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This study sought to develop polymer-lipid hybrid solid dispersions containing the poorly soluble drug lopinavir (LPV) by hot-melt extrusion (HME). Hence, the lipid and polymeric adjuvants were selected based on miscibility and compatibility studies. Film casting was used to assess the miscibility, whereas thermal, spectroscopic, and chromatographic analyses were employed to evaluate drug-excipient compatibility. Extrudates were obtained and characterized by physicochemical tests, including in vitro LPV dissolution. Preformulation studies led to select the most appropriate materials, i.e., the polymers PVPVA and Soluplus®, the plasticizers polyethylene glycol 400 and Kolliphor® HS15, phosphatidylcholine, and sodium taurodeoxycholate. HME processing did not result in LPV degradation and significantly increased entrapment efficiency (93.8% ± 2.8 for Soluplus® extrudate against 19.8% ± 0.5 of the respective physical mixture). LPV dissolution was also increased from the extrudates compared to the corresponding physical mixtures (p < 0.05). The dissolution improvement was considerably greater for the Soluplus®-based formulation (24.3 and 2.8-fold higher than pure LPV and PVPVA-based extrudate after 120 min, respectively), which can be attributed to the more pronounced effects of HME processing on the average size and LPV solid-state properties in the Soluplus® extrudates. Transmission electron microscopy and chemical microanalysis suggested that the polymer-lipid interactions in Soluplus®-based formulation depended on thermal processing., (© 2023. The Author(s), under exclusive licence to American Association of Pharmaceutical Scientists.)

The ground beetle, Neoaulacoryssus speciosus (Coleoptera: Carabidae) is of high relevance to field because it has been recorded as a pest of seeds and young plants of vegetables and other crops and a predator in agricultural crops, forest and weeds in Brazil. However, natural habitat changes are increasing agriculture and forest insect outbreaks in urban areas. A N. speciosus population outbreak occurred in October and November 2018 simultaneously in 12 neighboring municipalities at the beginning of the rainy season in the northern region of Minas Gerais State, Brazil. The objectives of this study were to report a sudden and simultaneous population outbreak of N. speciosus and to describe the factors of habitat change that could have contributed to this invasion in 12 municipalities in the northern region of Minas Gerais State in the Caatinga biome of Brazil. In addition, female and male genitals were described and illustrated, the scientific classification revised and common names of N. speciosus listed. Thousands of males and females of N. speciosus agglomerated in shady, humid places during the day and night for about 15 days. Neoaulacoryssus speciosus has been identified and illustrated, its scientific classification revised and four common names listed for this species.

Background: Digital technologies are becoming increasingly sophisticated and integrated, but the scope of the Fourth Industrial Revolution (4IR) is wider: technologies and innovations are diffusing faster and more widely. Despite the extensive use of telepharmacy technology by patients and pharmacists, there is an absence of research that examines the use of telepharmacy., Objective: To investigate the current evidence concerning the impact of the 4IR on the provision of pharmaceutical services., Methods: A scoping review based on the PCC (Population, Concept, and Context) mnemonic was conducted in Pubmed, Scopus and Web of Science. Population included pharmacist/clinical pharmaceutical services, the concept referred to the 4IR and the context was open. Electronic searches retrieved 8,694 articles that were screened by titles and abstracts. The search yielded 59 studies that were analyzed in terms of country of focus, publication year, type of publication, main technologies, types of pharmacy services, and the role of the Fourth Industrial Revolution., Results: Most of the included studies were conducted in the United States and were published between 2005 and 2021, with the highest number of publications in the year of 2021. The majority of included articles were observational studies. Technologies addressed in the studies were those within the scope of telepharmacy, and most of the articles discussed more than one pharmaceutical service, with medication therapy management as the most frequent., Conclusions: The Covid-19 pandemic accelerated the use of technologies, identifying promising niches for pharmacists in the field of Pharmaceutical Care. This should encourage pharmacists to fortify their capacity to adopt new technologies in the provision of pharmaceutical services. The frequent use of various technologies has been demonstrated, which indicates the exigency for further health education actions by the pharmacist., Competing Interests: Declarations of competing interest None., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Objective: To evaluate the follow-up of children diagnosed with deafness in neonatal hearing screening and risk factors for hearing loss., Methods: Quantitative, cross-sectional, and retrospective study to evaluate factors associated with hearing loss and the follow-up of cases of children diagnosed with audiological dysfunction, by analyzing electronic medical records of 5,305 children referred to a Specialized Center in Type I Rehabilitation, from January/2016 to February/2020, in the city of Manaus, Amazonas. The statistical study used Pearson's chi-square test and binary logistic regression in which odds ratio scans were obtained with reliability intervals of 95%., Results: Of the 5,305 children referred for the otoacoustic emission retest, 366 (6.9%) failed the retest. Children diagnosed with sensorineural hearing loss continued in the study, totaling 265 (72.4%). Only 58 (21.9%) children continued in the study to its end, of these 39 had received hearing aids at that point; and 16 (41%) had surgical indication for cochlear implants, of which only 3 (18.7%) had undergone surgery. Among the risk factors for hearing loss, we found 2.6 times more chance of failure in the otoacoustic emissions retest in those children who had a family history of hearing loss and ICU stay., Conclusion: Although the screening flow reaches a large part of live births, the dropout rates during the process are high, therefore, the socioeconomic and geographic characteristics of regions such as the Amazon should be considered as relevant factors to the evasion of rehabilitation programs of these children. Hospitalization in the neonatal ICU and family history of hearing loss in the investigations could be identified as the main and most important factors for alteration of the otoacoustic emissions retests.

