First phylogenetic analysis of dengue virus serotype 4 circulating in Espírito Santo state, Brazil, in 2013 and 2014.

The purpose of the present study was to reconstruct the phylogeny of dengue virus serotype 4 (DENV-4) that was circulating in Espírito Santo state, Brazil, in 2013 and 2014, and to discuss the epidemiological implications associated with this evolutionary hypothesis. Partial envelope gene of eight DENV-4 samples from Espírito Santo state were sequenced and aligned with 72 worldwide DENV-4 reference sequences from GenBank. A phylogenetic tree was reconstructed through Bayesian Inference and the Time of the Most Recent Common Ancestor was estimated. The study detected the circulation of DENV-4 genotype II in Espírito Santo state, which was closely related to strains from the states of Mato Grosso collected in 2012 and of São Paulo sampled in 2015. This cluster emerged around 2011, approximately 4 years after the entry of the genotype II in Brazil through its northern states, possibly imported from Venezuela and Colombia. This is so far the first phylogenetic study of the DENV-4 circulating in Espírito Santo state and shows the importance of an internal route of dengue viral circulation in Brazil to the introduction of the virus into this state.

Pre-transplant shedding of BK virus in urine is unrelated to post-transplant viruria and viremia in kidney transplant recipients.

BK virus-(BKV) associated nephropathy (BKVN) is a major cause of allograft injury in kidney transplant recipients. In such patients, subclinical reactivation of latent BKV infection can occur in the pre-transplant period. The purpose of this study was to determine whether urinary BKV shedding in the immediate pre-transplant period is associated with a higher incidence of viruria and viremia during the first year after kidney transplantation. We examined urine samples from 34 kidney transplant recipients, using real-time quantitative polymerase chain reaction to detect BKV. Urine samples were obtained in the immediate pre-transplant period and during the first year after transplant on a monthly basis. If BKV viruria was detected, blood samples were collected and screened for BKV viremia. In the immediate pre-transplant period, we detected BKV viruria in 11 (32.3%) of the 34 recipients. During the first year after transplantation, we detected BKV viruria in all 34 patients and viremia in eight (23.5%). We found no correlation between pre-transplant viruria and post-transplant viruria or viremia (p = 0.2). Although reactivation of latent BKV infection in the pre-transplant period is fairly common among kidney transplant recipients, it is not a risk factor for post-transplant BKV viruria or viremia.

[Predictors of cesarean delivery in the Kinkanda general hospital in Matadi (Democratic Republic of Congo)]. / Les predicteurs de l'accouchement par cesarienne a l'hopital general de reference de Kinkanda a Matadi (Republique Democratique du Congo).

OBJECTIVE: To identify predictors of cesarean delivery in the Kinkanda general hospital. METHOD: Cross-sectional study conducted from 1st January 2012 to 31st December 2012, on women in labor who delivered at the maternity of Kinkanda. Predictors of cesarean delivery were identified by using binary logistic regression. RESULTS: A uterine height of at least 33 cm at induction of labor (AOR 3.93 95% CI 2.04 to 7.59; p <0.001), primiparity (AOR 2.1 95%CI:1.06 to 4.15; p = 0.033), non-membership of the pregnant woman to this particular health center area (AOR 4.26 95%CI 2.16 to 8.42; p <0.001) and a weight of the newborn of at least 4000g (AOR 7.8 95%CI 2.3 to 26.44; p <0.001) were identified as risk factors for cesarean delivery. CONCLUSION: Primiparity, uterine height of at least 33 cm, living in another health area than the Matadi area and fetal macrosomia are the main predictors of cesarean delivery during the year 2012 at the Kinkanda general hospital.

OBJECTIF: Identifier les prédicteurs de l'accouchement par césarienne à l'Hôpital Général de Kinkanda. MÉTHODE: Etude transversale analytique réalisée du 01 janvier au 31 décembre 2012, auprès de parturientes ayant accouché à la maternité de l'Hôpital Général de Kinkanda. Les variables prédictives de l'accouchement par césarienne ont été mises en évidence à l'aide d'une régression logistique binaire. RÉSULTATS: Une hauteur utérine d'au moins 33 cm au déclenchement du travail (ORA : 3,93 IC95% : 2,04 ­ 7,59 ; p < 0,001), la primiparité (ORA :2,1 IC95% :1,06 ­ 4,15 ; p= 0,033), la non appartenance de la parturiente à la zone de santé (ORA : 4,26 IC95% : 2,16 ­ 8,42 ; p < 0,001) et le poids du nouveau- né avec au moins 4000g (ORA : 7,8 IC95% : 2,3 ­ 26,44 ; p< 0,001) ont été identifiés comme des facteurs prédictifs de l'accouchement par césarienne. CONCLUSION: La primiparité, la hauteur utérine d'au moins 33 cm, l'appartenance à une autre zone de santé que celle de l'Hôpital Général de Kinkanda et la macrosomie fœtale sont les principaux prédicteurs de la césarienne au cours de l'année 2012 à l'Hôpital Général de Référence de Kinkanda.

