Evaluation of berberine nanoparticles as a strategy to modulate acetylcholinesterase activity.

Researchers have concentrated efforts in the search for natural-based reversible inhibitors for cholinesterase enzymes as they may play a key role in the treatment of degenerative diseases. Diverse plant alkaloids can inhibit the action of acetylcholinesterase and, among them, berberine is a promising bioactive. However, berberine has poor water solubility and low bioavailability, which makes it difficult to use in treatment. The solid dispersion technique can improve the water affinity of hydrophobic substances, but berberine solid dispersions have not been extensively studied. Safety testing is also essential to ensure that the berberine-loaded solid dispersions are safe for use. This study investigated the effectiveness of berberine-loaded solid dispersions (SD) as inhibitors of acetylcholinesterase enzyme (AChE). Docking simulation was used to investigate the influence of berberine on AChE, and in vitro assays were conducted to confirm the enzymatic kinetics of AChE in the presence of berberine. Berberine SD also showed improved cytotoxic effects on tumoral cells when dispersed in aqueous media. In vivo assays using Allium cepa were implemented, and no cytotoxicity/genotoxicity was found for the berberine solid dispersion. These results suggest that berberine SD could be a significant step towards safe nanostructures for use in the treatment of neurodegenerative diseases.

Relationship between Biofilm Production and High Somatic Cell Count in Streptococcus agalactiae Isolated from Milk of Cows with Subclinical Mastitis.

Streptococcus agalactiae (S. agalactiae) is one of the main agents that causes mastitis in dairy cows, mainly inducing the subclinical form, which is characterized by a high somatic cell count (SCC). The aim of this study was to correlate the increase in SCC caused by S. agalactiae in cows with subclinical mastitis to the presence of genes related to adhesion and invasion in bovine mammary epithelial cells (BMEC) and biofilm formation. Considering the 145 isolates tested, 57.2% presented the capsular type Ia and 42.8% presented type III. We identified the virulence genes among the isolates and determined nine genetic profiles. The most common profile was identified in 69 isolates (47.5%): Ia, fbsA+, fbsB-, pI1-, pI2a-, pI2b+, and hylb+. All isolates produced biofilm, with 58.6% classified as strong producers, 29% as moderate producers and 12.4% as weak producers. No statistical correlation was found between the presence of virulence genes and increased SCC or biofilm production. However, biological evidence was observed between increased SCC and biofilm production. One isolate from each profile was randomly subjected to adhesion and invasion assays, and all of them adhered to BEMC, but none were able to invade. Our results showed that different genetic profiles do not provide advantages for bacteria to invade BMEC in vitro. In addition, biofilm production appears to be related to high SCC.

Genetic and Antimicrobial Resistance Profiles of Mammary Pathogenic E. coli (MPEC) Isolates from Bovine Clinical Mastitis.

Mammary pathogenic E. coli (MPEC) is one of the main pathogens of environmental origin responsible for causing clinical mastitis worldwide. Even though E. coli are strongly associated with transient or persistent mastitis and the economic impacts of this disease, the virulence factors involved in the pathogenesis of MPEC remain unknown. Our aim was to characterize 110 MPEC isolates obtained from the milk of cows with clinical mastitis, regarding the virulence factor-encoding genes present, adherence patterns on HeLa cells, and antimicrobial resistance profile. The MPEC isolates were classified mainly in phylogroups A (50.9%) and B1 (38.2%). None of the isolates harbored genes used for diarrheagenic E. coli classification, but 26 (23.6%) and 4 (3.6%) isolates produced the aggregative or diffuse adherence pattern, respectively. Among the 22 genes investigated, encoding virulence factors associated with extraintestinal pathogenic E. coli pathogenesis, fimH (93.6%) was the most frequent, followed by traT (77.3%) and ompT (68.2%). Pulsed-field gel electrophoresis analysis revealed six pulse-types with isolates obtained over time, thus indicating persistent intramammary infections. The genes encoding beta-lactamases detected were as follows: blaTEM (35/31.8%); blaCTX-M-2/blaCTX-M-8 (2/1.8%); blaCTX-M-15 and blaCMY-2 (1/0.9%); five isolates were classified as extended spectrum beta-lactamase (ESBL) producers. As far as we know, papA, shf, ireA, sat and blaCTX-M-8 were detected for the first time in MPEC. In summary, the genetic profile of the MPEC studied was highly heterogeneous, making it impossible to establish a common genetic profile useful for molecular MPEC classification. Moreover, the detection of ESBL-producing isolates is a serious public health concern.

