Relationship between clinical-epidemiological parameters and outcomes of patients with COVID-19 admitted to the intensive care unit: a report from a Brazilian hospital.

Background: People in low-income countries, especially those with low socio-economic conditions, are likelier to test positive for SARS-CoV-2. The unequal conditions of public health systems also increase the infection rate and make early identification and treatment of at-risk patients difficult. Here, we aimed to characterize the epidemiological profile of COVID-19 patients in intensive care and identify laboratory and clinical markers associated with death. Materials and methods: We conducted an observational, descriptive, and cross-sectional study in a reference hospital for COVID-19 treatment in the Southern Region of Bahia State, in Brazil, to evaluate the epidemiological, clinical, and laboratory characteristics of COVID-19 patients admitted to the intensive care unit (ICU). Additionally, we used the area under the curve (AUC) to classify survivors and non-survivors and a multivariate logistic regression analysis to assess factors associated with death. Data was collected from the hospital databases between April 2020 and July 2021. Results: The use of bladder catheters (OR 79.30; p < 0.0001) and central venous catheters (OR, 45.12; p < 0.0001) were the main factors associated with death in ICU COVID-19 patients. Additionally, the number of non-survivors increased with age (p < 0.0001) and prolonged ICU stay (p < 0.0001). Besides, SAPS3 presents a higher sensibility (77.9%) and specificity (63.1%) to discriminate between survivors and non-survivor with an AUC of 0.79 (p < 0.0001). Conclusion: We suggest that multi-laboratory parameters can predict patient prognosis and guide healthcare teams toward more assertive clinical management, better resource allocation, and improved survival of COVID-19 patients admitted to the ICU.

Cupuassu juice fermentation by Lactiplantibacillus plantarum Lp62 produces an antioxidant enriched probiotic beverage.

The present work aimed to produce a cupuassu juice (Theobroma grandiflorum) fermented by the probiotic bacterium Lactiplantibacillus plantarum Lp62 and to analyze its antioxidant potential, antimicrobial activity, and resistance to biological barriers. The fermented beverage showed an increase in the content of phenolics, flavonoids, and antioxidant potential. The culture showed antagonistic activity against pathogens, but this result was not observed when the juice was tested. The probiotic strain remained viable under refrigeration, even in an acidified environment, and survived simulated gastrointestinal transit in vitro. L. plantarum Lp62 showed 30% adherence to HT-29 intestinal cells and proved to be safe in terms of antibiotic resistance and production of virulence factors. Fermentation increased the functional characteristics of cupuassu juice. This drink proved to be a good vehicle for the delivery of the probiotic bacteria L. plantarum Lp62.

In vitro selection and characterization of probiotic properties in eight lactobacillus strains isolated from cocoa fermentation.

Traditionally, probiotic microorganisms are isolated from human and animal intestinal microbiota. However, the demand for diversification of biofunctional products has driven the search for new sources of probiotic candidates, such as fermented foods and vegetables. The present study found that strains isolated from the fermentation of fine cocoa from southern Bahia have biotechnological potential for use as a probiotic, since they showed capacity for self-aggregation and co-aggregation, antimicrobial activity against intestinal pathogens and resistance to gastrointestinal transits. Scores of importance for each property were established in order to more accurately assess the probiotic potential of the strains. The tests carried out contemplate the criteria previously established for the selection of probiotic candidates.

Antagonistic activity of Lactiplantibacillus plantarum 6.2 extracted from cocoa fermentation and its supernatant on Gardnerella vaginalis.

Search for alternative methods for the treatment of bacterial vaginosis has been growing, and probiotics being among them. The most well-known probiotic microorganisms are lactobacilli, which are naturally present in the vaginal microenvironment. Cocoa fermentation is a source of lactic acid bacteria, with lactobacilli being the most prominent. The aim of this study was to evaluate the antagonistic activity of Lactiplantibacillus plantarum 6.2 a strain of lactobacilli isolated from cocoa fermentation, and its cell-free supernatant on Gardnerella vaginalis. It was shown that Lpb. plantarum 6.2 and its supernatant, used at three concentrations, i.e., 40, 20 and 10 mg/mL, have a strong antagonistic activity against G. vaginalis, with a probable action of proteinaceous bacteriocins; the activity was lost after heat treatment. The ability to exclude and displace G. vaginalis from the adhesion site to vaginal HMVII epithelial cells was also demonstrated by the lactobacilli and the supernatant, with the latter showing a bactericidal effect. Thus, the Lpb. plantarum 6.2 strain presents itself as a good probiotic with potential to be used not only as a therapeutic alternative for vaginosis but also as a complement to existing therapies.

