Antiangiogenic properties of BthMP, a P-I metalloproteinase from Bothrops moojeni snake venom by VEGF pathway in endothelial cells.

Angiogenesis is a process that is controlled by a delicate combination of proangiogenic and antiangiogenic molecules and can be disrupted in various illnesses, including cancer. Non-cancerous diseases can also have an abnormal or insufficient vascular growth, inflammation and hypoxia, which exacerbate angiogenesis. These conditions include atherosclerosis, psoriasis, endometriosis, asthma, obesity and AIDS. Based on that, the present work assessed the in vitro and ex vivo antiangiogenic properties stemming from BthMP, a P-I metalloproteinase from Bothrops moojeni snake venom, via the VEGF pathway. BthMP at a concentration of 5 and 40 µg/mL showed no toxicity to endothelial cells (HUVEC) in the MTT assay and was not able to induce necrosis and colony proliferation. Interestingly, BthMP inhibited adhesion, migration and invasion of HUVECs in Matrigel and arrested in vitro angiogenesis by reducing the average number of nodules in toxin-treated cells by 9.6 and 17.32 at 5 and 40 µg/mL, respectively, and the number of tubules by 15.9 at 5 µg/mL and 21.6 at 40 µg/mL in a VEGF-dependent way, an essential proangiogenic property. Furthermore, BthMP inhibited the occurrence of the angiogenic process in an ex vivo aortic ring test by decreasing new vessel formation by 52% at 5 µg/mL and by 66% at 40 µg/mL and by increasing the expression of an antiangiogenic gene, SFLT-1, and decreasing the expression of the proangiogenic genes VEGFA and ANGPT-1. Finally, this toxin reduces the production of nitric oxide, a marker that promotes angiogenesis and VEGF modulation, and decreases the protein expression of VEGFA in the supernatant of the HUVEC culture by about 30 %. These results suggest that BthMP has a promising antiangiogenic property and proves to be a biotechnological mechanism for understanding the antiangiogenic responses induced by snake venom metalloproteinases, which could be applied to a variety of diseases that exhibit an imbalance of angiogenesis mechanisms.

Accuracy of surgeons for predicting radiological sarcopenia in colorectal cancer surgery.

BACKGROUND: This study explored the performance of surgeons for predicting radiological sarcopenia as accessed by psoas cross-sectional area in patients with colorectal cancer (CRC). METHODS: A cross-sectional study was carried out and a diagnostic accuracy strategy was applied using the radiologist team assessment as gold standard. RESULTS: Cohort analysis of 45 consecutive patients found that 31.1% had sarcopenia. Correlation of Total Psoas Index between radiologists and surgeons was very strong for the Junior and strong for the Senior surgeon, with a strong correlation between the surgeons. By the simplistic criterion, agreement between radiologists and surgeons was substantial for both the Junior and Senior surgeons, with a moderate level between the surgeons. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of Junior surgeon were 92.9%, 83.9%, 72.2%, 96.3%, and 86.7%, respectively. The corresponding results for the Senior surgeon were 78.6%, 90.3%, 78.6%, 90.3%, and 86.7%, respectively. We found no major differences on agreement levels and performance of surgeons using the composite criterion. CONCLUSIONS: Surgeons seem to be accurate for identifying radiological sarcopenia in patients with CRC. The simplistic criterion should be preferred since a composite criterion adds complexity without increasing accuracy or agreement levels.

Prevalence of Neisseria gonorrhoeae and Chlamydia trachomatis infections among adolescent men who have sex with men and transgender women in Salvador, Northeast Brazil.

Adolescent men who have sex with men (AMSM) and transgender women (ATGW) enrolled as part of the PrEP1519 study between April 2019 and February 2021 in Salvador were tested for Neisseria gonorrhoeae (NG) and Chlamydia trachomatis (CT) infections.We performed real-time polymerase chain reaction using oropharyngeal, anal, and urethral swabs; assessed factors associated with NG and CT infections using multivariable Poisson regression analysis with robust variance; and estimated the prevalence ratios (PRs) and 95% confidence intervals (95% CIs). In total, 246 participants were included in the analyses (median age: 18.8; IQR: 18.2-19.4 years). The overall oropharyngeal, anal, and urethral prevalence rates of NG were 17.9%, 9.4%, 7.6%, and 1.9%, respectively. For CT, the overall, oropharyngeal, anal, and urethral prevalence rates were 5.9%, 1.2%, 2.4%, and 1.9%, respectively. A low level of education, clinical suspicion of STI (and coinfection with Mycoplasma hominis were associated with NG infection. The prevalence of NG and CT, especially extragenital infections, was high in AMSM and ATGW. These findings highlight the need for testing samples from multiple anatomical sites among adolescents at a higher risk of STI acquisition, implementation of school-based strategies, provision of sexual health education, and reduction in barriers to care.

