Double-blind trial comparing two dosage schedules of beclomethasone dipropionate aerosol with a placebo in chronic bronchial asthma. Second report of the Brompton Hospital/Medical Research Council Collaborative Trial.

A follow-up study to 52 weeks is reported on 95 corticosteroid-dependent asthmatics who took part in a double-blind, controlled trial of beclomethasone dipropionate for 28 weeks. During the 28 weeks two dosage schedules (800 microgram and 400 microgram daily) were compared with a placebo. If the corticosteroid tablet dosage had been at least halved and the clinical condition was satisfactory, the allocated regimen was continued 'blind' for the second period (29-52 weeks); 51 patients continued for this period, 46 on beclomethasone dipropionate. If progress was unsatisfactory, known beclomethasone dipropionate 800 microgram daily was substituted, the supervision remaining unchanged, the clinical supervisors remaining unaware which schedule had been received during the first 28 weeks. Nearly all the patients who at least halved their corticosteroid tablet dosage by 28 weeks continued to do well up to 52 weeks on the allocated regimen. When the placebo patients with unsatisfactory progress at 28 weeks were changed to 800 microgram beclomethasone dipropionate they showed a substantial improvement. Patients changed from 400 to 800 microgram showed little improvement on average. Approximately 30% of the whole group of patients failed to have their dosage of corticosteroid tablets, despite receiving 800 microgram beclomethasone dipropionate daily for six or more months. The cumulative incidence of 'oral candidiasis' (as defined for this report) in placebo patients changed at 28 weeks to 800 microgram beclomethasone dipropionate daily was 30% at 36 weeks and 43% at 52 weeks. Of eight patients increased from 400 microgram to 800 microgram daily, one had 'candidiasis' by 36 weeks and three by 52 weeks. Oral candidiasis was often asymptomatic and never led to the discontinuation of the corticosteroid aerosol, although the dosage was reduced in nine of the 30 who received 800 microgram beclomethasone dipropionate during the first 28 weeks. The incidence of bacterial infections in patients on beclomethasone dipropionate was similar to that of the placebo group and systematic laboratory investigations revealed no increase in the rate of isolation of potential pathogens.


Tuberculosis in renal transplant recipients.

Of more than 400 patients who received renal transplants in Cambridge, five subsequently developed tuberculosis. All had pulmonary involvement. Radiological abnormalities included miliary opacities, bilateral lower zone shadowing or typical upper zone shadowing with cavitation. Lung biopsy was required for diagnosis in two cases. Three patients recovered after treatment with antituberculosis drugs and none has relapsed. One died during chemotherapy despite drug-susceptible organisms. The problems encountered during chemotherapy are discussed.


A comparison of oral choline theophyllinate and beclomethasone in severe perennial asthma in children.

Fourteen children with perennial asthma previously treated with beclomethasone dipropionate were studied for four months in a crossover trial to compare the efficacy of choline theophyllinate and beclomethasone dipropionate aerosol in the control of their symptoms. Assessments were made using twice daily peak flow measurements, together with daily records of symptom scores and use of extra salbutamol. Beclomethasone and oral choline theophyllinate together improved control of asthma in comparison with beclomethasone alone. Reduction of the dose of beclomethasone from 400 microgram/day caused worsening of asthma. Oral choline theophyllinate did not prevent this deterioration and therefore cannot replace beclomethasone for the long-term treatment of such asthmatic patients.


Two aminophylline preparations in nocturnal asthma.

In a double-blind trial the effects of aminophylline suppositories and slow-release aminophylline tablets were compared with similar placebo preparations in nine patients whose complaint was asthma which woke them from sleep at night. Forced expired volume in one second (FEV1) was measured in each patient at three-hourly intervals from 1900 hours to 0700 hours on four separate nights, the preparation being administered immediately after the 2200 hour measurement. On the nights when a placebo was administered the FEV1 showed a statistically significant fall between 1900 and 0400 hours. Aminophylline had a statistically highly significant effect in reducing or preventing this fall, suppositories and tablets being equally effective in this single-dose study.


Haemoptysis in eosinophilic granuloma.

Eosinophilic granuloma of bone may involve the lung in about 20% of cases (Lewis 1964); and may present in the lung without other organ involvement. Haemoptysis occurs in about 10% of cases for unknown reasons. I document here a case with typical presentation and course, but which was complicated by haemoptysis of increasing severity leading to death. The cause of haemoptysis was diagnosed at fibreoptic bronchoscopy.


Unusual course of beryllium lung disease.

The cases of two patients with beryllium disease of the lung are reported in which acute exposure produced immediate symptoms and progressed to chronic disease with no latent period.


Immune response to human keratoacanthoma.

Immune reactivity to human keratoacanthoma was investigated by microcytotoxicity tests and immunofluorescence. IgM and complement were consistently present in lesions; IgG and fibrin were infrequent. No evidence of in vivo bound immunoglobulin was found on the surface of keratoacanthoma cells by membrane immunofluorescence. Neither patients' sera nor peripheral blood leukocytes showed significant cytotoxicity against autochthonous tumour cells in microtitre assays. This study fails to support the view that regression of human keratoacanthoma is mediated by immunological mechanisms.


Architecture of the outer membrane of Escherichia coli K12. IV. Relationship between outer membrane particles and aqueous pores.

The hypothesis that intramembraneous particles, observed in the outer membrane of Escherichia coli by freeze-fracture electron microscopy, are the morphological representation of aqueous pores, was tested. A mutant which is deficient in five major outer membrane proteins, b, c, d, e and the phage lambda receptor protein, contains a largely decreased number of intramembraneous particles and also shows a greatly decreased rate of uptake of several solutes. In derivatives of this strain which contain only one of these proteins in large amounts a strong decrease of the number of intramembraneous particles is observed, which is accompanied by a complete restoration of the rate of uptake of those solutes which use pores in which the protein in question is involved. The results provide strong evidence for the notion that an individual pore contains only one protein species, a property which has been found earlier for individual particles. The observed correlation between particles and equeous pores strongly supports the hypothesis that the particles are the morphological representation of pores. Implications of this hypothesis for the structure of the particles are discussed.


Proteinases as probes of the structure of rat liver ribonucleoprotein particles carrying heterogeneous nuclear RNA.

Three proteolytic enzymes with different specificities produced essentially the same degradation pattern of the proteins of ribonucleoprotein particles. Proteins with apparent molecular weights of 144 000, 133 000 and 115 000 were hydrolyzed most readily which may indicate an exterior location in the ribonucleoprotein complex. The major particle proteins in the range of 30 000-42 000 daltons were much less susceptible to proteinases than the high molecular weight species. Proteins of 31 000, 33 000 and 42 000 daltons were resistant to proteolysis. The degradation rates of the proteins in intact ribonucleoprotein particles were considerably lower than rates obtained with the same proteins separated from the RNA. Incubation with trypsin and recentrifugation of polyparticles led to a shift in the sedimentation constant from 90 S to 0-20 S, corresponding to a decrease in the protein to RNA ratio from 4 to 1.3. No significant change in the small molecular weight RNA pattern was detected after trypsin digestion.


Subunit localizations of zinc(II) in DNA-dependent RNA polymerase from Escherichia coli B.

RNA Polymerase holoenzyme and core enzyme from Escherichia coli B have been shown to contain two zinc ions. Flameless atomic absorption spectroscopy of the isolated core subunits indicated that one zinc ion is localized on the beta subunit and the other is bound on the beta' subunit. Atomic fluorescence spectroscopy showed that prolonged dialysis of the metalloenzyme against 0.01 M o-phenanthroline resulted in the removal of both zinc(II) ions with accompanying loss of enzymatic activity. The activity of the apoenzyme was observed to be completely restored by readdition of zinc(II) and partially restored by cobalt(II).


Prediction of pairing schemes in RNA molecules-loop contributions and energy of wobble and non-wobble pairs.

Previously published models for predicting pairing schemes in RNA molecules, when applied to tRNA, give the clover leaf structure in only half the cases. We made a systematic investigation of the predictability of the clover leaf structure under various assumptions concerning the energetic contributions of single and double-stranded regions. We tested 21 different models and variants on a set of 100 tRNA sequences and many other variants on a smaller set of sequences. In our models we allowed not only G.C, A.U and G.U pairing, but also every other pair. Under conditions which are much less restrictive than those of previous attempts, we can nevertheless reach 90 per cent predictability for the clover leaf structure of tRNA. A most surprising and far-reaching result is that we can assign to C.G and C.C pairs binding energies quite close to the energies of G.U pairs, and still predict the clover leaf. The following ranking for non-complementary pairs was obtained : G.U, G.G and C.C, U.U, C.A, A.A and G.A, U.C. The main practical innovation which made possible the improvements in predictability are: i) not counting the stacking of base pairs separated by a bulge loop; ii) making the terminal C.C's in stems more stable than the terminal A.U's by merely -- 0.7 kcal; iii) replacing the distinction between G.C and A.U-closed loops by a distinction based on the presence of loop-favoring residues; iv) carefully adjusting the energetic balance between the various kinds of loops; v) narrowing the gap between the GC/GC and the GC/AU contributions; vi) using observations on nearest-neighbours in tRNA sequences to refine the contributions of G.U pairs.


Genetic and physical characterization of a segment of yeast mitochondrial DNA involved in the control of genetic recombination.

Genetic recombination between the 3 RIB (ribosomal) loci of yeast mitochondrial DNA is under the control of a mitochondrial locus named omega (with alleles omega+ and omega-) which is tightly linked to the RIBI locus. We have attempted to elucidate the molecular mechanisms(s) involved by using rho- mutants with similar (RIBI+ RIB2+ RIB3(0) genotype but different recombination properties in rho- x rho+ crosses. These were obtained through pedigree analysis and their mitochondrial DNAs were mapped on a high resolution physical map of the RIB section that had been built by analysis of thermal denaturation profiles and electron microscopy of partially denatured molecules. By comparison of physical and genetic data it can be shown that possession of the omega+ allele by the rho- cell is not sufficient for its expression in crosses, some additional DNA segments(s) in the ribosomal region being needed. This result and several features of the rho+ x rho- crosses are discussed in the light of current concepts in mitochondrial genetics of yeast and the recently discovered fact that omega+ and omega- strains differ by the presence of a 1000 base pairs insertion in the former.


Analysis of large specific T1 oligonucleotides of 17S and 25S ribosomal RNAs from Saccharomyces cerevisiae.