Bioprotective potential of lactic acid bacteria and their metabolites against enterotoxigenic Escherichia coli.

Escherichia coli is one of the main pathogens that impacts swine production. Given the need for methods for its control, the in vitro effect of lactic acid bacteria (LAB) and their metabolites against E. coli F4 was evaluated through cell culture and microbiological analysis. The strains Limosilactobacillus fermentum 5.2, Lactiplantibacillus plantarum 6.2, and L. plantarum 7.1 were selected. To evaluate the action of their metabolites, lyophilized cell-free supernatants (CFS) were used. The effect of CFS was evaluated in HT-29 intestinal lineage cells; in inhibiting the growth of the pathogen in agar; and in inhibiting the formation of biofilms. The bioprotective activity of LAB was evaluated via their potential for autoaggregation and coaggregation with E. coli. The CFS did not show cytotoxicity at lower concentrations, except for L. fermentum 5.2 CFS, which is responsible for cell proliferation at doses lower than 10 mg ml-1. The CFS were also not able to inhibit the growth of E. coli F4 in agar; however, the CFS of L. plantarum 7.1 resulted in a significant decrease in biofilm formation at a dose of 40 mg ml-1. Regarding LAB, their direct use showed great potential for autoaggregation and coaggregation in vitro, thus suggesting possible effectiveness in animal organisms, preventing E. coli fixation and proliferation. New in vitro tests are needed to evaluate lower doses of CFS to control biofilms and confirm the bioprotective potential of LAB, and in vivo tests to assess the effect of LAB and their metabolites interacting with animal physiology.

Citral modulates human monocyte responses to Staphylococcus aureus infection.

Staphylococcus aureus is a Gram-positive bacterium that is considered an important human pathogen. Due to its virulence and ability to acquire mechanisms of resistance to antibiotics, the clinical severity of S. aureus infection is driven by inflammatory responses to the bacteria. Thus, the present study aimed to investigate the modulating role of citral in inflammation caused by S. aureus infection. For this, we used an isolate obtained from a nasal swab sample of a healthy child attending a day-care centre in Vitória da Conquista, Bahia, Brazil. The role of citral in modulating immunological factors against S. aureus infection was evaluated by isolating and cultivating human peripheral blood mononuclear cells. The monocytes were treated with 4%, 2%, and 1% citral before and after inoculation with S. aureus. The cells were analysed by immunophenotyping of monocyte cell surface molecules (CD54, CD282, CD80, HLA-DR, and CD86) and cytokine dosage (IL-1ß, IL-6, IL-10, IL-12p70, IL-23, IFN-γ, TGF-ß, and TNF-α), and evaluated for the expression of 84 genes related to innate and adaptive immune system responses. GraphPad Prism software and variables with P values < 0.05, were used for statistical analysis. Our data demonstrated citral's action on the expression of surface markers involved in recognition, presentation, and migration, such as CD14, CD54, and CD80, in global negative regulation of inflammation with inhibitory effects on NF-κB, JNK/p38, and IFN pathways. Consequently, IL-1ß, IL-6, IL-12p70, IL-23, IFN-γ, and TNF-α cytokine expression was reduced in groups treated with citral and groups treated with citral at 4%, 2%, and 1% and infected, and levels of anti-inflammatory cytokines such as IL-10 were increased. Furthermore, citral could be used as a supporting anti-inflammatory agent against infections caused by S. aureus. There are no data correlating citral, S. aureus, and the markers analysed here; thus, our study addresses this gap in the literature.

Differentially expressed proteins in the interaction of Paracoccidioides lutzii with human monocytes.

