 (I;VII) two quite different types were obtained, both possibly originating with loss of the small VII-I translocation chromosome. One was isolated when the selective marker in repulsion to T1 (I;VII) was used and, without being homo- or hemizygous for the selective marker, it produced stable sectors homozygous for this marker...


Mutants of Saccharomyces cerevisiae resistant to the alpha mating-type factor. Mutants that are resistant to alpha-factor have been isolated from a mating-type haploid strains of yeast by direct selection on agar medium containing partially purified alpha-factor. All resistant mutants isolated were found to be sterile. They were characterized and compared with mutants previously isolated as non-mating. Among 93 able to mate at low frequency and to sporulate, none showed linkage to the mating-type locus. The results support the hypothesis that the response to alpha-factor by cells of mating-type a is essential for mating.


Genetic analysis of petite mutants of Saccharomyces cerevisiae: transmissional types. We have studied a number of petite [rho-] mutants of Saccharomyces cerevisiae induced in a wild-type strain of mitochondrial genotype [ome- CHL(R) ERY(S) OLI(S) (1, 2, 3) PAR(S)] by Berenil and ethidium bromide, all of which have retained two mitochondrial genetic markers, [CHL(R)] and [ERY(S)], but have lost all other known markers. Though stable in their ability to retain these markers in their genome, these mutants vary widely among themselves in suppressiveness and in the extent to which the markers are transmitted on crossing to a common wild-type tested strain. In appropriate crosses all of the strains examined in this study demonstrate mitochondrial polarity, and thus have also retained the [ome-] locus in a functional form; however, five different transmissional types were obtained, several of them quite unusual, particularly among the strains originally induced by Berenil. One of the most interesting types is the one that appears to reverse the parental genotypes with [CHL(R) ERY(S)] predominating over [CHL(S) ERY(R)] in the diploid [rho+] progeny, rather than the reverse, which is characteristic of analogous crosses with [rho+] or other petites. Mutants in this class also exhibited low or no suppressiveness. Since all of the petites reported here are derived from the same wild-type parent, and so have the same nuclear background, we have interpreted the transmissional differences as being due to different intramolecular arrangements of largely common retained sequences.


On the nature of cis-acting regulatory proteins and genetic organization in bacteriophage: the example of gene Q of bacteriophage lambda. We note the existence of a "partially cis-acting" regulatory protein of bacteriophage lambda: the product of the phage Q gene. We suggest that there may be a complete spectrum from "all cis" to "all trans" for such regulatory proteins. This behavior might arise because a DNA-binding protein either acts at a nearby (cis) site soon after synthesis or becomes "lost" for its trans activity on another genome through nonspecific interactions with DNA. Our proposed explanation provides one evolutionary basis for the linkage of genes for regulatory proteins and the sites at which such proteins act; it also suggests a possible rationale for the "metabolic instability" of certain regulatory proteins.


[An analysis of small differences between mice of lines C57BL/10 and 129 with regard to the chief system of tissue compatibility, H-2]. The differences in the H-2 system between mice of the C57BL/10 (H-2b) and 129 (H-2bc) were studied in crosses B10.D (H-2d) or recombinant strains R106 (KbSbDda), R107 (KbSbDd) and R101 (KdSdDb) with strains 129 or B10.129 (6M). Most skin grafts from the strain C57BL/10 were rapidly rejected in the F1 hybrids derived from the crosses with strains B10.D2, R106 and R107, thus indicating to the effect of the relatively strong H-locus either in the D region or in the Tla region of H-2b. Chronic responses were observed in the F1 hybrids derived from crosses with the strains R101 and R103, indicating to the weak effect of another locus. However all the skin grafts survived in the F1 hybrids with the strain HTG (KdSdDb). This weak H gene is apparently located to the left of the crossover in the recombinants R101 and R103, but to the right of the crossover in the HTG, i. e. between the loci Slp and H-2D. The new locus is tentatively designated as H-2W.


[Isolation and characteristics of UV-resistant revertants of recA-strains of Escherichia coli K-12]. The paper presents the results of a study of UV-resistant revertants of some recA strains induced by different mutagens. It is shown that some of produced revertants differ from original recA strains in some properties. It is established that all UV-resistant revertant fall into three phenotypic groups on their recombination proficiency in crosses with different donor strains (Hfr C, Hfr 3.0SO, F' W1655) and in their sensitivity to UV and gamma-rays. It is concluded that all UVr revertants are rec A+ and carried mutations either in the same recA gene (true reversion or intragenic suppression) or in genes closely linked with recA.


[Cyclic rearrangements in the incomplete genome of T4D phage transferred into a cell during a injections of DNA]. The genome structure of phage T4 chromosome fragments obtained by the method of DNA interrupted injection is studied directly. Anfragment population is shown by homogeneous in sizes. The fragment sizes are proved to the indentical to sizes. The fragment sizes are proved to be identical to sizes expected from the method conditions. The individual fragment genomes are shown to be cyclic gene permutations uniformly distributed on the map of physical distances between genes. The results suggest that any genetic marker has equal probabilities to be situated at the different distances from the individual particle of chromosome ends in a T4 D particles population.