The primary structure of 17S and 25S ribosomal RNAs from Saccharomyces cerevisiae has been analysed by two-dimensional fractionation of T1 oligonucleotides. This method consists of an electrophoresis at pH 3.5 followed by a homochromatography on DEAE-cellulose plates. After the second dimension, the large T1 oligonucleotides were hydrolyzed by pancreatic RNAse, followed by alkaline hydrolysis of the pancreatic products. By fractionating a mixture of tritiated HeLa cell ribosomal RNAs and 32 P yeast cell ribosomal RNAs, two autoradiographs were obtained; one corresponding to the 32P labelled material and the other to the tritiated labelled material. By superposition of the two autoradiographs, the mobility of the various T1 oligonucleotides can be accurately compared and it is shown that yeast 17S rRNA and human 18S rRNA have in common 5 large oligonucleotides and that yeast 25S rRNA and human 28S rRNA have 4 identical oligonucleotides.


Amino acid sequences of soluble tryptic peptides of chorismate mutase/prephenate dehydratase from Escherichia coli K12.

The amino acid sequences of 28 soluble tryptic peptides from chorismate mutase/prephenate dehydratase from Escherichia coli K12 have been determined. Together with the four unique cysteine-containing peptides sequenced by Gething and Davidson ((1976) Eur. J. Biochem. 71, 327-336) this accounts for approximately 75% of the total sequence expected for this protein. A high frequency of identify between some of the peptides suggests the possibility of gene duplication during the evolution of the structural gene for the enzyme.


Enhancement and suppression in production of IgM-antibody in mice treated with purified saponins.

Effects of purified saponins on antibody production against SRBC antigen were investigated in adult mice by using the direct hemolytic plaque technique of Cunningham and Szenberg. Intravenous injection of ginsenoside-Rb1, -Rc, senegin-III, -IV or platycodin-D caused increase in antibody-producing cells. However, the same treatment with ophiopogonin-D, ginsenoside-Rd or glycoside-H2 suppressed antibody production. Immuno-suppression due to the latter three substances seemed not to be associated with cytocidal effect. Ophiopogonin-B and platycodin-C had no effects.


Characterization of normal human cells by pyrolysis gas chromatography mass spectrometry.

Differentiation of normal human cells has been accomplished by pyrolysis gas chromatography mass spectrometry. Normal cells from human kidney, spleen, liver and brain tissues have been pyrolyzed and the products chromatographically separated and characterized by mass spectrometry. Molecular pyrolysis products giving rise to the characteristic pyro-mass chromatograms include, but are not limited to, alkenes, alkanes, nitriles and various ring compounds. Single ion mass chromatograms as well as multiple ion mass chromatograms have been used to explore the characteristic differences between various tissue materials. A dynamic computer methodology for comparing pyro-mass chromatograms has been developed for use in automatic identification and classification of the human cellular material.


Transcutaneous PO2 measurement on skin transplants.

Trnascutaneous PO2 measurements are suitable for an objective control of free skin transplants and pedicle flaps if used with standardized and controlled hyperemia. They are also suitable for monitoring the rejection of skin allografts and to investigate the influence of drugs on skin transplants in general.


Transcutaneous PO2 monitoring of flap circulation following surgery.

In a series of rabbit experiments, tcPO2 was found to be a very sensitive indicator of flap viability, reflecting minute-to-minute changes in physiology. It is safe, reproducible, and easily verified by comparing flap tcPO2 to a control site and by measuring response to increased FIO2. Transcutaneous PO2 measurements should prove a valuable tool in flap research and in monitoring pedicle flaps and microvascular tissue transfers.


[The release of proteinases from neutrophil leukocytes and macrophages].

The main source of proteinases that degrade tissue components in the course of inflammation are neutrophil leukocytes and macrophages. Both cells contain and/or produce a large variety of acid and neutral hydrolases including representatives of all four classes of tissue proteinases. Neutrophils are short-lived end cells with large enzyme stores (i.e. azurophil and specific granules) which they normally discharge in the course of phagocytosis. Macrophages are long-lived cells with a great potential for functional differentiation. In macrophages, proteinases and other lytic enzymes are released both by phagocytosis and by secretion (the latter particularly in activated macrophages) and replaced continuously by de novo synthesis. The acid and neutral proteinases of neutrophils and macrophages, their subcellular localization, their properties in relation to tissue damage and the mechanisms of their release are described.


[Regulatory effects of proteases in articular inflammation].

The role of proteases in the stimulation of cells participating in a chronic inflammatory reaction is reviewed. Several neutral proteases stimulate B lymphocytes to produce both specific and unspecific antibodies and such enzymes can replace, at least in vitro, T-helper cells. Proteases might influence macrophages as well, by a MIF-like activity, by increasing spreading in vitro or by liberating from C3 the C3b component which has an activating effect on these cells. Endocytosis of protease - alpha-2 macroglobulin complexes by macrophages induces production of neutral proteases as well. A stimulation of fibroblasts by proteases is possible but not yet clearly proven.


[Generalized periarthritis calcarea (generalized hydroxyapatite disease)].

The condition of generalized periarthritis calcarea (hydroxyapatite deposition disease) is characterised by multiple periarticular calcification which can be localised around practically any joint and also in proximity to the spine. This calcification consists of hydroxyapatite crystals which are responsible for the episodes of acute, subacute or chronic periarticular or articular inflammation so typical of the condition. Because of this one can classify periarthritis calcarea along with gout and chondrocalcinosis in the group of crystal deposition diseases. The actual cause of the calcification remains unknown but it is probable that, along with hereditary factors, disturbances in metabolism play an important role. The diagnosis of generalised periarthritis is made from the characteristic X-ray picture in conjunction with the clinical findings and, on occasion, the demonstration of hydroxyapatite crystals in the affected tissues. In the differential diagnosis gout, chondrocalcinosis, various inflammatory rheumatic conditions and septic arthritis must be excluded and various calcification processes, particularly interstitial calcinosis and lipocal cinogranulomatosis, must also be considered. Since the etiology of the calcification remains unknown to specific treatment is available. Symptomatic treatment with colchicine is mostly inadequate which is why one often has recourse to the use of non-steroid anti-inflammatory drugs and corticosteroids.


Depressive and extrapyramidal symptoms and clinical effects: a trial of fluphenazine versus flupenthixol in maintenance of schizophrenic out-patients.

Fifty-seven patients with a diagnosis of schizophrenia were started on either fluphenazine decanoate or flupenthixol decanoate injections in a double-blind trial just prior to discharge into the community. During the six month follow-up 30 per cent dropped out of the treatment. Of those observed for six months, 7 per cent relapsed, 54 per cent experienced depressive symptoms and 88 per cent extrapyramidial side-effects. Analysis of both clinical data and the ratings failed to discriminate between the two drugs.


A double-blind crossover trial of baclofen--a new treatment for the unstable bladder syndrome.

Forty patients with the unstable bladder syndrome have been treated in a double-blind crossover trial of baclofen 5 mg qds versus placebo. Baclofen significantly improved diurnal and nocturnal frequency of micturition and the severity of incontinence. Considerable improvement was also obtained with placebo.


Vesicoplication in the management of chronic urinary retention.

Eighteen patients have been treated by vesicoplication for low tension chronic urinary retention. The clinical presentation, operative technique and results in these 18 patients are discussed. Vesicoplication is shown to be a simple, safe and effective method of relieving symptoms in this group of patients.


Endourethral urethroplasty.

Following internal urethrotomy for treatment of strictures of the bulbous urethra, 11 patients had primary split skin grafts inlaid over the urethrotomy site. There were no operative complications and the results in 5 patients with a 9-month follow-up are good.


Impaired response of pancreatic polypeptide to hypoglycaemia: an early sign of autonomic neuropathy in diabetics.

The pancreatic polypeptide (PP) response to insulin-induced hypoglycaemia was studied in 18 juvenile diabetics and was calculated as the difference between the prestimulatory PP concentration and the maximal concentration measured. The response was severely impaired in patients with autonomic neuropathy (mean +/- SE of mean 22 +/- 12 pmol/l) as compared with patients without neuropathy (252 +/- 51 pmol/l). Patients whose diabetes was of only a few years' duration showed a normal PP response to hypoglycaemia, and the response diminished significantly with increasing duration of diabetes. The decreased PP response to hypoglycaemia was significantly correlated with an increased threshold of the sense of vibration, (rs = 0.86). These results suggest that impaired, secretion of PP may serve as an early sign of autonomic neuropathy in diabetes.


High affinity choline transport and acetylCoA production in brain and their roles in the regulation of acetylcholine synthesis.

This review describes recent advances made in the understanding of the regulation of acetylcholine synthesis in brain with regard to the availability of its two precursors, choline and acetylCoA. Choline availability appears to be regulated by the high affinity choline transport system. Investigations of the localization and inhibition of this system are reviewed. Procedures for measuring high affinity choline transport and their shortcomings are described. The kinetics and effects of previous in vivo and in vitro treatments on high affinity choline transport are reviewed. Kinetic and direct coupling of the transport and acetylation of choline are discussed. Recent investigations of the source of acetylCoA used for the synthesis of acetylcholine are reviewed. Three sources of acetylCoA have recently received support: citrate conversion catalyzed by citrate lyase, direct release of acetylCoA from mitochondria following its synthesis from pyruvate catalyzed by pyruvate dehydrogenase, and production of acetylCoA by cytoplasmic pyruvate dehydrogenase. Investigations indicating that acetylCoA availability may limit acetylcholine synthesis are reviewed. A model for the regulation of acetylcholine synthesis which incorporates most of the reviewed material is presented.


The effect of pantothenate on sulfate metabolism and sulfur isotope fractionation by Saccharomyces cerevisiae.

Growth of Saccharomyces cerevisiae in minimal salts-glucose-SO4(2-) medium with varying concentrations of pantothenate (0-1000 microgram/L) produced changes in the cellular lipid content and in the ratio of saturated to unsaturated fatty acids. Substantial differences in SO4(2-) diffusion were also observed with changes in pantothenate concentration. During sulfate reduction, the delta 34S value of the evolved sulfide varied with the pantothenate concentration ranging from -31% in the absence of pantothenate to 0% at 400-1000 microgram/L pantothenate. The isotope selectivity is related to the effect of pantothenate concentration on cellular metabolism.


A modified method for testing inositol assimilation by Cryptococcus species.

The proposed method for rapid testing of inositol assimilation incorporates shake cultures in an indicator-based broth containing inositol (1%), yeast nitrogen base (0.067%), bromocresol purple, and a heavy inoculum. Of 153 yeast isolates investigated, inositol assimilation was shown with the modified method, as also by the Adams-Cooper procedure, in all the 123 isolates, representing 11 species of Cryptococcus. The results were negative by both the methods in the remaining 30 isolates belonging to Candida, Rhodotorula, Torulopsis, Pichia, Saccharomyces, and Sporobolomyces. The modified method was found to be significantly more effective than the Adams-Cooper procedure; the results could be read within 36 h by the former as against 336 h by the latter method. The superiority of the modified method to the Adams-Cooper procedure is attributed to increased aeration in shake cultures, a heavier inoculum, and reduced concentration of yeast nitrogen base.


