Microbiological monitoring of coagulase-negative Staphylococcus in public drinking water fountains: Pathogenicity factors, antimicrobial resistance and potential health risks.

The presence of opportunistic bacteria such as coagulase-negative Staphylococcus (CoNS) in drinking water poses public health concerns because of its potential to cause human infection and due to its antimicrobial resistance (AMR) diversity. This study evaluated the occurrence, virulence markers and AMR of CoNS in 468 drinking water samples from 15 public fountains located in four urban parks of São Paulo city (Brazil). Out of 104 samples positive for the presence of Staphylococcus genus, we detected CoNS in 75 of them (16%), which did not meet the Brazilian sanitary standards for residual chlorine. All isolates were of concern to public health for being responsible for infection in humans from low to high severity, nine of them are considered the most of concern due to 63.6% being multiresistant to antimicrobials. The results demonstrated that CoNS in drinking water must not be neglected. It is concluded that the presence of resistant staphylococci in drinking water is a potential health risk, which urges feasible and quick control measures to protect human health, especially in crowded public places.

Pediatric postmortem CT angiography: validation of vascular access for PMCT angiography in stillbirths, babies and toddlers.

PURPOSE: The use of angiography in postmortem CT angiography (PMCTA) has several advantages. In adults, femoral vascular access is well established. Due to the small and specific anatomy in fetuses and infants, the technique has to be adapted, especially regarding the vascular access. The aim of this study was to evaluate vascular access for pediatric PMCTA (pedPMCTA). MATERIALS AND METHODS: Ten pedPMCTAs were performed in stillbirths, babies, and one toddler. A femoral approach by cannulation of the femoral artery and vein, an umbilical approach by cannulation of the umbilical vessels, and an intraosseous approach by an intraosseous needle were evaluated by handling and resulting imaging. RESULTS: The insertion of a cannula with a size of 18-20 G in the femoral vessels was possible in babies. An umbilical access with peripheral venous cannulas with a size of 14-20 G was feasible in stillbirths and newborns. An intraosseous access is advisable as equal alternative to umbilical and in cases where a femoral access is not possible. The most significant problem with the vascular access is the extravasation of contrast media, but this can be reduced significantly with practice. CONCLUSION: When performing pedPMCTA, an umbilical vascular access is recommended if an umbilical cord with open vessels is still present. Otherwise, a bone marrow access should be preferred in the presence of an arteriovenous shunt or if only the venous system needs to be shown. If that is not the case, the femoral access with the possibility to separate venous and arterial scan should be used.

Dietary administration of a commercial mixed-species probiotic improves growth performance and modulates the intestinal immunity of tilapia, Oreochromis niloticus.

The growth performance, immunological status, intestinal morphology and microbiology of tilapia, Oreochromis niloticus, were investigated after dietary administration of the commercial probiotic AquaStar(®) Growout. Tilapia (29.02 ± 0.33 g) were split into five treatments; control (CON), 1.5 g kg(-1) probiotic (PRO-1.5), 3 g kg(-1) probiotic (PRO-3), pulsed probiotic feeding (PRO-PULSE) or an initial probiotic feed followed by control feeding (PRO-INI). After six weeks of experimental feeding, fish fed PRO-3 displayed significantly higher final weight, weight gain and SGR compared to the CON or PRO-INI treatments. Supplementation of the probiotic at this dose induced an up-regulation of intestinal caspase-3, PCNA and HSP70 mRNA levels compared to the CON fed fish. Immuno-modulatory pathways were also affected; significantly higher expression of TLR2, pro-inflammatory genes TNFα and IL-1ß, and anti-inflammatory genes IL-10 and TGFß suggest that the probiotic may potentiate a higher state of mucosal tolerance and immuno-readiness. Histological appraisal revealed significantly higher numbers of intraepithelial leucocytes in the intestine of PRO-3 fed fish compared with treatments CON, PRO-PULSE and PRO-INI but not PRO-1.5. Additionally, fish receiving PRO-3 had a significantly higher abundance of goblet cells in their mid-intestine when compared with fish from all other treatments. Together, these data suggest that continuous provision of AquaStar(®) Growout at 3 g kg(-1) can improve tilapia growth and elevate the intestinal immunological status of the host.

