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The human skin microbiota is quantitatively dominated by Gram-positive bacteria, detected by both culture and metagenomics. However, metagenomics revealed a huge variety of Gram-negative taxa generally considered from environmental origin. For species affiliation of bacteria in skin microbiota, clones of 16S rRNA gene and colonies growing on diverse culture media were analyzed. Species-level identification was achieved for 81% of both clones and colonies. Fifty species distributed in 26 genera were identified by culture, mostly belonging to Actinobacteria and Firmicutes, while 45 species-level operational taxonomic units distributed in 30 genera were detected by sequencing, with a high diversity of Proteobacteria. This mixed approach allowed the detection of 100% of the genera forming the known core skin Gram-negative microbiota and 43% of the known diversity of Gram-negative genera in human skin. The orphan genera represented 50% of the current skin pan-microbiota. Improved culture conditions allowed the isolation of Roseomonas mucosa, Aurantimonas altamirensis and Agrobacterium tumefaciens strains from healthy skin. For proteobacterial species previously described in the environment, we proposed the existence of skin-specific ecotypes, which might play a role in the fine-tuning of skin homeostasis and opportunistic infections but also act as a shuttle between environmental and human microbial communities. Therefore, skin-associated proteobacteria deserve to be considered in the One-Health concept connecting human health to the health of animals and the environment.

The pneumococcal histidine triad protein D (PhtD) is believed to play a central role in pneumococcal metal ion homeostasis and has been proposed as a promising vaccine candidate against pneumococcal disease. To investigate for potential stabilizers, a panel of physiologically relevant metals was screened using the thermal shift assay and it was found that only Zn 2+ and Mn 2+ were able to increase PhtD melting temperature. Differential scanning calorimetry analysis revealed a sequential unfolding of PhtD and the presence of at least 3 independent folding domains that can be stabilized by Zn 2+ and Mn 2+ . UV spectroscopy and fluorescence quenching studies showed significant Zn 2+ -induced tertiary structure changes in PhtD characterized by decreased accessibility of inner tryptophan residues to the aqueous solvent. Isothermal titration calorimetry data show no apparent binding to Mn 2+ but revealed a Zn 2+ :PhtD exothermic interaction stoichiometry of 3:1 with strong enthalpic contribution, suggesting that 3 of the 5 histidine triads are accessible binding sites for Zn 2+ . Only Zn +2 , but not Mn +2 , was able to increase the thermal stability of PhtD in the presence of aluminum hydroxide adjuvant, making it a promising stabilizer excipient candidate in vaccine products containing PhtD., (Copyright © 2017 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.)

Background: Splenectomy is a common surgical procedure, but few reports focus on nontraumatic splenectomies. The aim of this study was to determine the predictors of morbidity and mortality of patients submitted to elective nontraumatic splenectomy., Methods: A descriptive cross-sectional study of 152 consecutive, nonselected, nontraumatic patients operated on by splenectomy between 1996 and 2010 was carried out. Clinical, laboratory, and surgical data, histological findings, perioperative mortality, and postoperative complications according to Clavien-Dindo classification, were recorded. Factors related to morbidity and mortality were analyzed., Results: Of the 152 patients (89 male and 63 female; mean age 49.8 ± 17.8 years), 74 (48.7%) were operated on for malignant hematologic disorders, 44 for benign hematologic process, and 34 for other nonhematologic disorders. The spleen was enlarged in 95 patients (62.5%) and 78 patients (51.3%) had hypersplenism. The overall complications rate was 40.1%: Grades I and II in 27 cases (17.7%), and Grades III and IV in 23 patients (15.1%). Perioperative mortality was 7.2% (11 patients). In univariate analysis, significant negative predictors for morbidity were age (p = 0.004), anemia (p = 0.03), leukocytosis (p = 0.016), and blood transfusions (p < 0.001). In the multivariate analysis, only the need for blood transfusion remained as an independent prognostic factor (p = 0.001). Related to mortality, negative prognostic factors were age (p = 0.003), leukocytosis (p = 0.048), American Society of Anesthesiologists (ASA) score (p < 0.001), blood transfusion (p < 0.001), pleural effusion (p = 0.031), and pneumonia (p = 0.001). Pneumonia remained an independent prognostic factor of mortality (p = 0.024)., Conclusion: Blood loss is the most important prognostic factor for postoperative complications after nontraumatic splenectomies. Pneumonia is the main prognosis factor for perioperative mortality., (Copyright © 2013. Published by Elsevier B.V.)