Short communication: Association between the accessory gene regulator (agr) group and the severity of bovine mastitis caused by Staphylococcus aureus.

Staphylococcus aureus can elicit mild to more severe degrees of mastitis in cattle, depending on the response of the host's immune system and the virulence factors of the specific isolate. Several virulence factors are controlled by a global regulatory system, designated accessory gene regulator (agr). Thus, the objective was to examine associations between different capsular and agr types and the severity of bovine mastitis caused by S. aureus. All isolates were obtained from bovine subclinical (n = 50), mild clinical (n = 73), and moderate clinical mastitis cases (n = 28). Isolates containing the agrI gene and lacking the agr locus (agr-) were more prevalent among subclinical than clinical mastitis cases, whereas isolates containing the agrII and agrIII genes were more prevalent among clinical mastitis cases. The capsular types 5 (cap5) and 8 (cap8) were found in 42 and 44%, respectively, of the isolates obtained from subclinical cases and in 38.6 and 58.4%, respectively, of those isolated from clinical mastitis cases. Capsular type was not associated with type of mastitis (subclinical, mild clinical, or moderate clinical). We found a strong association between agr type and type of mastitis, suggesting that knowledge of S. aureus genetic profiles could be an additional tool to control this disease.

First investigation of Staphylococcus argenteus in a Brazilian collections of S. aureus isolated from bovine mastitis.

BACKGROUND: Staphylococcus argenteus is a new specie positive coagulase staphylococci. We investigate the presence of S. argenteus in isolates previously classified as S. aureus, obtained from the milk of cows with mastitis in Brazil. RESULTS: Among 856 S. aureus tested in chocolate agar, tryptone soya agar and salt egg yolk agar, white or colorless colonies were observed in 185 (21.6%) isolates. Regarding the ctrOPQMN operon, 111 (60%) presented the complete cluster. Despite some missing genes in this cluster, the remaining strains (74) were confirmed as S. aureus using the nrps gene. CONCLUSIONS: As far as we know, this is the first review of S. aureus collection in Brazil and S. argenteus does not appear to be a significant problem in Brazilian herds.

Impacts of the COVID-19 pandemic on violence against women: reflections from the theory of human motivation from Abraham Maslow / Impactos de la pandemia de COVID-19 en la violencia contra las mujeres: reflexiones a partir de la teoría de la motivación humana de Abraham Maslow / Impactos da pandemia de COVID-19 na violência contra a mulher: reflexões a partir da teoria da motivação humana de Abraham Maslow

Introduction: The pandemic and social isolation scenario caused by the new Coronavirus (COVID-19) can directly affect the prevalence of domestic violence against women in Brazil. Objective: To reflect on the impacts of the COVID-19 pandemic on violence against women, through an analysis over the human motivation theory of Abraham Maslow. Method: This is a reflective study with documentary analysis carried out in June 2020, whose theoretical framework was Abraham Maslow's theory of basic human needs. Results: Hierarchical levels were described according to the Maslow pyramid and a parallel was drawn with data on violence against women before and during the COVID-19 pandemic period. Conclusion: The paths to services related to gender violence must be updated to reflect changes in available health services. The present study demonstrated that the COVID-19 pandemic can affect women who experience domestic violence at all hierarchical levels in the Maslow pyramid.

Introducción: El escenario de pandemia y aislamiento social causado por el nuevo Coronavirus (COVID-19) puede afectar directamente la prevalencia de la violencia doméstica contra las mujeres en Brasil. Objetivo: reflexionar sobre los impactos de la pandemia COVID-19 sobre la violencia contra las mujeres mientras an analysis de la Teoría de la Motivación Humana de Abraham Maslow. Método: Este es un estudio reflexivo con análisis documental realizado en junio de 2020, cuyo marco teórico fue la teoría de las necesidades humanas básicas de Abraham Maslow. Resultados: los niveles jerárquicos se describieron de acuerdo con la pirámide de Maslow y se trazó un paralelo con datos sobre la violencia contra las mujeres antes y durante el período de la pandemia COVID-19. Conclusión: los caminos hacia los servicios relacionados con la violencia de género deben actualizarse para reflejar los cambios en los servicios de salud disponibles. El presente estudio demostró que la pandemia de COVID-19 puede afectar a las mujeres que sufren violencia doméstica en todos los niveles jerárquicos de la pirámide de Maslow.  