Matrix metalloproteinase 9 -1562C/T polymorphism increased protein levels in patients with colorectal cancer in a sample from southeastern Brazil.

We examined whether the allelic and/or genotypic profile of locus -1562C/T of the matrix metalloproteinase (MMP-9) gene influences the protein expression levels of MMP-9 in patients with colorectal cancer (CRC) compared with controls. A total of 104 patients with CRC and 84 controls were evaluated. Peripheral blood was collected from both groups and DNA extraction was performed for -1562C/T genotyping; the plasma was used for MMP-9 quantification. The CT genotype was associated with increased MMP-9 expression (P = 0.0211). High levels of protein, independently of polymorphisms, were observed in the patient group (P < 0.0001) compared to controls. Mucinous tumors with signet ring cells were more frequent in females (P = 0.0177). Overall, patients older than 50 years showed a significant risk of developing CRC (P = 0.0001). MMP-9 plasma expression was increased in patients with CRC compared to controls, particularly in those with the heterozygous -1562CT genotype.

Modulation of the intestinal microbiota and morphology of tilapia, Oreochromis niloticus, following the application of a multi-species probiotic.

The intestinal microbiota and morphology of tilapia (Oreochromis niloticus) were investigated after the application of a multi-species probiotic containing Lactobacillus reuteri, Bacillus subtilis, Enterococcus faecium and Pediococcus acidilactici (AquaStar(®) Growout). Tilapia (55.03 ± 0.44 g) were fed either a control diet or a probiotic diet (control diet supplemented with AquaStar(®) Growout at 5 g kg(-1)). After four and eight weeks, culture-dependent analysis showed higher levels of lactic acid bacteria (LAB), enterococci and Bacillus spp. in the mucosa and digesta of fish fed AquaStar(®) Growout. At week four, polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) revealed a higher similarity within the probiotic fed replicates than replicates of the control group; after eight weeks, the compositional dissimilarity of the microbiome profiles between the groups was greater than the dissimilarities within each group (P < 0.05). High-throughput sequencing revealed that the probiotic treatment significantly reduced the number of operational taxonomic units and species richness in the digesta. Significantly higher proportions of reads belonging to Proteobacteria and Cyanobacteria were detected in the control group whereas the probiotic-fed fish displayed a significantly higher abundance of reads assigned to the Firmicutes (which accounted for >99 % of reads). Bacillus, Cetobacterium and Mycobacterium were the dominant genera in the digesta of control fish whereas Bacillus, Enterococcus and Pediococcus were the largest constituents in probiotic-fed fish. The addition of AquaStar(®) Growout to tilapia diets led to increased populations of intraepithelial leucocytes, a higher absorptive surface area index and higher microvilli density in the intestine. These data suggest that AquaStar(®) Growout can modulate both the intestinal microbiota and morphology of tilapia.

Disrupting membrane raft domains by alkylphospholipids.

Using phase contrast and fluorescence microscopy we study the influence of the alkylphospholipid, ALP, 10-(octyloxy) decyl-2-(trimethylammonium) ethyl phosphate, ODPC, in giant unilamellar vesicles, GUVs, composed of DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine), brain sphingomyelin (SM) and cholesterol (Chol). The results show that adding 100µM ODPC (below CMC) to the outer solution of GUVs promotes DOPC membrane disruption over a period of 1h of continuous observation. On the other hand, the presence of SM and Chol in homogeneous fluid lipid bilayers protects the membrane from disruption. Interestingly, by adding 100µM ODPC to GUVs containing DOPC:SM:Chol (1:1:1), which display liquid ordered (Lo)-liquid disordered (Ld) phase coexistence, the domains rapidly disappear in less than 1min of ODPC contact with the membrane. The lipids are subsequently redistributed to liquid domains within a time course of 14-18min, reflecting that the homogenous phase was not thermodynamically stable, followed by rupture of the GUVs. A similar mechanism of action is also observed for perifosine, although to a larger extent. Therefore, the initial stage of lipid raft disruption by both ODPC and perifosine, and maybe other ALPS, by promoting lipid mixing, may be correlated with their toxicity upon neoplastic cells, since selective (dis)association of essential proteins within lipid raft microdomains must take place in the plasma membrane.

Floristic diversity and edaphic filters in an urban forest under Cerrado domain, in Cuiabá, Central Western Brazil.