Nitrogen fixation associated with 'Park' Kentucky bluegrass (Poa pratensis L.).

Associative nitrogen fixation in Kentucky bluegrass (Poa pratensis L.) turfs inoculated with five nitrogen-fixing bacterial isolates was evaluated using the acetylene reduction assay and nitrogen accumulation as indicators of fixation. 'Park' and 'Nugget' Kentucky bluegrass turfs were grown in controlled environment chambers and inoculated with Klebsiella pneumoniae (W-2, W-6, and W-14), Erwinia herbicola (W-8), and Enterobacter cloacae (W-11). 'Park' inoculated with K. pneumoniae (W-6) had significant acetylene reduction activity using undisturbed turfs. Other treatments including turfs treated with heat-killed cells had no significant difference in acetylene reduction. In a second study, "Park' and 'South Dakota Certified' turfs were grown in a greenhouse and inoculated with K. pneumoniae (W-6) and E. herbicola (W-8). 'Park' inoculated with K. pneumoniae (W-6) had increased acetylene reduction activity rates and also a greater nitrogen accumulation in aerial tissues when compared to controls. Acetylene reduction activity was correlated (r = 0.92) to nitrogen accumulation. Other treatments did not effectively increase acetylene reduction activity or nitrogen accumulation.


Regulation of peptidoglycan biosynthesis in relA+ and relA- strains of Escherichia coli during diauxic growth on glucose and lactose.

In both relA+ and relA- derivatives, the biosynthesis of peptidoglycan, lipid intermediates, and nucleotide precursors abruptly halted at the onset of diauxic lag from glucose to lactose with a concomitant accumulation of guanosine 5'-diphosphate 3'-diphosphate (ppGpp). These results are consistent with the proposal that ppGpp is involved in inhibiting the incorporation of disaccharide-pentapeptide into peptidoglycan and in regulating nucleotide precursor synthesis.


Indigenous bacteria influence the number of Salmonella typhimurium in the ileum of gnotobiotic mice.

Gnotobiotic BALB/c mice associated with indigenous Lactobacillus, Bacteroides, and two fusiform-shaped Clostridium strains had fewer S. typhimurium present in the ileum 3 days after intragastric challenge with the pathogen than did similarly challenged germfree mice. Acetic and butyric acids were detected in the caecal contents of the gnotobiotic mice, but in smaller concentration than was present in conventionalized mice. No difference in the motility of the small intestine was detected between germfree, gnotobiotic, and conventionalized mice.


A method for establishing stable concentration gradients in agar suitable for studying chemotaxis on a solid surface.

A simple technique has been developed for establishing stable gradients of a substance in agar. The technique involves the creation of a spherically symmetric concentration profile in which concentration varies inversely with the distance from the source and is independent of the diffusion coefficient of the substance. It has been shown that the gradients established with this technique are stable for at least 190 h. and, on a theoretical basis, they can be kept stable for more than 1000 h. Time-variant gradients can also be established, if desired, using the same system and limiting either the source or the agar sink. It must be emphasized that a stable gradient cannot be obtained by using a shallow agar layer as a sink. The use of such conditions (e.g. the agar in a standard petri dish) can result only in time-variant gradients. The solution to the diffusion equation in a spherically symmetric system establishes the expected concentration profile, the basis for adjusting it, and the parameters that control the behavior of the system. Some useful applications for examining chemotaxis on a solid surface as well as possible further developments are discussed.


A temperature-gradient technique for the elimination of antibiotic resistance.

A technique was developed which facilitates the application of elevated temperatures to strains of unknown heat sensitivity during attempts to eliminate antibiotic resistance. Cultures were subjected to a temperature gradient and the lowest temperatures which inhibited growth were found to be highly efficient in the elimination of gentamicin resistance in several strains of Staphylococcus aureus and S. epidermidis.


[Methods to predict the percentage of degradation of fluorescence induced by laser impulses in the immunofluorescence FTA-ABS technic].

The force, diameter, and pulse time of laser rays in the FTA-ABS immunofluorescence technique permit us to calculate and to predict what the percentage of degradation of the emitted fluorescence will be for the operating conditions in which this degradation is produced.


Mycelial lipids as an aid in identifying rust fungi in culture-Cronartium fusiforme.

The sterol and fatty acid content of mycelium from germinating basidiospores of Cronartium fusiforme was determined. The mycelium contained stigmast-7-enol, fungisterol, and possibly stigmasta-5,7-dienol. No ergosterol was detected. The mycelium contained the expected fatty acids and low relative proportions of 9,10-epoxyoctadecanoic acid. The absence of ergosterol, and presence of the epoxy C18 acid and sterols typical of certain rust spores may be used for a relatively rapid confirmation of rust fungi in culture. Based on these chemical criteria, yeast-like cells isolated from the cultures of germinating basidiospores appear not to be C. fusiforme.


Purified viral neuraminidase vaccine to control influenza.

The control of influenza by immunoprophylaxis is difficult because of the antigenic mutability of the influenza virus and the unpredictability of its epidemiologic behaviour. The inactivated whole-virus vaccine currently used is not ideal. Vaccination with pure neuraminidase is suggested. The induced antineuraminidase antibody will restrict viral invasion. Mild illness may or may not occur. On subsequent exposure to influenza virus the individual will produce antihemagglutinin and antineuraminidase antibodies and will be resistant to both infection and illness. Since antigenic changes are less frequent in the viral neuraminidase than in the viral hemagglutinin, the vaccine would be usable for longer periods than the presently used inactivated whole-virus vaccine.


Yersinia pseudotuberculosis infection in a cat.

A diagnosis of Yersinia pseudotuberculosis in a cat was established by microbiological and histological means. Clinical signs were general malaise, anorexia and vomition. Significant laboratory findings included a serum glutamicpyruvic transaminase elevation and the presence of urine bilirubin and urobilinogen. Histological examination of liver biopsies revealed numerous varying-sized foci of caseation necrosis containing large numbers of Gram-negative coccobacilli. Bacteriological techniques, treatment and control measures are discussed.


Role of glycolytic flux in effect of glucose in decreasing fatty-acid-induced release of lactate dehydrogenase from isolated coronary ligated rat heart.

The mechanisms whereby glucose reduces fatty acid-induced release of enzyme from the coronary-ligated isolated perfused working rat heart are investigated.  Alterations in the tissue contents of ATP, phosphocreatine, or glycogen could be excluded as possible mechanisms for the beneficial effect of glucose in this system.  Provision of glycolytic ATP from increased glycolytic flux may be one important factor.


[Writing a computer program for the radioisotope exploration of cerebral blood flow (author's transl)].

After a brief review of the steps that lead to the first results in the radioisotope exploration of cerebral blood flow, the authors describe their own method utilizing a gamma ray camera and a computer. They outline the processes by which the data are acquired and elaborated. Last, they illustrate their own computer program in flow-chart form.


Preliminary 3-year results of 12 versus 6 cycles of surgical adjuvant CMF in premenopausal breast cancer.

In hope of reducing the duration of adjuvant treatment in premenopausal patients with operable breast cancer and histologically positive axillary lymph nodes, a prospective controlled study was started in September 1975. A total of 160 patients were randomized to receive 12 cycles of adjuvant CMF, while 165 were allocated to receive 6 cycles. At 3 years from radical mastectomy, the relapse-free survival was 85.4% in the 12-cycle group compared to 82.6% for the 6-cycle subset (P = 0.29). In both treatment groups, the incidence of treatment failure was directly proportional to the number of involved axillary nodes and to the clinical tumor size. Drug-induced amenorrhea as well as estrogen receptor status failed to significantly affect the results obtained. Treatment failures were more often documented in distant sites, while only 4--5% relapse was observed in local-regional areas. Overall survival was also similar for both treatment arms (86.2% vs. 85.1%, P = 0.49). Toxicity was moderate and reversible and no drug-induced neoplasms were so far observed. Present results achieved with 6 cycles of adjuvant CMF appear encouraging. However, they are still too preliminary to recommend the routine use of 6 cycles instead of the classical 12 cycles.


Use of VM-26 as a single agent in the treatment of renal carcinoma.

VM-26 was administered to 13 patients with metastatic adenocarcinoma of the kidney. Twelve of the 13 patients were assessable. The remaining patient was nonevaluable due to early death as defined in the protocol. No responses were observed in any of the 12 patients. Nonhematological toxicity was mild except for one instance of life-threatening hypotension, occurring during the infusion of VM-26. Significant leukopenia (less than 3000/mm3) occurred in 29% of courses and significant thrombocytopenia (less than 100,000/mm3) occurred in 14% of courses. Further studies using VM-26 in renal cell carcinoma do not seem warranted.


Nitrosoureas: a reappraisal of clinical trials.

The nitrosoureas (BCNU, CCNU, methyl CCNU) represent a new class of antineoplastic agents with a broad spectrum of antitumor activity. They are cell-cycle nonspecific cytotoxic agents. Postulated modes of action and pharmacology of these nitrosoureas are reviewed. Their therapeutic effectiveness as single agents and in combinations have been recognized in malignant lymphomas, multiple myeloma, melanoma, glioblastoma multiforme, gastric and colorectal carcionma, and small-cell carcinoma of the lung. The nitrosoureas are administered on an intermittent 6--8-week schedule because of delayed and frequently severe bone marrow toxicity which may be cumulative in nature.


Adriamycin, mitomycin C, and 5-fluorouracil in combination for advanced colorectal adenocarcinoma previously treated with 5-fluorouracil.

Twenty-one patients with progressing colorectal carcinoma previously treated with 5-FU were given a combination of 5-FU, adriamycin, and mitomycin C in an attempt to produce an objective response. No response was observed; 24% of patients had an arrest of their disease for a median of 18 weeks. 5-FU, adriamycin, and mitomycin C in combination failed to produce a response in patients with colorectal carcinoma previously treated with 5-FU.


cis-Platinum combination chemotherapy of bladder cancer: an update.