Introdução: O cenário de pandemia e isolamento social causado pelo novo Coronavírus (COVID-19) pode afetar diretamente a prevalência da violência doméstica contra a mulher no Brasil.  Objetivo: Refletir acerca dos impactos da pandemia de COVID-19 na violência contra a mulher, a partir da análise da teoria da motivação humana de Abraham Maslow. Método: Trata-se de um estudo do tipo reflexivo com análise documental realizada no mês de junho de 2020, cujo referencial teórico foi a teoria das necessidades humanas básicas de Abraham Maslow. Resultados: Foram descritos os níveis hierárquicos segundo a pirâmide de Maslow e traçado um paralelo com os dados de violência contra a mulher antes e durante o período da pandemia de COVID-19. Conclusão: Os caminhos para serviços referência à violência baseada em gênero devem ser atualizados para refletir as mudanças nas estruturas de acolhimento disponíveis. O presente estudo demonstrou que a pandemia da COVID-19 pode afetar a mulher que sofre violência doméstica em todos os níveis hierárquicos da pirâmide de Maslow.

Biofilm production under different atmospheres and growth media by Streptococcus agalactiae isolated from milk of cows with subclinical mastitis.

Different methods to analyze Streptococcus agalactiae biofilm formation have been investigated, but standardized protocols have not been developed. We compared S. agalactiae biofilm production among different atmospheres and growth media. Biofilm formation was studied in 32 isolates from bovine mastitis cases grown in Tryptone Soy Broth (TSB), Todd Hewitt Broth (THB), Luria Bertani Broth (LB) and Brain Heart Infusion (BHI), under two atmospheres, aerobic and 5% CO2. Regardless of the culture medium, growth under 5% CO2 resulted in a greater proportion of biofilm formation (65.63%), as compared with aerobic conditions (39.84%). Regardless of the atmosphere, the chances of biofilm formation were greater for isolates grown in TSB, as compared with THB [Odds ratio (OR) = 3.02], BHI (OR = 4.57), or LB (OR = 10.20). Thus, we suggest the use of 5% CO2 atmosphere and TSB in biofilm formation assays by Group-B streptococci (GBS) isolated from intramammary infections.

Efficacy of cefquinome and a combination of cloxacillin and ampicillin for treatment of dairy cows with Streptococcus agalactiae subclinical mastitis.

A randomized clinical trial was conducted to assess efficacy of intramammary cloxacillin and ampicillin (CLOXIMM), intramammary cefquinome (CEFIMM), and intramuscular cefquinome (CEFIM) to treat Streptococcus agalactiae intramammary infections (Trial 1). Subsequently, two treatment groups were extended to assess whether CLOXIMM was not inferior to CEFIMM (Trial 2). Nine farms were included in the study. Milk samples were collected from all quarters of all lactating cows for microbiological identification of S. agalactiae. Positive cows were randomly allocated into four groups: CLOXIMM, CEFIMM, CEFIM, or negative control (CONTROL). Study outcomes were bacteriological cure at 14 (CURE14), 21 (CURE21), and 14 and 21 (CURE1421) days after treatment onset, and somatic cell count. Logistic regression was used to estimate the odds of cure between each treatment and CONTROL. Non-inferiority analysis was performed considering a one-sided 95% confidence interval (CI) and non-inferiority margins (Δ) of 0.10, 0.15, 0.20, and 0.25. Adjusted S. agalactiae bacteriological cure for CLOXIMM, CEFIMM, CEFIM, and CONTROL was 86, 98, 55, and 25% at day 14; 82, 93, 52, and 0% at day 21; and 82, 92, 40, and 0% at days 14 and 21, respectively. Treatment with CLOXIMM and CEFIMM resulted in greater bacteriological cure rates, as compared with CEFIM or CONTROL, which does not justify the use of CEFIM in S. agalactiae eradication programs. The CURE14 difference between CEFIMM and CLOXIMM was of 12.1 percentage points (95% CI: 0.056-0.184). CLOXIMM was considered not inferior to CEFIMM for Δ = 0.20 or 0.25 and inconclusive for Δ = 0.10 or 0.15. Thus, it should be pondered by veterinarians whether an expected 12.1 (5.6-18.4) percentage points increase in cure rate would justify the use of a fourth-generation cephalosporin, as opposed to a combination of traditional IMM drugs (cloxacillin and ampicillin) to treat S. agalactiae subclinical mastitis.