We aimed to characterize the adult and regenerating tree components and their relationships with soil characteristics of a native vegetation remnant in Cuiabá, Mato Grosso, Brazil. The area of the fragment is stratified into "dry area" (lithic neosoil) and "damp area" (gleisoil). We conducted a forest inventory with a random distribution of 25 parcels. We analyzed the physical and chemical components of the soil. We evaluated the vegetation's horizontal structure, diversity, and sample sufficiency using the Bootstrap richness estimator. We classified the species according to dispersal syndrome and ecological group. Overall, we found 93 species in the adult layer and 70 species in the regenerating layer. The similarity dendrograms based on the two evaluated indices demonstrated the existence of the two initially stratified environments in both strata. The IndVal (%) indicated that the set of indicator species differed between the strata. Thereby, the fragment is in an intermediate stage of successional progression. PCA showed that plots in the wet area had higher pH values and Ca, Zn, and Fe levels, while plots in the dry area did not clearly distinguish, varying in terms of K, B, and organic matter content. In CCA, a set of species that occurred exclusively in the damp area showed a strong relationship with the analyzed variables. The area is a diverse ecosystem that efficiently provides ecosystem services to society and should be the subject of long-term conservation and research.

Dietary probiotic supplementation modulated gut microbiota and improved growth of juvenile rainbow trout (Oncorhynchus mykiss).

In aquaculture, infectious diseases are the major cause of economic losses. Probiotic supplementation may change the microbiota of the digestive tract and modulate the immune defences and nutritional performance. This study was conducted to evaluate the dietary supplementation of multi-species (A: Bacillus sp., Pediococcus sp., Enterococcus sp., Lactobacillus sp.) and single-species probiotics (B: Pediococcus acidilactici) on growth performance and gut microbiota of rainbow trout (Oncorhynchus mykiss). A basal diet was supplemented with probiotic A or B, at two concentrations each (A1, A2, B1 and B2) or not supplemented (control treatment). Diets were distributed to 30 groups of 20 fish, 3 times a day. The gut microbiota was analysed at the end of the feeding trial (96 days) with 16S rDNA denaturing gradient gel electrophoresis (16S-DGGE). Changes in gut microbial community were assessed by Shannon index (H´) and number of operational taxonomic units (OTUs). After 56 days of feeding, weight gain was significantly improved in fish fed diet A1 when compared to the control group. Dietary probiotic supplementation changed the gut microbial composition. Number of OTUs (R) was higher in fish fed A1 (multi-species at lower concentration) than in control group, while H´ was higher in fish fed A1, B1 and B2.

Additional activation of the AR gene may be involved in the development of the castration resistance phenotype in prostate cancer.

INTRODUCTION: Several studies have already shown that changes in the AR gene may be associated with a more aggressive disease phenotype and even castration-resistant prostate cancer. Thus, we investigated cytogenetic and molecular alterations linked to AR. MATERIALS AND METHODS: To evaluate AR methylation, we performed a cytogenetic-molecular analysis using fluorescence in situ hybridization that uses specific probes for the AR gene (Xq11.12) and the X chromosome centromere. For AR activity, we performed a qualitative analysis of human androgen receptor activity. To analyze the expression of AR in PC3 and LNCaP cell lines, we used qPCR assays. RESULTS: In the qPCR assay, we found downregulation of AR in the PC3 cell line compared with the LNCaP. We found the presence of X chromosome polysomy in PC-3 and LNCaP cell lines by FISH assay. In the HUMARA-Q assay, we found two X chromosomes/cell and the activity of both AR in the PC-3 cell line. In LNCaP cells, we found two X chromosomes/cell and methylation of only one AR. CONCLUSION: Castration-resistant prostate cancer phenotype represents a significant challenge in the setting of urological management. The X chromosomes and AR-linked alterations may contribute to a better understanding of the disease. However, further studies should be performed in an attempt to elucidate as much as possible the role of AR in the castration-resistant prostate cancer phenotype.

Interaction of 10-(octyloxy) decyl-2-(trimethylammonium) ethyl phosphate with mimetic membranes and cytotoxic effect on leukemic cells.

