An Inner Mitochondrial Membrane Microprotein from the SLC35A4 Upstream ORF Regulates Cellular Metabolism.

Upstream open reading frames (uORFs) are cis-acting elements that can dynamically regulate the translation of downstream ORFs by suppressing downstream translation under basal conditions and, in some cases, increasing downstream translation under stress conditions. Computational and empirical methods have identified uORFs in the 5'-UTRs of approximately half of all mouse and human transcripts, making uORFs one of the largest regulatory elements known. Because the prevailing dogma was that eukaryotic mRNAs produce a single functional protein, the peptides and small proteins, or microproteins, encoded by uORFs were rarely studied. We hypothesized that a uORF in the SLC35A4 mRNA is producing a functional microprotein (SLC35A4-MP) because of its conserved amino acid sequence. Through a series of biochemical and cellular experiments, we find that the 103-amino acid SLC35A4-MP is a single-pass transmembrane inner mitochondrial membrane (IMM) microprotein. The IMM contains the protein machinery crucial for cellular respiration and ATP generation, and loss of function studies with SLC35A4-MP significantly diminish maximal cellular respiration, indicating a vital role for this microprotein in cellular metabolism. The findings add SLC35A4-MP to the growing list of functional microproteins and, more generally, indicate that uORFs that encode conserved microproteins are an untapped reservoir of functional microproteins.

Type 2 vomeronasal receptor-A4 subfamily: Potential predator sensors in mice.

Sensory signals detected by olfactory sensory organs are critical regulators of animal behavior. An accessory olfactory organ, the vomeronasal organ, detects cues from other animals and plays a pivotal role in intra- and inter-species interactions in mice. However, how ethologically relevant cues control mouse behavior through approximately 350 vomeronasal sensory receptor proteins largely remains elusive. The type 2 vomeronasal receptor-A4 (V2R-A4) subfamily members have been repeatedly detected from vomeronasal sensory neurons responsive to predator cues, suggesting a potential role of this receptor subfamily as a sensor for predators. This review focuses on this intriguing subfamily, delving into its receptor functions and genetic characteristics.

Aflatoxins in maize flour produced in Mozambique and its risk assessment.

This study investigated the occurrence of aflatoxins (B1, B2, G1, and G2) in maize flour produced in Mozambique and to assess the associated carcinogenic risk. At different opportunities, 30 samples of maize flour were collected in five flour processing factories. These were determined by high-performance liquid chromatography (HPLC) with fluorescence detection. AFB1 concentrations ranged from 0.25 to 0.33 µg kg-1. The levels of total aflatoxins ranged from 0.55 to 1.05 µg kg-1, with a mean of 0.89 µg kg-1, for which maximum limits (MLs) are 10 and 4 µg kg-1 for Mozambique and the European Union, respectively. The calculated Margin of Exposure (MOE) for men and women was 243 and 231, respectively, so several folds below the risk cut-off level, indicating that consumption of such maize flour poses a potential risk of hepatocarcinoma related to aflatoxin exposure due to high intake of this food, a staple diet in most African countries.

Hypothalamic representation of the imminence of predator threat detected by the vomeronasal organ in mice.

Animals have the innate ability to select optimal defensive behavioral outputs with an appropriate intensity in response to predator threat in specific contexts. Such innate behavioral decisions are thought to be computed in the medial hypothalamic nuclei that contain neural populations directly controlling defensive behavioral outputs. The vomeronasal organ (VNO) is one of the major sensory input channels through which predator cues are detected with ascending inputs to the medial hypothalamic nuclei, especially to the ventromedial hypothalamus (VMH), through the medial amygdala (MeA) and bed nucleus of the stria terminalis (BNST). Here, we show that cat saliva contains predator cues that signal imminence of predator threat and regulate the robustness of freezing behavior through the VNO in mice. Cat saliva activates neurons expressing the V2R-A4 subfamily of sensory receptors, suggesting the existence of specific receptor groups responsible for freezing behavior induced by the predator cues. The number of VNO neurons activated in response to saliva correlates with the freshness of saliva and the intensity of freezing behavior, while the downstream neurons in the accessory olfactory bulb (AOB) and defensive behavioral circuit are quantitatively equally activated by fresh and old saliva. Strikingly, however, only the number of VMH neurons activated by fresh saliva positively correlates with the intensity of freezing behavior. Detailed analysis of the spatial distribution of fresh and old saliva-responding neurons revealed a neuronal population within the VMH that is more sensitive to fresh saliva than old saliva. Taken together, this study demonstrates that predator cues in cat saliva change over time and differentially activate the sensory-to-hypothalamus defensive behavioral pathway to modulate behavioral outputs.