The combination of cis-platinum (DDP), adriamycin, and 5-fluorouracil was evaluated in 44 patients with advanced urothelial cancer, 39 of whom were evaluable for response. There were 18 partial remissions (46.2%) and no complete responses. Remissions were clinically meaningful, but of short duration. Four patients had bulky pelvic disease that was made resectable by chemotherapy, but none of these patients remained disease-free. These results are not superior to that expected from DDP as a single agent. In addition, there appears to be little or no survival benefit associated with chemotherapy. DDP represents an advance in the treatment of bladder cancer, but additional active agents and innovative approaches are needed.


"CAMP" combination chemotherapy for unresectable non-oat cell bronchogenic carcinoma.

Experience with camp chemotherapy of 84 unresectable non-oat cell lung cancer patients is described. Fourteen (27%) of 51 patients with measurable lesions responded (3 CR, 11 PR) for a median response duration of 5 months with a range of 1 to 19+ months. Median survival of responders was 370 days, but response was not independent of performance status. Survival of patients without measurable lesions was less favorable than that of even nonresponders, suggesting prediction of response to CAMP by site of metastasis. Preliminary results of a randomized prospective trial of the CAMP combination versus CCNU as a single agent for patients with good performance status fail to show a difference in survival.


Treatment of advanced breast cancer with cyclophosphamide, 5-fluorouracil, and prednisone with and without methanol-extracted residue of BCG.

The value of immunotherapy as an adjuvant to chemotherapy for advanced breast cancer is an unsettled question. To clarify this issue, 71 women with measurable or evaluable metastatic breast cancer were randomized to receive cyclophosphamide, 5-fluorouracil, and prednisone (CFP) with or without methanol-extracted residue of Bacillus Calmette-Guerin (MER). The total regression rates were 52% (CFP) and 39% (CFP + MER), including complete regression rates of 13% (CFP) and 65% (CFP + MER). The median duration of regressions for CFP-treated patients was 257-261 days and for CFP + MER-treated patients was 385 days. The median time to progression was 248-261 days in the CFP group and 159 days in the CFP-MER group. Projected median survival for both treatment groups is 20 months. Immunotherapy (MER) as used in this study does not appear to augment regression rates or vurvival for patients with advanced breast cancer receiving CFP.


Whole abdominal irradiation for carcinoma of the ovary.

A selected group of 39 patients with ovarian carcinoma were treated by whole abdominal radiation therapy with a pelvic boost. No statement as to the efficacy of this treatment program can be made for patients with Stage I, II, and IV disease because of the small number of patients with those stages who were treated by this method. A surprising 30% 5-year survival was obtained, however, in patients with Stage III disease. The challenge of ovarian carcinoma lies in the advanced stage of disease in most patients at the time of diagnosis.


Detection of antigens and IgM antibodies for rapid diagnosis of viral infections: a WHO Memorandum.

This Memorandum describes recent progress in the development of simplified and rapid laboratory technology and reviews currently available methods for the direct detection of viral antigens in clinical material and/or quantification of specific IgM antibody in acute serum specimens. Particular emphasis is given to such infections as viral hepatitis, rotavirus gastroenteritis, viral infections of the skin, genital, and respiratory tracts, and rabies. In addition, the standardization and quality control of reagents are discussed and recommendations are made.


[The reaction of hyperbasophilic mononucleated cells during P. falciparum malaria; its role in the immune response].

In 34 patients with a primary attack of P. falciparum malaria, hyperbasophilic mononucleated cells (atypical lymphocytes) were found 22 times. These cells appeared around the 3rd or 4th day of fever and represented 4 to 10% of all white blood cells in 14 cases, 10 to 15% in 6 cases, and 15 to 21% in 2 cases. Their transitory appearance seems to reflect the involvement of cell-mediated immunity, and more specifically of K cell-related cytotoxicity.


Prednimustine in adult acute myeloid leukaemia.

In 23 patients with acute myeloid leukaemia (AML) and over the age of 64, four remissions (17%) were obtained with Prednimustine as a single drug. The daily dose was 24--60 mg orally. In 14 patients aged between 35 and 64 years who were treated with Prednimustine 60--80 mg daily and vincristine 2.25 mg i.v. every 7--10 days, six remissions were obtained (43%). Upon remission, patients were given ,0--40 mg of Prednimustine daily as maintenance therapy. Drug-induced pancytopenia preceding remission was not recorded in any patient. There were no side effects of major importance during maintenance therapy, and the median duration of remission was 8 months. It is concluded that the low toxicity of Prednimustine in normal bone marrow cells is of value, especially in elderly patients with AML.


Retinoids and cancer.

The early and recent investigations in the field of retinoids and cancer are reviewed. The retinoids, including natural vitamin A compounds and their synthetic analogs, present a new class of substances exerting a prophylactic and a therapeutic effect both in certain experimental tumor models and in certain clinical conditions of preneoplastic and neoplastic lesions. Because of a particular physiological mechanism of action, the retinoids offer a new approach to the cancer problem, which is different from those of surgery, X-ray therapy, conventional chemotherapy, and immunotherapy.


Resistance of essential fatty acid-deficient rats to endotoxic shock.

The role of lipids in the altered energy metabolism of shock remains to be delineated fully. During the course of our studies of endotoxic shock, the susceptibility of essential fatty acid (EFA)-deficient rats to endotoxin was evaluated. Intravenous administration of S. Salmonella enteritidis endotoxin (1 mg/100 gm) in normal male Long-Evans rats (7--8 weeks old) produced severe shock with an 88% mortality. In marked contrast, injection of this dose of endotoxin in EFA-deficient rats of the same age resulted in only an 18% mortality. The deficient state afforded significant protection to even supralethal doses of endotoxin (2 mg/100 gm). Evaluation of reticuloendothelial (RE) phagocytic activity with colloidal carbon did not reveal significant differences in RE clearance rates. Within five hours after induction of shock, however, plasma acid hydrolase activity of shocked control rats was approximately double that of the EFA-deficient group. Likewise, the endotoxin induced hypoglycemic response was milder in the EFA-deficient rats. The lower plasma glutamic-oxaloacetic transaminase activity and glutamic-pyruvic transaminase activity of the EFA-deficient group also indicated a maintenance of hepatic integrity. These observations suggest that essential fatty acids of their products (ie, prostaglandins) contribute to the pathogenesis of endotoxic shock.


Cardiovascular manifestations of acute antibiotic toxicity during E coli endotoxin shock in anesthetized dogs.

Surgically instrumented, pentobarbital-anesthetized dogs were used to examine the acute cardiovascular activities of gentamicin, tobramycin, sodium penicillin-G, and sodium cephalothin during a control state and during experimental circulatory shock induced by E coli endotoxin. Intravenous administration of 2.5, 5, 10, and 20 mg/kg gentamicin or tobramycin resulted in a transient (5--20-minute) state of cardiovascular depression, as reflected by dose-related decreases of systemic blood pressure, cardiac output, left ventricular pressure, dP/dt max, left ventricular contractile force, and dF/dt max; heart rate was affected little. Endotoxin produced a persistent state of circulatory depression characterized by hypotension, decreased cardiac output, arterial acidemia, and reduced indices of cardiac function. During endotoxin shock, the cardiovascular effects of gentamicin and tobramycin were relatively more pronounced (sometimes more than doubled) than effects observed during the control state. Equally large doses of penicillin or cephalothin, however, had no discernible circulatory effects in either control dogs or dogs subjected to endotoxin shock. Present data indicate that the cardiovascular toxicities of the aminoglycoside antibiotics gentamicin and tobramycin were augmented during experimental circulatory shock, and suggest the need for specific hemodynamic surveillance when intravenous administration of cardioactive antibiotics is required in patients with pre-existing circulatory dysfunction.


An 'artificial beta cell' for control of diabetes mellitus: effect on plasma glucagon levels.

We have investigated the use of a glucose-controlled insulin infusion system, or artificial beta cell. On the feedback day, mean plasma glucose was significantly effect of improved control on plasma glucagon levels. Five insulin-requiring diabetic subjects in stable control were hospitalized for two 24 h periods. During one, they were given their usual dose(s) of subcutaneous insulin. In the other, the 'feedback' day, insulin administration was under feedback control by the artificial beta cell. One the feedback day, mean plasma glucose was significantly-lower in all subjects. Variability in plasma glucose throughout the day was also significantly less on the feedback day. All five subjects showed a significant fall in serum immunoreactive glucagon levels on the feedback day, suggesting that the glucagon abnormalities of diabetes may be secondary to the insulin deficiency, rather than a second primary defect of diabetes.


In vivo anti-nuclear antibodies in epithelial biopsies in SLE and other connective tissue diseases.

In vivo anti-nuclear antibody (ANA) was observed by direct immunofluorescence microscopy in epithelial cell nuclei in forty-four biopsies from thirty-three patients. The tissue containing the ANA was macroscopically normal in twenty-seven patients. The thirty-three patients with in vivo biopsy ANA included twenty-three with SLE, three with mixed connective tissue disease, two each with multi-system Sjögren's syndrome, dermatomyositis, and progressive systemic sclerosis, and one with rheumatoid arthritis. Features of sicca syndrome were noted in seventeen patients. The patterns of the in vivo biopsy ANA in the thirty-three patients were speckled (21), homogeneous (6), nodular (2), and both speckled and homogeneous (4). Complement was not detected in the epithelial cell nuclei. Immunoglobulin(s) and/or complement were deposited along the dermoepidermal junction in thirty-two of the forty-four biopsies, and in dermal blood vessels in twenty-two biopsies. Each patient had serum ANA against rat liver substrate; twenty-seven had high titre ANA (1 in 1000 or greater). Elevated levels of DNA-binding were found in twenty patients (61%), but the level of DNA-binding did not correlate with the intensity of in vitro biopsy ANA staining. Serum antibody to ribonucleoprotein (RNP) was present in eight of the twenty-three patients tested (35%), all eight patients having clinical features of sicca syndrome. Hypocomplementaemia was found in thirteen patients (40%), all of whom had active SLE. In vivo biopsy ANA appears to be a real phenomenon of unknown aetiology, and not an artifact, which is found in some patients with active multisystem autoimmune disease, especially SLE.


Local immune response to Mycobacterium lepraemurium in C3H and C57Bl/6 mice.