Cross-Contamination and Biofilm Formation by Salmonella enterica Serovar Enteritidis on Various Cutting Boards.

Cross-contamination is one of the main factors related to foodborne outbreaks. This study aimed to analyze the cross-contamination process of Salmonella enterica serovar Enteritidis from poultry to cucumbers, on various cutting board surfaces (plastic, wood, and glass) before and after washing and in the presence and absence of biofilm. Thus, 10 strains of Salmonella Enteritidis were used to test cross-contamination from poultry to the cutting boards and from thereon to cucumbers. Moreover, these strains were evaluated as to their capacity to form biofilm on hydrophobic (wood and plastic) and hydrophilic materials (glass). We recovered the 10 isolates from all unwashed boards and from all cucumbers that had contacted them. After washing, the recovery ranged from 10% to 100%, depending on the board material. In the presence of biofilm, the recovery of salmonellae was 100%, even after washing. Biofilm formation occurred more on wood (60%) and plastic (40%) than glass (10%) boards, demonstrating that bacteria adhered more to a hydrophobic material. It was concluded that the cutting boards represent a critical point in cross-contamination, particularly in the presence of biofilm. Salmonella Enteritidis was able to form a biofilm on these three types of cutting boards but glass showed the least formation.

Development of a rapid and sensitive immunosensor for the detection of bacteria.

The presence of Gram-positive bacteria in foodstuffs is a chronic worldwide problem. Here, we present a cheap and simple colorimetric method for the detection of Lactobacillus species (spp.) and Staphylococcus aureus (S. aureus) using gold nanoparticles (AuNP) modified with monoclonal anti-Gram-positive bacteria to produce an immune-sensor. Detection is based on the fact that antibody-conjugated AuNPs can readily identify Gram-positive bacteria through antibody-antigen recognition, which results in a color change of AuNPs upon aggregation. The detection limit was 105CFU/ml in pure culture for Lactobacillus spp. and 120CFU/ml in pure culture for S. aureus. The method was applied successfully for detection of bacteria in samples of sugar cane, and agreed well with values obtained using other methods. These results suggested that the detection system could be used for the quantitative analysis of Gram-positive bacteria and might be applied potentially by the food industry.

Hygienic-sanitary quality of Minas fresh cheese sold in the city of Botucatu, São Paulo / Avaliação higiênico-sanitária de queijos Minas frescal comercializados na cidade de Botucatu, São Paulo

The Minas fresh cheese is a fresh and moist cheese and, therefore, has a short shelf-life. The use of raw milk as the main feedstock, the absence of ripening and the facility of contamination are factors that can compromise the product's microbiological quality. The aim of this study was to describe the hygienic-sanitary quality of 50 Minas fresh cheeses marketed in Botucatu city, São Paulo, Brazil, close to their production date, and another 50 units from the same lot exactly on the expiration date, according to the Brazilian Sanitary Surveillance Agency (ANVISA). We also searched for enterotoxins genes in Staphylococcus aureus and its in vitro expression. In 36% of the first analyzed samples, the count of coliforms at 45ºC was above the limit and, as for the second analysis, 44%. Regarding coagulase positive staphylococci, 10 (20%) samples showed concentration above the permitted by law next to the production date and 14% on the expiration date. Salmonella was only observed in one sample analyzed near the date of production, while L. monocytogenes only in one sample analyzed on the expiration day. We isolated three enterotoxigenic strains of S. aureus that produced Staphylococcal Enterotoxin B (SEB) and Staphylococcal Enterotoxin C (SEC) in vitro, highlighting the importance of proper storage of this product due to its potential to cause intoxication. Overall, the quality of Minas fresh cheese is still unsatisfactory, leading to risks to consumers' health.(AU)

