Bartonella spp. in bats from the Brazilian Amazon Forest.

Despite the great diversity of bats (64 species) in the State of Acre, northwestern Brazil, there are no studies on occurrence and diversity of Bartonella spp. in bats in this region. The present study investigated the occurrence and molecular identity of Bartonella spp. in spleen samples (n = 271) from bats of 30 different species from this region, within the Amazon biome. Twenty-one out of 208 (10.1%) samples positive in the PCR for the mammalian gapdh endogenous genes were positive in the qPCR for Bartonella spp. based on the nuoG gene. The two gltA Bartonella genotypes detected grouped with those previously identified in bats from other locations, expanding the diversity of genotypes associated with bats. This study provided the first molecular evidence of the presence of Bartonella spp. in bats in the state of Acre and in bats of the species Lophostoma silvicolum, Vampyressa thyone, Tonatia saurophila and Phyllostomus elongatus.

Novel Tick-Borne Anaplasmataceae Genotypes in Tropical Birds from the Brazilian Pantanal Wetland.

Despite numerous reports of Anaplasmataceae agents in mammals worldwide, few studies have investigated their occurrence in birds. The present study aimed to investigate the occurrence and molecular identity of Anaplasmataceae agents in birds from the Pantanal wetland, Brazil. Blood samples were collected from 93 different species. After DNA extraction, samples positive for the avian ß-actin gene were subjected to both a multiplex quantitative real-time (q)PCR for Anaplasma and Ehrlichia targeting the groEL gene and to a conventional PCR for Anaplasmataceae agents targeting the 16S rRNA gene. As a result, 37 (7.4%) birds were positive for Anaplasma spp. and 4 (0.8%) for Ehrlichia spp. in the qPCR assay; additionally, 13 (2.6%) were positive for Anaplasmataceae agents in the PCR targeting the 16S rRNA gene. The Ehrlichia 16S rRNA sequences detected in Arundinicola leucocephala, Ramphocelus carbo, and Elaenia albiceps were positioned closely to Ehrlichia sp. Magellanica. Ehrlichia dsb sequences detected in Agelasticus cyanopus and Basileuterus flaveolus grouped with Ehrlichia minasensis. The 16S rRNA genotypes detected in Crax fasciolata, Pitangus sulphuratus and Furnarius leucopus grouped with Candidatus Allocryptoplasma. The 23S-5S genotypes detected in C. fasciolata, Basileuterus flaveolus, and Saltator coerulescens were related to Anaplasma phagocytophilum. In conclusion, novel genotypes of Anaplasma, Ehrlichia, and Candidatus Allocryptoplasma were detected in birds from the Pantanal wetland.

Bartonella species in dromedaries and ruminants from Lower Shabelle and Benadir regions, Somalia.