Prevalence of Mollicutes among men who have sex with men and transgender women aged 15 to 19 years in Salvador, North-eastern Brazil.

BACKGROUND: Some species of Mollicutes have been associated with different pathologies of the urogenital tract in humans, with a high prevalence among adult men who have sex with men (MSM) and transgender women (TGW). However, few studies have been performed to investigate its prevalence among adolescents. In this study, we estimated the initial prevalence of Mycoplasma genitalium (MG), Mycoplasma hominis (MH), Ureaplasma urealyticum (UU), and Ureaplasma parvum (UP); the rate of misdiagnosis at different anatomical sites; and the associated factors with positive tests for Mollicutes among MSM and TGW aged 15 to 19 years enrolled in the PrEP1519 study. METHODS: PrEP-1519 is the first study to investigate the effectiveness of pre-exposure prophylaxis for human immunodeficiency virus among adolescent MSM and TGW aged 15 to 19 in Latin America. Oral, anal, and urethral swabs were taken from 246 adolescents upon enrolment in the study to detect MG, MH, UU, and UP by quantitative polymerase chain reaction (qPCR). Bivariate and multivariate analyses were conducted by Poisson regression and 95% confidence intervals (95% CI) were estimated. RESULTS: The prevalence of Mollicutes was 32.1%. UU was the most prevalent species (20.7%), followed by MH (13.4%), MG (5.7%), and UP (3.2%); 67.3% of the positive samples would have been missed if only urethral samples had been taken. Receptive anal sex (prevalence ratio [PR] = 1.79; 95% CI = 1.07-3.01) and clinical suspicion of sexually transmitted infection (PR = 1.62; 95% CI = 1.01-2.61) were factors associated with the detection of Mollicutes in general. Group sex (PR = 1.98; 95% CI = 1.12-3.50) and receptive anal sex (PR = 2.36; 95% CI = 0.95-5.86) were associated with the detection of Mycoplasma spp. No sociodemographic, clinical, or behavioural variable was significantly associated with the detection of Ureaplasma spp. CONCLUSIONS: A high prevalence of Mollicutes was observed among adolescent MSM and TGW, especially at extragenital sites. Further research is required to understand the epidemiological profile of high-risk adolescents in different regions and contexts, and to investigate the pathogenesis of Mollicutes in the oral and anal mucosa before routine screening can be recommended in clinical practice.

Estimates of life history parameters in a high latitude, arid-country vervet monkey population.

We present data on life history parameters from a long-term study of vervet monkeys in the Eastern Cape, South Africa. Estimates are presented of age at first conception for females and age at natal dispersal for males, along with the probability of survival to adulthood for infants born during the study, female reproductive life-span, reproductive output (including lifetime reproductive success for a subset of females), and inter-birth interval (IBI) duration. We also assess the effect of maternal age and infant survival on length of IBI. We then go on to compare life history parameters for our population with those from two East African populations in Kenya (Amboseli and Laikipia). We find there is broad consensus across the three populations, although mean infant survival was considerably lower for the two East African sites. Such comparisons must be made cautiously, however, as local ecology across the duration of the studies obviously has an impact on the estimates obtained. With this caveat in place, we consider that the concordance between values is sufficient to enable the values reported here to be used in comparative studies of primate life history, although data from habitats with higher rainfall and lower levels of seasonality are needed, and the results presented here should not be seen as canonical.

Host cell interactions of novel antigenic membrane proteins of Mycoplasma agalactiae.