[The mutagenic effect of new chemical substances. III. The mutagenic effect of several hydroxylamine derivatives]. The mutagenic effect of 21 new hydroxylamine derivatives was studied on biochemical mutants of Escherichia coli P-678 and Actinomyces rimosus 222. The Iver--Szybalsky method was used for the preliminary selection of mutagens. Dose-effect curves were plotted for the most efficient mutagens. Among the compounds studied 4 hydroxylamine derivatives were observed to have a mutagenic effect. The most efficient among them proved to be 0-(4-metoxybenzyl)-hydroxylamine and 0-(gamma-chlorocrotyl) hydroxylamine that induced reversions in the threonine locus of E. coli P-678 and in the lysine locus of Act. rimosus 222, being more efficient than hydroxylamine, the generally known mutagen. The dependence of the mutagenic activity of the compounds studied on their chemical structure is investigated.


[A genetic and physiologic study of mutations of T4 phage suppressing the lysis defect of gene stII mutants]. A new gene rVI phage T4B is found. Mutations rVI belong to the class of r-mutations of T4B. Mutations rVI do not show the lysis inhibition after superinfection and determine the r-morphology of plaques on Escherichia coli CR63 lawn. Mutations rVI are gene-specific suppressors of stII lysis defect, but do not suppress the e lysis defect. Alleles rVI and rVI+ are co-dominated with regard to the ability to suppress the stII lysis defect. The suggest the stoichiometric character of rVI gene function. Mutations rVI increase the permeability of infected cells before the lysis. The gene rVI located between the genes 56 and 39 on the T4B genetic map, close to the gene 56. The still mutations described earlier (Krylov, Yankovsky, 1973) suppress the lysis defect of the am z25 mutant in the gene e. The suppression is manifested when infected bacteria develope only in the liquid media.


[The influence of several prophages on the genetic behavior of Flexner dysentery bacteria]. In conjugation experiments of Escherichia coli K-12 Hfr strains and converted clones of Shigella flexneri var. y(-:3,4) that had acquired the capacity to synthesize type of antigens IV or V it is confirmed that the locus linked to lac-pro region in Sh. flexneri chromosome called Tp locus is a site of an attachment of prophages responsible for certain type specific antigens. Lac+ hybrids of the clone converted by phage IV lost the type specific antigen IV with the frequency comparable with the loss of the aforementioned antigen by wild strains of Sh. flexneri ser 4 in similar experiments carried out previously (90,8% and 97% respectively). Lac+ hybrids of the clones converted by phage V did not lose the type antigen that corresponded to the behaviour of wild strains of serotype 5b (V: 7,8) with double lysogenicity. It was shown that the clones converted by the aforementioned phage V had acquired an immunity not only to phage V, but to phage 7,8 as well. An independent segregation of immunity to phages V and 7,8 in lac+ hybrids of the converted clone was observed. It indicates that donor strain NTCC 595/52 carries two prophages, V and "defect" 7,8 (not expressing factor 7,8). A maintenance of antigen V observed in lac+ hybrids of the converted clone in this case confirms previous suggestions that an attachment of prophage 7,8 creates a state of some inhomology in this region disturbing the recombination process. The converting phage V isolated from the strain NTCC 595/52 is probably recombinant. An analysis of the hybrid classes allows to suggest the following approximate order of markers of the prophage on the chromosome of the converted clone: aV--imm V--imm 7,8--lac.


[The role of vegetative and reparative DNA synthesis in correcting single-strand breaks arising upon exposure of Escherichia coli to gamma rays]. It is shown that thymine starvation stops the growth of auxotrophic cultures at the moment of combining Okazaki fragments. Amino acid starvation prevents the mitosis in cells where DNA replication has completed. Therefore, it is easy to synchronize bacterial cells and to preserve them in two different states: when the chromosome is doubled or when the cells just divided and the chromosomes is single. The postirradiation repair, studied by measuring the number of single strand breaks, is going on with the same efficiency both in wild type cells and in different repair mutants. This suggests that sister chromatide exchange does not participate in postirradiation repair, especially in that affected by the rec-system. Some data are obtained that different pools of precursors are used by the cell in case of the repair synthesis of DNA and of vegetative replication.


[A change in the activity of the beta-phenylpropionic acid conversion enzyme in Escherichia coli under the influence of transmissive plasmids of Salmonella heidelberg]. The activity of the enzyme of the conversion of beta-phenylpropionic acid was studied in the strains of Escherichia coli serotypes 055:K59:H2 and O111:K58:H2 used as recipients via transmission of R- and Col-plasmids by conjugation in vitro. The activity of this enzyme was determined by modified method of Ben Hamida [3]. The wild type strain of Salmonella heidelberg carrying ColIb plasmid and preliminary obtained R1-19 plasmid from E. coli J 5-3 was used as a donor. The activity of the enzyme of conversion of beta-phenylpropionic acid in recombinants carrying R1-19ColIb plasmids was 3-5 times lower as compared with the original recipient. The colour reaction drived under the growth of original bacterial strains in the nutrient broth agar contained beta-phenylpropionic acid at the final concentration of 20 mg% was completely inhibited with that recombinants. The activity of this enzyme in recombinants carrying only R1-19 or ColIb plasmid remains unchanged.