BACKGROUND: Bartonellosis, caused by bacteria of the genus Bartonella, is a zoonotic disease with several mammalian reservoir hosts. In Somalia, a country heavily reliant on livestock, zoonotic diseases pose significant public health and economic challenges. To the best of our knowledge, no study has been performed aiming to verify the occurrence of Bartonella spp. in Somalia. This study investigated the occurrence and molecular characterization of Bartonella in dromedary (Camelus dromedarius, Linnaeus, 1758), cattle, sheep, and goats from Somalia. MATERIALS AND METHODS: 530 blood samples were collected from various animals (155 dromedary, 199 goat, 131 cattle, and 45 sheep) in Benadir and Lower Shabelle regions. DNA was extracted for molecular analysis, and a qPCR assay targeting the NADH dehydrogenase gamma subunit (nuoG) gene was used for Bartonella screening. Positive samples were also subjected to PCR assays targeting seven molecular markers including: nuoG, citrate synthase gene (gltA), RNA polymerase beta-subunit gene (rpoB), riboflavin synthase gene (ribC), 60 kDa heat-shock protein gene (groEL), cell division protein gene (ftsZ), and pap31 and qPCR targeting the 16-23S rRNA internal transcribed spacer (ITS) followed by Sanger sequencing, BLASTn and phylogenetic analysis. RESULTS: Out of 530 tested animals, 5.1% were positive for Bartonella spp. by the nuoG qPCR assay. Goats showed the highest Bartonella occurrence (17/199, 8.5%), followed by sheep (6/44, 6.8%), cattle (4/131, 3.1%), and dromedary (1/155, 1.9%). Goats, sheep, and cattle had higher odds of infection compared to dromedary. Among nuoG qPCR-positive samples, 11.1%, 14.8%, 11.1%, and 25.9% were positive in PCR assays based on nuoG, gltA, and pap31 genes, and in the qPCR based on the ITS region, respectively. On the other hand, nuoG qPCR-positive samples were negative in the PCR assays targeting the ribC, rpoB, ftsZ, and groEL genes. While Bartonella bovis sequences were detected in cattle (nuoG and ITS) and goats (gltA), Bartonella henselae ITS sequences were detected in dromedary, goat, and sheep. Phylogenetic analysis placed gltA Bartonella sequence from a goat in the same clade of B. bovis. CONCLUSION: The present study showed, for the first time, molecular evidence of Bartonella spp. in dromedary and ruminants from Somalia and B. henselae in sheep and goats globally. These findings contribute valuable insights into Bartonella spp. occurrence in Somali livestock, highlighting the need for comprehensive surveillance and control measures under the One Health approach.

Molecular evidence of Bartonella spp. in tropical wild birds from the Brazilian Pantanal, the largest wetland in South America.

Despite the worldwide occurrence of bartonellae in a broad range of mammal species, in which they usually cause a long-lasting erythrocytic bacteremia, few studies reported Bartonella spp. in avian hosts. The present work aimed to investigate the occurrence and molecular identity of Bartonella spp. infecting birds in the Pantanal wetland, central-western Brazil using a multigene approach. For this purpose, blood samples were collected from 517 individuals from 13 avian orders in the states of Mato Grosso and Mato Groso do Sul. DNA was extracted from avian blood and 500/517 (96.7%) samples were positive in a conventional PCR targeting the avian ß-actin gene. Nineteen (3.8%) out of 500 avian blood samples were positive in a qPCR assay for Bartonella spp. based on the nuoG gene. Among 19 avian blood DNA samples positive in the qPCR for Bartonella spp., 12 were also positive in the qPCR for Bartonella based on the 16S-23S RNA Intergenic region (ITS). In the PCR assays performed for molecular characterization, one 16S rRNA, three ribC, and one nuoG sequences were obtained. Based on BLASTn results, while 1 nuoG, 2 ribC, and 2 ITS sequences showed high identity to Bartonella henselae, one 16S rRNA and 2 ITS showed high similarity to Bartonella machadoae in the sampled birds. Bartonella spp. related to B. henselae and B. machadoae were detected, for the first time, in wild birds from the Brazilian Pantanal.

Acinetobacter baumannii Early-Onset Sepsis After Home Delivery Into Toilet Water.

Early-onset sepsis (EOS) is an important cause of morbidity and mortality in newborns, usually caused by pathogens acquired intrapartum. We present the case of a term neonate born by home delivery in the toilet, after an unsupervised pregnancy. He developed a culture-proven early-onset sepsis caused by Acinetobacter baumannii. This was the first case of neonatal sepsis by this pathogen in our unit. The microorganism was susceptible to all antibiotics tested. The neonate was treated empirically with ampicillin and cefotaxime and completed 21 days of directed therapy with meropenem, as meningitis could not be excluded. During the clinical course, the newborn developed severe and persistent thrombocytopenia and neutropenia. In this report, we discuss the etiology behind this clinical presentation. We intend to raise awareness for the consideration of Acinetobacter baumannii as a potential pathogen in EOS, particularly in the presence of adverse birth circumstances.