10-(Octyloxy) decyl-2-(trimethylammonium) ethyl phosphate (ODPC) is an alkylphospholipid that can interact with cell membranes because of its amphiphilic character. We describe here the interaction of ODPC with liposomes and its toxicity to leukemic cells with an ED-50 of 5.4, 5.6 and 2.9 microM for 72 h of treatment for inhibition of proliferation of NB4, U937 and K562 cell lines, respectively, and lack of toxicity to normal hematopoietic progenitor cells at concentrations up to 25 microM. The ED-50 for the non-malignant HEK-293 and primary human umbilical vein endothelial cells (HUVEC) was 63.4 and 60.7 microM, respectively. The critical micellar concentration (CMC) of ODPC was 200 microM. Dynamic light scattering indicated that dipalmitoylphosphatidylcholine (DPPC) liposome size was affected only above the CMC of ODPC. Differential calorimetric scanning (DCS) of liposomes indicated a critical transition temperature (T(c)) of 41.5 degrees C and an enthalpy (H) variation of 7.3 kcal mol(-1). The presence of 25 microM ODPC decreased T(c) and H to 39.3 degrees C and 4.7 kcal mol(-1), respectively. ODPC at 250 microM destabilized the liposomes (36.3 degrees C, 0.46 kcal mol(-1)). Kinetics of 5(6)-carboxyfluorescein (CF) leakage from different liposome systems indicated that the rate and extent of CF release depended on liposome composition and ODPC concentration and that above the CMC it was instantaneous. Overall, the data indicate that ODPC acts on in vitro membrane systems and leukemia cell lines at concentrations below its CMC, suggesting that it does not act as a detergent and that this effect is dependent on membrane composition.

Ingestion of a moderate dose of alcohol enhances physical exercise-induced changes in blood lactate concentration.

The consumption of alcoholic beverages influences carbohydrate and lipid metabolism, although it is not yet clear whether metabolism during physical exercise at different intensities is also affected. This was the objective of the present study. Eight young and healthy volunteers performed a treadmill test to identify the running speed corresponding to a lactate concentration of 4 mM (S4mM). At least 48 h later, they were subjected to two experimental trials (non-alcohol or alcohol) in which they performed two 1-km running sessions at the following intensities: 1) S4mM; 2) 15% above S4mM. In both trials, blood lactate, triglycerides, and glucose concentrations were measured before and after exercise. The acute alcohol intake increased triglycerides, but not lactate concentration under resting conditions. Interestingly, alcohol intake enhanced the exercise-induced increase in lactate concentration at the two intensities: S4mM (non-alcohol: 4.2±0.3 mM vs alcohol: 4.8±0.9 mM; P=0.003) and 15% above S4mM trial (P=0.004). When volunteers ingested alcohol, triglycerides concentration remained increased after treadmill running (e.g., at S4mM - at rest; non-alcohol: 0.2±0.5 mM vs alcohol: 1.3±1.3 mM; P=0.048). In contrast, glucose concentration was not modified by either alcohol intake, exercise, or their combination. We concluded that an acute alcohol intake changed lactate and lipid metabolism without affecting blood glucose concentration. In addition, the increase in lactate concentration caused by alcohol was specifically observed when individuals exercised, whereas augmented triglycerides concentration was already observed before exercise and was sustained thereafter.

The role of formulation and follicular pathway in voriconazole cutaneous delivery from liposomes and nanostructured lipid carriers.

In general, colloids provide increased cutaneous permeation of drugs. Still, skin interaction and main pathways for drug diffusion may vary depending on system and formulation characteristics. The knowledge of how different colloidal systems interact with biological membranes and the formulation impact on delivery is especially relevant for drugs that can be encapsulated in multiple nanosystems, as voriconazole (VOR). In here, we compared VOR release and permeation profile from liposomes (LP) and nanostructured lipid carriers (NLC) in aqueous colloidal dispersions and in gel formulations. Despite the controlled drug release provided by gel formulations, formulation only had a significant impact on drug skin accumulation from LP. The reduced mobility in gel formulations compromised follicle deposition and drug retention in the skin. Such a hypothesis was confirmed by permeation experiments evaluating follicle pathway influence. Follicular route also had an influence on delivery from NLC, which was only significant for total drug that reached the acceptor medium. These differences could be attributed to the mechanisms of colloid interaction with the skin and subsequent drug release. Follicle LP deposition and slow drug release leads to higher cutaneous amounts whilst NLC interaction with skin and fast drug release leads to fast drug diffusion and deeper penetration. By the low MIC50 values encountered against Trichophyton rubrum (∼ 0.001 µg/mL), permeated amounts could inhibit fungal growth, regardless the system. In conclusion, both LP and NLC seem to be valuable systems for cutaneous VOR delivery. Fluidic formulations could provide better efficiency for cutaneous drug delivery from LP.

