Presence of micronuclei and nuclear abnormalities in Caracara (Polyborus) plancus living in an airport area in southern Brazil.

The aviation sector is believed to be responsible for considerable environmental damage attributed to emission of a large number and amount of pollutants. Airports are often surrounded by forest fragments and humid areas that attract birds of prey and hence may potentially serve as useful bioindicators. The aim of the present study was to examine genotoxic potential in raptors exposed to airport pollution using the micronucleus (MN) test and morphological changes as evidenced by bilateral symmetry. This investigation was conducted at Salgado Filho International Airport of Porto Alegre - RS as well as in private and zoological breeding grounds. The presence of metals was measured in the blood cells of the collected birds. Seventeen birds (Caracara (Polyborus) plancus) were used in this study 11 from exposed and 6 from non-exposed group. The nuclear alterations clearly indicate that organisms exposed to airport pollution exhibited a significantly higher frequency of genetic damage compared to non-exposed birds. Further, manganese and chromium were detected exclusively in the blood of the exposed group. In contrast, the analysis of bilateral symmetry did not detect any significant morphologic differences between the two groups. Therefore, data indicate that blood genotoxic stress occurs in birds of prey living in civil aviation areas as evidenced by MN frequency increase and presence of manganese and chromium.

Effect of artificial eccentric occlusal interferences on masticatory performance: A randomised double-blind clinical trial.

BACKGROUND: Occlusal interferences lead to changes in mandibular kinematics to compensate and improve function. However, the effects of different types of eccentric disturbance on the comminution capacity are not known. OBJECTIVE: To determine the immediate effect of eccentric occlusal interferences on masticatory performance. METHODS: This crossover clinical trial included 12 healthy dentate subjects aged 25.2 ± 3.3 years who were randomly submitted to seven types of occlusal interference: unilateral and bilateral laterotrusive and mediotrusive, protrusive, dummy and control (no interference). The interference forms were planned in a semi-adjustable articulator, fabricated with composite resin and adhered to the mandibular first molars such that subjects' maximum intercuspation was maintained. Masticatory performance and the chewing rate during 20 cycles were evaluated during subjects' comminution of silicone test food under one interference condition per test day; the multiple sieve method was applied to the comminuted particles. The interference was removed upon test completion, and a 1-week washout period was applied between tests. RESULTS: Comminuted median particle sizes were larger under unilateral (4.94 ± 0.41 mm) and bilateral (4.81 ± 0.49 mm) laterotrusive, bilateral mediotrusive (4.65 ± 0.50 mm) and protrusive (4.83 ± 0.54 mm) interferences (p < .05) than under the control (4.01 ± 0.52 mm) and dummy (4.18 ± 0.58 mm) conditions (p < .05). Only unilateral and bilateral laterotrusive interferences narrowed the comminuted particle size dispersion (p < .05). The chewing rate did not differ among conditions (p = .1944). CONCLUSION: Artificial eccentric interferences had an immediate adverse effect on masticatory performance by resulting in larger comminuted particles. CLINICAL TRIAL REGISTRATION: Brazilian Registry of Clinical Trials (RBR-8g5zfg8).

Feeding protocol with live feed and inert diet to determine the ideal age/size for weaning mullet Mugil liza (Valenciennes, 1836).

The Lebranche mullet Mugil liza is a marine fish of great importance for artisanal and industrial fishing, as well as aquaculture. The use of live feeds during the larviculture phase of marine fish is a significant component of production costs. The present study evaluated the effects of the feeding transition on different larval stages of M. liza, from the combined supply of live feed (Brachionus rotundiformis + Artemia franciscana) and live + inert feed (Artemia franciscana + inert diet) until the weaning phase to only the inert diet. A total of 3240 M. liza larvae (weight 0.0133 ± 0.0062 g and length 0.793 ± 0.160 cm) were distributed among the 12 experimental units (n = 270), resulting in four groups with three replicates each. Treatment groups consisted of feed transition with A. franciscana (enriched metanauplii) to commercial inert feed starting weaning at four different larval ages: 28, 31, 34, and 37 days post hatching. Zootechnical performance indexes and intestinal histomorphometry were evaluated. Mortality, condition factor, and length variation coefficient did not show significant differences between treatments. Final weight, final length, weight gain, and length gain were significantly greater in larvae that started weaning at 31, 34, and 37 days post hatching. Weight coefficient of variation was significantly higher in larvae that started weaning at 28 days (67.51 ± 11.70) compared to 37 days (34.40 ± 7.30). In intestinal histology, villi height (180.3 ± 4.4) was significantly higher in larvae that started weaning at 37 days post hatching. Considering the evidence found in the present study, it is recommended to start weaning M. liza on the 31st day post-hatching, using a 2-day co-feeding protocol (31st and 32nd days). From the 33rd day after hatching, M. liza larvae can receive only commercial feed.