Analytical method for urinary homovanillic acid and 5-hydroxyindoleacetic acid levels using HPLC with electrochemical detection applied to evaluate children environmentally exposed to manganese.

Homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5-HIAA) are the urinary metabolites of dopamine (DA) and serotonin (5-HA), respectively. We aimed to develop an extraction method for the determination of HVA and 5-HIAA, using strong anionic exchange cartridges combined with HPLC with electrochemical detection, and apply it to measure the levels of HVA and 5-HIAA in children living near a ferro-manganese alloy plant in Simões Filho, Brazil. The validated method showed good selectivity, sensitivity, precision, and accuracy. The limits of detection (LOD) were 4 and 8 µmol/L for 5-HIAA and HVA, respectively, in urine. Recoveries ranged from 85.8 to 94%. The coefficients of determination (R2 ) of the calibration curves were greater than 0.99. Spot urine samples of 30 exposed children and 20 nonexposed ones were processed accordingly. The metabolite levels in exposed and reference children were within the physiological ranges. The medians (range) for 5-HIAA and HVA of the exposed ones were 36.4 µmol/L (18.4-58.0) and 32.9 µmol/L (

Oral supplementation of nicotinamide riboside alters intestinal microbial composition in rats and mice, but not humans.

The gut microbiota impacts systemic levels of multiple metabolites including NAD+ precursors through diverse pathways. Nicotinamide riboside (NR) is an NAD+ precursor capable of regulating mammalian cellular metabolism. Some bacterial families express the NR-specific transporter, PnuC. We hypothesized that dietary NR supplementation would modify the gut microbiota across intestinal sections. We determined the effects of 12 weeks of NR supplementation on the microbiota composition of intestinal segments of high-fat diet-fed (HFD) rats. We also explored the effects of 12 weeks of NR supplementation on the gut microbiota in humans and mice. In rats, NR reduced fat mass and tended to decrease body weight. Interestingly, NR increased fat and energy absorption but only in HFD-fed rats. Moreover, 16S rRNA gene sequencing analysis of intestinal and fecal samples revealed an increased abundance of species within Erysipelotrichaceae and Ruminococcaceae families in response to NR. PnuC-positive bacterial strains within these families showed an increased growth rate when supplemented with NR. The abundance of species within the Lachnospiraceae family decreased in response to HFD irrespective of NR. Alpha and beta diversity and bacterial composition of the human fecal microbiota were unaltered by NR, but in mice, the fecal abundance of species within Lachnospiraceae increased while abundances of Parasutterella and Bacteroides dorei species decreased in response to NR. In conclusion, oral NR altered the gut microbiota in rats and mice, but not in humans. In addition, NR attenuated body fat mass gain in rats, and increased fat and energy absorption in the HFD context.

Green tea actions on miRNAs expression - An update.