Background: The Creutzfeldt-Jakob disease (CJD) is a spongiform encephalopathy that manifests as a rapidly progressive dementia syndrome. Currently, CJD has no cure, and many patients die within the first year, but some drugs are being studied as options for managing this condition., Objective: To evaluate the effectiveness of pharmacological treatments offered to patients with CJD as a means to increase survival and reduce cognitive deterioration., Methods: A systematic review of the literature was performed using 4 independent reviewers and 1 extra reviewer to resolve possible divergences in the search and analysis of papers indexed in MedLINE (PubMed), SciELO and Lilacs databases. The Medical Subject Heading (MeSH) terms used were: prion diseases , Creutzfeldt-Jakob disease , pharmacologic therapy , therapeutics , quinacrine , doxycycline , flupirtine , and pentosan polysulfate , with the Boolean operators AND and OR . This search included controlled clinical trials, uncontrolled clinical trials, and case series published from the year 2000 onwards, in the English language., Results: A total of 85 papers were found using the descriptors used. At the end of the selection analyses, 9 articles remained, which were analyzed fully and individually., Conclusions: None of the drugs evaluated proved significantly effective in increasing survival in patients with CJD. Flupirtine appears to have a beneficial effect in reducing cognitive deterioration in patients with CJD. However, additional studies are needed to establish better evidence and therapeutic options for the management of patients with CJD., Competing Interests: The authors have no conflict of interests to declare., (Academia Brasileira de Neurologia. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/).)

Introduction: The goiter, a neglected heterogeneous molecular disease, remains a major indication for thyroidectomies in its endemic regions., Objectives: This study analyzed differential gene expression in surgical specimens diagnosed with multi nodular and compared the data to that of thyroid tissue without multinodular goiter from patients undergoing thyroidectomy in Manaus-AM, Brazil using RNA-seq technology., Methodology: The transcriptome information of the surgical specimen fragments with and without multinodular goiter was accessed by Illumina HiSeq 2000 New Generation Sequencing (NGS) using the RNA-seq NEBNext® Ultra™ RNA Library Prep Kit for Illumina®-#E7530L protocol and differential gene expression analysis., Results: Differences were found between the gene expression profiles of the diseased tissues and those of the healthy control tissues; at least 70 genes were differentially expressed. The HOTS gene was expressed only in multinodular goiter tissues (p < 0.05)., Conclusion: These results demonstrate that the gene expression profile of multinodular goiter is pro-tumoral and that HOTS can play a central role in multinodular goiter development., Competing Interests: The authors have declared that no competing interests exist.

The survival of children with diffuse intrinsic pontine glioma (DIPG) remains dismal, with new treatments desperately needed. In a prospective biopsy-stratified clinical trial, we combined detailed molecular profiling and drug screening in newly established patient-derived models in vitro and in vivo. We identified in vitro sensitivity to MEK inhibitors in DIPGs harboring MAPK pathway alterations, but treatment of patient-derived xenograft models and a patient at relapse failed to elicit a significant response. We generated trametinib-resistant clones in a BRAFG469V model through continuous drug exposure and identified acquired mutations in MEK1/2 with sustained pathway upregulation. These cells showed hallmarks of mesenchymal transition and expression signatures overlapping with inherently trametinib-insensitive patient-derived cells, predicting sensitivity to dasatinib. Combined trametinib and dasatinib showed highly synergistic effects in vitro and on ex vivo brain slices. We highlight the MAPK pathway as a therapeutic target in DIPG and show the importance of parallel resistance modeling and combinatorial treatments for meaningful clinical translation., Significance: We report alterations in the MAPK pathway in DIPGs to confer initial sensitivity to targeted MEK inhibition. We further identify for the first time the mechanism of resistance to single-agent targeted therapy in these tumors and suggest a novel combinatorial treatment strategy to overcome it in the clinic. This article is highlighted in the In This Issue feature, p. 587., (©2021 The Authors; Published by the American Association for Cancer Research.)