Association between molar hypomineralization, genes involved in enamel development, and medication in early childhood: A preliminary study.

BACKGROUND: Molar hypomineralization (MH) is defined as a multifactorial condition, and thus, its presence may be defined by interactions between environmental and genetic factors. AIM: To evaluate the association between MH, genes involved in enamel development, and the use of medication during pregnancy in early childhood. DESIGN: One hundred and eighteen children, 54 with and 64 without MH, were studied. The data collected included demographics, socioeconomic data, and the medical history of mothers and children. Genomic DNA was collected from saliva. Genetic polymorphisms in ameloblastin (AMBN; rs4694075), enamelin (ENAM; rs3796704, rs7664896), and kallikrein (KLK4; rs2235091) were evaluated. These genes were analyzed by real-time polymerase chain reaction using TaqMan chemistry. The software PLINK was used to compare allele and genotype distributions of the groups and to assess the interaction between environmental variables and genotypes (p < .05). RESULTS: The variant allele KLK4 rs2235091 was associated with MH in some children (odds ratio [OR]: 3.75; 95% confidence interval [CI] = 1.65-7.81; p = .001). Taking medications in the first 4 years of life was also associated with MH (OR: 2.94; 95% CI = 1.02-6.04; p = .041) and specifically in association with polymorphisms in ENAM, AMBN, and KLK4 (p < .05). The use of medications during pregnancy was not associated with MH (OR: 1.37; 95% CI = 0.593-3.18; p = .458). CONCLUSION: The results of this study suggest that taking medication in the postnatal period appears to contribute to the etiology of MH in some evaluated children. There may be a possible genetic influence of polymorphisms in the KLK4 gene with this condition.

A Novel Compound from the Phenylsulfonylpiperazine Class: Evaluation of In Vitro Activity on Luminal Breast Cancer Cells.

Breast cancer (BC) is the most common cancer in women, and is characterized by its histological and molecular heterogeneity. Luminal BC is an estrogen receptor-positive subtype, with varied clinical courses. Although BC patients are eligible for hormone therapy, both early and late relapses still occur, and thus there is a demand for new cytotoxic and selective treatment strategies for these patients. In the present study, inspired by the structure of phenylsulfonylpiperazine, a series of 20 derivatives were tested in bioassays against MCF7, MDA-MB-231 and MDA-MB-453 BC cells to discover new hit compounds. After 48 h of treatment, 12 derivatives impaired cell viability and presented significant IC50 values against at least one of the tumor lineages. Overall, the luminal BC cell line MCF7 was more sensitive to treatments. Compound 3, (4-(1H-tetrazol-1-yl)phenyl)(4-((4-chlorophenyl)sulfonyl)piperazin-1-yl)methanone, was the most promising, with IC50 = 4.48 µM and selective index (SI) = 35.6 in MCF7 cells. Compound 3 also presented significant antimigratory and antiproliferative activities against luminal BC cells, possibly by affecting the expression of genes involved in the epithelial-mesenchymal transition mechanism, upregulating E-Cadherin transcripts (CDH1). Our findings suggest that phenylsulfonylpiperazine derivatives are potential candidates for the development of new therapies, especially those targeting luminal BC.

Effects of evaporative cooling systems on the performance of lactating sows in a tropical climate.

