Nucleotide sequence of the murine leukaemia virus amphotropicstrain 4070A integrase (IN) coding region and comparativestructural analysis of the inferred polypeptide

Summary.  The complete nucleotide sequence of the integrase (IN) protein coding region of the murine leukaemia virus (MLV) amphotropic strain 4070A is presented. The sequence comprises 1,224 nucleotides, encoding a 408-residue polypeptide of M_r 46,312. Alignment of the inferred 4070A IN amino acid sequence with the IN proteins of other MLV showed that substitutions are confined largely to segments within the N- and C-terminal domains. In the N-terminal domain the majority of substitutions occur as contiguous 2- to 6-residue blocks, whereas in the C-terminal domain they occur as isolated entities except within a short segment characterized by deletions/insertions. Selection appears to act on the C-terminal 19 residues of IN rather than on the N-terminal residues of ENV (encoded by overlapping reading frames), suggesting a functional role for this segment. Phylogenetic analyses grouped the sequences into two clusters, one comprising IN from the amphotropic strain 4070A and three ecotropic MLV (CAS-BR-E, Moloney and Friend), the other consisting of IN from three ecotropic MLV (two radiation-induced viruses and AKV) and a mink cell focus-forming (MCF) MLV virus. The same dichotomy and cluster composition was obtained from analysis of the long terminal repeat (LTR) regions from these viruses (consistent with the functional interrelationship of IN and LTR) but not from analysis of envelope protein sequences (consistent with the functional independence of ENV proteins from both IN and LTR). Secondary structure predictions supported features determined from the catalytic domain of human immunodeficiency virus and avian sarcoma virus IN, and identified probable structures within the relatively long N- and C-terminal domains of MLV IN proteins. Received February 25, 1997 Accepted May 2, 1997     

Untersuchungen über das Verhalten der Glutamat-Oxalacetat-Transaminase (GOT) in Erythrocyten von gesunden Kindern und Kindern mit verschiedenen Anämieformen

Zusammenfassung Bei 102 hämatologisch gesunden Kindern und Erwachsenen wird in gewaschenen Erythrocyten die Aktivität der GOT bestimmt. Bei Bezug auf die Zellzahl (10⁹ Erythrocyten) ergibt sich dabei ein annähernd kontinuierlicher Aktivitätsabfall vom Neugeborenen- über das Säuglings- und Kindesalter zum Erwachsenenalter hin. Bei Bezug auf Hämoglobin- und Hämatokritwert zeigen sich davon zum Teil abweichende Befunde, was anscheinend mit der Hypochromie der roten Zellen in der zweiten Hälfte des 1. Lebensjahres zusammenhängt. Der Morbus haemolyticcus neonatorum weist ebenso wie die Neugeborenenhyperbilirubinämien anderer Ursache und die ätiologisch verschiedenartigen Anämien älterer Kinder eine Erhöhung der Erythrocyten-GOT auf. Diese ist als Ausdruck einer Verjüngung der Zellpopulation anzusehen. Der Vergleich der verschiedenen Altersgruppen sowie derjenige gesunder Neugeborener und Neugeborener mit Morbus haemolyticus neonatorum zeigt signifikante Unterschiede zwischen den mittleren GOT-Aktivitäten. Die Streubreite de Einzelwerte ist jedoch so groß, daß man im Einzelfall zu keiner klinisch verwertbaren Aussage kommt.

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Summary The activity of GOT was determined in washed erythrocytes in 102 cases of normal children and adults from the hematological point of view. Correlating the results to a constant number of cells (10⁹ erythrocytes) a contineous loss of activity was found with increasing age of the patient. There was a linear decrease of activity reaching from highest values in the newborn and medium levels in infants and children to low values in adults. Considering the hemoglobin and hematocrit the results were slightly different, which appears to be in connection with the hypochromic anemia in the second half of the first year of life. both the hemolytic disease and the hyperbilirubinemia of the newborn as well as the anemias of different etiology in older children show increased activity of the erythrocyte-GOT. This apparently is evidence for the fact that many more young cells are circulating in such patients than in normals. Comparing the results there are significant differences in the mean GOT-levels in the various age groups, in normal newborn and in newborn with hemolytic disease. The range of results, however, is so widespread that it is of no diagnostic value for individual clinical cases.

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Die Arbeit enthält die wichtigsten Ergebnisse der Dissertation von Herrn H. Ey, Freiburg i. Br.     