[The effect of DNA isolated from different sources and different physico-chemical conditions on Candida tropicalis strain D-2 his-]. Mutagenic effect of DNA, isolated from Escherichia coli, Actinomyces olivaceus and calf thymus, on hystidine-deficient strain of Candida tropicalis D-2 is demonstrated.


[The influence of adenine, adenosine and adenylic nucleotides on the mutagenic action of hydroxylamine]. The effect of some adenyl precurosors of DNA synthesis on the mutagenic activity oh hydroxylamine (HA) is studied. It is shown that the addition of adenine to a suspension of Escherichia coli Bcells increases the yield of mutants by more than two times as compared with HA alone. The effects of adenosine, AMP and dAMP are somewhat different. It is suggested that the increase of the HAmutagenic effect produced by the addition of adenine may be due to:1) the excess of the amount of adenylic precursors of DNA synthesis over guanilic ones, which promotes the erroneous base-pairing during the replication of the HA modified template; 2) the modification of adenylic precursors by HA into N6-oxy-dATP, and their incorporation into DNA. The mutagenic effect of N6-hydroxyadenosine, the product of the adenine modification by HA, in E. coli Bpur- was studied. The experiments showed that N6-hydroxyadenosine induced about 1% of mutations, a relatively low lethal effect (the cell survival was 80%), and provided a high mutagenic action of this compound.


[The influence of bacterial genes recB, recA, exr and polA on T4 phage DNA transfection of lysozyme spheroplasts of Escherichia coli]. The transfection efficiency of T4 phage DNA of Escherichia coli spheroplasts, carrying mutations in four genes, involved in recombination and reparation (recB, recA, exr, polA), is studied. RecB- and recA- mutations increased the phage yield during transfection, and exr- and polA- mutations decreased it. RecA- mutation is suggested to influence on spheroplasts membrane state, resulting in more efficient replication of transfecting phage DNA.


[Revertants of recA-strains of Escherichia coli K-12]. The paper presents the results of a study of revertants of strains AB2463 recA13, AB2465 rec15, No 41 rec16. It is shown that revertants are characterized as intermediate strains between recA and rec+ (on the level of recB, recC strains) on their recombination proficiency in crosses with Hfr, sensitivity to UV and gamma-rays and in F-heterogenote formed cultures on their capacity of the formation of recombinants between episome and chromosome and the capacity to chromosome mobilization. In the same time revertants do not differ from rec+ in the frequency of spontaneous and UV-induction of prophage constitution of recombinants formed in crosses, and in the synthesis of beta-galactosidase in postconjugation period. It is found that the mutations studied are mapped apparently in recA gene.


Gastric lesion in dermatitis herpetiformis. Five of 33 patients with dermatitis herpetiformis (DH) were found to have gastric parietal cell antibody in their sera, whereas it was not found in 30 healthy controls of comparable age distribution. Fifteen of the patients with DH underwent further studies to investigate the histological and functional state of their gastric mucosa. Atrophic gastritis was found in all five patients whose sera contained gastric parietal cell antibody and in three of 11 patients with no antibody in their sera. In addition, there was marked impairment of acid secretion in the DH group as a whole, but, apart from one patient with overt pernicious anaemia (PA), there was no evidence of malabsorption of B12.


Alcohol and absorption from the small intestine. 1. Impairment of absorption from the small intestine in alcoholics. An absorption screen was performed in 10 chronic alcoholic patients within a few days of admission due to an acute alcoholic episode. Impaired absorption of d-Xylose was noted in three patients and low leucocyte ascorbic acid and serum folate levels in five. No abnormality was detected in jejunal histology. The absorption of water and electrolytes from the jejunum was studied in these patients using a triple-lumen tube perfusion system. The mean rate of absorption of water in the alcoholic subjects (50-0 +/- 2-3 ml/h) was significantly lower (P less than 0-001) than the mean value in 14 healthy control subjects (205 +/- 15-9 ml/h). A significant reduction of Na+ and Cl-absorption was also demonstrated in the alcoholic subjects. These results indicate that patients with acute-on-chronic alcoholism may have a function impairment of water and electrolyte absorption from the jejunum. This may, in part, account for some of the nutritional deficiencies in such patients and for symptoms such as diarrhoea which may be present.


Impairment of jejunal absorption rate of carnosine by glycylglycine in man in vivo. Using a double-lumen tube jejunal perfusion system in vivo, the mutual effects of carnosine (beta-alanyl-L-histidine) and glycylglycine on their respective absorption rates have been studied in six Zambian African adults. Data on the effect of the constituent amino-acids of carnosine on glycylglycine absorption rate have similarly been obtained. The solutions infused in each subject contained (A) carnosine (50 mmol l.-1), (B) carnosine (50 mmol l.-1) and glycylglycine (50 mmol l.-1), (C) glycylglycine (50 mmol l.-1), and (D) glycylglycine (50 mmol l.-1), L-histidine (50 mmol l.-1) and beta-alanine (50 mmol l.-1). Glycylglycine produced a significant impairment in the mean rate of histidine absorption from carnosine (P less than 0-01). However, carnosine did not have a significant effect on the mean rate of glycine absorption from glycylglycine. Mean rate of histidine absorption from solution D was significantly higher than that from solution A (P less than 0-01). Mean rate of glycine absorption from glycylglycine was not significantly different during infusion of solutions B, C, and D. The results are consistent with the concept that carnosine on glycylglycine is probably because the affinity of mechanism; the lack of influence of carnosine on glycylglycine is probably because the affinity of carnosine for the dipeptide uptake mechanism is relatively low. A gross difference has been shown between mean absorption rate of histidine from free L-histidine (solution D) (25-8 mmol h-1) and when it is given in the form of carnosine in the presence of another dipeptide (solution B) (8-7 mmol h-1); that emphasizes the complexity of amino acid and peptide interaction during absorption, which must be important in nutrition.