This study evaluated litter performance, behavioral and physiological parameters, and milk characteristics of sows submitted to different thermal environments. Fifty sows were distributed in a completely randomized design with two treatments: an evaporative cooling system (ECS) and a conventional system (CS). Sow and its litter were characterized as an experimental unit. The animals were weighed at equalization and weaning. Feed intake, milk production, and bromatological characteristics of milk were measured; the sows respiratory rate, rectal, and surface temperature were monitored. Litter uniformity was determined at equalization and weaning. Behaviors of the sows and litters were monitored for 24 h on the 7th and 15th day of lactation. Temperature and relative humidity inside the maternity was 25.00 to 28.00 °C and 26.00 to 32.55 °C and 30.00 to 70.00% and 70.00 to 88.00%, respectively, considering ECS and CS. Nutritional quality of the milk remained stable during lactation in both systems evaluated. ECS improved the average weight of the piglets, weaning weight, and daily milk production by 0.038, 0.699, and 2.31 kg/day, respectively. Sows housed in the ECS had a reduction in physiological parameters and, increase in inactive alert behavior (1.79 percentage points) and breastfeeding behavior. Piglets showed a decrease of 2.43% in the range of feedings at night and 0.15% during the day. ECS provided better comfort to the sows at the expense of the CS and, consequently, better litter performance.

Zika Virus affects neurobehavioral development, and causes oxidative stress associated to blood-brain barrier disruption in a rat model of congenital infection.

Zika virus (ZIKV) is a mosquito-borne flavivirus associated with several neurodevelopmental outcomes after in utero infection. Here, we studied a congenital ZIKV infection model with immunocompetent Wistar rats, able to predict disabilities and that could pave the way for proposing new effective therapies. We identified neurodevelopmental milestones disabilities in congenital ZIKV animals. Also, on 22nd postnatal day (PND), blood-brain barrier (BBB) proteins disturbances were detected in the hippocampus with immunocontent reduction of ß_Catenin, Occludin and Conexin-43. Besides, oxidative stress imbalance on hippocampus and cortex were identified, without neuronal reduction in these structures. In conclusion, even without pups' microcephaly-like phenotype, congenital ZIKV infection resulted in neurobehavioral dysfunction associated with BBB and oxidative stress disturbances in young rats. Therefore, our findings highlighted the multiple impact of the congenital ZIKV infection on the neurodevelopment, which reinforces the continuity of studies to understand the spectrum of this impairment and to provide support to future treatment development for patients affected by congenital ZIKV.

Dry tobacco leaves: an in vivo and in silico approach to the consequences of occupational exposure.

Exposure of tobacco workers handling dried tobacco leaves has been linked to an increased risk of toxicity and respiratory illness due to the presence of nicotine and other chemicals. This study aimed to evaluate the DNA damage caused by the exposure of tobacco growers during the dry leaf classification process and the relation to cellular mechanisms. A total of 86 individuals participated in the study, divided into a group exposed to dry tobacco (n = 44) and a control group (n = 42). Genotoxicity was evaluated using the alkaline comet assay and lymphocyte micronucleus (MN) assay (CBMN-Cyt), and measurement of telomere length. The levels of oxidative and nitrosative stress were evaluated through the formation of thiobarbituric acid reactive species, and nitric oxide levels, respectively. The inorganic elements were measured in the samples using particle-induced X-ray emission method. The combination of variables was demonstrated through principal component analysis and the interactions were expanded through systems biology. Comet assay, MN, death cells, thiobarbituric acid reactive species, and nitrosative stress showed a significant increase for all exposed groups in relation to the control. Telomere length showed a significant decrease for exposed women and total exposed group in relation to men and control groups, respectively. Bromine (Br) and rubidium (Rb) in the exposed group presented higher levels than control groups. Correlations between nitrate and apoptosis; Br and MN and necrosis; and Rb and telomeres; besides age and DNA damage and death cells were observed. The systems biology analysis demonstrated that tobacco elements can increase the nuclear translocation of NFKB dimers inducing HDAC2 expression, which, associated with BRCA1 protein, can potentially repress transcription of genes that promote DNA repair. Dry tobacco workers exposed to dry leaves and their different agents showed DNA damage by different mechanisms, including redox imbalance.

Polyethylenimine as a Versatile Simultaneous Reducing and Stabilizing Agent Enabling One-Pot Synthesis of Transition-Metal Nanoparticles: Fundamental Aspects and Practical Implications.