BACKGROUND: Fungi of the genus Paracoccidioides are the etiological agents of paracoccidioidomycosis, a highly prevalent mycosis in Latin America. Infection in humans occurs by the inhalation of conidia, which later revert to the form of yeast. In this context, macrophages are positioned as an important line of defense, assisting in the recognition and presentation of antigens, as well as producing reactive oxygen species that inhibit fungal spreading. AIMS: The objective of this study was to identify differentially expressed proteins during the interaction between Paracoccidioides lutzii Pb01 strain and human U937 monocytes. METHODS: Two-dimensional electrophoresis, combined with mass spectrometry, was used to evaluate the differential proteomic profiles of the fungus P. lutzii (Pb01) interacting with U937 monocytes. RESULTS: It was possible to identify 25 proteins differentially expressed by Pb01 alone and after interacting with U937 monocytes. Most of these proteins are directly associated with fungal metabolism for energy generation, such as glyceraldehyde-3-phosphate dehydrogenase, and intracellular adaptation to monocytes. Antioxidant proteins involved in the response to oxidative stress, such as peroxiredoxin, cytochrome, and peroxidase, were expressed in greater quantity in the interaction with monocytes, suggesting their association with survival mechanisms inside phagocytic cells. We also identified 12 proteins differentially expressed in monocytes before and after the interaction with the fungus; proteins involved in the reorganization of the cytoskeleton, such as vimentin, and proteins involved in the response to oxidative stress, such as glioxalase 1, were identified. CONCLUSIONS: The results of this proteomic study of a P. lutzii isolate are novel, mimicking in vitro what occurs in human infections. In addition, the proteins identified may aid to understand fungal-monocyte interactions and the pathogenesis of paracoccidioidomycosis.

Lactiplantibacillus plantarum strains isolated from spontaneously fermented cocoa exhibit potential probiotic properties against Gardnerella vaginalis and Neisseria gonorrhoeae.

BACKGROUND: Probiotics are important tools in therapies against vaginal infections and can assist traditional antibiotic therapies in restoring healthy microbiota. Recent research has shown that microorganisms belonging to the genus Lactobacillus have probiotic potential. Thus, this study evaluated the potential in vitro probiotic properties of three strains of Lactiplantibacillus plantarum, isolated during the fermentation of high-quality cocoa, against Gardnerella vaginalis and Neisseria gonorrhoeae. Strains were evaluated for their physiological, safety, and antimicrobial characteristics. RESULTS: The hydrophobicity of L. plantarum strains varied from 26.67 to 91.67%, and their autoaggregation varied from 18.10 to 30.64%. The co-aggregation of L. plantarum strains with G. vaginalis ranged from 14.73 to 16.31%, and from 29.14 to 45.76% with N. gonorrhoeae. All L. plantarum strains could moderately or strongly produce biofilms. L. plantarum strains did not show haemolytic activity and were generally sensitive to the tested antimicrobials. All lactobacillus strains were tolerant to heat and pH resistance tests. All three strains of L. plantarum showed antimicrobial activity against the tested pathogens. The coincubation of L. plantarum strains with pathogens showed that the culture pH remained below 4.5 after 24 h. All cell-free culture supernatants (CFCS) demonstrated activity against the two pathogens tested, and all L. plantarum strains produced hydrogen peroxide. CFCS characterisation in conjunction with gas chromatography revealed that organic acids, especially lactic acid, were responsible for the antimicrobial activity against the pathogens evaluated. CONCLUSION: The three strains of L. plantarum presented significant probiotic characteristics against the two pathogens of clinical importance. In vitro screening identified strong probiotic candidates for in vivo studies for the treatment of vaginal infections.

Immobilization of PR4A3 enzyme in pluronic F127 polymeric micelles against colorectal adenocarcinoma cells and increase of in vitro bioavailability.

Traditional therapy for malignant neoplasms involving surgical procedures, radiotherapy and chemotherapy aims to kill neoplastic cells, but also affects normal cells. Therefore, exogenous proteases are the target of studies in cancer therapy, as they have been shown to be effective in suppressing tumors and reducing metastases. Pluronic F127 (F127) is a copolymer of amphiphilic blocks that has shown significant potential for drug administration, as it is capable of incorporating hydrophobic drugs and self-assembling in micrometers of nanometric size. This study investigated the effects of immobilization of the alkaline protease PR4A3 with pluronic F127 micelles on the enzyme-induced cytotoxicity. Protease immobilization was demonstrated through UV-visible and circular dichroism (CD) spectroscopies, as the enzyme interacts with the polymeric micelle of Pluronic F127 without changing its secondary structure. In addition, the immobilized form of the enzyme showed greater bioavailability after passing through the simulated gastrointestinal transit. Cell viability was assessed using the tetrazoic methylthiazole (MTT) assay. The results open perspectives for new research and development for PR4A3 in the treatment of colorectal carcinoma.