Molecular evidence of Bartonella spp. in wild lowland tapirs (Tapirus terrestris), the largest land mammals in Brazil.

The genus Bartonella (Hyphomicrobiales: Bartonellaceae) encompasses facultative intracellular α-proteobacteria that parasite erythrocytes and endothelial cells from a wide range of vertebrate hosts and can cause disease in animals and humans. Considering the large diversity of vertebrate species that may act as reservoirs and arthropod species that may be associated with Bartonella transmission, the exposure of animals and humans to these microorganisms is likely underestimated. The present study aimed to investigate the occurrence of Bartonella sp. in wild tapirs (Tapirus terrestris; Perissodactyla: Tapiridae) from two biomes in Brazil: Pantanal and Cerrado. Ninety-nine GPS-monitored wild tapirs were sampled in Pantanal (n = 61/99) and Cerrado (n = 38/99). A qPCR (quantitative real-time polymerase chain reaction) assay targeting the nuoG gene was used for the screening for Bartonella spp. DNA. Positive samples were additionally subjected to conventional PCR assays targeting five molecular markers (ribC, gltA, rpoB, groEL, ITS). Eight (8/99; 08,08%) animals were positive in the qPCR assay for Bartonella spp.: 7 from Cerrado (7/8; 87.5%) and 1 from Pantanal (1/8; 12.5%). The 5 Bartonella ribC sequences obtained from tapirs' blood samples grouped together with Bartonella henselae obtained from cats, humans, wild felids and Ctenocephalides felis (Siphonaptera: Pulicidae) fleas. To the best of author's knowledge, this is the first report of Bartonella sp. in Tapirus terrestris. This finding contributes to the understanding of the occurrence of B henselae in wild mammals from Brazil as well as expands the knowledge regarding the potential vector-borne pathogens that may affect wild tapis from Cerrado and Pantanal biomes.

Prevention of risky sexual behavior in primary healthcare: a best practice implementation project.

INTRODUCTION AND OBJECTIVES: Sexually transmitted infections are a public health problem affecting 45% of adolescents and young adults worldwide. The evidence suggests that primary care settings are uniquely positioned to provide an opportunity for these preventive interventions. The aim of this project is to improve nurses' interventions for preventing risky sexual behaviors in adolescents attending nursing consultations in a primary healthcare unit. METHODS: An audit and feedback were conducted by the JBI Model and Implementation Framework. Five audit criteria representing best practice recommendations for preventing risky sexual behaviors in adolescents were used. Barriers to compliance with the best practices were identified, and strategies were adopted to overcome them. A follow-up audit was conducted using the same approach as the baseline audit. RESULTS: Compliance rates improved in four criteria from baseline audit to follow-up audit. CONCLUSION: Through auditing and feedback, evidence-based interventions were implemented to prevent sexual risk behavior in adolescents in primary care settings. Further best practice implementation projects should be conducted to improve adolescent health outcomes.

Molecular detection and characterization of vector-borne agents in common opossums (Didelphis marsupialis) from northeastern Brazil.

