Gene-corrected p.A30P SNCA patient-derived isogenic neurons rescue neuronal branching and function.

Parkinson's disease (PD) is characterised by the degeneration of A9 dopaminergic neurons and the pathological accumulation of alpha-synuclein. The p.A30P SNCA mutation generates the pathogenic form of the alpha-synuclein protein causing an autosomal-dominant form of PD. There are limited studies assessing pathogenic SNCA mutations in patient-derived isogenic cell models. Here we provide a functional assessment of dopaminergic neurons derived from a patient harbouring the p.A30P SNCA mutation. Using two clonal gene-corrected isogenic cell lines we identified image-based phenotypes showing impaired neuritic processes. The pathological neurons displayed impaired neuronal activity, reduced mitochondrial respiration, an energy deficit, vulnerability to rotenone, and transcriptional alterations in lipid metabolism. Our data describes for the first time the mutation-only effect of the p.A30P SNCA mutation on neuronal function, supporting the use of isogenic cell lines in identifying image-based pathological phenotypes that can serve as an entry point for future disease-modifying compound screenings and drug discovery strategies.

Vibrio Species in an Urban Tropical Estuary: Antimicrobial Susceptibility, Interaction with Environmental Parameters, and Possible Public Health Outcomes.

The genus Vibrio comprises pathogens ubiquitous to marine environments. This study evaluated the cultivable Vibrio community in the Guanabara Bay (GB), a recreational, yet heavily polluted estuary in Rio de Janeiro, Brazil. Over one year, 66 water samples from three locations along a pollution gradient were investigated. Isolates were identified by MALDI-TOF mass spectrometry, revealing 20 Vibrio species, including several potential pathogens. Antimicrobial susceptibility testing confirmed resistance to aminoglycosides, beta-lactams (including carbapenems and third-generation cephalosporins), fluoroquinolones, sulfonamides, and tetracyclines. Four strains were producers of extended-spectrum beta-lactamases (ESBL), all of which carried beta-lactam and heavy metal resistance genes. The toxR gene was detected in all V. parahaemolyticus strains, although none carried the tdh or trh genes. Higher bacterial isolation rates occurred in months marked by higher water temperatures, lower salinities, and lower phosphorus and nitrogen concentrations. The presence of non-susceptible Vibrio spp. was related to indicators of eutrophication and sewage inflow. DNA fingerprinting analyses revealed that V. harveyi and V. parahaemolyticus strains non-susceptible to antimicrobials might persist in these waters throughout the year. Our findings indicate the presence of antimicrobial-resistant and potentially pathogenic Vibrio spp. in a recreational environment, raising concerns about the possible risks of human exposure to these waters.

Not That Close to Mommy: Horizontal Transmission Seeds the Microbiome Associated with the Marine Sponge Plakina cyanorosea.

Marine sponges are excellent examples of invertebrate-microbe symbioses. In this holobiont, the partnership has elegantly evolved by either transmitting key microbial associates through the host germline and/or capturing microorganisms from the surrounding seawater. We report here on the prokaryotic microbiota during different developmental stages of Plakina cyanorosea and their surrounding environmental samples by a 16S rRNA metabarcoding approach. In comparison with their source adults, larvae housed slightly richer and more diverse microbial communities, which are structurally more related to the environmental microbiota. In addition to the thaumarchaeal Nitrosopumilus, parental sponges were broadly dominated by Alpha- and Gamma-proteobacteria, while the offspring were particularly enriched in the Vibrionales, Alteromonodales, Enterobacterales orders and the Clostridia and Bacteroidia classes. An enterobacterial operational taxonomic unit (OTU) was the dominant member of the strict core microbiota. The most abundant and unique OTUs were not significantly enriched amongst the microbiomes from host specimens included in the sponge microbiome project. In a wider context, Oscarella and Plakina are the sponge genera with higher divergence in their associated microbiota compared to their Homoscleromorpha counterparts. Our results indicate that P. cyanorosea is a low microbial abundance sponge (LMA), which appears to heavily depend on the horizontal transmission of its microbial partners that likely help the sponge host in the adaptation to its habitat.

Mitochondria interaction networks show altered topological patterns in Parkinson's disease.

Mitochondrial dysfunction is linked to pathogenesis of Parkinson's disease (PD). However, individual mitochondria-based analyses do not show a uniform feature in PD patients. Since mitochondria interact with each other, we hypothesize that PD-related features might exist in topological patterns of mitochondria interaction networks (MINs). Here we show that MINs formed nonclassical scale-free supernetworks in colonic ganglia both from healthy controls and PD patients; however, altered network topological patterns were observed in PD patients. These patterns were highly correlated with PD clinical scores and a machine-learning approach based on the MIN features alone accurately distinguished between patients and controls with an area-under-curve value of 0.989. The MINs of midbrain dopaminergic neurons (mDANs) derived from several genetic PD patients also displayed specific changes. CRISPR/CAS9-based genome correction of alpha-synuclein point mutations reversed the changes in MINs of mDANs. Our organelle-interaction network analysis opens another critical dimension for a deeper characterization of various complex diseases with mitochondrial dysregulation.