Somatic mutations in ACVR1 are found in a quarter of children with diffuse intrinsic pontine glioma (DIPG), but there are no ACVR1 inhibitors licensed for the disease. Using an artificial intelligence-based platform to search for approved compounds for ACVR1 -mutant DIPG, the combination of vandetanib and everolimus was identified as a possible therapeutic approach. Vandetanib, an inhibitor of VEGFR/RET/EGFR, was found to target ACVR1 ( K d = 150 nmol/L) and reduce DIPG cell viability in vitro but has limited ability to cross the blood-brain barrier. In addition to mTOR, everolimus inhibited ABCG2 (BCRP) and ABCB1 (P-gp) transporters and was synergistic in DIPG cells when combined with vandetanib in vitro . This combination was well tolerated in vivo and significantly extended survival and reduced tumor burden in an orthotopic ACVR1 -mutant patient-derived DIPG xenograft model. Four patients with ACVR1 -mutant DIPG were treated with vandetanib plus an mTOR inhibitor, informing the dosing and toxicity profile of this combination for future clinical studies. SIGNIFICANCE: Twenty-five percent of patients with the incurable brainstem tumor DIPG harbor somatic activating mutations in ACVR1 , but there are no approved drugs targeting the receptor. Using artificial intelligence, we identify and validate, both experimentally and clinically, the novel combination of vandetanib and everolimus in these children based on both signaling and pharmacokinetic synergies. This article is highlighted in the In This Issue feature, p. 275 ., (©2021 The Authors; Published by the American Association for Cancer Research.)

The role of PPM1D mutations in de novo gliomagenesis has not been systematically explored. Here we analyze whole genome sequences of 170 pediatric high-grade gliomas and find that truncating mutations in PPM1D that increase the stability of its phosphatase are clonal driver events in 11% of Diffuse Midline Gliomas (DMGs) and are enriched in primary pontine tumors. Through the development of DMG mouse models, we show that PPM1D mutations potentiate gliomagenesis and that PPM1D phosphatase activity is required for in vivo oncogenesis. Finally, we apply integrative phosphoproteomic and functional genomics assays and find that oncogenic effects of PPM1D truncation converge on regulators of cell cycle, DNA damage response, and p53 pathways, revealing therapeutic vulnerabilities including MDM2 inhibition., (© 2022. The Author(s).)

The production of lignin nanoparticles (LNPs) has emerged as a way to overcome the highly variable and complex molecular structure of lignin. It can offer morphological control of the lignin polymer, allowing the formation of stable LNP dispersions in aqueous media, while increasing the potential of lignin for high-value applications. However, the polydispersity and morphology of LNPs varies depending on the lignin grade and preparation method, and a systematic comparison using different technical lignins is lacking. In this study, it was attempted to find a green fabrication method with a distinct solvent fractionation of lignin to prepare LNPs using three different technical lignins as starting polymers: BLN birch lignin (hardwood, BB), alkali Protobind 1000 (grass, PB), and kraft LignoBoost (softwood, LB). For that, three anti-solvent precipitation approaches to prepare LNPs were systematically compared: 70 % aqueous ethanol, acetone/water (3 : 1) and NaOH as the lignin solvent, and water/aqueous HCl as the anti-solvent. Among all these methods, the acetone/water (3 : 1) approach allowed production of homogeneous and monodisperse LNPs with a negative surface charge and also spherical and smooth surfaces. Overall, the results revealed that the acetone/water (3 : 1) method was the most effective approach tested to obtain homogenous, small, and spherical LNPs from the three technical lignins. These LNPs exhibited an improved stability at different ionic strengths and a wider pH range compared to the other preparation methods, which can greatly increase their application in many fields, such as pharmaceutical and food sciences., (© 2021 The Authors. ChemSusChem published by Wiley-VCH GmbH.)