Opossums are synanthropic marsupials able to interchange among wild, peri­urban and urban environments, playing an epidemiologically important role as hosts for emerging pathogens and ectoparasites of relevance in public health. The present study aimed to detect and molecularly characterize vector-borne agents in a population of common opossums (Didelphis marsupialis) from the Island of São Luís do Maranhão, northeastern Brazil. Of the 45 animals analyzed, one (2.22%) was positive in the nested PCR assay based on the 18S rRNA gene of piroplasmids. The obtained sequence was phylogenetically positioned in a clade containing sequences of Babesia sp. previously detected in Didelphis aurita, Didelphis albiventris and associated ticks from Brazil. Eight (17.77%) samples were positive in PCR for Ehrlichia spp. based on the dsb gene; four samples were sequenced and positioned into a new clade, sister to E. minasensis and Ehrlichia sp. clade detected in Superorder Xenarthra mammals. No samples tested positive in the screening PCR assays based on the 16S rRNA gene of Anaplasma spp. Two samples were positive in the qPCR for Bartonella spp. based on the nuoG gene. Seven animals (15.56%) were positive in the nPCR based on the 16S rRNA gene of hemoplasmas. Of these, three were positive in a PCR based on the 23S rRNA gene. The phylogenies based on both 16S rRNA and 23S rRNA genes corroborated to each other and positioned the sequences in the same clade of hemoplasmas previously detected in D. aurita and D. albiventris sampled in Brazil. Finally, three (6.66%) animals were positive in the PCR for Hepatozoon spp.; the obtained 18S rRNA sequence was positioned into the H. felis clade.The present study showed, for the first time, the circulation of piroplasmids, Hepatozoon spp., Ehrlichia spp., hemoplasmas and Bartonella spp. in D. marsupialis sampled in northeastern Brazil, with description of putative novel genotypes of Ehrlichia and Hepatozoon and copositivity by different vector-borne agents. The present work consolidates the "South American Marsupialia" piroplasmid clade, adding one more genotype of Babesia sp. to this clade.

Multi-locus Sequencing Typing of Bartonella henselae isolates reveals coinfection with different variants in domestic cats from Midwestern Brazil.

Bartonella henselae is a zoonotic pathogen responsible for causing Cat Scratch Disease (CSD) and other clinical manifestations in humans. Domestic cats are the main reservoirs of this Bartonella species. Previous studies have suggested that certain genotypes of B. henselae seem to be more associated with human infections. The present study aimed to genotype B. henselae isolates from domestic cats' blood samples in the state of Goiás, midwestern Brazil. The association of quantitative real-time PCR (qPCR) based on the nuoG gene from Bartonella spp. of blood samples, before and after incubation in pre-enrichment liquid medium (BAPGM) and isolation on chocolate agar, showed a positivity frequency of 42% (42/100) for Bartonella spp. Twelve B. henselae isolates obtained on agar chocolate from six cats' blood samples (two isolates from each animal) were characterized by Multi-locus Sequencing Typing (MLST) and revealed to belong to Sequence Types ST1 and ST5. One of the cats (1/6) presented both STs, demonstrating that domestic cats can be coinfected with different variants of B. henselae. The STs detected in this study are distributed worldwide and have already been detected in humans with clinical manifestations of bartonellosis. This is the first report of the zoonotic variants ST1 and ST5 of B. henselae in domestic cats from Brazil.

First Report of Bartonella spp. in Marsupials from Brazil, with a Description of Bartonella harrusi sp. nov. and a New Proposal for the Taxonomic Reclassification of Species of the Genus Bartonella.

The genus Bartonella (Rhizobiales: Bartonellaceae) encompasses facultative intracellular Gram-negative alphaproteobacteria that parasitize mainly erythrocytes and endothelial cells, as well as macrophages, monocytes and dendritic cells. Although they can infect numerous mammal species and arthropod vectors worldwide, reports of Bartonella infections in marsupials are scarce. In fact, such agents have only been detected in marsupials and/or associated ectoparasites in Australia and the United States of America until the present moment. The present study aimed to isolate and characterize molecularly, morphologically and phenotypically Bartonella infecting free-living marsupials sampled in the Brazilian Pantanal, the largest wetland in South America. Two marsupials were captured in December 2018 and six marsupials in February 2019, totaling eight small mammals sampled: five (62.5%) Thylamys macrurus and three (37.5%) Monodelphis domestica. All blood samples were submitted to qPCR for Bartonella spp. based on the nuoG gene, a pre-enrichment liquid culture and a chocolate agar solid culture. Bartonella sp. was isolated from 3 T. macrurus and one M. domestica. One Bartonella isolate obtained from a T. macrurus blood sample (strain 117A) that showed to be closely related to the Bartonella vinsonii complex and Bartonella machadoae was selected for whole genome sequencing using a hybrid approach based on Illumina NovaSeq and Nanopore sequencing platforms. This strain showed a genome of 2.35 Mbp, with an average C + G content of 38.8%, coding for 2013 genes, and a 29 kb plasmid with an average C + G content of 34.5%. In addition, this strain exhibited an average nucleotide identity (ANI) of 85% with Bartonella species belonging to the B. vinsonii group and 91% with B. machadoae. Phylogenomic analysis based on 291 protein coding genes shared by the genomes of 53 Bartonella species positioned this strain closely to B. machadoae. This new isolated species was named Bartonella harrusi sp. nov., which was characterized as having small capnophilic, microaerophilic and aerobic rods with an absence of pili and flagella. In conclusion, the present work describes the biochemical, phenotypic and genomic characteristics of Bartonella harrusi, a new species isolated from the T. macrurus blood samples of the Brazilian Pantanal. Finally, a review of the taxonomic classification of members of the genus Bartonella is proposed, based on the ANI values accessed by whole genome sequencing analyses.