BACKGROUND: Mycoplasma agalactiae is the main etiological agent of Contagious Agalactia syndrome of small ruminants notifiable to the World Organization for Animal Health. Despite serious economic losses, successful vaccines are unavailable, largely because its colonization and invasion factors are not well understood. This study evaluates the role of two recently identified antigenic proteins (MAG_1560, MAG_6130) and the cytadhesin P40 in pathogenicity related phenotypes. RESULTS: Adhesion to HeLa and sheep primary mammary stromal cells (MSC) was evaluated using ELISA, as well as in vitro adhesion assays on monolayer cell cultures. The results demonstrated MAG_6130 as a novel adhesin of M. agalactiae whose capacity to adhere to eukaryotic cells was significantly reduced by specific antiserum. Additionally, these proteins exhibited significant binding to plasminogen and extracellular matrix (ECM) proteins like lactoferrin, fibrinogen and fibronectin, a feature that could potentially support the pathogen in host colonization, tissue migration and immune evasion. Furthermore, these proteins played a detrimental role on the host cell proliferation and viability and were observed to activate pro-apoptotic genes indicating their involvement in cell death when eukaryotic cells were infected with M. agalactiae. CONCLUSIONS: To summarize, the hypothetical protein corresponding to MAG_6130 has not only been assigned novel adhesion functions but together with P40 it is demonstrated for the first time to bind to lactoferrin and ECM proteins thereby playing important roles in host colonization and pathogenicity.

Causal analysis as a bridge between qualitative and quantitative research.

Yarkoni argues that one solution is to abandon quantitative methods for qualitative ones. While we agree that qualitative methods are undervalued, we argue that both are necessary for thoroughgoing psychological research, complementing one another through the use of causal analysis. We illustrate how directed acyclic graphs can bridge qualitative and quantitative methods, thereby fostering understanding between different psychological methodologies.

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.

Antimicrobial activity of Lactobacillus fermentum TcUESC01 against Streptococcus mutans UA159.

Dental caries is a multifactorial chronic-infection disease, which starts with a bacterial biofilm formation caused mainly by Streptococcus mutans. The use of probiotics has shown numerous health benefits, including in the fight against oral diseases. Strains of Lactobacillus fermentum have already shown probiotic potential against S. mutans, but there are still few studies. Thus, the aim of our study was to evaluate the antimicrobial activity of the inoculum and metabolites produced by L. fermentum TcUESC01 against S. mutans UA159. For this, a growth curve of L. fermentum was performed and both the inoculum and the metabolites formed in the fermentation were tested against the growth of S. mutans UA159 in agar diffusion tests, and only its metabolites were tested to determine the minimum inhibitory concentration, minimal bactericidal concentration and inhibition of cell adhesion. Inhibition of biofilm formation, pH drop and proton permeability were also tested with the metabolites. The zone of inhibition began to be formed at 14 h and continued until 16 h. The inoculum containing L. fermentum also showed zone of inhibition. The MIC for the metabolites was 1280 mg/mL and the MBC was obtained with a concentration higher than the MIC equal to 5120 mg/mL. Half of the MIC concentration (640 mg/mL) was required to inhibit S. mutans adhesion to the surface of the microplates. In the biofilm analyzes, the treatment with the metabolites in the tested concentration was not able to reduce biomass, insoluble glucans and alkali soluble compared to the control biofilm (p > 0.05). The metabolites also did not affect acid production and acid tolerance of S. mutans cells in biofilms compared to saline group (p > 0.05). Lactic acid (50.38%) was the most abundant organic acid produced by L. fermentum. This is the first report showing that the metabolites produced by the Lactobacillus fermentum TcUESC01 have a potential to be used as an antimicrobial agent against S. mutans, showing anti-adherence and bactericidal activity against planktonic cells of S. mutans. Thus, further studies should be carried out in order to better understand the antimicrobial activity of metabolites of L. fermentum TCUESC01.

Immunoinformatics and analysis of antigen distribution of Ureaplasma diversum strains isolated from different Brazilian states.