Fibreoptic endoscopy and the use of the Sengstaken tube in acute gastrointestinal haemorrhage in patients with portal hypertension and varices. The value of emergency upper gastrointestinal fibre-endoscopy, followed where required by the use of a modified Sengstaken tube, was studied during 84 episodes of acute bleeding in 75 patients who had evidence of portal hypertension with varices. The portal hypertension was due to alcoholic cirrhosis in 80% and to cryptogenic cirrhosis in 9% of the patients. By definition, varices were present in all patients, but in only 66% of episodes were the varices the cause of the bleed. The correct diagnosis of the source of bleeding was made at endoscopy in 89%. A Boyce modification of the Sengstaken-Blakemore tube was passed in 73% of the episodes of variceal bleeding. It effectively stopped the bleeding primarily in 85% of patients but was successful as a final definitive measure only in 46%. Furthermore, only 40% of the patients in whom the tube was passed, survived. Mortality rate could be related to the severity of the bleed and to hepatocellular dysfunction. Survival increased from 23% in those patients with jaundice, ascites, and encephalopathy on admission to 92% in those without these manifestations. The in-hospital survival rate was 52% in patients bleeding from varices and 64% in those bleeding from other causes, with an overall survival rate of 56%, indicating the poor prognosis in cirrhotic patients with gastrointestinal bleeding, irrespective of the cause.


Continuous monitoring of the effect of pentagastrin on gastric emptying of solid food in man. By continuous monitoring of a solid meal labelled with a radiopharmaceutical it has been possible to determine the effects of drugs on gastric emptying and motility during a single study. Predictably hyoscine delayed, and bethanechol increased, the rate of gastric emptying. Pentagastrin initially produced marked antral activity resulting in a physiological stricture and subsequent delay in the overall rate of gastric emptying. Fundal motility was unaffected though reflux from the antrum occurred.


Induction of pyloric hypertrophy by pentagastrin. An animal model for infantile hypertrophic pyloric stenosis. Administration of pentagastrin in depot form to 20 pregnant bitches produced pyloric hypertrophy in about 28% and gastroduodenal ulceration in about 16% of their pups. The two lesions were not necessarily found in the same individuals. Histological appearances of the pylorus in affected pups closely resembled those of human infantile pyloric stenosis.


Determination of liver volume by the use of a gamma camera connected with 1600 or 4096 channel analyser in diffuse liver diseases. A method of in vivo calculation of liver volume using a gamma camera and a 1600 or 4096 channel analyser is described. Topographically, the maximum thickness giving the highest counts in matrix data in the right anterior oblique projection is measured as the maximum breadth in the right posterior oblique projection. Then the total counts given by the liver can be transformed to the volume. The necropsy criteria indicated that the errors were between --11-9 +/- 14-7% in 13 cases by the use of 198Au-colloid. In 20 selected normal control subjects, the mean of measured liver volumes was 779 +/- 99 cm3/m2 of surface area. The volumetric calculation using 99mTc-Sn-colloid or 99mTc-phytate was compared with 198Au-colloid in the same patients. The results with 99mTc-Sn-colloid were well correlated with those with 198Au-colloid, but the results with 99mTc-phytate and 198Au-colloid were almost equal. It was concluded that, excluding rare severe cases of liver cirrhosis, liver volume can be measured with reasonable accuracy.


Relationship between bile acid malabsorption and pancreatic insufficiency in cystic fibrosis. Bile acid loss (mg/m2 24h) in the stools of 43 cystic fibrosis (CF) children with pancreatic insufficiency was 751-1 +/- 48-3, while that of six without clinical evidence of pancreatic disease (133-4 +/- 15-9) did not differ from values in 25 controls (109-8 +/- 9-8). There was a good correlation between the degree of bile acid (BA) and fat sequestration. Concomitant changes in bile acid and fat loss were observed in the one group of six patients studied on and off pancreatic enzymes as well as in a second group of seven children treated with pancreatic supplements and maintained on a normal diet followed by a low fat diet supplemented with medium chain triglycerides. Administration of NA bicarbonate led to a significant decrease in fat loss (15-8 +/- 2-7 leads to 10-3 +/- 1-9) without any simultaneous change in bile acid excretion (533-1 +/- 58-3 leads to 500-4 +/- 58-6). Qualitative bile acid patterns in controls, in infants after an ileal resection, and in patients with CF or with coeliac disease showed that the percentage of primary BA followed closely the total amount excreted except in situations where antibiotics were administered. The exact mechanism for the increased loss of BA in CF is unknown. It is found in all age groups and is related to the presence and degree of pancreatic insufficiency. The possibility that unhydrolysed triglycerides may interfere with the intestinal absorption of bile acid needs further confirmation.