Compounds derived from plants have been widely studied in the context of metabolic diseases and associated clinical conditions. In this regard, although the effects of Camellia sinensis plant, from which various types of teas, such as green tea, originate, have been vastly reported in the literature, the mechanisms underlying these effects remain elusive. A deep search of the literature showed that green tea's action in different cells, tissues, and diseases is an open field in the research of microRNAs (miRNAs). miRNAs are important communicator molecules between cells in different tissues implicated in diverse cellular pathways. They have emerged as an important linkage between physiology and pathophysiology, raising the issue of polyphenols can act also by changing miRNA expression. miRNAs are short, non-coding endogenous RNA, which silence the gene functions by targeting messenger RNA (mRNA) through degradation or translation repression. Therefore, the aim of this review is to present the studies that show the main compounds of green tea modulating the expression of miRNAs in inflammation, adipose tissue, skeletal muscle, and liver. We provide an overview of a few studies that have tried to demonstrate the role of miRNAs associated with the beneficial effects of compounds from green tea. We have emphasized that there is still a considerable gap in the literature investigating the role and likely involvement of miRNAs in the extensive beneficial health effects of green tea compounds already described, indicating miRNAs as potential polyphenols' mediators with a promising field to be investigated.

A class of anti-inflammatory lipids decrease with aging in the central nervous system.

Lipids contribute to the structure, development, and function of healthy brains. Dysregulated lipid metabolism is linked to aging and diseased brains. However, our understanding of lipid metabolism in aging brains remains limited. Here we examined the brain lipidome of mice across their lifespan using untargeted lipidomics. Co-expression network analysis highlighted a progressive decrease in 3-sulfogalactosyl diacylglycerols (SGDGs) and SGDG pathway members, including the potential degradation products lyso-SGDGs. SGDGs show an age-related decline specifically in the central nervous system and are associated with myelination. We also found that an SGDG dramatically suppresses LPS-induced gene expression and release of pro-inflammatory cytokines from macrophages and microglia by acting on the NF-κB pathway. The detection of SGDGs in human and macaque brains establishes their evolutionary conservation. This work enhances interest in SGDGs regarding their roles in aging and inflammatory diseases and highlights the complexity of the brain lipidome and potential biological functions in aging.

Análise de lacunas e perspectivas sobre programas de doação de corpos: relato de experiência no Brasil / Gap analysis and perspectives on body donation programs: experience report in Brazil

Resumo: Introdução: As recomendações internacionais reforçam o estímulo à prática da doação voluntária para a disponibilidade de corpos destinados à formação de profissionais de saúde, substituindo o sistema de corpos não reclamados nas instituições de ensino superior (IES). Relato de experiência: Este trabalho reuniu informações sobre programas de doação de corpos (PDC) brasileiros, analisou lacunas e fatores impeditivos à doação do corpo, e propôs estratégias para ampliar as informações sobre o cadastro de participantes em PDC e a efetivação da doação. Discussão: No Brasil existem 39 PDC em funcionamento - seis no Nordeste, quatro no Centro-Oeste, um no Norte, 11 no Sul e 17 no Sudeste - e duas comissões de distribuição de corpos para ensino. O cadastro como participante no PDC pode agilizar as providências documentais, mas não garante a doação na ocasião da morte. No geral, as IES recebem o corpo doado e a documentação necessária. Quando o óbito acontece no município da IES, é possível ter o translado do corpo providenciado por parcerias com funerárias e prefeitura. Conclusão: Embora a legislação tenha regulamentado desde 2002 a doação do próprio corpo em vida, é necessária a implementação de campanhas de esclarecimento à população geral sobre essa possibilidade, além de mecanismos que possam promover a celeridade nos prazos de para obtenção das documentações com a finalidade de doação de corpo para fins científicos, a isenção das taxas cartoriais e a viabilização do translado do corpo doado por carro de funerária das prefeituras, sem custo. O apoio da IES por meio do cadastro do PDC como atividade de extensão universitária colabora para assegurar aspectos éticos relacionados à participação dos doadores no programa em vida, no que se refere ao tratamento com respeito e dignidade dos doadores e de suas famílias.