BACKGROUND: Ureaplasma diversum has numerous virulence factors that contribute to pathogenesis in cattle, including Lipid-associated membrane proteins (LAMPs). Therefore, the objectives of this study were to evaluate in silico important characteristics for immunobiological applications and for heterologous expression of 36 LAMPs of U. diversum (UdLAMPs) and, also, to verify by conventional PCR the distribution of these antigens in strains of Brazilian states (Bahia, Minas Gerais, São Paulo, and Mato Grosso do Sul). The Manatee database was used to obtain the gene and peptide sequences of the antigens. Similarity and identity studies were performed using BLASTp and direct antigenicity was evaluated by the VaxiJen v2.0 server. Epitope prediction for B lymphocytes was performed on the BepiPred v2.0 and CBTOPE v1.0 servers. NetBoLApan v1.0 was used to predict CD8+ T lymphocyte epitopes. Subcellular location and presence of transmembrane regions were verified by the software PSORTb v3.0.2 and TMHMM v2.2 respectively. SignalP v5.0, SecretomeP v2.0, and DOLOP servers were used to predict the extracellular excretion signal. Physico-chemical properties were evaluated by the web-software ProtParam, Solpro, and Protein-sol. RESULTS: In silico analysis revealed that many UdLAMPs have desirable properties for immunobiological applications and heterologous expression. The proteins gudiv_61, gudiv_103, gudiv_517, and gudiv_681 were most promising. Strains from the 4 states were PCR positive for antigens predicted with immunogenic and/or with good characteristics for expression in a heterologous system. CONCLUSION: These works contribute to a better understanding of the immunobiological properties of the UdLAMPs and provide a profile of the distribution of these antigens in different Brazilian states.

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.

Ureaplasma diversum protein interaction networks: evidence of horizontal gene transfer and evolution of reduced genomes among Mollicutes.

Ureaplasma diversum is a member of the Mollicutes class responsible for urogenital tract infection in cattle and small ruminants. Studies indicate that the process of horizontal gene transfer, the exchange of genetic material among different species, has a crucial role in mollicute evolution, affecting the group's characteristic genomic reduction process and simplification of metabolic pathways. Using bioinformatics tools and the STRING database of known and predicted protein interactions, we constructed the protein-protein interaction network of U. diversum and compared it with the networks of other members of the Mollicutes class. We also investigated horizontal gene transfer events in subnetworks of interest involved in purine and pyrimidine metabolism and urease function, chosen because of their intrinsic importance for host colonization and virulence. We identified horizontal gene transfer events among Mollicutes and from Ureaplasma to Staphylococcus aureus and Corynebacterium, bacterial groups that colonize the urogenital niche. The overall tendency of genome reduction and simplification in the Mollicutes is echoed in their protein interaction networks, which tend to be more generalized and less selective. Our data suggest that the process was permitted (or enabled) by an increase in host dependence and the available gene repertoire in the urogenital tract shared via horizontal gene transfer.

Anti-inflammatory activity of ß-caryophyllene combined with docosahexaenoic acid in a model of sepsis induced by Staphylococcus aureus in mice.

BACKGROUND: Sepsis is a set of serious organic manifestations caused by an infection, whose progression culminates in exacerbated inflammation and oxidative stress, poor prognosis, and high hospital costs. Antioxidants used against sepsis have been evaluated, including essential oils such as ß-caryophyllene (BCP), and polyunsaturated fatty acids, such as docosahexaenoic acid (DHA). The aim of this study was to evaluate the anti-inflammatory activity of the association of these two compounds. RESULTS: Treatment with BCP-DHA, at a dose of 200 µL/animal, significantly inhibited the migration of neutrophils in a Cg-induced peritonitis model. After Staphylococcus aureus infection, in the groups treated with BCP-DHA there was a significant decrease in the total and differential count of leukocytes, increased expression of cytokines TNF-α and IFN-γ in treated groups, an increase of IL-4 and IL-5 in B/D and B/D + SA groups, and an augmentation of IL-6 and IL-12 groups in B/D + SA groups. Histological and bacterial analysis revealed lower neutrophil migration and lower bacterial load in the infected and treated groups. CONCLUSION: In general, the BCP-DHA association presented anti-inflammatory activity against two different models of acute inflammation and infection, showing promising potential as a therapeutic adjuvant in sepsis. © 2019 Society of Chemical Industry.

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.

Inhibition of Staphylococcus aureus biofilm by Lactobacillus isolated from fine cocoa.