Técnica de inseminação artificial por tração cervical e função luteal em ovelhas Santa Inês após estro sincronizado com cloprostenol ou progestágeno-eCG / Technique of artificial ins emination by cervical traction and luteal function in Santa Inês ewes after synchronized estrus with cloprostenol or progestagen-eCG

Avaliou-se a influência do protocolo de sincronização de estro na execução da técnica de inseminação artificial por tração cervical (ITC) e subsequente perfil sérico de progesterona em ovelhas Santa Inês. Um grupo foi submetido a duas aplicações de cloprostenol com sete dias de intervalo (PGF2α, n=43), outro grupo foi sincronizado com MAP-eCG (P4-eCG, n=40) e ovelhas ao estro natural (EN, n=10) formaram grupo controle. Com as ovelhas contidas em estação, foi realizada a técnica ITC utilizando um aplicador metálico e mandril. Foram avaliadas a profundidade de penetração e intensidade de reação da ovelha ao manejo da técnica. Dosagens de estradiol e progesterona foram feitas antes e depois da ITC/estro natural, respectivamente. Os níveis de estradiol foram superiores com P4-eCG vs PGF2α (P < 0,05), mas não houve influência do protocolo na execução da técnica. Ocorreram 83,5% de deposição intrauterina do sêmen, o tempo médio foi de 32 segundos e 90,7% das ovelhas apresentaram reação fraca ou moderada durante o manejo. Os níveis de progesterona foram semelhantes entre ITC e estro natural. Independente do protocolo hormonal, a técnica ITC em ovelhas Santa Inês mantidas em estação mostrou-se viável no tempo de penetração da cérvice e deposição intrauterina do sêmen, não comprometendo a função luteal subsequente.(AU)

The influence of estrus synchronization protocol was evaluated on the artificial insemination technique by cervical traction (ICT) and subsequent progesterone serum profile in Santa Inês ewes. One group was submitted to two applications of cloprostenol seven days apart (PGF2α, n=43), another group was synchronized with MAP-eCG (P4-eCG, n=40) and ewes at natural estrus (NE, n=10) formed a control group. With the ewe contained in the stand position, the ICT technique was performed using a metal applicator and mandrel. Depth of penetration and reaction intensity of the ewe to the handling of the technique were evaluated. Dosages of estradiol and progesterone were performed before and after ICT / natural estrus, respectively. Estradiol levels were higher with P4-eCG vs PGF2α(AU)

Técnica de inseminação artificial por tração cervical e função luteal em ovelhas Santa Inês após estro sincronizado com cloprostenol ou progestágeno-eCG / Technique of artificial ins emination by cervical traction and luteal function in Santa Inês ewes after synchronized estrus with cloprostenol or progestagen-eCG

Avaliou-se a influência do protocolo de sincronização de estro na execução da técnica de inseminação artificial por tração cervical (ITC) e subsequente perfil sérico de progesterona em ovelhas Santa Inês. Um grupo foi submetido a duas aplicações de cloprostenol com sete dias de intervalo (PGF2α, n=43), outro grupo foi sincronizado com MAP-eCG (P4-eCG, n=40) e ovelhas ao estro natural (EN, n=10) formaram grupo controle. Com as ovelhas contidas em estação, foi realizada a técnica ITC utilizando um aplicador metálico e mandril. Foram avaliadas a profundidade de penetração e intensidade de reação da ovelha ao manejo da técnica. Dosagens de estradiol e progesterona foram feitas antes e depois da ITC/estro natural, respectivamente. Os níveis de estradiol foram superiores com P4-eCG vs PGF2α (P < 0,05), mas não houve influência do protocolo na execução da técnica. Ocorreram 83,5% de deposição intrauterina do sêmen, o tempo médio foi de 32 segundos e 90,7% das ovelhas apresentaram reação fraca ou moderada durante o manejo. Os níveis de progesterona foram semelhantes entre ITC e estro natural. Independente do protocolo hormonal, a técnica ITC em ovelhas Santa Inês mantidas em estação mostrou-se viável no tempo de penetração da cérvice e deposição intrauterina do sêmen, não comprometendo a função luteal subsequente.