Abstract: Introduction: International recommendations reinforce the practice of voluntary donation for providing bodies for the training of health professionals, replacing the system of unclaimed bodies in medical courses (MC). Experience report: This work gathered information on Brazilian body donation programs (BDP), analyzed gaps and difficulties to the effectiveness of body donation, and proposed strategies to expand the information offered on the registration of participants in BDP and the effectiveness of the donation. Discussion: In Brazil, there are 39 BDP - six in the Northeast, four in the Midwest, one in the North, 11 in the South, and 17 in the Southeast - and two committees for body donation. Documents may be easily available through registration during life as a participant in the BDP, but it does not guarantee donation. In general, Brazilian's institutions receive the donated body along with the documentation. When the death occurs in the same city of the institution, it is possible to have the transfer of the body provided by partnerships with funeral services and the city hall. There is no fee to be paid or received either for registration during life or for donation after death. Conclusion: Although the legislation has regulated the donation of body in life since 2002, it is necessary to implement campaigns to inform about the donation, in addition to mechanisms that can promote speed in the deadlines for obtaining documentation for the purpose of donating body for scientific purposes, the exemption of notary fees and the possibility of transporting the body donated with no fees by funeral home of municipal governments. The MC support through the registration of the PDC as an extension activity collaborates to ensure ethical aspects related to the participation of donors in the living program, with regard to the treatment with respect and dignity of donors and their families.

Extracting Thyroid Nodules Characteristics from Ultrasound Reports Using Transformer-based Natural Language Processing Methods.

The ultrasound characteristics of thyroid nodules guide the evaluation of thyroid cancer in patients with thyroid nodules. However, the characteristics of thyroid nodules are often documented in clinical narratives such as ultrasound reports. Previous studies have examined natural language processing (NLP) methods in extracting a limited number of characteristics (<9) using rule-based NLP systems. In this study, a multidisciplinary team of NLP experts and thyroid specialists, identified thyroid nodule characteristics that are important for clinical care, composed annotation guidelines, developed a corpus, and compared 5 state-of-the-art transformer-based NLP methods, including BERT, RoBERTa, LongFormer, DeBERTa, and GatorTron, for extraction of thyroid nodule characteristics from ultrasound reports. Our GatorTron model, a transformer-based large language model trained using over 90 billion words of text, achieved the best strict and lenient F1-score of 0.8851 and 0.9495 for the extraction of a total number of 16 thyroid nodule characteristics, and 0.9321 for linking characteristics to nodules, outperforming other clinical transformer models. To the best of our knowledge, this is the first study to systematically categorize and apply transformer-based NLP models to extract a large number of clinical relevant thyroid nodule characteristics from ultrasound reports. This study lays ground for assessing the documentation quality of thyroid ultrasound reports and examining outcomes of patients with thyroid nodules using electronic health records.

Factors related to the quality of life of mothers of children with Congenital Zika Virus Syndrome.

OBJECTIVE: To relate sociodemographic and clinical aspects to the quality of life of mothers of children with Congenital Zika Virus Syndrome. METHOD: Cross-sectional analytical study, conducted in a rehabilitation center in Goiânia, central region of Brazil. A sociodemographic and clinical profile questionnaire and the World Health Organization Quality of Life were used. Data analysis was performed using the Mann-Whitney tests; Kruskal-Wallis, followed by Dunnett's Post hoc; and Spearman's correlation. RESULTS: The sample consisted of 30 mothers of children with Congenital Zika Virus Syndrome, with a mean age of 30.57 ± 6.67 years. Mothers who had a partner, leisure activity, who used a car as a means of transport and who had a child with congenital syndrome with microcephaly had a better quality of life (p<0,05). CONCLUSION: The quality of life of these mothers was related to sociodemographic and clinical aspects, which suggests the need for more specific public policies for this population.

Reporting ChAracteristics of cadaver training and sUrgical studies: The CACTUS guidelines.