Sex differences in the association between social capital and healthcare use-Results from the Survey of Health, Ageing and Retirement in Europe (SHARE).

Females and males frequently report substantial differences in social capital indicators and may use healthcare distinctly. Nevertheless, the potential effect of sex on the relation between social capital and healthcare use remains unclear. This study aims to quantify the association between different indicators of individuals' social capital and healthcare use, according to sex. Data were retrieved from the Sixth Wave of the Survey of Health, Ageing and Retirement in Europe (SHARE), which was conducted in 2015, and included 68,188 participants from 18 countries. Adjusted odds ratios (AOR) and 95% confidence intervals (95%CI) were computed using logistic regression. Overall, males and females with smaller social networks, those who live alone or with any other relatives besides their partners, and those whose first close confidant was a family member or a neighbour reported fewer contacts with medical doctors or nurses, as well as with dentists or dental hygienists. Amongst females, participation in educational or training courses (AOR = 1.67, 95%CI:1.40-2.00; p for interaction = 0.035) and sport, social or any other club (AOR = 1.79, 95%CI:1.58-2.02; p for interaction = 0.043) was associated with a more frequent contact with dentists or dental hygienists. Females who participated in volunteer or charity work (AOR = 0.76, 95%CI:0.64-0.91; p for interaction = 0.042) and political or community-related organisations (AOR = 0.72, 95%CI:0.52-1.00; p for interaction = 0.030) were less likely to report the use of polypharmacy. This outcome was more frequently observed amongst females who referred feelings of severe loneliness (AOR = 1.44, 95%CI:1.22-1.68; p for interaction < 0.001). Social capital is associated with healthcare use distinctively amongst males and females. Increasing opportunities for social participation may improve healthcare use, particularly amongst females.

Bartonella machadoae sp. nov. isolated from wild rodents in the Pantanal wetland.