Magnetic resonance diffusion tensor imaging for the pedunculopontine nucleus: proof of concept and histological correlation.

The pedunculopontine nucleus (PPN) has been proposed as target for deep brain stimulation (DBS) in patients with postural instability and gait disorders due to its involvement in muscle tonus adjustments and control of locomotion. However, it is a deep-seated brainstem nucleus without clear imaging or electrophysiological markers. Some studies suggested that diffusion tensor imaging (DTI) may help guiding electrode placement in the PPN by showing the surrounding fiber bundles, but none have provided a direct histological correlation. We investigated DTI fractional anisotropy (FA) maps from in vivo and in situ post-mortem magnetic resonance images (MRI) compared to histological evaluations for improving PPN targeting in humans. A post-mortem brain was scanned in a clinical 3T MR system in situ. Thereafter, the brain was processed with a special method ideally suited for cytoarchitectonic analyses. Also, nine volunteers had in vivo brain scanning using the same MRI protocol. Images from volunteers were compared to those obtained in the post-mortem study. FA values of the volunteers were obtained from PPN, inferior colliculus, cerebellar crossing fibers and medial lemniscus using histological data and atlas information. FA values in the PPN were significantly lower than in the surrounding white matter region and higher than in areas with predominantly gray matter. In Nissl-stained histologic sections, the PPN extended for more than 10 mm in the rostro-caudal axis being closely attached to the lateral parabrachial nucleus. Our DTI analyses and the spatial correlation with histological findings proposed a location for PPN that matched the position assigned to this nucleus in the literature. Coregistration of neuroimaging and cytoarchitectonic features can add value to help establishing functional architectonics of the PPN and facilitate neurosurgical targeting of this extended nucleus.

Asynchronous expression of myeloid antigens in leukemic cells in a PML/RARalpha transgenic mouse model.

Acute promyelocytic leukemia (APL) is characterized by the expansion of blasts that resemble morphologically promyelocytes and harbor a chromosomal translocation involving the retinoic acid receptor alpha (RARalpha) and the promyelocytic leukemia (PML) genes on chromosomes 17 and 15, respectively. The expression of the PML/RARalpha fusion gene is essential for APL genesis. In fact, transgenic mice (TM) expressing PML/RARalpha develop a form of leukemia that mimics the hematological findings of human APL. Leukemia is diagnosed after a long latency (approximately 12 months) during which no hematological abnormality is detected in peripheral blood (pre-leukemic phase). In humans, immunophenotypic analysis of APL blasts revealed distinct features; however, the precise immunophenotype of leukemic cells in the TM model has not been established. Our aim was to characterize the expression of myeloid antigens by leukemic cells from hCG-PML/RARalpha TM. In this study, TM (N = 12) developed leukemia at the mean age of 13.1 months. Morphological analysis of bone marrow revealed an increase of the percentage of immature myeloid cells in leukemic TM compared to pre-leukemic TM and wild-type controls (48.63 +/- 16.68, 10.83 +/- 8.11, 7.4 +/- 5.46%, respectively; P < 0.05). Flow cytometry analysis of bone marrow and spleen from leukemic TM identified the asynchronous co-expression of CD34, CD117, and CD11b. This abnormal phenotype was rarely detected prior to the diagnosis of leukemia and was present at similar frequencies in hematologically normal TM and wild-type controls of different ages. The present results demonstrate that, similarly to human APL, leukemic cells from hCG-PML/RARalpha TM present a specific immunophenotype.