INTRODUCTION: Recent systematic reviews highlighted increasing use of cadaveric models in the surgical training, but reports on the characteristics of the models and their impact on training are lacking, as well as standardized recommendations on how to ensure the quality of surgical studies. The aim of our survey was to provide an easy guideline that would improve the quality of the studies involving cadavers for surgical training and research. METHODS: After accurate literature review regarding surgical training on cadaveric models, a draft of the CACTUS guidelines involving 10 different items was drawn. Afterwards, the items were improved by questionnaire uploaded and spread to the experts in the field via Google form. The guideline was then reviewed following participants feedback, ergo, items that scored between 7 and 9 on nine-score Likert scale by 70% of respondents, and between 1 and 3 by fewer than 15% of respondents, were included in the proposed guideline, while items that scored between 1 and 3 by 70% of respondents, and between 7 and 9 by 15% or more of respondents were not. The process proceeded with Delphi rounds until the agreement for all items was unanimous. RESULTS: In total, 42 participants agreed to participate and 30 (71.4%) of them completed the Delphi survey. Unanimous agreement was almost always immediate concerning approval and ethical use of cadaver and providing brief outcome statement in terms of satisfaction in the use of the cadaver model through a short questionnaire. Other items were subjected to the minor adjustments. CONCLUSION: 'CACTUS' is a consensus-based guideline in the area of surgical training, simulation and anatomical studies and we believe that it will provide a useful guide to those writing manuscripts involving human cadavers.

Prevalence of Depression in Pregnant Women with Bariatric Surgery History and Associated Factors.

OBJECTIVE: To analyze the prevalence and factors associated with depressive symptoms among Brazilian pregnant women with history of bariatric surgery (BS). METHODS: This is a cohort study with 247 women who got pregnant after BS. Based on data collection via Google Form, the recruitment of participants occurred in Facebook groups for 13 months. All of them answered a form with Informed Consent, a general data protocol and the Brazilian version of the Depression, Anxiety and Stress Scale-21. Descriptive and inferential analysis were performed, and a binary logistic regression model was tested to predict the factors associated with depressive symptoms. RESULTS: The prevalence of depressive symptoms was 32.8%, noted as being higher in the first (40.6%) and third (34.3%) gestational trimesters. Significative associations were found between depression and marital status (p = 0.000), planned pregnancy (p = 0.001), desired pregnancy (p = 0.004) and psychiatric history (p = 0.000). Women who were not married (odds ratio, OR = 3,38; p = 0.002) and had a psychiatric history (OR = 2.70; p = 0.102) had higher chances of showing depression symptoms; while planned and desired pregnancy showed as protective factors to the symptoms of depression. CONCLUSION: These findings highlight the importance of psychological assistance for pregnant women with history of BS, to prevent development of mental disorders and their outcomes for maternal-child health.

OBJETIVO: Analisar a prevalência e fatores associados à sintomatologia depressiva entre gestantes brasileiras com histórico de cirurgia bariátrica. MéTODOS: Trata-se de um estudo de coorte com 247 mulheres que engravidaram após uma Cirurgia Bariátrica. Baseado em coleta via Google Form o recrutamento das gestantes ocorreu em grupos do Facebook durante 13 meses. Todas responderam a um formulário com o Termo de Consentimento Livre e Esclarecido, um protocolo de dados gerais e a versão brasileira da Depression, Anxiety and Stress Scale-21. Análises, descritiva e inferencial foram realizadas, e um modelo de regressão logística binária foi testado para predizer os fatores associados à sintomatologia depressiva. RESULTADOS: A prevalência da sintomatologia depressiva foi de 32,8%, sendo notada como maior no primeiro (40,6%) e terceiro (34,3%) trimestres gestacionais. Associações significativas foram encontradas entre sintomatologia depressiva e estado civil (p = 0,000), gestação planejada (p = 0,001), gestação desejada (p = 0,004) e histórico psiquiátrico (p = 0,000). Mulheres que não eram casadas (OR = 3,38; p = 0,002) e tinham um histórico psiquiátrico (OR = 2,70; p = 0,102) apresentaram mais chances de manifestarem sintomas de depressão; enquanto gestação planejada e desejada mostraram-se como fatores protetivos aos sintomas de depressão. CONCLUSãO: Os achados ressaltam a importância do acompanhamento psicológico para gestantes com histórico de cirurgia bariátrica para prevenção do desenvolvimento de transtornos mentais e seus desfechos para a saúde materno-infantil.