It has been estimated that 75% of emerging infectious diseases comprise zoonoses, whose majority have free-living animals as reservoirs and are mainly transmitted by arthropod vectors. Although rodents represent important Bartonella reservoirs, there are few studies on the genotypic characterization of Bartonella species commonly found in this taxon and from different Brazilian biomes. Therefore, the present study aimed to investigate the occurrence, isolate and molecularly, morphologically and phenotypically characterize a new Bartonella species infecting free-living rodents sampled in the Brazilian Pantanal, the largest wetland in South America. For this purpose, 129 free-living rodents (79 Thrichomys fosteri, 4 Clyomys laticeps, and Oecomys mamorae) were captured. While blood samples were collected from 57 T. fosteri, 4 C. laticeps and 32 O. mamorae; spleen samples were collected from 22 T. fosteri and 14 O. mamorae. Blood and spleen samples were submitted to a qPCR for Bartonella spp. targeting the nuoG gene, using DNA samples extracted directly from blood/spleen, after passage in pre-enrichment liquid culture, and from colonies obtained from solid culture on chocolate agar. Combining all techniques, occurrence of 24.8% for Bartonella sp. was found among the sampled rodents. One Bartonella isolate (strain 56A) obtained from a T. fosteri's blood sample was closely related to the Bartonella vinsonii complex and selected for Whole Genome Sequencing (WGS) hybrid approach using Illumina NovaSeq and Nanopore sequencing platforms. This strain exhibits a circular 2.7 Mbp genome with an average C+G content of 39% and encoding to 2239 genes. In the phylogenomics based on 291 shared protein-coding genes, this strain was positioned in a unique clade, closely related to Bartonella vinsonii subsp. vinsonii, B. vinsonii subsp. berkhoffii and B. visonii subsp. arupensis. An Average Nucleotide Identity of 85% was found between the obtained isolate and Bartonella species belonging to B. vinsonii complex. These findings supported the separation of this strain, now formally named as Bartonella machadoae sp. nov., from the Bartonella vinsonii complex. In addition, Bartonella machadoae sp. nov. was characterized by capnophilic, microaerophilic and aerobic small rods with absence of pili and flagella. Phylogenetic and distance analyses based on five concatenated molecular markers suggest that Bartonella machadoae may parasite rodents from different Brazilian biomes. In conclusion, we described biochemical, phenotypic and genomic characteristics of Bartonella machadoae nov. sp. isolated from blood samples of T. fosteri rodents from the Brazilian Pantanal.

Aryl Hydrocarbon Receptor and Cysteine Redox Dynamics Underlie (Mal)adaptive Mechanisms to Chronic Intermittent Hypoxia in Kidney Cortex.

We hypothesized that an interplay between aryl hydrocarbon receptor (AhR) and cysteine-related thiolome at the kidney cortex underlies the mechanisms of (mal)adaptation to chronic intermittent hypoxia (CIH), promoting arterial hypertension (HTN). Using a rat model of CIH-HTN, we investigated the impact of short-term (1 and 7 days), mid-term (14 and 21 days, pre-HTN), and long-term intermittent hypoxia (IH) (up to 60 days, established HTN) on CYP1A1 protein level (a sensitive hallmark of AhR activation) and cysteine-related thiol pools. We found that acute and chronic IH had opposite effects on CYP1A1 and the thiolome. While short-term IH decreased CYP1A1 and increased protein-S-thiolation, long-term IH increased CYP1A1 and free oxidized cysteine. In addition, an in vitro administration of cystine, but not cysteine, to human endothelial cells increased Cyp1a1 expression, supporting cystine as a putative AhR activator. This study supports CYP1A1 as a biomarker of obstructive sleep apnea (OSA) severity and oxidized pools of cysteine as risk indicator of OSA-HTN. This work contributes to a better understanding of the mechanisms underlying the phenotype of OSA-HTN, mimicked by this model, which is in line with precision medicine challenges in OSA.

Genetic diversity and Multilocus Sequence Typing Analysis of Bartonella henselae in domestic cats from Southeastern Brazil.