The large surface area of metallic nanoparticles provides them with particular optical, chemical, and biological properties, accordingly enabling their use in a wide array of applications. In this regard, facile and fast synthetic approaches are desirable for ready-to-use functional materials. Following early investigations focused on the direct synthesis of polymer-coated gold nanoparticles, we herein demonstrate that such a strategy can be used to manufacture different types of d-block transition-metal nanoparticles via a one-pot method in aqueous media and mild temperature conditions. Gold (Au3+), palladium (Pd2+), and silver (Ag+) ions could be reduced using only polyethylenimine (PEI) or PEI derivatives acting simultaneously as a reducing and stabilizing agent and without the aid of any other external agent. The process gave rise, for instance, to Pd urchin-like nanostructures with a large surface area which confers to them outstanding catalytic performance compared to AuNPs and AgNPs produced using the same strategy. The polymer-stabilized AgNPs were demonstrated to be biocide against a variety of microorganisms, although AuNPs and PdNPs do not hold such an attribute at least in the probed concentration range. These findings may provide significant advances toward the practical, facile, and ready-to-use manufacturing of transition-metal nanoparticles for a myriad of applications.

Effects of calcium chloride substitution on the physicochemical properties of Minas Frescal cheese.

The aim in this research paper was to investigate the effect of using calcium monophosphate (MCP) and MCP mixed with commercial phosphates salts, in total or partial replacement of calcium chloride (CaCl2) in the manufacture of Minas Frescal cheese. Initially, model cheeses were made to perform the rheological analysis during the coagulation process. Of these, the five best treatments were chosen to carry out the production of Minas Frescal cheese, used only CaCl2 and MCP, and partial replacements of MCP + polyphosphate, MCP + potassium monophosphate (MKP) and MCP. The cheeses showed no significant difference in physicochemical composition, yield and syneresis, however, the cheese with partial replacement of CaCl2 by MCP + polyphosphate and MCP + MKP showed the highest hardness values, like the control. This demonstrates that it is possible to replace calcium chloride without significant changes in the physicochemical characteristics and yield of Minas Frescal cheese, and it is still possible to modulate the hardness of the cheese produced according to the type of calcium/phosphate source used. This allows the industry to replace the source of calcium in the manufacture of Minas Frescal cheese according to the desired hardness.

COVID-19 hospitalized patients and oral changes: a case-control study.

OBJECTIVES: To investigate the association between hospitalization for COVID-19 and oral changes, and to evaluate whether oral changes can indicate a higher risk of disease progression to death. MATERIALS AND METHODS: This case-control study analyzed patients hospitalized (university hospital), including those in intensive care unit and clinical wards. The study group comprised 69 COVID-19 positive patients (PCR-test), while the control group included 43 COVID-19 negative patients. A dentist performed oral evaluations, and salivary samples were collected for calcium, phosphatase, and pH analysis. Sociodemographic data, hospitalization information, and hematological test results were collected from electronic-medical records. The presence of oral changes was assessed using chi-square tests, and the predicted risk of death was analyzed using binary logistic regression. RESULTS: COVID-19 positive patients had a significantly higher prevalence of oral changes compared to COVID-19 negative patients. The presence of any oral changes in COVID-19 positive patients indicated a 13-fold higher risk of mortality. "Bleeding ulcers," "pressure ulcers," and "angular cheilitis" were significantly associated with hospitalization for COVID-19. CONCLUSION: There may be an association between hospitalization for COVID-19 and the development of oral changes, including bleeding ulcers, pressure ulcers. and angular cheilitis. These oral changes may serve as potential indicator for disease progression an increased risk of death. CLINICAL RELEVANCE: COVID-19 hospitalized patients have a higher prevalence of oral changes, which indicate an increased risk of mortality. Oral medicine staff should be included in multidisciplinary teams to detect and treat these oral changes promptly.

Establishment and Comprehensive Molecular Characterization of an Immortalized Glioblastoma Cell Line from a Brazilian Patient.

Glioblastoma (GBM) is the most common and aggressive primary brain tumor in adults, with few effective treatment strategies. The research on the development of new treatments is often constrained by the limitations of preclinical models, which fail to accurately replicate the disease's essential characteristics. Herein, we describe the obtention, molecular, and functional characterization of the GBM33 cell line. This cell line belongs to the GBM class according to the World Health Organization 2021 Classification of Central Nervous System Tumors, identified by methylation profiling. GBM33 expresses the astrocytic marker GFAP, as well as markers of neuronal origin commonly expressed in GBM cells, such as ßIII-tubulin and neurofilament. Functional assays demonstrated an increased growth rate when compared to the U87 commercial cell line and a similar sensitivity to temozolamide. GBM33 cells retained response to serum starvation, with reduced growth and diminished activation of the Akt signaling pathway. Unlike LN-18 and LN-229 commercial cell lines, GBM33 is able to produce primary cilia upon serum starvation. In summary, the successful establishment and comprehensive characterization of this GBM cell line provide researchers with invaluable tools for studying GBM biology, identifying novel therapeutic targets, and evaluating the efficacy of potential treatments.