Factors related to the quality of life of mothers of children with Congenital Zika Virus Syndrome / Factores relacionados con la calidad de vida de madres de niños con Síndrome Congénito del Virus Zika / Fatores relacionados à qualidade de vida de mães de crianças com Síndrome Congênita do Zika Virus

ABSTRACT Objective: To relate sociodemographic and clinical aspects to the quality of life of mothers of children with Congenital Zika Virus Syndrome. Method: Cross-sectional analytical study, conducted in a rehabilitation center in Goiânia, central region of Brazil. A sociodemographic and clinical profile questionnaire and the World Health Organization Quality of Life were used. Data analysis was performed using the Mann-Whitney tests; Kruskal-Wallis, followed by Dunnett's Post hoc; and Spearman's correlation. Results: The sample consisted of 30 mothers of children with Congenital Zika Virus Syndrome, with a mean age of 30.57 ± 6.67 years. Mothers who had a partner, leisure activity, who used a car as a means of transport and who had a child with congenital syndrome with microcephaly had a better quality of life (p<0,05). Conclusion: The quality of life of these mothers was related to sociodemographic and clinical aspects, which suggests the need for more specific public policies for this population.

RESUMEN Objetivo: Relacionar aspectos sociodemográficos y clínicos con la calidad de vida de madres de niños con síndrome congénito del virus del Zika. Método: Estudio analítico transversal, realizado en un centro de rehabilitación en Goiânia, región central de Brasil. Se utilizó un cuestionario de perfil sociodemográfico y clínico y el World Health Organization Quality of Life. El análisis de los datos se realizó mediante las pruebas de Mann-Whitney; Kruskal-Wallis, seguida de Posthoc de Dunnett; y correlación de Spearman. Resultados: La muestra estuvo conformada por 30 madres de niños con síndrome congénito del virus del Zika, con una edad promedio de 30.57 ± 6.67 años. Las madres que tenían pareja, actividad de ocio, que usaban el automóvil como medio de transporte y que tenían un hijo con síndrome congénito con microcefalia tenían una mejor calidad de vida (p <0,05). Conclusión: La calidad de vida de estas madres se relacionó con aspectos sociodemográficos y clínicos, lo que sugiere la necesidad de políticas públicas más específicas para esta población.

RESUMO Objetivo: Relacionar aspectos sociodemográficos e clínicos com a qualidade de vida de mães de crianças com Síndrome Congênita do Zika Vírus. Método: Estudo transversal analítico, realizado em um centro de reabilitação de Goiânia, região central do Brasil. Foram utilizados um questionário de perfil sociodemográfico e clínico e o World Health Organization Quality of Life. A análise dos dados foi realizada por meio dos testes Mann-Whitney; Kruskal-Wallis, seguido do Post hoc de Dunnett e correlação de Spearman. Resultados: A amostra foi constituída de 30 mães de crianças com Síndrome Congênita do Zika Vírus, com média de idade de 30,57 ± 6,67 anos. As mães que tinham companheiro, atividade de lazer, que utilizavam carro como meio de transporte e que tinham filho com a síndrome congênita com microcefalia, tiveram melhor qualidade de vida (p<0,05). Conclusão: A qualidade de vida dessas mães teve relação com aspectos sociodemográficos e clínicos, o que sugere necessidade de mais políticas públicas específicas para esta população.

Assessment of potters' occupational exposure to lead and associated risk factors in Maragogipinho, Brazil: preliminary results.

BACKGROUND AND PURPOSE: Lead (Pb) is used as a flux in the glazing process of pottery utensils in several regions of the world. It can affect the nervous and hematopoietic systems; in addition, it is classified as a probable human carcinogen. This work aims to evaluate Pb exposure of potters and describe the main determinants of elevated blood Pb (PbB) levels in this group of workers. METHODS: This is a cross-sectional study with potters of Maragogipinho Village, Bahia, Brazil, of both sexes, aged 16-72 years (n = 85). Non-exposed workers of the same age range residing in the urban area of Aratuípe town were also recruited (n = 50). We evaluated Pb dust deposition rates (PbDrt) in pottery workshops and PbB levels. All Pb measurements were performed by Graphite Furnace Atomic Absorption Spectrometry. RESULTS: The median of PbB (min-max) and geometric mean (SD) PbDrt for the exposed group were 7.9 (0.9-49.8) µg/dL and 1463 (± 290,000) µg/m2/30 days, respectively. For the control group, levels were 1.5 (0.1-19.8) µg/dL and 82 (46) µg/m2 30 days, respectively. CONCLUSION: The data found showed an excessive exposure among artisans, exceeding occupationally safe levels and those reported in the literature. It is important to implement occupational hygiene measures and improvements of the working conditions of these labors, especially the replacement of lead oxide in the pottery-glazing process.