Bartonella henselae is the causative agent for the infectious disease Cat Scratch Disease (CSD), which can be fatal. Domestic and wild felines are known to be its main mammal reservoirs. The present study aimed to investigate the occurrence and genetic diversity of Bartonella spp. in cats sampled in São Paulo (SP) and Minas Gerais (MG) States, Southeastern Brazil. Based on a quantitative real-time PCR (qPCR) assay, a Bartonella sp. nuoG gene fragment was detected in 39.9% (122/306) of the blood samples (46/151 cats of SP; 76/155 cats of MG). The blood samples were submitted to a pre-enrichment culture technique that allowed the detection of 12 additional positive samples, which showed to be negative in the qPCR using DNA blood samples as templates. Furthermore, five B. henselae isolates were obtained from qPCR-negative samples for both blood and pre-enrichment culture. Seven out of 24 Ctenocephalides felis fleas were positive for Bartonella spp. in the qPCR assay; 4/7 positive fleas were collected from Bartonella-negative cats. Twenty-three rpoB B. henselae cloned sequences were obtained from nine cats' blood samples, showing the occurrence of 13 different genotypes. Median-joining network and SplitsTree distance analysis showed that the obtained sequences represented distinct B. henselae genotypes when compared to those previously deposited in GenBank. Intra-host diversity was found, since different rpoB genotypes of B. henselae were detected in individual single cats. Bartonella henselae isolates showed two allelic profiles (ST37 in cats from MG state and ST9 in SP state) by MLST (Multilocus Sequence Typing) based on sequencing of eight molecular markers. The present study is the first molecular report of Bartonella sp. in cats from Minas Gerais State. In summary, this body of work showed the occurrence of different B. henselae rpoB genotypes at an intra-reservoir host level. Based on qPCR from blood samples and pre-enrichment liquid culture and isolation, occurrence of 33.1% (50/151) and 56.8% (88/155) for Bartonella sp. was found in cats from SP and MG states, respectively. Two different allelic profiles of B. henselae were found in cats from the states of São Paulo (ST9) and Minas Gerais (ST37), suggesting a clonal evolution of Bartonellae in a certain geographical region.

A Mechanistic-Based and Non-invasive Approach to Quantify the Capability of Kidney to Detoxify Cysteine-Disulfides.

Our general goal was to non-invasively evaluate kidney tubular dysfunction. We developed a strategy based on cysteine (Cys) disulfide stress mechanism that underlies kidney dysfunction. There is scarce information regarding the fate of Cys-disulfides (CysSSX), but evidence shows they might be detoxified in proximal tubular cells by the action of N-acetyltransferase 8 (NAT8). This enzyme promotes the addition of an N-acetyl moiety to cysteine-S-conjugates, forming mercapturates that are eliminated in urine. Therefore, we developed a strategy to quantify mercapturates of CysSSX in urine as surrogate of disulfide stress and NAT8 activity in kidney tubular cells. We use a reduction agent for the selective reduction of disulfide bonds. The obtained N-acetylcysteine moiety of the mercapturates from cysteine disulfides was monitored by fluorescence detection. The method was applied to urine from mice and rat as well as individuals with healthy kidney and kidney disease.

The 2-hydroxy-nevirapine metabolite as a candidate for boosting apolipoprotein A1 and for modulating anti-HDL antibodies.

The antiretroviral nevirapine (NVP) is associated to a reduction of atherosclerotic lesions and increases in high-density lipoprotein (HDL)-cholesterol. Despite being a hepatotoxic drug, which forbids its re-purposing to other therapeutic areas, not all NVP metabolites have the same potential to induce toxicity. Our aim was to investigate the effects of NVP and its metabolites in an exploratory study, towards the identification of a candidate to boost HDL. A pilot prospective (n = 11) and a cross-sectional (n = 332) clinical study were performed with the following endpoints: HDL-cholesterol and apolipoprotein A1 (ApoA1) levels, anti-HDL and anti-ApoA1 antibodies titers, paraoxonase, arylesterase and lactonase activities of paraoxonase-1, and NVP's metabolite profile. NVP treatment increased HDL-cholesterol, ApoA1 and paraoxonase-1 activities, and lowered anti-HDL and anti-ApoA1 titers. In the prospective study, the temporal modulation induced by NVP was different for each HDL-related endpoint. The first observation was a decrease in the anti-HDL antibodies titers. In the cross-sectional study, the lower titers of anti-HDL antibodies were associated to the proportion of 2-hydroxy-NVP (p = 0.03). In vitro models of hepatocytes were employed to clarify the individual contribution of NVP's metabolites for ApoA1 modulation. Long-term incubations of NVP and 2-hydroxy-NVP in the metabolically competent 3D model caused an increase in ApoA1 reaching 43 % (p < 0.05) and 86 % (p < 0.001), respectively. These results support the contribution of drug biotransformation for NVP-induced HDL modulation, highlighting the role of 2-hydroxy-NVP as ApoA1 booster and its association to lower anti-HDL titers. This biotransformation-guided approach allowed us to identify a non-toxic NVP metabolite as a candidate for targeting HDL.