Although cystic echinococcosis (CE) has been a recognized public health problem in Portugal, molecular data regarding the types and prevalence of infecting strains of its etiological agent (Echinococcus granulosus) are still scarce. Over the last years we have been evaluating the prevalence of CE in the country, and in this report we determined the parasite genotypes infecting sheep, goats, cattle and human in Portugal, based on 209 hydatid cysts recovered from liver (n=96), lung (n=95), pancreas (n=17) and kidney (n=1) samples obtained between 2008 and 2011. Protoscoleces or germinal layers were collected from individual cysts, DNA was extracted, and a part of the mitochondrial DNA encoding the cytochrome c oxidase subunit 1 was amplified by PCR. Overall, the results confirm the overall dominance of the G1-G3 cluster of strains, which are particularly prevalent in southern Portugal in livestock ruminants. Unexpectedly, one parasite sequence with cattle origin was found to correspond to E. granulosus G7 genotype (also known as E. intermedius), here reported for the first time in bovine, in Portugal., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Acute mesenteric ischemia (AMI) is a catastrophic surgical condition, especially in older patients with multiple comorbidities. The aim of this study was to evaluate the impact of comorbidity on perioperative mortality and overall survival in patients surgically treated for AMI. A series of 186 consecutive patients (106 men and 80 women) who underwent surgery because of AMI in a university tertiary care center between 1990 and 2006 were retrospectively studied. The Charlson Comorbidity Index (CCI) score, unadjusted and adjusted by age, was preoperatively calculated in each patient. Perioperative mortality and overall survival were also recorded. The association between unadjusted and adjusted by age CCI and perioperative mortality and overall survival were analyzed. The mean age of the studied population was 72.1 years (SE +/-13.7 years). Hospital mortality was 64.5%. One-year, 3-year, and 5-year overall estimated survival by the Kaplan-Meier method after surgery for AMI was 26%, 23% and 21%, respectively. Perioperative mortality was not related to the unadjusted preoperative CCI (p = 0.093). Nevertheless, a statistically significant association was found between mortality and preoperative adjusted CCI (p = 0.007). Likewise, CCI unadjusted was almost related to overall survival (p = 0.055), but the values of the categorized CCI adjusted by age showed a statistically significant difference in overall survival (p = 0.012). In multivariate analysis, CCI adjusted by age remained independent prognostic factor of mortality. Comorbidity adjusted by age may play a role as a predictive factor for perioperative mortality and long-term survival in patients operated on for AMI.

A methodology using interactive computerized microscopy (ICM) was developed to quantify in the mesenteric arterial bed the morphometric changes associated with diabetes and the influence of treatment with SR 49059, an antagonist of vasopressin V1a receptors. Four groups of rats were studied: untreated normal (N) or streptozotocin- (60 mg/kg i.v.) induced diabetic (D), and treated (0.4 mg/g SR 49059 included in food) normal (NT) or diabetic (DT) animals. Treatment was initiated 4 days after diabetes induction and continued for 3 weeks. Nested (hierarchical) analysis of variance of ICM data was performed on raw diameter or after logarithmic normalization of area and nuclei values. Diabetes was associated with an increase in arterial diameters, and in total vessel, wall, media, adventitia, and lumen areas. The same parameters, with the exception of the lumen, were also increased in DT as compared to D. The number of nuclei in the media or adventitia was increased in D as compared to N, and in DT as compared to D. In summary, ICM is allowed to further characterize the vascular mesenteric changes and describe for the first time the enlargement of adventitia associated with diabetes. Our study also suggested that the blockade of Via receptors is unable to prevent diabetes-related vascular changes, although the slight increase in food intake associated with SR 49059 treatment may have had an indirect influence on angiopathy development.

The cellular localization of peripheral-type benzodiazepine receptors (PBRs) was characterized in several human blood cell subpopulations including erythrocytes, platelets, monocytes and polymorphonuclear neutrophils (PMN), B, NK, T8 and T4-cells. Pharmacological properties of the PBR were established by binding studies and PBR mRNA expression was measured by quantitative polymerase chain reaction based method. These data clearly indicate 1) the PBR is pharmacologically homogeneous in the various types of blood cells, 2) the rank order of PBR cell density is monocytes = PMN > lymphocytes >> platelets > erythrocytes, 3) the PBR appears to be transcriptionally regulated since mRNA levels are roughly correlated with PBR density.