A Method to Induce Brown/Beige Adipocyte Differentiation from Murine Preadipocytes.

Adipocytes exhibit different morphological and functional characteristics, depending on their anatomical location, developmental origin, and stimulus. While white adipocytes tend to accumulate energy as triglycerides, brown and beige adipocytes tend to direct carbon sources to fuel thermogenesis. White and beige adipocytes originate from common progenitor cells, which are distinct from brown adipocyte precursors. Having a method to study white vs. beige vs. brown adipocyte differentiation may help to unveil the mechanisms driving distinct adipogenic programs. Preadipocytes can be cultured and differentiated in vitro using a combination of compounds to stimulate adipogenesis. Here, we describe and compare protocols designed to stimulate adipocyte differentiation and induce brown/beige-like or white-like characteristics in differentiating adipocytes. The protocols consist in exposing murine preadipocytes to pharmacological stimuli aimed at triggering adipogenesis and inducing (or not) a thermogenic gene expression program. After 8 days of differentiation with a pro-browning cocktail, immortalized preadipocytes isolated from interscapular brown fat (9B cells) or inguinal white fat (9W cells) from the same mouse expressed higher levels of brown/beige adipocyte markers (e.g., Ucp1) and pan-adipocyte differentiation markers (e.g., Pparg, Cebpa and aP2) when compared to the same cells differentiated with a cocktail that lacked brown/beige adipogenic inducers (i.e., rosiglitazone, T3, and indomethacin). Consistent with a higher thermogenic potential of brown vs. beige adipocytes, differentiated 9B cells expressed higher Ucp1 levels than differentiated 9W cells. This simple protocol may help researchers to understand mechanisms of adipogenesis and how adipocytes become thermogenic.

Zinc-protoporphyrin determination by HPLC with fluorescence detection as a biomarker of lead effect in artisanal pottery workers.

Lead (Pb) exposure compromises heme synthesis by inhibiting ferrochelatase, forming zinc-protoporphyrin (ZnPP). This study aims to validate a method for the determination of ZnPP by HPLC with fluorescence detection and apply this method to evaluate the extent of Pb exposure of artisanal pottery workers. Extraction procedures were tested using both nonacid and acid liquid-liquid extraction. The former presented a better chromatogram and recovery results. The validated method yielded a good resolution of ZnPP and its free form peaks with acceptable precision and accuracy. Total run time was 15 min and ZnPP peak retention time was 5.6 min. We applied this method to evaluate 39 potters (90% male), mean age 40 years (9-80). The medians (ranges) of blood lead, ZnPP and hemoglobin were 16.0 µg/dl (2.2-71.5), 12.6 µg/dl (4.6-279.8) and 15.1 g/dl (11.0-17.8), respectively. Significant differences were observed for blood lead according to gender, age range (>40 years), direct handling of lead oxide and years of occupation. Significantly higher levels of ZnPP were observed in male potters involved with lead glazing activity. The validated method was shown to be simple with one-step nonacid extraction, good sensitivity, reproducibility and accuracy. Our data shows that these laborers are dangerously exposed to Pb, reflecting the effect on the heme synthesis.

Enoxacin induces oxidative metabolism and mitigates obesity by regulating adipose tissue miRNA expression.