O queijo Minas frescal é muito apreciado no Brasil. Por ser fresco e úmido, seu tempo de prateleira é curto. A utilização de leite cru como matéria prima, a não maturação do queijo e a facilidade de contaminação durante seu processamento são fatores que comprometem a qualidade microbiológica do produto. Este trabalho verificou a qualidade higiênico-sanitária de 50 queijos Minas frescal comercializados na cidade de Botucatu, São Paulo, Brasil, em datas próximas a sua fabricação e na data de expiração da validade, utilizando os parâmetros propostos pela RDC nº 12 da Agência Nacional de Vigilância Sanitária (ANVISA). Cepas de S. aureus foram submetidas à pesquisa de genes codificadores de produção de enterotoxinas clássicas e, também, sua produção in vitro. Encontraram-se coliformes a 45ºC, acima do permitido por legislação, em 36% das amostras durante a primeira análise e, na segunda, em 44%. Quanto à contagem de estafilococos coagulase positiva, 10% não atendiam ao padrão exigido por lei na data de produção, e 14%, na data de expiração da validade. Salmonella foi apenas observada em uma amostra próxima à data de produção, enquanto L. monocytogenes somente em uma amostra na data de expiração da validade. Observou-se produção de Staphylococcal Enterotoxin B (SEB)e Staphylococcal Enterotoxin C (SEC) in vitro por três cepas de S. aureus. Ressalta-se, em decorrência disso, a importância de armazenar corretamente esse alimento, por seu potencial de causar intoxicação. Concluiu-se que a qualidade higiênico-sanitária desses queijos Minas frescais foi insatisfatória, implicando riscos para a saúde do consumidor.(AU)

Presence of Immune Evasion Cluster and Molecular Typing of Methicillin-Susceptible Staphylococcus aureus Isolated from Food Handlers.

The hands and noses of food handlers colonized by Staphylococcus aureus are an important source of food contamination in restaurants and food processing. Several virulence factors can be carried by mobile elements in strains of S. aureus, including the immune evasion cluster (IEC). This gene cluster improves the capacity of S. aureus to evade the human immune response. Many studies have reported the transmission of strains between animals and humans, such as farm workers that have close contact with livestock. However, there are few studies on the transmission between food and food handlers. The aim of this study was to detect the IEC and the mecA gene in strains isolated from food handlers and to type these strains using the spa typing method. Thirty-five strains of S. aureus isolated from the noses and hands of food handlers in three different kitchens were analyzed for the presence of the mecA gene and IEC and by spa typing. All strains were negative for the mecA gene, and the presence of IEC was observed in 10 (28.6%) strains. Fifteen different spa types were observed, with the most frequent being t127 (42.85%) and t002 (11.42%). Strains from the two most prevalent spa types and a novel spa type were typed by multilocus sequence typing. spa types t127, t002, and t13335 were determined to be multilocus sequence types (ST) ST-30, ST-5, and ST-45, respectively. The food handlers may have been contaminated by these strains of S. aureus through food, which is suggested by the low frequency of IEC and by ST that are observed more commonly in animals.

Posterior Reversible Encephalopathy Syndrome: Clinical Differences in Patients with Exclusive Involvement of Posterior Circulation Compared to Anterior or Global Involvement.