Antileishmanial Activity, Toxicity and Mechanism of Action of Complexes of Sodium Usnate with Lanthanide Ions: Eu(III), Sm(III), Gd(III), Nd(III), La(III) and Tb(III).

Leishmaniases are neglected diseases with limited therapeutic options. Diffuse cutaneous leishmaniasis can occur in Brazil due to Leishmania amazonensis. This study details the antileishmanial activity and cytotoxicity of complexes of sodium usnate (SAU) with lanthanide ions ([LnL3 (H2O)x] (Ln = La(III), Nd(III), Gd(III), Tb(III), Eu(III) and Sm(III); L = SAU). All lanthanide complexes were highly active and more potent than SAU against L. amazonensis promastigotes and intracellular amastigotes (Pro: IC50 < 1.50 µM; Ama: IC50 < 7.52 µM). EuL3·3H2O and NdL3·3H2O were the most selective and effective on intracellular amastigotes, with a selectivity index of approximately 7.0. In silico predictions showed no evidence of mutagenicity, tumorigenicity or irritation for all complexes. Treatment with EuL3·3H2O triggered NO release even at the lowest concentration, indicating NO production as a mechanism of action against the parasite. Incubating promastigotes with the lanthanide complexes, particularly with SmL3·4H2O and GdL3·3H2O, led to a change in the mitochondrial membrane potential, indicating the ability of these complexes to target this essential organelle. The same complexes caused cell death through cell membrane disruption, but their relationship with early or late apoptotic processes remains unclear. Thus, the inclusion of lanthanide ions in SAU improves selectivity with a promising mechanism of action targeting the mitochondria.

Is the regulation by miRNAs of NTPDase1 and ecto-5'-nucleotidase genes involved with the different profiles of breast cancer subtypes?

Breast cancer (BC) is a public health problem worldwide, causing suffering and premature death among women. As a heterogeneous disease, BC-specific diagnosis and treatment are challenging. Ectonucleotidases are related to tumor development and their expression may vary among BC. miRNAs may participate in epigenetic events and may regulate ectonucleotidases in BC. This study aimed to evaluate the expression of ectonucleotidases according to BC subtypes and to predict if there is post-transcriptional regulation of them by miRNAs. MCF 10A (non-tumorigenic), MCF7 (luminal BC), and MDA-MB-231 (triple-negative BC - TNBC) breast cell lines were used and ENTPD1 (the gene encoding for NTPDase1) and NT5E (the gene encoding for ecto-5'-nucleotidase) gene expression was determined. Interestingly, the expression of ENTPD1 was only observed in MCF7 and NT5E was lower in MCF7 compared to MDA-MB-231 cell line. ATP, ADP, and AMP hydrolysis were observed on the surface of all cell lines, being higher in MDA-MB-231. Like qPCR, the activity of AMP hydrolysis was also lower in the MCF7 cells, which may represent a striking feature of this BC subtype. In silico analyses confirmed that the miRNAs miR-101-3p, miR-141-3p, and miR-340-5p were higher expressed in MCF7 cells and targeted NT5E mRNA. Altogether, data suggest that the regulation of NT5E by miRNAs in MCF7 lineage may direct the molecular profile of luminal BC. Thus, we suggest that the roles of ecto-5'-nucleotidase and the aforementioned miRNAs must be unraveled in TNBC to be possibly defined as diagnostic and therapeutic targets.

Synthesis and biological evaluation of 4-hydroxy-methylpiperidinyl-N-benzyl-acylarylhydrazone hybrids designed as novel multifunctional drug candidates for Alzheimer's disease.