Ingestion of a moderate dose of alcohol enhances physical exercise-induced changes in blood lactate concentration

The consumption of alcoholic beverages influences carbohydrate and lipid metabolism, although it is not yet clear whether metabolism during physical exercise at different intensities is also affected. This was the objective of the present study. Eight young and healthy volunteers performed a treadmill test to identify the running speed corresponding to a lactate concentration of 4 mM (S4mM). At least 48 h later, they were subjected to two experimental trials (non-alcohol or alcohol) in which they performed two 1-km running sessions at the following intensities: 1) S4mM; 2) 15% above S4mM. In both trials, blood lactate, triglycerides, and glucose concentrations were measured before and after exercise. The acute alcohol intake increased triglycerides, but not lactate concentration under resting conditions. Interestingly, alcohol intake enhanced the exercise-induced increase in lactate concentration at the two intensities: S4mM (non-alcohol: 4.2±0.3 mM vs alcohol: 4.8±0.9 mM; P=0.003) and 15% above S4mM trial (P=0.004). When volunteers ingested alcohol, triglycerides concentration remained increased after treadmill running (e.g., at S4mM - at rest; non-alcohol: 0.2±0.5 mM vs alcohol: 1.3±1.3 mM; P=0.048). In contrast, glucose concentration was not modified by either alcohol intake, exercise, or their combination. We concluded that an acute alcohol intake changed lactate and lipid metabolism without affecting blood glucose concentration. In addition, the increase in lactate concentration caused by alcohol was specifically observed when individuals exercised, whereas augmented triglycerides concentration was already observed before exercise and was sustained thereafter.

Diets Containing α-Linolenic (ω3) or Oleic (ω9) Fatty Acids Rescues Obese Mice From Insulin Resistance.

Subclinical systemic inflammation is a hallmark of obesity and insulin resistance. The results obtained from a number of experimental studies suggest that targeting different components of the inflammatory machinery may result in the improvement of the metabolic phenotype. Unsaturated fatty acids exert antiinflammatory activity through several distinct mechanisms. Here, we tested the capacity of ω3 and ω9 fatty acids, directly from their food matrix, to exert antiinflammatory activity through the G protein-coupled receptor (GPR)120 and GPR40 pathways. GPR120 was activated in liver, skeletal muscle, and adipose tissues, reverting inflammation and insulin resistance in obese mice. Part of this action was also mediated by GPR40 on muscle, as a novel mechanism described. Pair-feeding and immunoneutralization experiments reinforced the pivotal role of GPR120 as a mediator in the response to the nutrients. The improvement in insulin sensitivity in the high-fat substituted diets was associated with a marked reduction in tissue inflammation, decreased macrophage infiltration, and increased IL-10 levels. Furthermore, improved glucose homeostasis was accompanied by the reduced expression of hepatic gluconeogenic enzymes and reduced body mass. Thus, our data indicate that GPR120 and GPR40 play a critical role as mediators of the beneficial effects of dietary unsaturated fatty acids in the context of obesity-induced insulin resistance.

IR studies on carrageenan of Ahnfeltia concinna, a marine red alga.

The principal polysaccharide of Ahnfeltia concinna gave IR absorptions at 1240, 930, 845-850, and 805 cm-1, all typical of a carrageenan. The polysaccharide was separated into soluble and insoluble fractions with potassium chloride.

A method for separating sex-linked imperfect albino (S*ALS) and nonalbino embryos before hatch.

The feasibility of using the sex-linked gene for imperfect albinism (S*ALS) to sex chicks during incubation by candling was studied. With this technique, the dark eye of nonalbino embryos can be positively identified. Two trials were performed. In a first trial, 66.5 and 89.5% of the 254 nonalbino and 210 albino chicks produced in four hatches were correctly identified by candling at 7 d of incubation. Of 191 eggs predicted to be nonalbinos, 22 were albinos, resulting in an overall accuracy of 88.5% for identification of nonalbino embryos. In a second trial, the accuracy of the technique from 7 to 10 d of incubation was evaluated. Increased age resulted in a tendency for lower accuracy, but candling at 8, 9, or 10 d of incubation allowed identification of a greater (P < 0.05) proportion of the nonalbino population than at 7 d of incubation. Candling at 8 d of incubation allowed identification of nonalbinos and albinos with an accuracy of 81.3 and 84.9%, respectively, suggesting that the ALS gene could be used to sex chicks during incubation when used in a sex-linked cross. This technique may prove advantageous to the laying industry because of savings of incubator and hatcher space. The males of commercial layer lines are normally killed at hatch. Reducing the number that hatch by eliminating them before 10 d of incubation could diminish animal welfare concerns.