Subcutaneous footpad inoculation of living M. lepraemurium (L.MLM) induced, in high responder C57Bl/6 mice, a local granulomatous reaction associated with the production of effector cells which stopped the multiplication of bacilli in the draining popliteal node with the concurrent development of 24--48 hr delayed type hypersensitivity (DTH). The thymus-dependent local reaction did not occur after the injection of heat-killed M. lepraemurium (HK.MLM) or after the inoculation of L.MLM in nude mice. However, HK.MLM injection interfered with the onset of the local reaction and enhanced acid-fast bacteria (AFB) counts in the draining node. In low responder C3H mice, L.MLM produced a local and delayed footpad swelling but no restriction of bacilli multiplication in the draining lymph node was observed. This unresponsiveness was not due to an overloading of the inoculum dose since doses ranging from 3 x 10(4) to 3 x 10(7) MLM did not produce any granulomatous local reaction as in C57Bl/6 mice. The injection of dead bacilli in the contralateral footpad of subcutaneously (s.c.) infected C3H mice revealed Arthus-like and 18--24 hr delayed reactions. When 10(6) L.MLM per mouse were injected intravenously (i.v.), systemic infection, measured in the spleen, was found to be less restricted in C57Bl/6 than in C3H mice. Moreover, in C57Bl/6 mice low doses of L.MLM injected i.v. delayed the local reaction at first, then enhanced footpad swelling and AFB counts in the draining nodes, indicating some acquired defect of peripheral immunity. When a high dose of L.MLM (2 x 10(8)/mouse) was injected i.v., C57Bl/6 mice died sooner than C3H mice, indicating certain discrepancies between local resistance and systemic susceptibility.


Hyperbaric oxygen and chronic osteomyelitis.

Forty patients with chronic osteomyelitis were treated with hyperbaric oxygen as an adjunct to surgical therapy and antibiotics and followed for an average of 2 years with a recurrence rate of 15%. The mechanism of action of hyperbaric oxygen in osteomyelitis is probably an indirect one of improving local vascularity and potentiating phagocytosis. Many of these patients represent a refractory group with poor prognosis due to the etiology of the infection, site of involvement, and duration of infection prior to treatment. The recurrence rate following this mode of treatment seems to be primarily related to inadequate surgical management. There was no definite correlation between the site of the infection of pathologic organism and recurrence. Although this is a preliminary report, the results are encouraging. Hyperbaric oxygen may be indicated as an adjunct to good surgical and medical management, particularly in patients with refractory chronic osteomyelitis and in whom ablative surgery is under consideration as the only other means of controlling the infection.


Results of Harrington instrumentation in the treatment for severe scoliosis.

Between July 1963 and December 1974, we surgically treated 207 patients who had severe scoliosis (curves greater than 90 degrees), 196 of whom were eligible for inclusion in this study. No patient was treated preoperatively with a cast or traction. Upon statistical analysis, age and preoperative curve magnitude proved to be significant variables relative to the amount of surgical correction obtained and the maintenance of that correction; sex and etiology of the scoliosis were not shown to affect either surgical outcome or maintenance of correction. A comparative analysis showed our results to be as good as those reported by others using various preoperative regimens of casting or traction or both. Therefore, we find that the preoperative application of traction to patients with severe scoliosis yields no better correction than the use of Harrington instrumentation and fusion alone.


Leg lengthening in achondroplastic children.

Bilateral simultaneous elongation of the lower limbs was performed in 3 achondroplastic children. A total of 10 bones were elongated with Wagner's distraction apparatus, mainly by a modified Wagner technique. Maximal elongation of 34 to 44% of the original bone length resulted in relatively minor complications. Elongation of the lower limbs in achondroplastic children improves trunk-to-lower-limb proportions.


Infantile cortical hyperostosis with unusual clinical manifestations.

Infantile cortical hyperostosis may mimic a variety of serious conditions in early infancy, such as trauma, vitamin deficiencies, infection, or tumors, In a patient in good general condition there are typical roentgenological features often with involvement of the mandible that are diagnostic of the condition. In 3 cases of infantile cortical hyperostosis (Caffey's disease), one had the clinical picture of Erb's palsy and subsequently developed hyperostosis of the scapula. The second, had a "pulled elbow" and radial nerve palsy and later developed hyperostotic changes in the humerus and radius on the same side. The third was referred for treatment of congenital torticollis and found to develop hyperostosis of the clavicle on the same side. The recognition of this condition obviates unnecessary prolonged investigation and treatment.


An analysis of topic areas and topic trends in the Community Mental Health Journal from 1965 through 1977.

A content analysis of all (N = 604) published articles in the Community Mental Health Journal from 1965 through 1977 was performed (a) to provide a cross-sectional view of the community mental health field, and (b) to identify topic trends over time. Sixty-one content categories were used to classify topic areas. Results revealed major and growing emphases on organizing for the delivery and measuring of the effects of mental health service systems. Topics found to be increasing in relative frequency over time included training, measurement and research methodology, and intervention; those decreasing included planning and needs assessment, therapy, and suicide. Topic areas were compared with key characteristics of the field. Topic trends were interpreted in terms of open systems theory.


The nature of needs assessment in community mental health.

Confusion in defining needs assessment is discussed. A tripartite model of needs assessment is proposed: an identification of a problem; a statement about priorities; and a chosen solution. The parts can be used independently or linked together to plan new or altered services. Various needs assessment techniques are examined for their usefulness in each area. Two examples of needs assessment processes are given.


Paradoxes of psychotherapy. In honor and memory of Ludwig Binswanger.

Existential analysis has made us face the paradoxes, if not antinomies, in psychotherapy that we did not seem to be aware of. Existential philosophy is a highly elaborate, technical and linguisticly difficult field. In psychiatry, existential analysis of turned out to be just a set of hallowed commonplaces. As philosophy, its intent is the discovery of the grounds for our existence and in this way it seems to approach theology. In some instances it may turn into atheistic arrogance. With the stern zeal of philosophy and the pessimistic theology of existentialism, one hardly knows where to begin with psychotherapy. Often it fascinates the mediators, the writers who seem to offer us patterns helpful in our daily work. The effort, to take a radical stand to try to rewrite psychiatry, seems to be justified. And, if existential analysis hardly succeeds in bringing back into 'existence' (Dasein) some of our disturbed patients, it has aroused a review of our stale and abused terms and concepts. Psychiatry cannot exist, disciplined and sensible, without the enlightenment which philosophy offers. Some new departures could be attempted again. But if philosophy is not a way of talking and teaching but a way of living, is the psys goal. If he failed, his work remains a lighthouse in our time, warning us on our voyage.


Self-portraits and selected indices of psychopathology of a group of heroin-addicted patients.

This paper describes a self-portrait scoring system and compares the self-portraits of two groups of adults. The scoring system effectively differentiated between the two groups of subjects which were known to differ on several observable behavioral characteristics. It correlated well with the ratings obtained from counselors but not with a self-report inventory of anxiety. A self-portrait is a single manifestation of an individual's self-concept. It presents only a fraction of the hypothetical universe of one's behavioral manifestations. In addition, every single observation is affected by the place, time, and the presence of the observer. However, if these isolated observations can be studied systematically, they can contribute to a reliable and a valid index of the self-concept. This paper presents one means of achieving that end.


Enrichment methods for recovery of Yersinia enterocolitica from foods and raw milk.

Modified Rappaport broth used directly and in a two-step procedure with preliminary cold enrichment was evaluated for recovery of Yersinia enterocolitica from laboratory-inoculated samples of ground meat, and naturally-occurring strains from raw milk. The use of modified Rappaport broth generally improved recovery from both inoculated meats and raw milk, but results were not consistent between serotypes used for inoculation and between meat samples. The overall isolation rate for naturally-occurring strains of Y. enterocolitica in producer samples of raw milk was 18.3%. The majority of 98 isolates from milk were indole-positive (98%), lecithinase-positive (87%), hydrolyzed esculin (98%), and fermented salicin (100%) and rhamnose (53%). Thirty-two isolates (33%) were serotypable, representing eleven different serotypes with 0:4 occurring most frequently followed by 0:5 and 0:6,30.


Testing for the recovery of Yersinia enterocolitica in foods and their ability to invade HeLa cells.

Tests have demonstrated that current enrichment methods may fail to detect low levels of some clinical strains of Yersinia enterocolitica inoculated into meats and oysters and need improvement. The results of cold enrichment with phosphate and tripolyphosphate buffers were very poor and the sensitive clinical strain failed to grow in Wauters' MgCl2 broth. Twenty typical clinical strains of Y. enterocolitica belonging to Nilehn's biotypes 2, 3, and 4 had the ability to invade HeLa cells tissue culture but none of the 52 atypical clinical and food strains belonging to Nilehn's biotype 1 or rhamnose-positive were invasive. Although temperature sensitive, this test is reproducible and useful for screening noninvasive isolates.


Human gastrointestinal infections by Yersinia enterocolitica.

In an average-sized peripheral hospital in Belgium, Y. enterocolitica has been isolated from 1.1% of stool cultures, using an unmodified routine technique. In a group of unselected patients undergoing appendicectomy, 1.3% of patients had a positive culture, either from the mesenteric lymph nodes or the appendix, or from both sources. In the same group of patients, 1.7% of cultures grew a Salmonella. In more than half of the patients, the diagnosis of mesenteric adenitis was made during the surgical intervention. The clinical picture of Yersinia infection was greatly influenced by the age of the patients. Bacteriological (serotypes, antibiotic sensitivity, associated enteric pathogens), epidemiological (sex distribution, seasonal influence and secondary infection among family contacts) and clinical data confirm previous experience with human yersiniosis in this country.


Zigzag incisions for improved exposure and scarring.

Incisions for elective orthopedic procedures frequently result in scars that are unesthetic (wide and hypertrophied) and impeded function (are inelastic). The best cicatrix is obtained when the operation can be performed through an incision that follows the lines of skin tension (transverse exposure). If longitudinal exposure is required to gain access to the deeper structures, the best esthetic and functional result is obtained by the zigzag incision. Moreover, the zigzag incision affords a wider exposure than a long straight incision, thus greatly facilitating the operative procedure.


Local antifertility effect of luteinizing hormone-releasing hormone (LRH).

Injection of luteinizing hormone-releasing hormone (LRH) into the right uterine horn of the rat on D5 produced a dose related inhibition of pregnancy in the treated horn. The number of viable fetuses as well as the uterine weight was reduced while the number of placental sites was increased. The contralateral uninjected horn was unaffected. These data demonstrate that LRH can inhibit pregnancy in the rat through an extrapituitary effect.


The origin of the osteoclast.