Lignin is an abundant natural feedstock that offers great potential as a renewable substitute for fossil-based resources. Its polyaromatic structure and unique properties have attracted significant research efforts. The advantages of an enzymatic over chemical or thermal approach to construct or deconstruct lignins are that it operates in mild conditions, requires less energy, and usually uses non-toxic chemicals. Laccase is a widely investigated oxidative enzyme that can catalyze the polymerization and depolymerization of lignin. Its dual nature causes a challenge in controlling the overall direction of lignin-laccase catalysis. In this Review, the factors that affect laccase-catalyzed lignin polymerization were summarized, evaluated, and compared to identify key features that favor lignin polymerization. In addition, a critical assessment of the conditions that enable production of novel lignin hybrids via laccase-catalyzed grafting was presented. To assess the industrial relevance of laccase-assisted lignin valorization, patented applications were surveyed and industrial challenges and opportunities were analyzed. Finally, our perspective in realizing the full potential of laccase in building lignin-based materials for advanced applications was deduced from analysis of the limitations governing laccase-assisted lignin polymerization and grafting., (© 2021 The Authors. ChemSusChem published by Wiley-VCH GmbH.)

Fibrodysplasia ossificans progressiva (FOP) and diffuse intrinsic pontine glioma (DIPG) are debilitating diseases that share causal mutations in ACVR1, a TGF-β family type I receptor. ACVR1 R206H is a frequent mutation in both diseases. Pathogenic signaling via the SMAD1/5 pathway is mediated by Activin A, but how the mutation triggers aberrant signaling is not known. We show that ACVR1 is essential for Activin A-mediated SMAD1/5 phosphorylation and is activated by two distinct mechanisms. Wild-type ACVR1 is activated by the Activin type I receptors, ACVR1B/C. In contrast, ACVR1 R206H activation does not require upstream kinases, but is predominantly activated via Activin A-dependent receptor clustering, which induces its auto-activation. We use optogenetics and live-imaging approaches to demonstrate Activin A-induced receptor clustering and show it requires the type II receptors ACVR2A/B. Our data provide molecular mechanistic insight into the pathogenesis of FOP and DIPG by linking the causal activating genetic mutation to disrupted signaling., (© 2021 The Authors. Published under the terms of the CC BY 4.0 license.)

The effects of resistance training (RT) associated with calcium β-hydroxyβ-methylbutyrate (CaHMB) supplementation on the body composition and gene expression of cytokines related to skeletal muscle hypertrophy and adipose tissue metabolism were studied in rats. Male Wistar rats were divided into four groups of 12 animals: sedentary control (SC); sedentary supplemented (SS); resistance training control (RTC) and resistance training supplemented (RTS). Rats from RTC and RTS groups were submitted to an RT programme and those from SS and RTS groups received 1 mL of CaHMB (320 mg kg -1 day -1 ) by gavage, for 8 weeks. We evaluated: body composition; plasma lipid profile; the gene expression of interleukin (IL)-6, IL-10, IL-15 and fibronectin type III domain-containing protein 5 (FNDC-5) in skeletal muscle, and IL-6, mitochondrial uncoupling protein 1 (UCP-1) in white adipose tissue (WAT); and the concentration of irisin in WAT. Compared to RTC alone, the combination of CaHMB with RT (RTS) further reduced abdominal circumference (5.3%), Lee index (2.4%), fat percentage (24.4%), plasma VLDL cholesterol (16.8%) and triglycerides (17%) and increased the gene expression of FNDC-5 (78.9%) and IL-6 (47.4%) in skeletal muscle and irisin concentration (26.9%) in WAT. Neither RT nor CaHMB affected the protein percentage or the gene expression of IL-6 and UCP-1 in WAT and IL-10, IL-15 in skeletal muscle. In conclusion, CaHMB supplementation increased the beneficial effects of RT on body fat reduction and was associated with muscular genic expression of IL-6 and FNDC-5 and irisin concentration in WAT, despite the lack of change in protein mass and maximal strength., Competing Interests: Juliano Magalhães Guedes, Maria do Carmo Gouveia Pelúzio, John A. Rathmacher, Tiago Ferreira Leal, Miguel Araújo Carneiro Júnior, Diego Milhomem de Carvalho, Leandro Licursi de Oliveira and Antônio José Natali declare that they have no conflict of interest., (Copyright © 2020 Termedia.)