Serum lysozyme levels in Crohn's disease and ulcerative colitis. Serum lysozyme levels were determined in healthy volunteers, patients with Crohn's disease, and patients with ulcerative colitis. The mean concentration in Crohn's disease was significantly greater than in the other groups. In patients with Crohn's disease, as well as in patients with ulcerative colitis, the lysozyme levels correlated with the severity of the disease process and with the extent of the lesions: the more severe the disease and the more extensive the involvement, the higher the lysozyme levels. However, the lysozyme values of the different groups overlapped considerably. Our results indicate that lysozyme determinations have only limited discriminative value for the diagnosis of Crohn's disease and for determining the severity and the extent of the disease process in the individual patient.


Influence of autonomic nerves on the internal and sphincter in man. The internal and sphincter receives its parasympathetic nerve supply from the sacral outflow and its sympathetic supply from the thoracicolumbar outflow of the spinal cord. In order to investigate the influence of the tonic discharge of these nerves, eight healthy subjects receiving high spinal anaesthesia (T 6-T 12) and five receiving low spinal anaesthesia (L 5-S 1) were examined. Continuous recordings of anal pressure and electromyographic activity from the external sphincter were obtained during rest and during expansion of the ampulla recti by means of an air-filled balloon. The results were compared with those obtained in an earlier study from 10 subjects with a bilateral pudendal block which paralysed the striated sphincter muscles without affecting the autonomic nerve supply to the internal sphincter. Anal pressure at rest decreased significantly more with high spinal anaesthesia (32 +/- 3-2 mm Hg) than with low (11 +/- 7-1 mm Hg) or with pudendal block (10 +/- 3-9 mm Hg) and the relaxations of the internal sphincter induced by rectal distension were somewhat smaller with high spinal anaesthesia. However, the remaining anal pressure at maximal relaxation, induced by a substantial rectal distension, was essentially the same with the three forms of anaesthesia. It is concluded that, at rest, there is a tonic excitatory sympathetic discharge to the internal anal sphincter in man. However, this seems to be without excitatory effect when the sphincter is relaxed after a substantial rectal distension. Furthermore, the results indicate that at rest there is no tonic parasympathetic discharge affecting the sphincter tone.


[Sulpiride in the management of gastroduodenal ulcers. A comparative double-blind study]. Sulpiride (Dogmatil) is a new type of drug acting on the hypothalamus resulting in a physiological increase in amplitude and frequency of gastric motor activity, reduction of signs of abnormal activity and pyloric spasm and an increase in the rate of gastric emptying. Gastric secretion is for all practical purposes unchanged, either in quantity or in acid content. The effects of Sulpiride on the duodenum are similar through some hypotony may be seen. The motor effects of Sulpiride do not relate to its effectiveness in treatment of gastroduodenal ulceration and experiments have shown that this activity can be explained by the reduction of capillary vaso-constriction in local blood circulation at the site of experimentally-induced ulcers, producing a vaso-dilatation assisting healing. This study objectively evaluates the activity of Sulpiride in gastro-duodenal ulcers. 64 patients were studied under  strict criteria and acute hemorragic, multiple or abnormally situated ulcers as well as those associated with other infections were eliminated. The dosage was uniform, two ampoules daily of 100 mgs i.m. of Sulpiride or placebo for 12 days, followed by 3 capsules daily of 50 mgs of Sulpiride or placebo for 48 days. No other treatment was given and no dietery restrictions were imposed, though alcohol and Aspirin and similar drugs were restricted. Of the 64 patients, 26 (18 men, 8 women, aged from 21 to 80 years) had gastric ulcers, 14 were treated with Sulpiride, 12 with placebo. 38 cases (29 men and 9 women, aged between 21 and 70 years) had duodenal ulcers, 18 were treated with Sulpiride, 20 with placebo. Full clinical, biological, radiological and endoscopic tests were carried out and a full questionnaire was completed. Highly significant differences in favour of Sulpiride were shown in both gastric and duodenal ulcer cases, maximum from the 20th day than less significative radiological results appeared. Similar remarks apply to endoscopic results. Sulpiride was extremely well tolerated, though amenorrhea (1) or galactorrhea (2) were noted without causing inconvenience to the patient. The author believes that the clinical effectiveness of Sulpiride in the treatment of gastro-duodenal ulcers is incontestable showing maximum results in ambulatory patients from the 20th day. A large number of patients would probably have produced statistically significant differences in radiological and endoscopic evidence of healing.