The influence of estrus synchronization protocol was evaluated on the artificial insemination technique by cervical traction (ICT) and subsequent progesterone serum profile in Santa Inês ewes. One group was submitted to two applications of cloprostenol seven days apart (PGF2α, n=43), another group was synchronized with MAP-eCG (P4-eCG, n=40) and ewes at natural estrus (NE, n=10) formed a control group. With the ewe contained in the stand position, the ICT technique was performed using a metal applicator and mandrel. Depth of penetration and reaction intensity of the ewe to the handling of the technique were evaluated. Dosages of estradiol and progesterone were performed before and after ICT / natural estrus, respectively. Estradiol levels were higher with P4-eCG vs PGF2α

Reproductive performance and luteal function of Santa Ines ewes inseminated by cervical retraction with fresh or frozen semen / Desempenho reprodutivo e função luteal de ovelhas Santa Inês inseminadas por retração cervical com sêmen fresco ou congelado

The artificial insemination (AI) by cervical retraction using fresh or commercially available frozen semen was evaluated. Santa Inês ewes (n=151) were assigned in five groups. Natural mating (NM) composed the control group. Four groups were submitted to hormonal treatment and timed insemination by cervical retraction (CRI) or laparoscopy (LAI), using fresh or frozen semen. To perform CRI, the ewes were kept in standing position. The time required to penetrate the cervical canal and to perform CRI were recorded; local deposition of semen and the reaction intensity of the ewe (weak, moderate or strong) were measured. Blood samples were collected to perform the progesterone dosage at days 0, 3, 5, 12 and 17 (day 0 = AI/NM). Non-return to estrus rate (NRE), pregnancy (at D35) and fertility (birth / mated ewes) were evaluated. Mean time for cervical penetration and CRI were 52.8 ± 21.2s and 3:26min ± 47s, respectively. Reaction intensity to CRI was weak or moderate in 92.3% of the ewes. Serum levels of progesterone after CRI, LAI or NM were similar. NRE of the inseminated ewes were similar, except for CRI with frozen semen, which was lower (P<0.05). Fertility rates were similar between CRI and LAI (35.4% and 42.2%, respectively). Artificial insemination by cervical retraction with ewes in a standing position is practical, and does not change the progesterone profile, providing reproductive rates similar to laparoscopy. However, the use of this technique along with commercial frozen semen presents low fertility rate.(AU)

Avaliou-se a inseminação artificial por retração cervical usando sêmen fresco ou congelado disponível comercialmente. Ovelhas Santa Inês foram divididas em cinco grupos. A monta natural (MN) correspondeu ao grupo controle. Quatro grupos foram submetidos a um tratamento hormonal e inseminação artificial por retração cervical (CRI) ou laparoscopia (LAI) usando sêmen fresco ou congelado. Para a CRI, ovelhas foram mantidas em estação. O tempo para penetrar o canal cervical e realizar a CRI foi mensurado, bem como o local de deposição do sêmen e intensidade de reação da ovelha (fraca, moderada, forte). Amostras de sangue foram coletadas para dosagem de progesterona aos dias 0, 3, 5, 12 e 17 (dia 0 = inseminação ou monta). Foram avaliadas as taxas de não retorno ao estro (NRE), gestação (ao dia 35) e fertilidade (nascimento / ovelhas acasaladas). O tempo médio para penetração cervical foi de 52,8 ± 21,2s e para a CRI foi de 3:26min ± 47s. A intensidade de reação à CRI foi fraca ou moderada em 92,3% das ovelhas. Os níveis séricos de progesterona após CRI, LAI e MN foram similares. NRE das ovelhas inseminadas foi similar, exceto em ovelhas com CRI e sêmen congelado, que foi inferior (P<0,05). As taxas de fertilidade foram semelhantes entre CRI e LAI (35,4% e 42,2%, respectivamente). A inseminação artificial por retração cervical com ovelhas em estação se mostrou prática e não alterou o perfil de progesterona, possibilitando índices reprodutivos similares à laparoscopia. No entanto, o uso desta técnica com sêmen disponível comercialmente apresentou baixa taxa de fertilidade.(AU)

Fermentation in fine cocoa type Scavina: Change in standard quality as the effect of use of starters yeast in fermentation.

In the present work we aimed to demonstrate the influence of inoculum starter in support high quality fermentation. Cocoa fermentations were performed in wooden boxes and eight yeasts strains were used in separated fermentations of fine cocoa, type Scavina, as starter inoculum. Temperature, pH, titirable acidity, reducing sugar and free amino acids were evaluated during or after fermentation. The influence of starters yeasts on the decrease of acidity, sugar concentration and free amino acids was significant. The strains Candida parapsilosis, Torulaspora delbrueckii and Pichia kluyveri showed greater changes in the reducing sugar and free amino acids in fermented cocoa beans. These results indicate the ability of yeast used as inoculum starter to modify the end condition and further enhance the quality of fine cocoa beans.