Background: The use of liquid biopsy is of potential high importance for children with high grade (HGG) and diffuse midline gliomas (DMG), particularly where surgical procedures are limited, and invasive biopsy sampling not without risk. To date, however, the evidence that detection of cell-free DNA (cfDNA) or circulating tumor DNA (ctDNA) could provide useful information for these patients has been limited, or contradictory., Methods: We optimized droplet digital PCR (ddPCR) assays for the detection of common somatic mutations observed in pediatric HGG/DMG, and applied them to liquid biopsies from plasma, serum, cerebrospinal fluid (CSF), and cystic fluid collected from 32 patients., Results: Although detectable in all biomaterial types, ctDNA presented at significantly higher levels in CSF compared to plasma and/or serum. When applied to a cohort of 127 plasma specimens from 41 patients collected from 2011 to 2018 as part of a randomized clinical trial in pediatric non-brainstem HGG/DMG, ctDNA profiling by ddPCR was of limited use due to the small volumes (mean = 0.49 mL) available. In anecdotal cases where sufficient material was available, cfDNA concentration correlated with disease progression in two examples each of poor response in H3F3A &#95;K27M-mutant DMG, and longer survival times in hemispheric BRAF &#95;V600E-mutant cases., Conclusion: Tumor-specific DNA alterations are more readily detected in CSF than plasma. Although we demonstrate the potential of the approach to assessing tumor burden, our results highlight the necessity for adequate sample collection and approach to improve detection if plasma samples are to be used., (© The Author(s) 2021. Published by Oxford University Press, the Society for Neuro-Oncology and the European Association of Neuro-Oncology.)

This study demonstrates the potential of feruloylated arabinoxylan (AX) from wheat bran for the preparation of bioactive barrier films with antioxidant properties. We have comprehensively evaluated the influence of the structural features and chemical acetylation of feruloylated AX extracted by subcritical water on their film properties, in comparison with alkaline extracted AX and a reference wheat endosperm AX. The degree of substitution (DS) of AX had a large influence on film formation, higher DS yielded better thermal and mechanical properties. The barrier properties of AX films were significantly enhanced by external plasticization by sorbitol. Chemical acetylation significantly improved the thermal stability but not the mechanical or barrier properties of the films. The presence of bound ferulic acid in feruloylated AX films resulted in higher antioxidant activity compared to external addition of free ferulic acid, which demonstrates their potential use in active packaging applications for the preservation of oxygen-sensitive foodstuff., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Introduction: Anemia in obese patients is common and multifactorial and is also a complication of bariatric surgery. The aim of this study is to establish which variables are associated with a higher prevalence of post-bariatric surgery anemia., Methods: Retrospective cohort study involving 1999 patients submitted to bariatric surgery with a follow-up period of 4 years. Anthropometric, laboratorial parameters and the presence of comorbidities were evaluated before surgery and during follow-up. Patients were divided into two groups, according to whether they developed anemia, or not. Groups were compared using independent sample T-tests or Chi-squared tests, as appropriate. Univariate binary logistic regression models were used to test the association between the diagnosis of anemia during follow-up and all the possible explanatory variables. Independent variables with an association with the outcome (p < 0.1) were included in multivariate binary logistic regression models, as well as possible confounders (age, BMI, and supplementation)., Results: Anemia was diagnosed in 24.4% of the patients. Females present a two-fold increased risk of developing anemia (OR 2.11, 95% CI 1.48-3.01, p < 0.001). Patients subjected to gastric sleeve and gastric band surgery present approximately half the risk of anemia when compared with gastric bypass surgery (RYGB) (OR 0.46, 95% CI 0.35-0.59, p < 0.001; OR 0.51, 95% CI 0.36-0.72, p < 0.001). None of the other variables evaluated showed association with the outcome., Conclusion: In our study, the only factors associated with an increased risk of developing anemia were female gender and RYBG surgery. Prospective studies evaluating the risk factors for anemia in patients undergoing bariatric surgery are needed.

Membrane bioreactor (MBR) has been widely employed for industrial effluent treatment, as its higher efficiency in removing pollutants makes effluent reuse more feasible. However, membrane fouling remains as a limiting factor for its greater diffusion. This work performed a sensitivity analysis study to investigate the effects of analytical and operating variables on membrane permeability. The case study is a MBR treating oil refinery effluents. After the identification and validation of a predictive neural model for permeability, sensitivity analysis methods based on both connection weights and variable disturbances were used to quantify and rank the variables influence. A comprehensive analysis showed that Suspended solids and Days between cleanings exerted greater effects on permeability, whereas sludge filterability and sludge temperature were less significant. In sequence, a specific analysis revealed distinct dynamics in MBR operation given different solids concentrations. For instance, from higher solids concentrations, among all the evaluated parameters, only COD presented low significance to the permeability. This evidence suggests that permeability is more susceptible to variations when operating with higher concentrations of Suspended solids. The global result of this study contributes to more efficient MBR operations since distinct relations with permeability imply different effects on membrane fouling.