BACKGROUND: Biofilm production represents an important virulence and pathogenesis factor for Staphylococcus aureus. The formation of biofilms on medical devices is a major concern in hospital environments, as they can become a constant source of infection. Probiotic bacteria, such as Lactobacillus fermentum and L. plantarum, have been found to inhibit biofilm formation; however little is known about the underlying mechanism. In this study, we tested the activity of supernatants produced by L. fermentum TCUESC01 and L. plantarum TCUESC02, isolated during the fermentation of fine cocoa, against S. aureus CCMB262 biofilm production. We measured inhibition of biofilm formation in vitro and analyzed biofilm structure by confocal and electronic microscopy. Additionally, we quantified the expression of S. aureus genes icaA and icaR involved in the synthesis of the biofilm matrix by real-time PCR. RESULTS: Both Lactobacillus supernatants inhibited S. aureus growth. However, only L. fermentum TCUESC01 significantly reduced the thickness of the biofilm, from 14 µm to 2.83 µm (at 18 mg∙mL-1, 90 % of the minimum inhibitory concentration, MIC), 3.12 µm (at 14 mg∙mL-1, 70 % of the MIC), and 5.21 µm (at 10 mg∙mL-1, 50 % of the MIC). Additionally, L. fermentum TCUESC01 supernatant modulated the expression of icaA and icaR. CONCLUSIONS: L. fermentum TCUESC01 reduces the formation of S. aureus biofilm under subinhibitory conditions. Inhibition of biofilm production probably depends on modulation of the ica operon.

Sepsis induced by Staphylococcus aureus: participation of biomarkers in a murine model.

BACKGROUND: This study aimed to evaluate the role of biomarkers in the pathophysiological process induced by a Staphylococcus aureus strain obtained in a hospital environment. For this, we intraperitoneally inoculated groups of male BALB/c mice with S. aureus, using a clinical isolate (CI) of S. aureus. MATERIAL/METHODS: Mice were divided into groups according to time of euthanasia (24, 48, 72, 96, 120, 144, and 168 hours of infection). After being euthanized, blood samples were collected for quantification of microorganisms and leukocytes, as well as measurement of biomarkers of tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), C-reactive protein (CRP), and Procalcitonin (PCT) by ELISA. Heart, kidneys, and lungs were removed for histopathological analysis, assessment of biomarkers of tissue expression by RT-PCR (polymerase chain reaction with reverse transcriptase), and quantification of microorganisms by real-time quantitative PCR (real-time PCR). RESULTS: The animals infected at between 120 hours and 168 hours had the highest blood levels of S. aureus. We observed that infection promoted increases in the levels of circulating neutrophils and monocytes. However, there was a reduction of circulating neutrophils and monocytes after 96 hours of infection. The infected mice also had increased levels of blood lymphocytes. In this model of infection with S. aureus, IL-6, CRP, and PCT demonstrated greater fidelity as markers of infection, since serum levels were elevated and lowered along with the number of circulating neutrophils and monocytes after resolution of the infection. The lungs showed hyperemia, with enlargement of the alveolar septa. On the other hand, infection with S. aureus did not promote visible change in histological tissue in the heart and kidneys. CONCLUSIONS: In this model of infection with S. aureus, IL-6, CRP, and PCT demonstrated greater fidelity as markers of infection, since serum levels were elevated and lowered along with the number of circulating neutrophils and monocytes after resolution of the infection. We believe our results may provide a better understanding of the pathophysiology, as well as aid in the search for a more reliable method of diagnosis.

Role of the aryl hydrocarbon receptor in the immune response profile and development of pathology during Plasmodium berghei Anka infection.

Infection with Plasmodium falciparum may result in severe disease affecting various organs, including liver, spleen, and brain, resulting in high morbidity and mortality. Plasmodium berghei Anka infection of mice recapitulates many features of severe human malaria. The aryl hydrocarbon receptor (AhR) is an intracellular receptor activated by ligands important in the modulation of the inflammatory response. We found that AhR-knockout (KO) mice infected with P. berghei Anka displayed increased parasitemia, earlier mortality, enhanced leukocyte-endothelial cell interactions in the brain microvasculature, and increased inflammation in brain (interleukin-17 [IL-17] and IL-6) and liver (gamma interferon [IFN-γ] and tumor necrosis factor alpha [TNF-α]) compared to infected wild-type (WT) mice. Infected AhR-KO mice also displayed a reduction in cytokines required for host resistance, including TNF-α, IL-1ß, and IFN-γ, in the brain and spleen. Infection of AhR-KO mice resulted in an increase in T regulatory cells and transforming growth factor ß, IL-6, and IL-17 in the brain. AhR modulated the basal expression of SOCS3 in spleen and brain, and P. berghei Anka infection resulted in enhanced expression of SOCS3 in brain, which was absent in infected AhR-KO mice. These data suggest that AhR-mediated control of SOCS3 expression is probably involved in the phenotype seen in infected AhR-KO mice. This is, to our knowledge, the first demonstration of a role for AhR in the pathogenesis of malaria.