The origin of the osteoclast has long been controversial. Various approaches such as traditional light microscopy, electron microscopy, microcinephotography, autoradiography, parabiosis, quail-chick nuclear marker, etc, have been used in attempts to resolve the problem. There are 2 major concepts: the view of skeletal (local) origin vs, that of extraskeletal origin (from blood-borne cells), and recently the latter view appears to have prevailed. Electron micrographic studies revealed that osteocytes and preosteoblasts can merge with pre-existing osteoclasts. Several studies by autoradiography and by quail-chick nuclear marker strongly supported the concept of extraskeletal origin. Although an alternative interpretation of the experimental results of Crelin and Koch supports the concept of skeletal origin, the majority of various experimental results by other authors would be in accordance with the following supposition: the osteoclast originates from a hematopoietic cell line (mononuclear phagocyte system) of the bone marrow which is essentially of extrinsic origin, but in the process of increasing the number of nuclei of the osteoclast, osteoblastic cells as well as osteoclastic cells can merge with the preosteoclasts or osteoclast. However, several problems regarding the origin of the osteoclast including the relation between the osteoclast and other giant cells, still remain unsolved.


The pathogenesis of Yersinia enterocolitica gastroenteritis.

Of nine Y. enterocolitica serotypes examined by us, only type 3 was shown to be enteropathogenic. 3 of 16 strains recovered from clinical material, produced a heat-stable toxin similar to that reported in E. coli. A fourth isolate produced histological changes resembling those associated with salmonellae. The mechanism whereby the majority of Y. enterocolitica strains produce diarrhoea is uncertain. Our isolates may have lost a plasmid essential for enteropathogenicity, or they may resemble the 'classical' enteropathogenic strains of E. coli for which no satisfactory experimental model exists. Further studies on freshly isolated strains are in progress in order to determine the prevalence of enterotoxigenic Y. enterocolitica.


Detection of enterotoxigenic and invasive strains of Yersinia enterocolitica.

Membrane filtrates and heavy cell suspensions of 11 strains of Yersinia enterocolitica isolated from a recent epidemic were examined respectively for enterotoxin activity and tissue invasiveness. Although no heat-labile enterotoxins were detected, some strains were found to possess heat-stable enterotoxins and to have the ability to penetrate the conjunctival epithelia of guinea pig eyes. These activites were detected only in cultures grown at 25 degrees C and not at 36 degrees C.


Biochemical, pathogenicity and toxicity studies of type III strains of Yersinia pseudotuberculosis isolated from the cecal contents of pigs.

Recent investigations have shown that the pig is a symptomless intestinal carrier of Y. pseudotuberculosis, the great majority of the strains isolated belonging to type III. Although these type III strains have a world-wide distribution there is little evidence to suggest that they are transferred readily to other animals and man. For that reason we examined the biochemical, pathogenic, immunogenic and exotoxin-producing properties of 24 type III strains isolated from the cecal contents of apparently healthy pigs. For comparison, we also examined type III strains from other sources, type I strains from normal pigs and isolates belonging to the other serotypes of Y. pseudotuberculosis. Biochemical reactions of type III strains were in accordance with data in the literature except for maltose and melibiose. Type III porcine strains were uniformly avirulent and atoxic but possessed a considerable degree of immunogenicity. On the other hand, type III strains isolated from aborted lambs and calves, although atoxic, were virulent for the guinea pig, as were also type I strains from healthy pigs. The pathogenic mechanism of Y.pseudotuberculosis, which is not unlike that found in E. coli, is discussed.


The molecular taxonomy of Yersinia enterocolitica.

The stretches of Yersinia enterocolitica strain Ye 62 DNA which show base sequence homology with Escherichia coli strain B DNA were removed from a preparation of tritiated Y. enterocolitica DNA by DNA reassociation techniques. This DNA preparation was then used in reassociation reactions with DNA from various selected strains of Yersinia and coliform bacteria. The results suggest that the bulk of the genes, which are in common between Y. enterocolitica and E. coli, are different from those shared by Y. enterocolitica and other species of Yersinia.


Structure and function of plasmid ColE1 and related plasmids.

Analysis of plasmid ColE1, its naturally occurring relatives ColK and CloDF13, and a wide range of ColE1 derivatives containing either insertions or deletions of genetic material has allowed localization on the ColE1 genome of DNA sequences responsible for colicin E1 synthesis, immunity to colicin killing, conjugal mobility and incompatibility. We have examined incompatibility between pairs of ColE1 derivatives ranging in size from 2.6 to 13.8 Md. Though all the plasmids tested exerted ColE1 incompatibility, a definite pattern was observed regarding the dominant plasmid in any pair tested (i.e. the plasmid that displaces the other from a heterozygote). Usually the larger plasmid is displaced. We conclude that loci for incompatibility reside within 0.7 kb of the ColE1 replication region. A model is presented to explain both the incompatibility data and the observation that the fraction of total DNA occurring as ColE1-like plasmid in a cell is approximately constant. Transposons Tn1 and Tn3 (3.2 Md; Apr and approximately 85% homologous), Tn501 (5.5 Md; Hgr), and Tn7 (9.3 Md; Tpr Smr) can all be transposed into ColE1. Though all have closely related. Tn501 and Tn7 do not complement transposition of Tn3 transposition defective deletions. A Tn3-specified 19,000 dalton protein is absent in one particular class of transposition-defective deletion.


Replication and gene functions of the bacteriocinogenic plasmid CloDF13.

The replication and genetic constitution of plasmid CloDF13 was studied using mutants of CloDF13 obtained by NTG mutagenesis, insertion of the ampicillin transposon Tn901, or deletion of particular CloDF13 DNA regions. Analysis of the polypeptides encoded by these mutant plasmids enabled us to locate six genes on the CloDF13 physical map. These genes cover about 60% of the coding capacity of CloDF13. A large part of the CloDF13 genome (about 30%) is involved in the conjugal transfer of this plasmid. This transfer region codes for at least two polypeptides, polypeptide B (61,000 daltons) and C (24,000 daltons). Those CloDF13 DNA regions responsible for the synthesis of the cloacin protein and immunity protein were also mapped on the plasmid genome. In addition we were able, using a copy mutant of CloDF13, CloDF13-cop3, to locate those DNA sequences involved in interaction with male-specific RNA phages and bacteriophage P1. For replication of CloDF13, two regions are essential. One region, from 43% to 64%, affects the stability of CloDF13-cop3 plasmids. In the case of the CloDF13-cop3 mutant, deletion of this region results in the generation of multimeric plasmid molecules accompanied by an impaired segregation of plasmid DNA molecules to daughter cells. The second region, from about 1.8% to 11.5%, contains an origin of replication as well as well as DNA sequences involved in the control of CloDF13 replication. The replication of CloDF13 starts at about 3% on the CloDF13 physical map and proceeds unidirectionally counter-clockwise.


Deletions, rearrangements and tandem duplications of recombinant plasmids containing yeast ribosomal DNA.

Recombinant DNA plasmids, formed by the insertion of yeast ribosomal DNA into Escherichia coli plasmids pSC101 or pMB9, underwent deletions, rearrangements and tandem duplications. Independently derived deletion products of plasmids, constructed using pMB9 as a vector, are indistinguishable from each other. These deletion products from multimers composed of tandem repeats. In at least one case a plasmid, constructed by inserting an SmaI fragment of yeast ribosomal DNA into pSC101, underwent deletion and rearrangement to form a product in which a segment, consisting of part of the pSC101 sequence and part of the yeast ribosomal DNA sequence, was duplicated to form a tandem repeat. Deletion and rearrangement take place in Rec+, recA- and recB- recC- cells. The rate of deletion in Rec+ cells is higher than in recA- cells. The rate of deletion in minicell-producing, X-ray resistant strains is much higher than in other Rec+ strains.


The persistence of R-plasmid-carrying E. coli in a married couple, one of whom was receiving antibiotics.

The aerobic Gram-negative intestinal flora of two individuals, husband and wife, has been followed for about 20 months. The wife was receiving prolonged tetracycline treatment for acne during the first year of the study and was found to carry a large proportion of tetracycline resistant E. coli in her faecal flora even after the tetracycline treatment had ended. A brief therapeutic course of ampicillin during the period when no tetracycline was being taken resulted in the temporary disappearance of the tetracycline resistant flora, but this returned, even though no tetracycline was being taken, as soon as the ampicillin ended. The husband took no antibiotics during the period under study but was frequently found to excrete the same resistant E. coli as his wife. Moreover, the plasmids carried by the tetracycline resistant strains in the wife and the husband were often indistinguishable. This suggests that R-plasmids may spread from people under treatment to close relatives who have not been treated.


Resistance transfer factor in E. coli strains isolated from human and water sources.

Conjugation experiments with a recipient E. coli strain F-lac-NaR were performed in order to determine the extent to which transferable drug resistance is prevalent among Gram-negative bacteria isolated from the following sources: healthy infants both treated and untreated with antibiotic drugs; healthy adults previously exposed to antibiotics; water sources; and nosocomial infections. Transferable drug resistance was demonstrated in 56.6% of E. coli isolates from the antibiotic-treated infant group, in 53.2% of isolates from untreated infants, and in 43% of isolates from the adult group. R factor was carried by only 8.3% of the strains isolated from water sources, and by 26.7% of the nosocomial strains. A positive correlation between transferable antibiotic drug resistance and resistance to mercury ions was found in donor and conjugate strains. Acridine and several of its derivatives were found to be active in preventing transfer of R factor.


Recombination between the plasmid prophages P1 and P7 and the E. coli chromosome.

The prophages of the related temperate phages P1 and P7, which normally exist as plasmid DNA, suppress E. coli dnaA(Ts) by integrating into the host chromosome. Integratively suppressed strains may either be capable of producing phage or may have prophage deletions. In strains containing non-defective prophages, the location of the site on the prophage used for integrative recombination was identified by use of restriction analysis and DNA-DNA hybridization techniques. At least seven different integration sites were found on the prophage; the site used most often may be at the 'end' of the genetic map generated by vegetative phage crosses. For suppression of P1 and P7, the sites on the host chromosome utilized for prophage integration are not distributed randomly.


Correlations of the fibrinogen level with the blood pressure and with the plasma renin activity in hypertensive patients. Preliminary communication.

The blood pressure, the plasma renin activity and the fibrinogen level were examined in 44 hypertensive patients. It was found that the fibrinogen level was significantly elevated compared to the control group, but even so remained below the upper limit of the normal value. The increase was even more pronounced in hypertension associated with renal parenchymatous disease, and most marked in malignant hypertension. A significant linear correlation was found between plasma renin activity and the fibrinogen level, between the systolic pressure and the fibrinogen level, and between the diastolic pressure and the fibrinogen level. Questions connected with these correlations and relating to the pathogenesis are discussed.


Serum levels of IgM, IgD, IgA and IgG before and during rejection of renal allotransplants.