Untersuchungen über den Gasaustausch bei Narkosebronchoskopien mit Muskelrelaxantien

Ohne Zusammenfassung     

Effect of a High Fat Diet on Quantitative Features of Adipocytes in the Omentum: An Experimental, Stereological and Ultrastructural Study

Background: Omental adipose tissue specimens of female rats that were fed a high fat (HF) diet were evaluated stereologically and histopathologically. To our knowledge, there is no stereological study on numerical density, nuclear height and volume of adipocytes in omental adipose tissue in the female rat fed a HF diet in the literature. Method: 20 female Spraque Dawley rats were used in the study. 10 of the animals were fed HF diet consisting of 30% of calories from fat for 3 months. The remaining 10 rats, the control group, were fed a normal diet. After the experimental procedure, all animals were anesthetized and omental adipose tissues in the same area were dissected and fixed for the histochemical process using a mixture of 3% glutaraldehyde and 1% osmium tetraoxide in 0.1 M phosphate buffer. After embedding of tissues in araldite CY 212, semi-thin and thin sections were cut. The semi-thin sections were stained with toluidine blue. The physical dissector counting method was used for estimation of numerical density and nuclear height of adipocytes. Cavalieri principle was used for the estimation of adipocyte volume; volume fraction approach was applied to find the volume fraction of adipose tissue components. Results: The mean numerical density of adipocytes in the HF diet group was significantly higher than the control. The mean nuclear height of adipocytes was also very high in the HF diet group. The volume fraction of adipose mass was increased whereas the extracellular matrix volume fraction was reduced in the HF diet group compared to the controls. The mean volume of adipocytes in the HF diet group was also significantly higher than in the control group. At the light microscopy level, it was found that adipocytes were enlarged and gaining irregular shape in the HF diet group. Thicker basal lamina and electron dense lipid content were also found in this group at the electron microscopy level. Conclusion: Lipid content and number of adipocytes in the adipose tissue of HF diet rats were higher than in the controls. Thus, HF diet induces increase in body weight via both hypertrophy and hyperplasia of adipocytes.     

Operative failure during ureteroscopic pneumatic lithotripsy: factors affecting successful outcome

AIM: We aimed to evaluate the predictive factors that would in turn indicate stone migration and the effects of these factors on the ultimate success of the intervention. METHOD: Patients were divided into two groups with respect to the migration of the stone treated. Group I: patients demonstrating stone migration during manipulation; group II: no migration of the stones noted. In the second phase of the study, the results of ureteroscopic management in 433 patients were evaluated with respect to the success rates obtained. Parameters such as stone size, stone burden, experience of the surgeon, length of the ureter proximal to the stone treated, and lastly transverse diameter of the ureter were noted in all patients as possible risk factors for stone migration. RESULTS: Statistical analysis of ureteroscopic success in all patients revealed that there was a meaningful correlation with respect to the length of the proximal ureteral portion (p < 0.0001) and surgeon's experience (p = 0.004). p value for the correlation between stone burden and operative success was 0.056. There was no significant correlation between stone size (p = 0.51), ureter diameter (p = 0.78), and operative success. CONCLUSION: Stones that are close to the renal pelvis and treated by inexperienced physicians are the ones most likely to migrate to the renal pelvis during manipulation with pneumatic lithotripsy.     

Comparison of genetic groups determined by molecular and immunological analyses of giardia isolated from animals and humans in switzerland and australia

Nine axenic isolates of Giardia originating from four different host species in Switzerland were subjected to genetic analysis using the polymerase chain reaction (PCR) to amplify segments of genes encoding different trophozoite variant surface proteins (VSPs). Three genotypes were identified on the basis of product yield, size, and restriction-fragment-length polymorphisms (RFLPs). Five G. duodenalis isolates (01, Bl, B2, B3-1A1 and Cl — from a sheep, three calves and a dog, respectively) were classified as belonging to genetic group I of Andrews et al. (1989). DNA amplified from the VSP genes tsp11, tsa417 and vsp1267 of these isolates was indistinguishable in size and restriction characteristics from that amplified from group-I Giardia isolated from humans in Australia. One human-derived Swiss isolate (H2-17A1), typed as belonging to genetic group II, yielded a vsp1267-specific PCR product that was indistinguishable by size or restriction sites from the equivalent 1.6-kb product amplified from human-derived Australian group-II organisms. This isolate also yielded 1.8-kb tspll and 0.52-kb tsa417/tsp11-like PCR products possessing RFLPs typical of group-II organisms. Three isolates (O2-4A1, 03 and H3-15K2 — originating from two sheep and a human, respectively) represent a novel genotype that is closely related to genetic groups I and II. These three isolates exhibited identical RFLPs in their tspll PCR products and failed to yield a vsp1267 PCR product. An antiserum specific for the 90-kDa VSP of the sheep-derived clone O2-4A1 reacted strongly by immunofluorescence and on Western blots with surface proteins from the 02, 03 and H3 isolates only — consistent with the genetic classification determined above. The data provide no evidence for the occurrence of hostspecific genotypes.