Biometrical implications of factorial experiments for the study of lymphocyte mitogenic response.

Lymphocyte responsiveness to mitogens is affected by a number of experimental variables; the latter may be conveniently studied several at a time, following a factorial design that allows the complete evaluation of the effects of each variable and of their interplays. Statistical inferences may be drawn through an analysis of variance, provided that the data conform to the underlying model, i.e. that the effects are additive and that the errors are normally distributed and homoscedastic. Thymidine incorporation data should first be transformed to log cpm; the effects on the metameter may then be assumed to be additive, and their errors approach normality; the error variances may be affected by the experimental variables involved, so that the homogeneity of the error must be checked before relying on the pooled estimate of the error for parametric tests of significance. The above considerations have been illustrated with factorial experiments on [3H]TdR incorporation by rabbit spleen cells stimulated with varying amounts of PHA-P or PHA-M, under a variety of culture conditions.

Age-related changes in the prevalence of precipitating antibodies to BK virus in infants and children.

Nearly 1000 sera from children were tested by immunoelectro-osmophoresis against BK virus, and age-specific prevalence rates were estimated from birth until the age of 12 years. Declining rates during the first 12 months showed the waning of passive immunity, which at birth reflects the mother's immune status. The changes of prevalence suggested that the peak incidence of primary infections occurred at about 2 years, with an estimated peak annual rate of 24.6%.

Immunoelectroosmophoresis for the human papovavirus BK.

The reaction between BK virus and its antibodies may well be visualized with immunoelectroosmophoresis (IEOP). Serum antibody levels may be compared or graded by reacting them against a continuous antigen front. In seroepidemiologic surveys IEOP compares favourably with hemagglutination inhibition in terms of sensitivity, practicability and reproducibility. Technical conditions are described which are suitable for antigen detection, quantification and identification.

The Reoviridae family.

The members of the Reoviridae family are extremely varied in host ranges and have such diverse natural histories that it is compelling to conclude that their structural asset and replication strategy are uniquely successful in evolutionary terms. It follows that their study addresses fundamental aspects of virology, besides the ones which are customary with important pathogens affecting humans, animals and plants. We deal here with the present taxonomy of the family Reoviridae and of its genera, two of which (Orbivirus and Rotavirus) are treated separately in this issue. Along with a cursory presentation of the remaining genera, we give a concise update of recent findings on the genus Orthoreovirus.

[Multivariate analysis of aminoacyl-tRNA synthetase activities of liver and hepatomas (author's transl)]. / Analisi multivariata delle attività aminoacil-tRNA sintetasiche del fegato e degli epatomi

Data on the aminoacyl-tRNA synthetase activities in normal liver and in three transplantable rat hepatomas, viz Yoshida's AH 130 and Morris's 5123C and 7793, were studied by means of factor analysis, a powerful technique of multivariate analysis particularly suitable foridentifying factors which theoretically may account for the pattern of interrelations between several variables. The analysis has been performed on 51 patterns of enzyme activities, covering 17 out of the 20 aminoacyl-tRNA synthetases; 80% on average of the total variability of the 17 enzyme activities may be accounted for by the first 4 factors extracted. The enzymes seem to fall into different groups, depending on their relationship with the factors thus identified. The results suggest that enzymes belonging to the same group share a common control mechanism, and are independent of the enzymes belonging to different groups, both in normal liver and in hepatomas.

[Role of viruses in acute gastroenteritis]. / Inquadramento etiopatogenetico delle enteriti acute virali.

In a two years screening carried out on Florentine children we confirmed that Rotaviruses are the most important ethiological agents of acute gastroenteritis in infants less than two years old, irrespective of sex. We had evidence that Rotavirus diarrhea does not occur with the same incidence every year. Rotavirus gastroenteritis is characterized by profuse diarrhea, vomiting, fever and often by respiratory symptoms. Severe complications have not been observed.

Nosocomial diarrhoeas in a surgical division hyperendemic for Clostridium difficile: epidemiologic aspects emerging from an analysis of clinical records.

Having previously shown that Clostridium difficile was responsible for an intense and protracted endemic of nosocomial diarrhoeas in the surgical division of a Tuscan hospital, we started a retrospective analysis on all records from the affected division, to cover a period of 15 months. A statistical description is given of a large series of nosocomial diarrhoeas, as well as direct estimates of their incidence rates in selected high risk subgroups. The situation described is epidemiologically unique, and its study tells how an endemic may reach a steady state of high nosocomial morbidity.

[Pyocin typing of 111 strains of Pseudomonas aeruginosa isolated in Tuscany, using the Gillies and Govan method]. / Tipizzazione piocinica secondo Gillies e Govan di 111 stipiti di "Pseudomonas aeruginosa" isolati in Toscana.