Generation of two iPS cell lines (HIHDNDi001-A and HIHDNDi001-B) from a Parkinson's disease patient carrying the heterozygous p.A30P mutation in SNCA.

Dermal fibroblasts from a patient carrying a heterozygous c.88G > C mutation in the SNCA gene that encodes alpha-synuclein were reprogrammed to pluripotency by retroviruses. This pathogenic mutation generates the p.A30P form of the alpha-synuclein protein leading to autosomal dominantly inherited Parkinson's disease (PD). Two clonal iPS cell lines were generated (A30P-3 and A30P-4) and characterised by validating the silencing of viral transgenes, the expression of endogenous pluripotency genes, directed differentiation into three germ layers in-vitro and a stable molecular genotype. These iPSC lines will serve as a valuable resource in determining the role of the p.A30P SNCA mutation in PD pathogenesis.

Isolation and prospection of diazotrophic rhizobacteria associated with sugarcane under organic management.

Microorganisms associated with organic management are essential in nutrient transformation and release for plant use. The present study aimed to isolate, identify and characterize plant growth promoting diazotrophic rhizobacteria associated with sugarcane under organic management. Rhizospheres of organic sugarcane varieties IAC 911099 and CTC4 were sampled and inoculated onto nitrogen free NFb and Burk media. The isolated microorganisms were screened in vitro concerning their ability to produce plant growth promoting factors. Eighty-one bacteria were isolated; 45.6% were positive for the nifH gene and produced at least one of the evaluated plant growth promotion factors. The production of indole-3-acetic acid was observed in 46% of the isolates, while phosphate solubilization was observed in 86.5%. No isolates were hydrogen cyanide producers, while 81% were ammonia producers, 19% produced cellulases and 2.7%, chitinases. Microorganisms belonging to the Burkholderia genus were able to inhibit Fusarium moniliforme growth in vitro. Plant growth promoting microorganisms associated with organic sugarcane, especially belonging to Burkholderia, Sphingobium, Rhizobium and Enterobacter genera, can be environmentally friendly alternatives to improve sugarcane production.

Atitude do Enfermeiro Relativamente à Sexualidade dos Adolescentes / Nurses' Attitude Regarding Adolescents' Sexuality

O presente trabalho teve como objetivo geral conhecer as atitudes que os enfermeiros a trabalhar em Cuidados de Saúde Primários têm relativamente à sexualidade dos adolescentes. Para tal, recorreu-se a uma entrevista semiestruturada a 49 enfermeiros a exercerem em 13 Centros de Saúde da Unidade Local de Saúde da Guarda, com uma idade mínima de 21 anos e uma máxima de 54 anos, correspondendo-lhe uma idade média de 43,54 anos (±8,33 anos). Quanto ao tempo de serviço, obteve-se um tempo de serviço médio de 20,49 anos (±6,512 anos). Grosso modo, o tempo de serviço corresponde ao tempo de serviço no atual Centro de Saúde. Os resultados revelaram que os enfermeiros apresentam uma atitude muito positiva em relação às questões de sexualidade dos adolescentes. A maioria referiu que os adolescentes não abordam por vontade própria as questões afetivas ou relacionais, o que, segundo alguns entrevistados, se poderá dever ao facto de este ser um assunto que, por norma, é partilhado com os pares e não com os profissionais de saúde, ou porque os seus relacionamentos são maioritariamente esporádicos e transitórios e não efetivos. Referiram também que há falta de maturidade em questões de sexualidade por parte dos adolescentes, independentemente do sexo. A esmagadora maioria dos enfermeiros referiu que são poucos os adolescentes que procuram o Centro de Saúde por questões de índole sexual e os que o fazem são motivados pela procura de métodos contracetivos e por gravidez indesejada. Sobressaiu que o grupo etário que mais procura os serviços de saúde por problemas de índole sexual é o de adolescentes maiores de 15 e menores de 18 anos, sobretudo do sexo feminino. A maioria dos enfermeiros considera que as crenças e os valores veiculados pela sociedade influenciam a liberdade sexual dos adolescentes, bem como as fontes de informação, sobretudo a internet, levando-os a procurar mais os enfermeiros por motivos de contraceção. São de opinião de que não deve haver um estereótipo de idade para o início da vida sexual, que esta deve ter início quando o adolescente se sentir preparado, não obstante a necessidade de experimentação própria da idade. Foram unânimes ao considerarem que a maior parte das vezes os adolescentes iniciam a sua vida sexual por uma questão de pressão social e porque outros já o fizeram com idades anteriores à sua. A esmagadora maioria dos enfermeiros sente-se confortável em abordar questões de natureza sexual com os adolescentes, alegando que esta abordagem requer uma adaptação das intervenções para satisfazer as necessidades de cada adolescente. A generalidade dos enfermeiros afirmou que a promoção da saúde sexual na adolescência contribui para o bem-estar global do adolescente no presente e no futuro.