First evidence of aryl hydrocarbon receptor as a druggable target in hypertension induced by chronic intermittent hypoxia.

BACKGROUND AND PURPOSE: Obstructive sleep apnea (OSA) is associated to a high prevalence of resistant arterial hypertension (HTN) justifying the research on novel targets. Chronic intermittent hypoxia (CIH) is a key feature in the development of OSA comorbidities, including HTN. EXPERIMENTAL APPROACH: We used a rat model of CIH-induced HTN to disclose the hypothesis that the aryl hydrocarbon receptor (AHR) is activated by CIH once it shares the same binding partner of HIF-1α and promotes pro-oxidant, pro-inflammatory (NF-kB) and pro-fibrotic events in common with CIH. KEY RESULTS: Upon established hypertension (21 days exposure to CIH), we observed an increase in Cyp1a1 mRNA in kidney cortex (6-fold), kidney medulla (3-fold) and liver (3-fold), but not in other tissues. Increased renal expression of Ahr and markers of inflammation (Rela), epithelial to mesenchymal transition markers, the rate-controlling step of gluconeogenesis, Pepck1, and members of HIF-pathway, namely, Hif3a were also observed. Daily administration (14 days) of AHR antagonist, CH-223191 (5 mg.kg-1.day-1, gavage), simultaneously to CIH prevented the increase in systolic blood pressure (SBP) by 53 ± 12% and in diastolic blood pressure (DBP) by 44 ± 16%. Moreover, its administration (14 days) upon already established HTN reversed the increase in SBP by 52 ± 12%. CONCLUSION AND IMPLICATIONS: CIH caused an activation of AHR signaling particularly in the kidney and its pharmacological blockade had a significant impact reverting already established HTN. This first evidence inspires innovative research opportunities for the understanding and treatment of this particular type of HTN.

Andiroba oil (Carapa guianensis Aubl) shows cytotoxicity but no mutagenicity in the ACPP02 gastric cancer cell line.

Andiroba (Carapa guianensis Aubl) is an Amazonian plant whose oil has been widely used in traditional medicine for various purposes, including anti-inflammation. Research reports indicate that the oil can confer antitumor activity due to the presence of fatty acids, which can directly influence cell death mechanisms. Thus, andiroba oil (AO) has gained interest for its potential to be used in antineoplastic therapies. Here, we report an in vitro analysis of the cytotoxic and mutagenic potential of AO in the gastric cancer cell line, ACP02. Cell survival was assessed by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay, differential staining with ethidium bromide and acridine orange assessed apoptosis-necrosis, and mutagenesis was assessed by the micronucleus test. The apolar oil was first diluted in 0.1% dimethyl sulfoxide (DMSO) and then further diluted to six concentrations (0.01, 0.1, 1, 10 and 100 µg/mL and 1 mg/mL) in RPMI medium. Controls included RPMI alone (negative control) and 0.1% DMSO diluted in medium (vehicle control). The MTT test showed that AO significantly reduced cell viability (P < .05) only when the highest tested concentration was applied for 48 hours. The apoptosis/necrosis test showed that the highest concentration of AO induced cell death by apoptosis at 24 and 48 hours. There was no statistically significant increase in the frequency of micronuclei. The ability of the AO to decrease the viability of ACP02 cells via apoptosis, without exerting mutagenic effects, suggests that the oil could be useful as an alternative therapeutic agent for primary tumors of stomach cancer.