Pseudomonas aeruginosa strains from Tuscany have been typed for pyocine production against Govan and Gillies' extended set of indicator strains; results have been expressed following a binary code. Among 111 strains only 6 could not be typed; the remaining 105 strains (94.6%) presented 59 different patterns, 44 of which were represented by one isolate only. The most commonly encountered pattern (n. 8045, corresponding to Govan and Gillies' subtype 1e) was shared by 13 strains. Isolates from outpatients or from the environment tended to a uniform distribution of pyocine patterns; those from two different hospitals tended to group into distinct subtypes.

Epidemiological studies of Pseudomonas aeruginosa infections in leg ulcers.

A 6-month prospective study was carried out in an angiology ward on patients with leg ulcers. Pseudomonas aeruginosa was isolated from about half of the samples from the patients. Of the isolates, 85 were serotyped, fingerprinted as to passive pyocin sensitivity and characterized for antibiotic susceptibility pattern. According to all criteria, the isolates belonged to six strains, two of which were clearly epidemic: one infected 11 patients yielding 52 identical isolates, the other infected 4 patients (19 isolates).

Tentative evidence of AIDS-associated biotype of Mycobacterium kansasii.

Previous studies revealed heterogeneous behavior within the species Mycobacterium kansasii against commercially available DNA probes (Accuprobe M. kansasii culture identification test; Gen-Probe); several isolates, conventionally identified as M. kansasii, failed in fact to hybridize. Looking for a possible association with phenotypic features, we tested a fully characterized panel of 69 clinical isolates of M. kansasii (19 of which were Accuprobe negative) with a semiquantitative micromethod which tests for 19 enzymatic activities (Api Zym; BioMérieux). The strains were from 25 hospitals in 18 Italian towns; 20 isolates came from human immunodeficiency virus type 1-positive patients who fulfilled the Centers for Disease Control criteria for AIDS diagnosis. On the basis of the whole set of phenotypic traits, our strains clustered in two groups, allowing the differentiation of biotypes within the species. There was a perfect association between biotype 2 and hybridization failures with Accuprobe and a very significant association between this novel biotype 2 and AIDS status, which suggests that it differs in virulence.

[Serodiagnosis of rotavirus infections in gastroenteritis of infants]. / Sierologia diagnostica di infezioni da rotavirus in casi di gastroenterite infantile.

We report the use of the line immunoelectroosmophoresis (LIEOP), a modification of the immunoelectroosmophoresis (IEOP), for the serodiagnosis of rotavirus infection, in comparison with the complement fixation test. We tested 17 paired and 29 single sera from hospitalized children affected by acute gastroenteritis; rotavirus had been searched in the stools both by electron microscopy and IEOP. The LIEOP showed seroconversion in all the 11 cases with positive stools and also in 5 out of 5 patients whose stool sample had been negative. Either we had not been able to detect the virus, or there was an intervening (nosocomial) rotavirus infection. All sera which were reactive by complement fixation with bovine rotavirus were also reactive by LIEOP. Some of the sera which reacted by LIEOP, both with the homologous and the heterologous antigen, were negative by complement fixation.

Surface glycosaminoglycans and calcium distribution in 3T3 cells.

Resting 3T3 cells have relatively more sulphated glycosaminoglycans and Ca2+ in their cell coat than do growing or SV40-virus-transformed cells. It is suggested that the different Ca2+-binding capacity controls cellular activities by affecting the distribution of Ca2+ between the intra- and ecto-cellular compartments.

Burden of unidentifiable mycobacteria in a reference laboratory.

Modern identification techniques at the genomic level have greatly improved the taxonomic knowledge of mycobacteria. In adjunct to nucleic acid sequences, mycobacterial identification has been endorsed by investigation of the lipidic patterns of unique mycolic acids in such organisms. In the present investigation, the routine use of high-performance liquid chromatography (HPLC) of mycolic acids, followed by the sequencing of the 16S rRNA, allowed us to select 72 mycobacterial strains, out of 1,035 screened, that do not belong to any of the officially recognized mycobacterial species. Most strains (i.e., 47) were isolated from humans, 13 were from the environment, 3 were from animals, and 9 were from unknown sources. The majority of human isolates were grown from the respiratory tract and were therefore most likely not clinically significant. Some, however, were isolated from sterile sites (blood, pleural biopsy, central venous catheter, or pus). Many isolates, including several clusters of two or more strains, mostly slow growers and scotochromogenic, presented unique genetic and lipidic features. We hope the data reported here, including the results of major conventional identification tests, the HPLC profiles of strains isolated several times, and the whole sequences of the 16S rRNA hypervariable regions of all 72 mycobacteria, may encourage reporting of new cases. The taxonomy of the genus Mycobacterium is, in our opinion, still far from being fully elucidated, and the reporting of unusual strains provides the best background for the recognition of new species. Our report also shows the usefulness of the integration of novel technology to routine diagnosis, especially in cases involving slow-growing microorganisms such as mycobacteria.