In recipients of renal allotransplants the relationship of IgM, IgA and IgG serum levels to 26 rejections was investigated, as well as the relationship of IgD and 22 rejection. For IgM and IgG a significant decline was found in the whole group (p less than 0.05) and in the sub-group of rejections after a period longer than one month following transplantation (p less than 0.05 in IgM and p less than 0.02 in IgD). As compared with values before rejection, a decline of IgM occurred in 11 rejections which amounted to more than 20% and a decline of IgD in 10 rejections by more than 50%. Changes of the serum levels of IgA and IgG were not statistically significant (p greater than 0.05). In recipients of renal allotransplants serum immunoglobulin (Ig) levels of the main classes were investigated for two reasons: (1) as part of a check-up of the condition of the graft and screening of rejection and (2) to detect possible deficiencies of humoral immunity with increased liability to infections. As regards monitoring of the risk of the rejection crisis, hitherto assembled experience did not provide an unequivocal answer in which class of Ig the greatest change may be expected, whether a rise or fall, and it is not even clear whether investigations of serum Ig levels will be a positive contribution. There is only agreement on the point that IgA levels are not related to rejection; according to some authors this applies also to IgM and IgG (8, 11). Other authors (1, 15, 17) appreciate in conjunction with rejections the importance of high IgM and IgG levels, while other workers observed a significant decline of IgM (13, 16, 19). IgD levels were investigated in our previous work on the relationship of renal functions and serum Ig levels in recipients of renal allotransplants (7); we found no reports on the relationship between IgD and rejection.


Cellular sources of thymic hormone (thymosin) in human thymus.

Antiserum against partially purified calf thymosin (5) was used for the localisation of this complex material in human thymus for comparison with calf thymi. Antithymosine IgG produced with human thymosin 5 by comparative immunoelectrophoresis one line identical with one of the lines of calf thymosin 5. Immunofluorescent evidence suggests the presence of the material in question, which also contains a biologically active principle, mainly in the epithelial cells of the thymus cortex. The contents of the traced material, however, fluctuates substantially between individual thymic lobules.


NGF and anti-NGF: evidence against effects on fiber growth in locus coeruleus from cultures of perinatal CNS tissues.

The present study examines whether the developing noradrenergic neurons of locus coeruleus depend on endogenous nerve growth factor (NGF) for nerve fiber production and if exogenous NGF stimulates fiber growth in this nucleus, using a collagen gel tissue culture technique. Lucus coeruleus from perinatal rat brain was used in three culture experiments: (1) lucus coeruleus, parietal cerebral cortex, and the superior cervical ganglion, prepared from newborn rats and cultured in different sectors of the same dishes; (2) locus coeruleus and parietal cerebral cortex from 17-day-old rat fetuses cultured in the same manner, and (3) locus coeruleus from 17-day-old rat fetuses co-cultured with spinal, sympathetic and ciliary ganglia from 8-day chick embryos. Experiments 1 and 2 were run with and without addition of NGF and anti-NGF, experiment 3 with and without anti-NGF. Total fiber production in all cultured tissues was evaluated daily by dark field and phase contrast microscopy during 4 days. Adrenergic nerve fiber production was then studied in the same locus coeruleus and superior cervical ganglia  from the rats by Falck-Hillarp fluorescence histochemistry. Locus coeruleus and cortex cerebri from fetal rats produced dense fiber halos in culture. Locus coeruleus from newborn rats produced considerably less fibers, newborn cortex only few fibers. Superior cervical ganglia from the same newborn animals produced no or almost no fibers. Addition of NGF was not able to stimulate fiber growth in locus coeruleus nor in cortex cerebri as observed both in the living cultures and by fluorescence microscopy. Likewise, addition of anti-NGF did not affect fiber production in the CNS areas. The negative results with NGF on newborn locus coeruleus and cortex cerebri were in sharp contrast to the strong, highly significant fiber growth response demonstrated by the superior cervical ganglion from the same animals cultured in the same dishes. The third experiment tested whether locus coeruleus in tissue culture contained or produced nerve growth factor or any one of the three chick embryo ganglia. No response whatsoever in these three ganglia was observed. It is concluded that the developing locus coeruleus area does not contain or produce NGF, does not depend on NGF for fiber production, and is not stimulated by exogenous NGF.


Experience with a radioimmunoassay of luteinizing hormone-releasing hormone (LH-RH) in biological material of man.

A radioimmunoassay of LH-RH with a sensitivity of 7.8 pg/ml is described. Labelling and purification techniques, methods for extraction of LH-RH and separation techniques of bound and free labelled hormone are compared. Determination of LH-RH levels in serum after administration of synthetic LH-RH by different routes and measurements of endogenous LH-RH levels in serum of normal subjects and patients with different endocrine diseases as well as in cerebrospinal fluid of normal men are performed. The measurement of exogenously administered LH-RH in serum reflects the disappearance of synthetic LH-RH from peripheral circulation in dependence upon the kind of administration route. The level of endogenous LH-RH was found to be under the limit of the assay in all samples of cerebrospinal fluid and of serum of normal male subjects. The results obtained in the patient groups show that the radioimmunological estimation of endogenous LH-RH in peripheral body fluids does not reflect the hypophysiotrophic role of this hypothalamic peptide.


Reduced aldosterone and sodium excretion in endurance-trained athletes before and during immersion.

Aldosterone excretion (AE) and plasma renin activity (PRA) were measured in eight untrained (UT) and eight endurance-trained (TR) male subjects before and during 4 h head-out immersion to study the mechanism of reduced renal sodium excretion in athletes. AE was significantly lower before immersion, and decreased less during immersion, in TR than in UT. Fractional sodium excretion, too, was lower and increased less during immersion in TR than in UT. PRA decreased in the water bath in all subjects (p less than 0.001) with no significant difference between the groups. During immersion, plasma sodium concentration oscillated whereas potassium concentration showed a temporary rise in TR (p less than 0.001). The attenuated response of AE in TR may be due partly to this increase of plasma potassium concentration. The generally reduced aldosterone release in TR might be caused by a training induced adaptation of the adrenals to corticotropin. The lowered renal sodium excretion of TR in spite of the decreased AE suggests an intensified aldosterone effect in these subjects, diminishing the salt loss during exercise.


[Excretion of catecholamines in juvenile diabetes mellitus].

Twenty four hours' urinary excretion of catecholamines was examined in 30 children suffering from diabetes and in 30 healthy children. The results showed that in children suffering from diabetes, excretion of adrenaline was increased and that of noradrenaline decreased when compared with healthy children. Taking into consideration changes in the sympatho-adrenergic system activity in cases with different degrees of compensation of metabolic processes during insulinization, it should be stressed that in children with ketoacidosis, excretion of catecholamines was still different in comparison with control group. On the other hand in children with compensate diabetes after insulin therapy, that differences were little. Marked increase in free adrenaline excretion observed in children with ketoacidosis and its normalization by effective insulinization seems to support the diabetogenic and ketogenic role of this neurohormone. Decrease in free noradrenaline excretion in children suffering from juvenile diabetes appears to suggest that there is a diminished sensitivity of the sympathetic system resulting probably from specific autonomic neuropathy accompanying this disease.


Effects of insulin on hepatic lipid synthesis in alloxan diabetic rats.

In vitro hepatic synthesis of lipids starting from 1-(14)C-acetate was studied in rats made diabetic by subcutaneous alloxan administration (175 mg/kg b.w.). A second group of diabetic rats was treated with lente insulin. In the alloxan-treated rats, a decrese was observed in hepatic incorporation of 1-(14)C-acetate into phospholipids, triglycerides and esterified cholesterol; there was an increased incorporation into nonesterified fatty acids (NEFA) and free cholesterol. Insulin administration restored lipid synthesis values to normal. On histologic examination, an intranuclear glycogenesis was observed in the hepatocytes of the alloxan-treated rats, along with severe hepatic necrosis; the latter however, only in rats sacrified on the 3rd day. Hepatic steatosis with small, medium and large droplets was present in the insulin-treated rats; signs of cellular degeneration were less evident.


Xenogeneic bone marrow stimulating effect in vitro on antibody-producing cells.

It was described earlier that in the tissue cultures of immune mouse LN cells the number of antibody producing cells was increased 2-3-fold when syngeneic of allogeneic nonimmune BM cells were included in the cultures. In these experiments, parallel mixed cultures were set up of mouse immune LN cells with non-immune BM cells of either syngeneic or xenogeneic origin (rat, pig and chicken). Xenogeneic BM also increased the number of PFC in the mixed cultures. The intensity of the stimulation effected by xenogeneic BM of all three species tested was comparable to that induced by syngeneic BM. The number of PFC in all types of the mixed cultures was significantly higher (P less than 0.005) than in the LN cell cultures alone. The differences between the effect of syngeneic and xenogeneic BM and between the xenogeneic BM cells of different origin were not statistically significant (P less than 0.05).


Allyl alcohol-induced changes in the rat exocrine pancreas.

A histologic, histochemical and electron microscopic study was carried out on the rat exocrine pancreas 6-96 h after a single allyl alcohol intake. Histologically, acidophilia, necrosis and vacuolation of the pancreatic acinar cells were observed. Histochemically, a low but evident alcohol dehydrogenase activity could focally be demonstrated in the pancreatic acinar cell cytoplasm of the experimental and control rats. By electron microscopy, cytoplasmic lipid droplets, mitochondrial degeneration and necrosis were found in the acinar cells. Focal cytoplasmic degradation and vacuolation were seen in acinar, centroacinar and ductular cells of the pancreas. The findings are regarded as the consequence of the toxic action or acrolein that originates after allyl alcohol oxidation by alcohol dehydrogenase. Analogies between the liver and the pancreas in their response to acute allyl alcohol and chronic ethanol intake are discussed.


Age at diagnosis and seasonal variation in the onset of insulin-dependent diabetes in Chile (Southern hemisphere).

The age at diagnosis, sex, and the date of both the diagnosis (month--year) and the onset of symptoms of 525 insulin-depdendent diabetic patients, under 30 years of age at the time of diagnosis, were collected retrospectively from hospital records. The patients came from Chilean cities located in the Central Zone: Santiago (87%) and Temuco--Concepcion (13%). The age at diagnosis showed a bimodal distribution up to 19 years of age, with the main peak at 12 years and another at 6 years. The female peak preceded the male peak by 1 to 2 years. The onset of symptoms showed a seasonal variation, with the highest frequency in winter and the lowest in spring (P less than 0.01). There was also a significant variation in the time of diagnosis, especially in the 0--19 age group: most cases were diagnosed in winter (June, July) with the lowest number in summer (December).


Diabetes: the genetic connections.