INTRODUCTION: Posterior reversible encephalopathy syndrome (PRES) is a clinical-radiologic syndrome not yet fully understood and characterized by transient neurologic symptoms in addition to typical radiological findings. There are only a few articles that describe the clinical differences between patients with PRES that involve carotid and vertebrobasilar circulations. Our study aims to further evaluate the differences between predominantly anterior and posterior circulation PRES. METHODS: We review 54 patients who had received the diagnosis of PRES from 2009 to 2015. The patients were divided into 2 groups: (1) exclusively in posterior zones; and (2) anterior plus posterior zones or exclusively anterior zones. Several clinical characteristics were evaluated, including the following: age, sex, previous diseases, the neurologic manifestations, the highest blood pressure in the first 48 hours of presentation, highest creatinine level during symptoms, and the neuroimaging alterations in brain magnetic resonance imaging. RESULTS: Mean age at diagnosis was 28.5 years old (9 men and 45 women) and mean systolic blood pressure among patients with lesions only in posterior zones was 162.1 mmHg compared to 179.2 mmHg in the anterior circulation. The most common symptoms in the 2 groups were headache and visual disturbances. DISCUSSION: PRES may have several radiological features. A higher blood pressure seems to be 1 of the factors responsible for developing widespread PRES, with involvement of carotid vascular territory. This clinical-radiological difference probably occurs because of the larger number of autonomic receptors in the carotid artery in comparison to the vertebral-basilar system.

Efeito de clorexidina em gel ou dentifrício nos níveis salivares de estreptococos do grupo mutans em crianças do distrito do Saí(São Francisco do Sul, SC, Brasil) / Effect of chlorhexidine gel or toothpaste in the salivary count of mutans streptococciin children of Saí district (São Francisco do Sul, SC, Brazil)

Introdução: A cárie dentária é a doença mais prevalente no ser humano, sendo caracterizada pela destruição progressiva dos dentes afetados. Para que ocorra são necessários um hospedeiro susceptível,uma microbiota bucal cariogênica, principalmente os estreptococos do grupo mutans (EGM), e um substrato adequado, os quais devem estar presentes por um período de tempo suficiente. Dos agentes antimicrobianos disponíveis para uso dentário, a clorexidina é um dos mais eficazes em reduzir a proporção de alguns microrganismos,especialmente os EGMs. Objetivo: O objetivo do estudo foi comparar aeficácia de duas formas de clorexidina a 1% (gel e dentifrício) aplicadas em escolares, sob escovação supervisionada, fora do ambiente deconsultório odontológico, na redução dos níveis salivares de EGM.Material e métodos: A amostra foi composta de 30 crianças e separada em dois grupos: grupo A (dentifrício) e grupo B (gel). As escovaçõesforam realizadas uma vez por semana durante quatro semanas consecutivas. Anteriormente à primeira escovação e 30 minutos após a escovação da quarta semana, realizou-se a coleta salivar. Foram preparadas diluições seriadas decimais das amostras de saliva atéa diluição 10-3. Amostras das diluições foram semeadas em meio de cultivo ágar Mitis Salivarius bacitracina sacarose (MSBS), seletivo aocrescimento de EGM. Resultados e conclusão: Após contagem das unidades formadoras de colônia de EGM, foi realizada a média das quadruplicatas para obter o valor de crescimento. Apenas o grupo dentifrício apresentou redução nos níveis salivares de EGM, não sendo estatisticamente significativa pelo teste t de Student (alfa = 0,05).

Introduction: Dental caries is the most prevalent disease affecting humans and it is characterized by the progressive destruction ofthe affected teeth. A susceptible host, a cariogenic oral microflora(mainly mutans streptococci) and a suitable substrate are necessary conditions for the development of caries. Chlorhexidine is one of the most effective antimicrobial agents available for dental use in reducing the proportion of some microorganisms, especially the mutans streptococci. Objective: The aim of this study was to compare theeffectiveness of two manipulated forms of chlorhexidine 1% (gel andtoothpaste) in reducing the salivary levels of mutans streptococci,which are used by schoolchildren in supervised toothbrushing, outsidethe dental clinic. Material and methods: The sample of 30 childrenwas divided into two groups: group A (toothpaste) and group B (gel).Applications were performed once a week for 4 weeks. Saliva samples were collected prior to the first brushing and 30 minutes after the lastone. Serial decimal dilutions of saliva samples were prepared up to10-3 dilution, and these samples were inoculated in Mitis Salivarius bacitracin sucrose agar, selective for growth of mutans streptococci.Results and conclusion: After counting the colony forming units of mutans streptococci, the average of quadruplicates was calculated so to obtain the growth value. Only the toothpaste group showed adecrease of the salivary levels of mutans streptococci although it wasnot statistically significant by Student t test (alpha = 0.05).