The search for new drug candidates against Alzheimer's disease (AD) remains a complex challenge for medicinal chemists due to its multifactorial pathogenesis and incompletely understood physiopathology. In this context, we have explored the molecular hybridization of pharmacophore structural fragments from known bioactive molecules, aiming to obtain a novel molecular architecture in new chemical entities capable of concomitantly interacting with multiple targets in a so-called multi-target directed ligands (MTDLs) approach. This work describes the synthesis of 4-hydroxymethyl)piperidine-N-benzyl-acyl-hydrazone derivatives 5a-l, designed as novel MTDLs, showing improved multifunctional properties compared to the previously reported parent series of N-benzyl-(3-hydroxy)piperidine-acyl-hydrazone derivatives 4. The new improved derivatives were studied in silico, regarding their mode of interaction with AChE enzyme, and in vitro, for evaluation of their effects on the selective inhibition of cholinesterases, cellular antioxidant, and neuroprotective activities as their cytotoxicity in human neuroblastoma (SH-SY5Y) cells. Overall, compound PQM-181 (5 k) showed the best balanced selective and non-competitive inhibition of AChE (IC50 = 5.9 µM, SI > 5.1), with an additional antioxidant activity (IC50 = 7.45 µM) against neuronal t-BOOH-induced oxidative stress and neuroprotective ability against neurotoxicity elicited by both t-BOOH and OAß1-42, and a moderate ability to interfere in Aß1-42 aggregates, with low cytotoxicity and good predictive druggability properties, suggesting a multifunctional pharmacological profile suitable for further drug development against AD.

Neighborhood factors associated with leisure-time physical activity in a Brazilian city.

The purpose of this study was to estimate the association between environmental variables and leisure-time physical activity levels in an adult population of a medium-sized city in Brazil. This is a cross-sectional study with a sample of 965 adults (20-59 years). Individual, socioeconomic, demographic and behavioral data were obtained through a population-based survey. Environmental data were collected through direct observation and subsequently georeferenced. The outcome variable was leisure-time physical activity level, dichotomized as insufficiently active (<150 min/week) and active (≥150 min/week). Physical activity-related facilities were categorized as public or private, and food outlets as healthy, unhealthy or mixed. Monthly per capita income and crime rate by census tract were used to categorize the social environment. Statistical analysis was performed using binary logistic regression (generalized estimating equation models). A direct association was found between the availability of public (adjusted OR = 2.16; 95% CI = 1.44-3.25) and private (adjusted OR = 1.21; 95% CI = 1.02-1.44) physical activity facilities in the census tract and leisure-time physical activity. A greater availability of mixed food establishments was associated with higher leisure-time physical activity levels (adjusted OR = 1.10; 95% CI = 1.004-1.22). Environmental characteristics can contribute together to leisure-time physical activity and could be the focus of policies aimed at promoting physical activity in middle-income countries.

This research estimated how leisure-time physical activity relates to environmental aspects in an adult population of a medium-sized city in Brazil. Nine hundred sixty-five adults (20­59 years) participated in a survey to give individual, socioeconomic, demographic and behavioral information. Researchers also directly observed the environment to collect data and subsequently georeferenced it, to further apply regression models (statistical analysis). Being active during leisure, (i.e. practicing 150 min/week or more), was related to having more public (city squares and outdoor gyms, walking areas and university campus) and private physical activity facilities (clubs, courts, sports halls, soccer fields and gyms), as well as having greater availability of mixed food establishments (restaurants, bakeries, supermarkets and hypermarkets). Therefore, environmental characteristics can contribute together to leisure-time physical activity and could be the focus of policies aimed at promoting physical activity in middle-income countries.

Adapting an evidence-supported optimization program for mental health and sport performance in collegiate athletes to fit youth from ethnic/racial minority and low-income neighborhoods: A National Institutes of Health stage model feasibility study.

The current study addresses the need to empirically develop effective mental health interventions for youth from ethnic/racial minority and low-income neighborhoods. Using Stage Model evaluation methods supported by the National Institutes of Health in the US to address underutilization of mental healthcare among racial/ethnic minority youth, this feasibility study demonstrates empirical adaptation of an innovative sport-specific psychological intervention for use in youth from ethnic/racial minority and low-income neighborhoods. An international group of professionals familiar with sport performance and mental health intervention serving the target population experientially examined the adapted intervention protocols in workshops and provided feedback. Survey results indicated the professionals found the intervention components were easy to administer and likely to be safe, enjoyable, engaging and efficacious for youth mental health and sport performance. The protocols were revised based on feedback from these professionals and the intervention was examined in a case trial involving an Asian American youth who evidenced Social Anxiety Disorder. Case study results indicated the intervention could be implemented with integrity, and severity of psychiatric symptoms and factors interfering with sport performance decreased after intervention implementation. The participant's relationships with family, coaches and teammates were also improved.