Insulin dependent (IDD) and non-insulin dependent diabetes (NIDD) are separate disorders. Twin studies show that IDD cannot be entirely due to genetic causes as concordance is no more than about 50%, but there is some inherited predisposition to it as shown by HLA patterns. NIDD, on the other hand, is predominantly due to genetic causes since identical twins are nearly always concordant. Many cases of NIDD show chlorpropamide alcohol flushing (CPAF), a dominantly inherited feature which may precede the appearance of diabetes and thus act as a genetic marker for this type of diabetes. Diabetics who show chlorpropamide acohol flushing are less likely to develop retinopathy than those who do not. Genetic factors must therefore affect the incidence and severity of diabetic retinopathy. Chlorpropamide alcohol flushing is due to sensitivity to enkephalin. Enkephalin and other opioids affect carbohydrate metabolism and insulin release. It is possible therefore that they act as neurotransmitters and cause NIDD by a sympathetically mediated effect on the liver and pancreas--in other words, that as far as NIDD is concerned Claude Bernard's views on the cause of diabetes may have been right.


Identification of a lobe in the adult human pancreas rich in pancreatic polypeptide.

Systematic sampling of human necropsy pancreases has revealed that pancreatic polypeptide (PP) cells are not distributed equally in the gland. PP-cells are the most abundant cell type in the posterior part of the pancreatic head while they are scarce or absent in the remainder of the gland. The PP-rich part of the head can be separated by blunt dissection from the pancreas as a discrete lobe. This lobe probably originates from the ventral pancreatic bud during embryogenesis. A quantitative study of the immunofluorescent endocrine cell types (insulin, glucagon, somatostatin and pancreatic polypeptide cells) in PP-rich and PP-poor regions of pancreases in 8 subjects with ages ranging from 33 fetal weeks to 80 years, showed that the proportions of the cell types were different in youngs and adults.


Apparent discrepancy between the insulin secretory responses in vivo and in vitro in carbohydrate-intolerant Sand rats.

A subpopulation (n = 27) of normoglycaemic Sand rats was characterized as carbohydrate-intolerant by intraperitoneal glucose loading. Five of these animals did not show any rise in peripheral insulin concentrations when injected with glucose. However, when isolated by collagenase digestion their islets still exhibited a significant enhancement of insulin secretion in response to glucose, glyceraldehyde, mannose and theophylline. The in vitro secretory responses were comparable to those of islets from carbohydrate-tolerant Sand rats. The results underline the importance of the natural environment for the B-cell response in vivo.


Chemical diabetes in the adult rat as the spontaneous evolution of neonatal diabetes.

Injection of streptozotocin in newborn rats induced a severe diabetic syndrome on day 4 after birth, with acute hyperglycaemia and glycosuria. Over the next 3 weeks spontaneous recovery occurred as attested by normal basal blood glucose and plasma insulin levels. Recovery was, however, incomplete in the adult since a definite impairment in insulin release and glucose disposal was observed. This state was characterized by the following features: 1) a 72% decrease in pancreatic insulin stores without change in pancreatic glucagon stores; 2) a slight but consistent elevation of blood glucose in the fasted and fed basal states and especially of blood glucose 90 min after an IV glucose load (2 g/kg) performed under pentobarbitone anaesthesia; 3) a considerable decline in the glucose-induced insulin release with a decrease in the maximal response. Both early and late phases of insulin release were impaired, as indicated by in vivo glucose infusion experiments. Basal plasma glucagon levels were normal. Over a period of 12 months with a normal laboratory diet no aggravation of the chemical diabetic state was observed. This new experimental syndrome is a potentially interesting model for the study of the influence of environmental factors on the development of overt diabetes.


Relationship of binding to internatlization of 125I-insulin in isolated rat hepatocytes.

By quantitative electron microscopic autoradiographic technique, we have previously shown that 125I-insulin initially localizes to the plasma membrane of isolated rat hepatocytes and is subsequently internalized in a limited region of the peripheral cytoplasm. In the present study, we have shown that when cells are incubated at 20 degrees C, steady state binding is reached by 60 minutes and maintained up until 120 minutes of incubation while at 37 degrees C steady state binding is reached by 10 minutes and maintained for 30 minutes. Under both of these conditions, internalization of the labelled material occurs as a constant function of the binding. These data suggest that under normal conditions the binding of the ligand is an important rate limiting determinant of the internalization process.


Streptozotocin diabetes: a glucoreceptor dysfunction affecting D cells as well as B and A cells.

Somatostatin release from the isolated pancreas of 3 normal and 6 streptozotocin diabetic dogs has been measured in response to various stimuli to determine whether abnormalities in somatostatin release are present in the diabetic pancreas. Simultaneous measurement of glucagon secretion was also made. In the pancreas from normal dogs increases in perfusate glucose from 25 to 200 mg/100 ml induced a 2--3 fold increase in somatostatin release and a two thirds decrease in glucagon secretion. In contrast, in the diabetic pancreas glucose caused no change in the secretion of the two hormones. In the diabetic pancreas addition of insulin to the perfusate (25,000 microU/ml) for periods from 10 to 75 minutes aimed at restoring normal extracellular insulin levels in the islets failed to restore either somatostatin or glucagon secretion to normal. In contradistinction to the lack of effect of glucose, the somatostatin and glucagon responses to arginine (5 mmol/l), isoproterenol (2 ng/ml) and calcium (5 mmol/l) were normal in the diabetic pancreas. The data suggests the presence of a selective glucoreceptor abnormality of the D as well as of B and A cells in the streptozotocin diabetic dog.


Antitumor activity of cell-wall skeleton of Propionibacterium acnes C7 in mice and guinea pigs.

The antitumor activity of the cell-wall skeleton (CWS) of Propionibacterium acnes C7 was examined by using transplantable tumors in syngeneic mice and in guinea pigs, and autochthonous tumors in mice. P. acnes-CWS was shown to suppress the growth of fibrosarcomas, EL4 leukemia, and MH134 hepatoma in syngeneic mice, and to regress the established tumors of a fibrosarcoma (MC104) in C57BL/6J mice, and a hepatoma (line-10) in strain-2 guinea pigs. The oil-attached P. acnes-CWS mixed with fructose mycolate was effective for suppression of the autograft of fibrosarcoma in mice. The repeated intralesional injections of suspension of P. acnes-CWS in phosphate-buffered saline was effective for prolongation of survival period of mice bearing 3-methylcholanthrene-induced fibrosarcoma. The test results on the cell fractions of P. acnes indicated that the CWS, but not the cytoplasmic or glucan fraction, of P. acnes had anti-tumor activity. The activation of peritoneal macrophages of mice was observed when P. acnes-CWS, but not the cytoplasmic fraction, was injected intraperitoneally 4 days before. The relationship between the cytolytic activity of peritoneal macrophages and antitumor activities of P. acnes-CWS was also discussed.


Carcinogenicity and mutagenicity of 4-nitropyridine 1-oxide derivatives.

Carcinogenicity and mutagenicity of 4-nitropyridine 1-oxide (4-NPO) and 7 kinds of its alkyl derivatives were tested on mice and on Salmonella typhimurium strains and Escherichia coli strains. 3-Methyl-4-NPO is the most potent carcinogen, followed by 3-ethyl-4-NPO and then 4-NPO. Mutagenicity was most potent in 3-methyl, 2,3-dimethyl, and 2,5-dimethyl derivatives, moderate in 4-NPO and 2-methyl and 2,6-dimethyl derivatives, and to a least extent in 3,5-dimethyl-4-NPO. Structure-mutagenicity relationship was discussed on the basis of molecular mechanism of the carcinogenesis of 4-nitroquinoline 1-oxide. Quantitative relationship between mutagenicity and carcinogenicity was not strictly found among the compounds examined.


Realignment of the genetic map of the terminus of the rIIB cistron of bacteriophage T4.

Duplication end-point mapping in the rIIB cistron indicates that the order of the BS-B10b segments is the inverse of that presented in Benzer's (1961) genetic maps. This findings is supported by two- and three-factor crosses and the phenotypes of rII deletions extending into the D region.


Regional specificity of illegitimate recombination by the translocatable ampicillin-resistance element Tn1 in the genome of phage P22.

Insertions of the translocatable ampicillin-resistance element Tn1 were selected in the genome of the temperate Salmonella phage P22 by growing the phage on hosts carrying the resistance plasmid RP4. Insertions of Tn1 into phage P22 are rare (10(-10) per phage) and nonrandomly distributed in the P22 genome. They are found mainly in the vicinity of the P22 ant gene. Insertions within the ant gene are found at many (at least 15) genetically separable sites, are found equally frequently in both orientations and cause irreversible loss of gene function. Some insertions in ant appear to be associated with an adjecent deletion. Prophage deletions were derived from P22::Tn1 phages by two methods. Low multiplicity transductants have nonrandomly distributed endpoints. One end is at or very near the site of the Tn1 insertion, and the other is in the vicinity of gene 12; however, there are many genetically distinguishable endpoints within gene 12. Prophage deletions selected as survivors of induction of a P22Ap mnt-ts lysogen have similarly nonrandom endpoints, with the Tn1-distal end frequently near the ant gene, as well as gene 12. Physical analysis of several prophage deletions suggests that the Tn1 is intact to the resolution of DNA electron microscopy and that the deletions begin at the end of the Tn1 insertion. These results suggest that illegitimate recombination associated with Tn1 shows regional specificity (i.e., preference for some large areas of the P22 genome over other areas), but that within these regions is quite nonspecific.


Mating-type effect on cis mutations leading to constitutivity of ornithine transaminase in diploid cells of Saccharomyces cerevisiae.

Cis-acting regulatory mutations have been isolated that affect L-ornithine transaminase (OTAse), an enzyme catalyzing the second step of arginine breakdown in yeast. These mutations lead to constitutive synthesis of OTAse at various levels. Two different types of mutations have been recovered, both of which are tightly linked to the structural gene (cargB) for this enzyme. One type behaves as a classical operator-constitutive mutation similar to the cargB+O---1 mutation previously described (DUBOIS et al. 1978). The second type is peculiar in two respects: the higher level of constitutive OTAse synthesis and the expression of constitutivity in diploid cells. These mutations are designated cargB+Oh. They behave as usual operator-constitutive mutations in diploid strains homozygous for mating type (a/a or alpha/alpha), but the constitutivity is strongly reduced in a/alpha diploid cells.


Escherichia coli K-12 auxotrophs induced by insertion of the transposable element Tn5.

The sites of insertion of the transposable kanamycin-neomycin resistance-determining element, Tn5, in the E. coli K-12 chromosome were assessed in a collection of over 300 auxotrophs. Although mutations in at least 45 different cistrons were obtained, the distribution of insertion sites was not completely random: proA or proB; cysG; and cysH, cysD or cysC mutants were found in excess.