Reproductive performance and luteal function of Santa Ines ewes inseminated by cervical retraction with fresh or frozen semen / Desempenho reprodutivo e função luteal de ovelhas Santa Inês inseminadas por retração cervical com sêmen fresco ou congelado

The artificial insemination (AI) by cervical retraction using fresh or commercially available frozen semen was evaluated. Santa Inês ewes (n=151) were assigned in five groups. Natural mating (NM) composed the control group. Four groups were submitted to hormonal treatment and timed insemination by cervical retraction (CRI) or laparoscopy (LAI), using fresh or frozen semen. To perform CRI, the ewes were kept in standing position. The time required to penetrate the cervical canal and to perform CRI were recorded; local deposition of semen and the reaction intensity of the ewe (weak, moderate or strong) were measured. Blood samples were collected to perform the progesterone dosage at days 0, 3, 5, 12 and 17 (day 0 = AI/NM). Non-return to estrus rate (NRE), pregnancy (at D35) and fertility (birth / mated ewes) were evaluated. Mean time for cervical penetration and CRI were 52.8 ± 21.2s and 3:26min ± 47s, respectively. Reaction intensity to CRI was weak or moderate in 92.3% of the ewes. Serum levels of progesterone after CRI, LAI or NM were similar. NRE of the inseminated ewes were similar, except for CRI with frozen semen, which was lower (P<0.05). Fertility rates were similar between CRI and LAI (35.4% and 42.2%, respectively). Artificial insemination by cervical retraction with ewes in a standing position is practical, and does not change the progesterone profile, providing reproductive rates similar to laparoscopy. However, the use of this technique along with commercial frozen semen presents low fertility rate.

Avaliou-se a inseminação artificial por retração cervical usando sêmen fresco ou congelado disponível comercialmente. Ovelhas Santa Inês foram divididas em cinco grupos. A monta natural (MN) correspondeu ao grupo controle. Quatro grupos foram submetidos a um tratamento hormonal e inseminação artificial por retração cervical (CRI) ou laparoscopia (LAI) usando sêmen fresco ou congelado. Para a CRI, ovelhas foram mantidas em estação. O tempo para penetrar o canal cervical e realizar a CRI foi mensurado, bem como o local de deposição do sêmen e intensidade de reação da ovelha (fraca, moderada, forte). Amostras de sangue foram coletadas para dosagem de progesterona aos dias 0, 3, 5, 12 e 17 (dia 0 = inseminação ou monta). Foram avaliadas as taxas de não retorno ao estro (NRE), gestação (ao dia 35) e fertilidade (nascimento / ovelhas acasaladas). O tempo médio para penetração cervical foi de 52,8 ± 21,2s e para a CRI foi de 3:26min ± 47s. A intensidade de reação à CRI foi fraca ou moderada em 92,3% das ovelhas. Os níveis séricos de progesterona após CRI, LAI e MN foram similares. NRE das ovelhas inseminadas foi similar, exceto em ovelhas com CRI e sêmen congelado, que foi inferior (P<0,05). As taxas de fertilidade foram semelhantes entre CRI e LAI (35,4% e 42,2%, respectivamente). A inseminação artificial por retração cervical com ovelhas em estação se mostrou prática e não alterou o perfil de progesterona, possibilitando índices reprodutivos similares à laparoscopia. No entanto, o uso desta técnica com sêmen disponível comercialmente apresentou baixa taxa de fertilidade.

Biotechnological potential of mangrove sediments: Identification and functional attributes of thermostable and salinity-tolerant ß-glucanase.

Microorganisms native to mangroves are expected to contain enzymes capable of hydrolyzing different carbon sources. However, most of these microorganisms aren't cultivable; hence, alternative techniques as metagenomics are tools for studying and obtaining some of the natural genomes, genes and enzymes of biotechnological interest. The ß-glucanase was produced using a metagenomic clone of mangrove sediments and detected by functional screening on carboxymethylcellulose substrate. The enzyme was purified by cation exchange chromatography. The peptides detected by mass spectrometry showed 20% identity with the polypeptide deduced from the genomic fragment sequenced. The ORF identified as BglfosD9 possessed 729 bp and the encoded protein showed predicted MW and pI of 28kD and 6.8, respectively. The enzyme was active in a wide range of pH (5-10) with optimum pH at 8, had relative activity greater than 50% at all temperatures tested (5-90 °C), was stable at temperatures of 5, 50 and 90 °C and showed excellent relative activity at high NaCl concentrations. This ß-glucanase also showed high relative activity in the presence of SDS and it could hydrolyze ß-glucan, CMC and Avicel as substrates. These findings support the idea of a new thermostable and active enzyme at basic pH from metagenomic library of mangrove sediment.