Fluorescence lifetime imaging is able to recognize different hematopoietic precursors in unstained routine bone marrow films.

Fluorescence lifetime imaging (FLIM) has been used in living cells to measure metabolic activity and demonstrate cell differentiation. The aim of this study was to investigate whether the FLIM technique could be able to demonstrate cell maturation during myelopoiesis and erythropoiesis in unlabeled routine bone marrow (BM) preparations. Air-dried, unstained smears of BM aspiration samples of 32 patients without BM disease and a normal morphology on May-Grünwald-Giemsa (MGG) stained smears entered the study. FLIM images were captured with a Zeiss LSM 780 NLO multiphoton microscope equipped with a Becker & Hickl SPC-830 TCSPC FLIM module and HPM-100-40 hybrid detector. The samples were irradiated by two-photon excitation at 800 nm with a titanium-sapphire laser of the LSM 780 NLO. FLIM images were compared with those obtained by autofluorescence high resolution imaging. FLIM images of unstained smears were highly contrasted. Different cell types could be easily recognized as they were similar to those seen in MGG stained preparations. Cytoplasm of cells from the erythroid lineage revealed relatively short fluorescence lifetimes due to the presence of hemoglobin, and therefore could easily be distinguished from granulocytic precursors. Nuclear fluorescence lifetimes of all cell types were higher than those of the corresponding cytoplasm. So, FLIM of unstained BM smears obtained under routine real-life conditions permits an easy identification of BM cells, by highlighting differences of their physicochemical properties.

The role of two-component regulatory systems in environmental sensing and virulence in Salmonella.

Adaptation to environments with constant fluctuations imposes challenges that are only overcome with sophisticated strategies that allow bacteria to perceive environmental conditions and develop an appropriate response. The gastrointestinal environment is a complex ecosystem that is home to trillions of microorganisms. Termed microbiota, this microbial ensemble plays important roles in host health and provides colonization resistance against pathogens, although pathogens have evolved strategies to circumvent this barrier. Among the strategies used by bacteria to monitor their environment, one of the most important are the sensing and signalling machineries of two-component systems (TCSs), which play relevant roles in the behaviour of all bacteria. Salmonella enterica is no exception, and here we present our current understanding of how this important human pathogen uses TCSs as an integral part of its lifestyle. We describe important aspects of these systems, such as the stimuli and responses involved, the processes regulated, and their roles in virulence. We also dissect the genomic organization of histidine kinases and response regulators, as well as the input and output domains for each TCS. Lastly, we explore how these systems may be promising targets for the development of antivirulence therapeutics to combat antibiotic-resistant infections.

The virulence of Salmonella Enteritidis in Galleria mellonella is improved by N-dodecanoyl-homoserine lactone.

Salmonella is a food and waterborne pathogen responsible for outbreaks worldwide, and it can survive during passage through the stomach and inside host phagocytic cells. Virulence genes are required for infection and survival in macrophages, and some are under the regulation of the quorum sensing (QS) system. This study investigated the influence of the autoinducer 1 (AI-1), N-dodecanoyl-homoserine lactone (C12-HSL), on the virulence of Salmonella PT4 using Galleria mellonella as an infection model. Salmonella PT4 was grown in the presence and absence of C12-HSL under anaerobic conditions for 7 h, and the expression of rpoS, arcA, arcB, and invA genes was evaluated. After the inoculation of G. mellonella with the median lethal dose (LD50) of Salmonella PT4, the survival of bacteria inside the larvae and their health status (health index scoring) were monitored, as well as the pigment, nitric oxide (NO), superoxide dismutase (SOD), and catalase (CAT) production. Also, the hemocyte viability, the induction of caspase-3, and microtubule-associated light chain 3 (LC3) protein in hemocytes were evaluated. Salmonella PT4 growing in the presence of C12-HSL showed increased rpoS, arcA, arcB, and invA expression and promoted higher larvae mortality and worse state of health after 24 h of infection. The C12-HSL also increased the persistence of Salmonella PT4 in the hemolymph and in the hemocytes. The highest pigmentation, NO production, and antioxidant enzymes were verified in the larva hemolymph infected with Salmonella PT4 grown with C12-HSL. Hemocytes from larvae infected with Salmonella PT4 grown with C12-HSL showed lower viability and higher production of caspase-3 and LC3. Taken together, these findings suggest that C12-HSL could be involved in the virulence of Salmonella PT4.