Apoptosis in HEp-2 cells infected with Ureaplasma diversum.

BACKGROUND: Bacterial pathogens have many strategies for infecting and persisting in host cells. Adhesion, invasion and intracellular life are important features in the biology of mollicutes. The intracellular location of Ureaplasma diversum may trigger disturbances in the host cell. This includes activation or inhibition of pro and anti-apoptotic factors, which facilitate the development of host damage. The aim of the present study was to associate U. diversum infection in HEp-2 cells and apoptosis induction. Cells were infected for 72hs with four U. diversum clinical isolates and an ATCC strain. The U. diversum invasion was analyzed by Confocal Laser Scanning Microscopy and gentamicin invasion assay. The apoptosis was evaluated using pro-apoptotic and anti-apoptotic gene expression, and FITC Annexin V/Dead Cell Apoptosis Kit. RESULTS: The number of internalized ureaplasma in HEp-2 cells increased significantly throughout the infection. The flow cytometry analysis with fluorochromes to detect membrane depolarization and gene expression for caspase 2, 3 and 9 increased in infected cells after 24 hours. However, after 72 hours a considerable decrease of apoptotic cells was observed. CONCLUSIONS: The data suggests that apoptosis may be initially induced by some isolates in association with HEp-2 cells, but over time, there was no evidence of apoptosis in the presence of ureaplasma and HEp-2 cells. The initial increase and then decrease in apoptosis could be related to bacterial pathogen-associated molecular pattern (PAMPS). Moreover, the isolates of U. diversum presented differences in the studied parameters for apoptosis. It was also observed that the amount of microorganisms was not proportional to the induction of apoptosis in HEp-2 cells.

A practitioner's field guide to the behaviour settings method.

Since the 1950s, Roger Barker's theory of behaviour settings has been useful for a wide number of disciplines. Few realize, however, that behaviour settings theory is also a methodology. Barker fully describes how to identify, describe and measure behaviour settings in his seminal book Ecological psychology: concepts and methods for studying the environment of human behavior (1968), and this method is further delineated in Phil Schoggen's Behavior settings: a revision and extension of Roger G. Barker's ecological psychology (1989). Nevertheless, beyond these two (rather expensive) books there are few other resources available to twenty-first century researchers who wish to systematically describe and measure behaviour in its ecological context using the principles of behaviour settings theory. In this article, I offer a practitioner's field guide to implementing the behaviour settings method, which includes a contemporary illustration of defining a behaviour setting using a recent observational study of an art gallery in Lethbridge, Canada. I discuss how researchers can use Barker's original methodology to determine what is a behaviour setting and how to define its boundaries, and I suggest best practices, offering practitioners the tools to replicate Barker's procedures. This article is part of the theme issue 'People, places, things, and communities: expanding behaviour settings theory in the twenty-first century'.

People, places, things and communities: expanding behaviour settings theory in the twenty-first century.

Trends and developments in recent behavioural and cognitive sciences demonstrate the need for a well-developed theoretical and empirical framework for examining the ecology of human behaviour. The increasing recognition of the role of the environment and interaction with the environment in the organization of behaviour within the cognitive sciences has not been met with an equally disciplined and systematic account of that environment (Heft 2018 Ecol. Psychol. 30, 99-123 (doi:10.1080/10407413.2018.1410045); McGann 2014 Synth. Philos. 29, 217-233). Several bodies of work in behavioural ecology, anthropology and ecological psychology provide some frameworks for such an account. At present, however, the most systematic and theoretically disciplined account of the human behavioural ecosystem is that of behaviour settings, as developed by the researchers of the Midwest Psychological Field Station (see Barker 1968 Ecological psychology: concepts and methods for studying the environment of human behavior). The articles in this theme issue provide a critical examination of these theoretical and methodological resources. The collection addresses their theoretical value in connecting with contemporary issues in cognitive science and research practice in psychology, as well as the importance of the methodological specifics of behaviour settings research. Additionally, articles diagnose limitations and identify points of potential extension of both theory and methods, particularly with regard to changes owing to the advance of technology, and the complex relationship between the individual and the collective in behaviour settings work. This article is part of the theme issue 'People, places, things, and communities: expanding behaviour settings theory in the twenty-first century'.