Serum insulin changes in arteriosclerotic breeder female rats versus non-arteriosclerotic virgin rats. Immunoreactive serum insulin (IRI), glucose, free fatty acids (FFA), and corticosterone (Cmpd. B) were measured in non-arteriosclerotic virgin versus arteriosclerotic, breeder female rats exhibiting normal 4-day cycles. In the arteriosclerotic breeder females, the normal increase in IRI during proestrus and estrus did not occur. The IRI levels during these times were comparable to the minimal levels found during metestrus in the non-arteriosclerotic virgin rats and were even lower than the IRI levels or ovariectomized rats. Since both young and old non-arteriosclerotic virgin rats had similar IRI patterns, the lack of cyclic insulin change in the arteriosclerotic breeder rats is not considered to be age-related. Instead, these changes are ascribed to hypothalamic-pituitary-adrenal hyperactivity associated with repeated reproductive activity and the development of arteriosclerosis.


Therapy-orientated diagnosis of secondary amenorrhoea. A programme for investigation of secondary amenorrhoea is described, which is based upon a functional classification of the endocrine causes of anovulation. All diagnostic enquiries not directly yielding a therapeutic dividend are excluded from this protocol. Three main categories of amenorrhoea are recognised: primary ovarian disease, hyperprolactinaemia and disorders of gonadotrophin secretion. There are subdivisions of each category, each with its own special diagnostic and therapeutic implications. Initial investigations take 6 weeks to complete and include measurements of follicle-stimulating hormone and prolactin, pituitary radiology and assessment of oestrogen production and of the response to clomiphene. The results of analysis of 75 consecutive cases of amenorrhoea and of 18 patients treated for infertility are presented and a cost benefit analysis applied.


Cell mediated immunity and the inflammatory system. In this review we have considered the evidence for the existence of lymphokines and have focused on the specific mechanisms by which lymphokines modify the behavior of the various inflammatory cells. These mechanisms are based for the most part on in vitro observations. We have therefore discussed in detail data that document an in vivo role for various lymphokines. This evidence is based on experiments falling into two broad categories: the identification of lymphokines in tissue extracts and serum, and the demonstration of various biologic activities of exogenous lymphokines administered locally or systemically. The details of these kinds of experiments have been presented throughout the preceding discussion. The demonstration of lymphokines in vivo adds a new dimension to our ability to study human disease. The detection of migration inhibition factor in serum, for example, is technically simpler than studies of lymphocyte reactivity in patients with altered cellular immunity. Migration inhibition factor has already been found in the serum in several clinical settings, as described. The capacity of exogenous lymphokines for suppressing manifestations of delayed hypersensitivity may provide an explanation for the altered immunoreactivity often observed in some of  those diseases. A final comment is in order regarding the significance of lymphokine production in the overall biologic scheme of things. In previous sections we noted that although activation of lymphocytes for lymphokine production by specific antigen is a property of T cells, B cells may be so activated nonspecifically by certain mitogens. Moreover, it has been shown that migration inhibition factor, or substances with similar biologic and physicochemical properties, may be found in certain replicating cultures of nonlymphoid cells. Also, we have recently shown that migration inhibitory activity, as well as certain other lymphokine-like activities such as macrophage and lymphocyte chemotaxis, may appear following the in vitro or in vivo infection of nonlymphoid as well as lymphoid cells by certain viruses. All these results suggest that lymphokine production, rather than "merely" representing an effector mechanism for cell mediated immunity and for certain kinds of helper functions in antibody synthesis, represents a general biologic phenomenon that may play a role in various aspects of host defense. Thus, such mediator substances should be more properly called "cytokines." Lymphokines represent a restricted set of cytokines made by one class of cells (lymphocytes) activated in certain unique ways. In this view the lmyphocyte has acquired some specialized means for triggering such production, not available to other cells.


Current taxonomy and identification of nonfermentative gram negative bacilli. In recent years significant advances have been made in the characterization and taxonomy of nonfermentative bacilli as well as in the recognition of their pathogenic potential for man. In addition, certain other infrequently isolated bacteria that may be fermentative have been recognized, and improved methods for their identification have been developed. This discussion attempts to present a practical approach to the identification of these micro-organisms in the clinical microbiology laboratory.


Specimen collection and transport.

Specimen collection and transport. The approach to the collection and transport of meaningful and accurate microbiologic specimens depends upon the nature of the suspected infectious entity and upon the location of the pathogenic process, as well as the possession of appropriate instrumentation and containers. New information and newly developed collection-transport materials have improved the potential for the successful collection and isolation of pathogenic micro-organisms.


Depression of the plaque-forming cells to sheep red blood cells by the new-born larvae of Trichinella spiralis. The number of plaque-forming cells (PFC) per spleen is reduced in mice experimentally infected with 200 Trichinella larvae at 14 and 28 days after infection, whereas no difference is shown at days 7 and 56. The worms of the three different phases of the life cycle of Trichinella have been isolated and kept alive in vitro at 37 degrees in the inner compartment of a Marbrook chamber. The outer compartment of the chamber contained normal spleen cells and sheep erythrocytes. After 4 days, the spleen cells were removed and assayed for PFC. The results indicate that the new-born larvae are capable of producing substances which can diffuse through a Millipore filter and affect the spleen cells forming antibody to sheep erythrocytes. The suppression of antibody to sheep red blood cells in mice infected with Trichinella is a transitory phenomenon and can be related to the migrating phase of the life cycle of the parasite.