Toxoplasma gondii induces extracellular traps release in cat neutrophils.

Neutrophils respond differently to violations of the body's physiological barriers during infections. Extracellular traps comprise one of the mechanisms used by these cells to reduce the spread of pathogens to neighboring tissues, as well as ensure a high concentration of antimicrobial agents at the site of infection. To date, this innate defense mechanism has not been previously demonstrated in neutrophils of cats exposed to Toxoplasma gondii. The aim of this study was to characterize the in vitro release of neutrophil extracellular traps (NETs) when neutrophils isolated from cats were exposed to T. gondii. First, cellular viability was tested at different time points after parasite exposure. The production of reactive oxygen species (ROS) and lactate dehydrogenase and the amount of extracellular DNA were quantified. In addition, the number of parasites associated with neutrophils was determined, and the observed NETs formed were microscopically characterized. Results showed that (i) in culture, neutrophils isolated from cats presented diminished cellular viability after 4 h of incubation, and when neutrophils were incubated with T. gondii, they displayed cytotoxic effects after 3 h of interaction; (ii) neutrophils were able to release structures composed of DNA and histones, characterized as NETs under optical, immunofluorescence, and electron scanning microscopy, when stimulated with T. gondii; (iii) only 11.4% of neutrophils were able to discharge NETs during 3 h of incubation; however, it was observed through extracellular quantification of DNA that this small number of cells were able to display different behavior compared to a negative control (no parasite) group; (iv) significant differences in ROS production were observed in neutrophils exposed to T. gondii. In conclusion, our results showed that neutrophils isolated from cats exposed to T. gondii release structures composed of DNA and histones, similar to what has already been described in other neutrophil species infected with the parasite.

Administration of Lactobacillus plantarum Lp62 to dam rats at the end of delivery and during lactation affects TGF-ß1 level and nutritional milk composition, and body weight of pups.

PURPOSE: Lactobacillus plantarum Lp62 is a lactic acid bacteria strain that has been isolated from cocoa beans and exhibited probiotic potential. The influence of oral administration of L. plantarum Lp62 on the growth of rat's pups; on yield, cytokines and milk composition was studied. METHODS: Lactobacillus plantarum Lp62 is a lactic acid bacteria strain that has been isolated from cocoa beans. It was administered daily by gavage to Wistar rats (n = 8), from the 7th day before delivery and for 20 days during lactation, in a concentration of 1.44 × 109 CFU/rat. The dam and pups were weighed and milk was collected at 12th and 19th day for determination of protein, triglycerides, cholesterol and lactose by colorimetric assays. TGF-ß1 milk levels were analyzed by ELISA. The mammary glands of rats were removed for histological analysis. To detect statistical differences between the groups, tests of mean differences at a significance level of 5% was performed. RESULTS: Supplementation with L. plantarum L62 resulted in significant higher weight of pups (p < 0.05), with similar weight on dams (p > 0.05). The milk yield was not altered by L. plantarum treatment, but the levels of protein, triglycerides and cholesterol were increased (p < 0.05), with no difference in lactose concentration (p > 0.05). Levels of TGF-ß1 were higher in the milk of L. plantarum treatment (p < 0.05). CONCLUSIONS: The treatment of dams at the end of pregnancy and lactation with L. plantarum Lp62 increased nutritional content of milk, probably contributing to the higher weight of the pups. The higher levels of TGF-ß1 in the milk, could promote immune benefits to the pups. Further studies in this field are needed to prove the potential use of L. plantarum Lp62 as a probiotic.

Potential of Maintaining a Healthy Vaginal Environment by Two Lactobacillus Strains Isolated from Cocoa Fermentation.