This study aimed to know the attitudes that nurses working in primary health care have regarding adolescent sexuality. For this, we used a semistructured interview with 49 nurses to exercise in 13 Health Centers Health in the Local Health Unit of the Guarda, with a minimum age of 21 years and a maximum of 54 years, corresponding to an average age of 43.54 years (±8.33 years). As to the time, we obtained an average service time of 20.49 years (±6.512 years). Roughly speaking, the service time is the time of service in the current Health Centre. The results showed that nurses have a very positive attitude towards sexuality issues of adolescents. Most said that teenagers do not address by will own the affective and relational issues, which, according to some respondents, may be due to the fact that this is a subject that, as a rule, is shared with peers and not with the professionals health, or because their relationships are mostly sporadic and transient and ineffective. Also mentioned that there is a lack of maturity in sexuality issues by adolescents, regardless of gender. The overwhelming majority of nurses said that few teenagers seeking health center for sexual issues and those that do are motivated by the demand for contraception and unwanted pregnancy. It stood out that the age group that seeks health care for sexual problems is to teenagers over 15 and under 18, especially women. Most nurses believes that the beliefs and the values conveyed by society influence the sexual freedom of adolescents, as well as the sources of information, especially the internet, leading them to seek more nurses for contraceptive reasons. Are of the opinion that there should be an age stereotype to the onset of sexual life, this should begin when the adolescent feel prepared, despite the need to own the old trial. They were unanimous in considering that most of the time the teens start their sexual life as a matter of social pressure and because others have done with previous ages to yours. The overwhelming majority of nurses feel comfortable in addressing sexual issues with teenagers, claiming that this approach requires an adaptation of interventions to meet the needs of each adolescent. The majority of nurses said that promoting sexual health in adolescence contributes to the adolescent's overall well-being now and in the future

HCV viremia drives an increment of CD86 expression by myeloid dendritic cells.

The host immune response, including innate and adaptive immunity, plays a critical role in determining the outcome of viral infection. Nevertheless, little is known about the exact reasons for the failure of the host immune system in controlling hepatitis C virus (HCV) infection. Impairment of dendritic cells (DCs) function is probably one of the mechanisms responsible for immune evasion of HCV. In this study, the frequency and phenotype of DCs subsets were analyzed in three groups: HCV-infected individuals who developed viral persistence (1), HCV-infected individuals who spontaneously cleared the virus (2) and HCV-seronegative uninfected subjects (3). The results showed that the frequency of DCs subsets was not statistically significant between groups. Plasmacytoid DCs circulating exhibited an immature phenotype characterized by low expression of CD86. On the other hand, CD86 expression in myeloid DCs was significantly higher in chronic infected individuals compared to healthy controls (P=0.037). A positive correlation was observed between CD86(+) myeloid DC (mDC) and HCV viral load (r=0.4121, P=0.0263). These results suggest that HCV did not have an inhibitory effect on mDC maturation and the HCV viremia drives the increase of CD86 expression in mDC. The regulation of DCs maturation and migration lies at the level of intracellular signaling. HCV can activate or block intracellular signaling pathways and alter DC function. In conclusion, the present study suggests that imbalance of DC maturation by the virus represents a mechanism of evasion of the immune system despite the fact that HCV viremia appears to exert a "stimulatory" effect on cell-surface immune phenotype.

Lower numbers of circulating Natural Killer T (NK T) cells in individuals with human T lymphotropic virus type 1 (HTLV-1) associated neurological disease.

Human T lymphotropic virus type 1 (HTLV-1) infects 10-20 million people worldwide. The majority of infected individuals are asymptomatic; however, approximately 3% develop the debilitating neurological disease HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). There is also currently no cure, vaccine or effective therapy for HTLV-1 infection, and the mechanisms for progression to HAM/TSP remain unclear. NK T cells are an immunoregulatory T cell subset whose frequencies and effector functions are associated critically with immunity against infectious diseases. We hypothesized that NK T cells are associated with HAM/TSP progression. We measured NK T cell frequencies and absolute numbers in individuals with HAM/TSP infection from two cohorts on two continents: São Paulo, Brazil and San Francisco, CA, USA, and found significantly lower levels when compared with healthy subjects and/or asymptomatic carriers. Also, the circulating NK T cell compartment in HAM/TSP subjects is comprised of significantly more CD4(+) and fewer CD8(+) cells than healthy controls. These findings suggest that lower numbers of circulating NK T cells and enrichment of the CD4(+) NK T subset are associated with HTLV-1 disease progression.