Induction of delayed-type hypersensitivity with BCG-associated proteins. Microgram quantities of bovine serum albumin (BSA) and hen egg albumin (OA) associate spontaneously with living BCG in aqueous suspension. The association is stable insofar as bound protein cannot be readily dissociated from the organism by repeated washing. Association of BSA or OA is dependent upon the concentration of specific protein or competing protein in the incubation mixture, pH and the amount of protein already bound to the organisms. Washed BCG "complexes" containing either BSA or OA are potent inducers of delayed-type hypersensitivity (DTH). The same soluble proteins are far less effective inducers of DTH, even when injected with BCG. The capacity of OA-BCG complexes to provoke a cell-mediated response seems to be related, at least in part, to "stabilization" of the antigen on an appropriate carrier and its concentration at the site of BCG injection.


Increased survival in calves of Escherichia coli K-12 carrying an Ent plasmid. Escherichia coli K-12 strains with and without an Ent plasmid were fed to calves, and the survival of each was monitored by viable bacterial counts of the feces. The E. coli K-12 strain carrying the Ent plasmid survived in the calves at significantly higher levels and for a longer period of time than the E. coli F(-) stain.


Effect of luteinizing hormone-releasing hormone (LH-RH) on the multiunit activity of the arcuate nucleus in the proestrous rats. Multiple Unit Activity (MUA) of the hypothalamic arcuate nucleus as indicated by discriminated spikes with window setting between 18-30 uV, the integrated MUA and the cortical EEG of the urethane anaesthetized rats on the proestrous day was recorded with the help of a polygraph. Infusion of LH-RH in the 3rd ventricle markedly increased the arcuate nucleus MUA. The effect started within minutes of infusion, peak came after 20-30 minutes and the activity remained increased for 3-4 hours. Saline infusion in the 3rd ventricle was ineffective. Feedback action of CSF-LH-RH to the arcuate nucleus for the regulation of LH-RH is suggested.


Antibacterial activity in colostrum and milk associated with protection of piglets against enteric disease caused by K88-positive Escherichia coli. Piglets suckled by dams that had been vaccinated with K88 antigen were significantly more resistant to deaths caused by neonatal diarrhea after challenge with a large dose of a K88-positive enteropathogenic strain of Escherichia coli than piglets suckled by control dams. The factors most likely to be involved in protection of the piglets were investigated by comparing the antibacterial activities of serum and mammary secretions from the two groups of dams. Vaccination stimulated the production of K88 antibodies, which were associated with anti-adhesive activity directed against the adhesive properties of the K88 antigen, and of O8 antibodies; the latter antibodies were attributed to traces of O8 antigen in the vaccine. Neutralizing activity against heat-labile enterotoxin was present in several dams before vaccination but was not stimulated by bacteriostatic activities were similar in serum and mammary secretions from both groups of dams and appeared to play no significant role in the protective after parturition were atrributed to exposure of the dams to the challenge strain excreted by the piglets. It was concluded that neutralization of the adhesive properties of K88 antigen by K88 antibodies in colostrum and in milk contributed significantly to the protection of piglets from vaccinated dams. However, the contribution of antibacterial activities associated with the greater levels of O8 antibodies in colstrum from the vaccinated group cannot be entirely excluded.


Virulent Treponema pallidum: aerobe or anaerobe. Substrate degradation and protein synthesis served as indicators of metabolism in virulent Treponema pallidum. Opitmal metabolic activity in these spirochetes was observed at 10 to 20% O2 concentrations, with markedly reduced activity at higher or lower O2 levels or under anaerobiosis; alternate functioning electron acceptors that might substitute for O2 were not found. Carbon monoxide and cyanide at concentrations that inactivate cytochrome oxidase were not effective metabolic poisons for T. pallidum, although Micrococcus lutea, a strict aerobe with cytochrome-dependent respiration, was inhibited under similar experimental conditions. Motility of virulent T. pallidum was vigorous in the presence of O2 and sluggish or inhibited in its absence, reinforcing the role of O2 in T. pallidum metabolism.


The role of nerves in the production of cardiac arrest during surgical anaesthesia. The role of cardiac nerves in the production of cardiac arrest during surgical anaesthesia on coronary ligated hypoxic heart has been studied. When atropinished coronary ligated dogs were exposed to hypoxia the terminal event was a cardiac asystole in 88% of the dogs. In propranolol treated dogs, or in dogs where sympathetic ganglia upto T6 were bilaterally removed earlier, coronary ligation and hypoxia produced ventricular extrasystoles, ventricular tachycardia and repeated sinus arrest followed by ventricular fibrillation. The possibility of the origin of arrythmia from the damaged myocardium, and the presence of an intact vagus in the production of ventricular fibrillation has been discussed.