Bacteria in the genera Mycoplasma and Ureaplasma do not have cell walls and therefore interact with host cells through lipid-associated membrane proteins (LAMP). These lipoproteins are important for both surface adhesion and modulation of host immune responses. Mycoplasma and Ureaplasma have been implicated in cases of bacterial vaginosis (BV), which can cause infertility, abortion, and premature delivery. In contrast, bacteria of the genus Lactobacillus, which are present in the vaginal microbiota of healthy women, are thought to inhibit local colonization by pathogenic microorganisms. The aim of the present study was to evaluate the in vitro interactions between lipoproteins of Mycoplasma and Ureaplasma species and vaginal lineage (HMVII) cells and to study the effect of Lactobacillus isolates from cocoa fermentation on these interactions. The tested Lactobacillus strains showed some important probiotic characteristics, with autoaggregation percentages of 28.55% and 31.82% for L. fermentum FA4 and L. plantarum PA3 strains, respectively, and percent adhesion values of 31.66 and 41.65%, respectively. The two strains were hydrophobic, with moderate to high hydrophobicity values, 65.33% and 71.12% for L. fermentum FA4 and L. plantarum PA3 in toluene. Both strains secreted acids into the culture medium with pH=4.32 and pH=4.33, respectively, and showed antibiotics susceptibility profiles similar to those of other lactobacilli. The strains were also able to inhibit the death of vaginal epithelial cells after incubation with U. parvum LAMP from 41.03% to 2.43% (L. fermentum FA4) and 0.43% (L. plantarum PA3) and also managed to significantly decrease the rate of cell death caused by the interaction with LAMP of M. hominis from 34.29% to 14.06% (L. fermentum FA4) and 14.61% (L. plantarum PA3), thus demonstrating their potential for maintaining a healthy vaginal environment.

In Vitro Activity of Lactobacilli with Probiotic Potential Isolated from Cocoa Fermentation against Gardnerella vaginalis.

Study of the probiotic potential of microorganisms isolated from fermented foods has been increasing, especially studies related to lactobacilli. In intestinal models, lactobacilli have demonstrated beneficial properties, such as anti-inflammatory activity and increased antibody production, but the molecular mechanisms involving probiotic and antagonistic action as well as their effect on human vaginal cells have not yet been fully elucidated. The aim of this study was to evaluate the functional and antagonistic properties of three strains of lactobacilli isolated from cocoa fermentation (Lactobacillus fermentum 5.2, L. plantarum 6.2, and L. plantarum 7.1) against Gardnerella vaginalis. Our results show that the lactobacilli have potential use as probiotics, since they have high hydrophobicity and autoaggregation properties and effectively adhere to vaginal cells. Metabolites secreted into the culture medium and whole cells of the strains under study are capable of interfering with the growth of G. vaginalis to different degrees. The elucidation of the antagonistic mechanisms as well as their effect on human cells may be useful in the development of a product containing such microorganisms or products secreted by them.

Co-infection of sexually transmitted pathogens and Human Papillomavirus in cervical samples of women of Brazil.

BACKGROUND: Some sexually transmitted infectious agents, such as Chlamydia trachomatis and Herpes simplex, cause local inflammation, and could contribute to Human Papillomavirus (HPV) and cervical lesion progression. Thus, the aim of this study was to determine any association between the presence of microorganisms of gynecological importance, sexual behavior, clinical and demographical variables to the development and progress of cervical lesions. METHODS: One hundred and thirty-two women between 14 and 78 years and living at Vitória da Conquista, Bahia, Brazil, were included (62 individuals with cervical lesions and 70 without lesions). They answered a questionnaire to provide data for a socioeconomic and sexual activity profile. Samples of cervical swabs were collected and analyzed by PCR to detect genital microorganisms and HPV. Quantitative PCR was used to detect and quantify Ureaplasma urealyticum and Ureaplasma parvum. Univariate and multiple logistic regression were performed to measure the association with the cervical lesions, and an odds ratio (OR) with 95% confidence intervals (95%CI) were calculated. The Mann-Whitney U test was also used to compare the microorganism load in the case and control groups. The significance level was 5% in all hypotheses tested. RESULTS: Cervical lesions were associated with: women in a stable sexual relationship (OR = 14.21, 95%CI = 3.67-55.018), positive PCR for HPV (OR = 16.81, 95%CI = 4.19-67.42), Trichomonas vaginalis (OR = 8.566, 95%CI = 2.04-35.94) and Gardnerella vaginalis (OR = 6.13, 95%CI = 1.53-24.61), adjusted by age and qPCR for U. parvum. U. parvum load showed a statistical difference between the case and control groups (p-value = 0.002). CONCLUSION: Variables such as stable relationship, HPV, T. vaginalis, G. vaginalis were associated with cervical lesions in epidemiological studies. U. parvum load was higher in woman with cervical lesions compared with women without lesions. Additional studies are needed to better understand the role of these factors in cervical lesion development.