MicroRNAs (miRNAs) have been implicated in oxidative metabolism and brown/beige adipocyte identity. Here, we tested whether widespread changes in miRNA expression promoted by treatment with the small-molecule enoxacin cause browning and prevent obesity. Enoxacin mitigated diet-induced obesity in mice, and this was associated with increased energy expenditure. Consistently, subcutaneous white and brown adipose tissues and skeletal muscle of enoxacin-treated mice had higher levels of markers associated with thermogenesis and oxidative metabolism. These effects were cell autonomous since they were recapitulated in vitro in murine and human cell models. In preadipocytes, enoxacin led to a reduction of miR-34a-5p expression and up-regulation of its target genes (e.g., Fgfr1, Klb, and Sirt1), thus increasing FGF21 signaling and promoting beige adipogenesis. Our data demonstrate that enoxacin counteracts obesity by promoting thermogenic signaling and inducing oxidative metabolism in adipose tissue and skeletal muscle in a mechanism that involves, at least in part, miRNA-mediated regulation.

Small and mighty: adaptation of superphylum Patescibacteria to groundwater environment drives their genome simplicity.

BACKGROUND: The newly defined superphylum Patescibacteria such as Parcubacteria (OD1) and Microgenomates (OP11) has been found to be prevalent in groundwater, sediment, lake, and other aquifer environments. Recently increasing attention has been paid to this diverse superphylum including > 20 candidate phyla (a large part of the candidate phylum radiation, CPR) because it refreshed our view of the tree of life. However, adaptive traits contributing to its prevalence are still not well known. RESULTS: Here, we investigated the genomic features and metabolic pathways of Patescibacteria in groundwater through genome-resolved metagenomics analysis of > 600 Gbp sequence data. We observed that, while the members of Patescibacteria have reduced genomes (~ 1 Mbp) exclusively, functions essential to growth and reproduction such as genetic information processing were retained. Surprisingly, they have sharply reduced redundant and nonessential functions, including specific metabolic activities and stress response systems. The Patescibacteria have ultra-small cells and simplified membrane structures, including flagellar assembly, transporters, and two-component systems. Despite the lack of CRISPR viral defense, the bacteria may evade predation through deletion of common membrane phage receptors and other alternative strategies, which may explain the low representation of prophage proteins in their genomes and lack of CRISPR. By establishing the linkages between bacterial features and the groundwater environmental conditions, our results provide important insights into the functions and evolution of this CPR group. CONCLUSIONS: We found that Patescibacteria has streamlined many functions while acquiring advantages such as avoiding phage invasion, to adapt to the groundwater environment. The unique features of small genome size, ultra-small cell size, and lacking CRISPR of this large lineage are bringing new understandings on life of Bacteria. Our results provide important insights into the mechanisms for adaptation of the superphylum in the groundwater environments, and demonstrate a case where less is more, and small is mighty.

Esthetic perception of facial profile changes after extraction and nonextraction Class II treatment.

This retrospective study evaluated facial profile pleasantness determined by two protocols of Class II treatment. The sample comprised facial profile silhouettes obtained retrospectively from the pretreatment (T1) and posttreatment (T2) cephalograms of 60 patients (42 males and 18 females) divided into two groups. One group of 30 patients (mean age of 12.84 years) was treated with the extraction of maxillary first premolars (mean treatment time of 2.7 years), and the other group of 30 patients (mean age of 12.81 years) was treated with a mandibular advancement appliance (Forsus) (mean treatment time of 2.49 years). The facial profile silhouettes (T1 and T2) were randomly distributed in an album containing one patient per sheet. The examiners consisted of 60 orthodontists and 60 lay individuals, who analyzed the profiles in regard to facial pleasantness, using the Likert scale. A comparison between stages T1 and T2 of the two treatment protocols and between the examiners was performed by mixed-design analysis of variance at a significance level of 5%. The results demonstrated a significant difference between T1 and T2 (greater scores for T2 compared to T1), and between lay individuals and orthodontists (orthodontists assigned higher scores), but with no significant difference between the treatment protocols. Both protocols produced positive effects on the facial profile esthetics, from the standpoint of lay individuals and orthodontists.