Failure of endotoxin to increase nonspecific resistance to infection of lipopolysaccharide low-responder mice. In vitro and in vivo responses to lipopolysaccharide (LPS) and various other bacterial immunostimulants were compared in c3H/He low-responder mice. The principal findings were as follows. (i) Their splenic lymphocytes were stimulated by various gram-negative mitogens such as an Escherichia coli peptidoglycan, a detoxified derivative of LPS, and even endotoxins extracted by trichloroacetic acid that are known to contain protein; spleen cells of these mice were also transformed by two other B-cell mitogens extracted from acid-fast organisms. (ii) Their macrophages were refractory to LPS and weakly responsive to a mycobacterial prepartion. (iii) LPS failed to elicit nonspecific resistance in these mice against Klebsiella pneumoniae infection. (iv) Endotoxin extracted by trichloroacetic acid and a mycobacterial preparation that could increase nonspecific resistance to infection in other strains did not protect C3H/He mice against a challenge by K. pneumoniae, although both prepartions could evoke nonspecific responses of B cells in this low-responder subline.


Inhibitory action of D-galactose on phagocyte metabolism and function. To account for enhanced susceptibility to infection among  galactosemics, the acute effects of D-galactose on metabolic and functional activities of phagocytic cells in vitro were investigated. Human and guinea pig polymorphonuclear leukocytes (PMN) when incubated in medium containing 30 mM galactose displayed substantially less killing of Escherichia coli than when incubated in medium with 5 mM glucose. Impaired bactericidal activity was dependent upon galactose concentration but could be partially averted by supplementing the galactose-containing medium with 15 mM glucose. Phagocytic activities of guinea pig PMN and peritoneal macrophages were assayed by following ingestion of 32P-labeled E. coli and were also depressed by elevated galactose. Galactose was readily epimerized to glucose by resting PMN, and this conversion was stimulated by phagocytosis. Incubation of macrophages in the presence of galactose resulted in depletion of intracellular levels of adenosine 5' -triphosphate as well as other metabolities.


Serum chemotactic inhibitory activity: heat activation of chemotactic inhibition. Serum chemotactic inhibitory activity (CIA) was studied in 46 patients with various systemic diseases, using a system consisting of normal human leukocytes as indicator cells and 10% fresh normal serum as a control chemotactic attractant. It was shown, as previously reported, that an association exists between CIA and skin test anergy. Heat treatment of sera at 56 C for 30 min increased both the incidence and the degree of chemotactic inhibition observed in these patients. The effects of heat treatment of sera containing CIA on other chemotactic attractants (C3a, bacteria-derived chemotactic factor (BF),  and casein) are shown. Before heat treatment, some sera suppressed chemotaxis mediated by BF in the absence of suppression of normal serum-mediated chemotaxis, indicating the possible involvement of more than one system of inhibition. Multiple systems were further supported by data indicating that room temperature incubation resulted in a loss of CIA as measured by normal serum-mediated chemotoxis with no apparent decrease in the inhibition of BF -mediated chemotaxis. Separation of sera containing CIA by Sephadex G-200 showed chemotactic inhibitory activity to be increased in both the void volume region. Experiments showed that heat treating before separation resulted in similar increases in both peaks, implying the presence of an antagonist to CIA. Experiments demonstrating that sera containing CIA do not suppress casein-mediated chemotaxis by means of an irreversible inactivation of chemotactic factor are included along with experiments demonstrating a cellular mode of action. The possible presence of two systems of chemotactic inhibition, one acting directly upon chemotactic factors and one interacting with the responding cell, are discussed.


In vivo and in virto cellular responses to cytoplasmic and cell wall antigens of Histoplasma capsulatum in artificially immunized or infected guinea pigs. Guinea pigs were infected with different doses of yeasts of Histoplasma capsulatum or artifically immunized with several concentrations of unextracted yeast cell walls, and then tested in vivo and in vitro for cell-mediated responses to various subcellular fractions of the fungus. Three types of cell-mediated responses were measured, viz., skin test activity, production of migration inhibition factor, and lymphocyte transformation. Positive cutaneous reactions were elicited in animals immunized with 100 or 1,000 mug of cell walls when such animals were skin-tested with cell wall glycoprotein of soluble cytoplasmic substances, whereas animals immunized with 2,000 mug of cell walls did not react significantly greater than unsensitized animals when skin-tested with the same antigens. Histoplasmin did not elicit cutaneous sensitivity in guinea pigs infected with the smallest inoculum, 6 X 10(5) yeast cells, or in animals immunized with cell walls, regardless of the concentration of cell walls used as immunogen. However, hypersensitivity to H. capsulatum could be detected with cytoplasmic substances in animals infected with 6X 10(5). In guinea pigs infected with larger doses, i.e., 10 X 10(7), 15 X10(7), or 20 X 10(7), hypersensitivity could be detected with histoplasmin, cell wall glycoprotein, a ribosome-rich fraction, and soluble cytoplasmic substances. Both cell wall glycoprotein and soluble cytoplasmic substances were functional in migration inhibition factor assays with peritoneal exudate cells from animals immunized with 100 or 1,000 mug of cell walls. The transformation of lymphocytes from infected and artificially immunized guinea pigs in the presence of cell wall glycoprotein and soluble cytoplasmic substances was variable and unpredictable, the lymphocytes from some animls within a given group transforming and those from other animals showing no evidence of stimulation. Moreover, the level of stimulation could not be correlated with the degree of dermal hypersensitivity. These findings suggest that cell wall glycoprotein, and the fractions containing ribosomes and soluble cytoplasmic substances, could be useful antigens in assays for cellular immunity, and warrant further investigation with respect to specificity and active components.