Infant high-grade gliomas appear clinically distinct from their counterparts in older children, indicating that histopathologic grading may not accurately reflect the biology of these tumors. We have collected 241 cases under 4 years of age, and carried out histologic review, methylation profiling, and custom panel, genome, or exome sequencing. After excluding tumors representing other established entities or subgroups, we identified 130 cases to be part of an "intrinsic" spectrum of disease specific to the infant population. These included those with targetable MAPK alterations, and a large proportion of remaining cases harboring gene fusions targeting ALK ( n = 31), NTRK1/2/3 ( n = 21), ROS1 ( n = 9), and MET ( n = 4) as their driving alterations, with evidence of efficacy of targeted agents in the clinic. These data strongly support the concept that infant gliomas require a change in diagnostic practice and management. SIGNIFICANCE: Infant high-grade gliomas in the cerebral hemispheres comprise novel subgroups, with a prevalence of ALK, NTRK1/2/3, ROS1 , or MET gene fusions. Kinase fusion-positive tumors have better outcome and respond to targeted therapy clinically. Other subgroups have poor outcome, with fusion-negative cases possibly representing an epigenetically driven pluripotent stem cell phenotype. See related commentary by Szulzewsky and Cimino, p. 904 . This article is highlighted in the In This Issue feature, p. 890 ., (©2020 American Association for Cancer Research.)

Background: In current management of type 2 diabetes (T2DM), cardiovascular and renal prevention have become important targets to be achieved. In this context, a joint panel of four endocrinology societies from Brazil and Portugal was established to develop an evidence-based guideline for treatment of hyperglycemia in T2DM., Methods: MEDLINE (via PubMed) was searched for randomized clinical trials, meta-analyses, and observational studies related to diabetes treatment. When there was insufficient high-quality evidence, expert opinion was sought. Updated positions on treatment of T2DM patients with heart failure (HF), atherosclerotic CV disease (ASCVD), chronic kidney disease (CKD), and patients with no vascular complications were developed. The degree of recommendation and the level of evidence were determined using predefined criteria., Results and Conclusions: In non-pregnant adults, the recommended HbA 1c target is below 7%. Higher levels are recommended in frail older adults and patients at higher risk of hypoglycemia. Lifestyle modification is recommended at all phases of treatment. Metformin is the first choice when HbA 1c is 6.5-7.5%. When HbA 1c is 7.5-9.0%, dual therapy with metformin plus an SGLT2i and/or GLP- 1 RA (first-line antidiabetic agents, AD1) is recommended due to cardiovascular and renal benefits. If an AD1 is unaffordable, other antidiabetic drugs (AD) may be used. Triple or quadruple therapy should be considered when HbA 1c remains above target. In patients with clinical or subclinical atherosclerosis, the combination of one AD1 plus metformin is the recommended first-line therapy to reduce cardiovascular events and improve blood glucose control. In stable heart failure with low ejection fraction (< 40%) and glomerular filtration rate (eGFR) > 30 mL/min/1.73 m 2 , metformin plus an SGLT-2i is recommended to reduce cardiovascular mortality and heart failure hospitalizations and improve blood glucose control. In patients with diabetes-associated chronic kidney disease (CKD) (eGFR 30-60 mL/min/1.73 m 2 or eGFR 30-90 mL/min/1.73 m 2 with albuminuria > 30 mg/g), the combination of metformin and an SGLT2i is recommended to attenuate loss of renal function, reduce albuminuria and improve blood glucose control. In patients with severe renal failure, insulin-based therapy is recommended to improve blood glucose control. Alternatively, GLP- 1 RA, DPP4i, gliclazide MR and pioglitazone may be considered to reduce albuminuria. In conclusion, the current evidence supports individualizing anti-hyperglycemic treatment for T2DM., Competing Interests: Competing interestsMarcello Casaccia Bertoluci: has received grants and fees from Astra-Zeneca, Novo Nordisk, Boehringer Ingelheim, Sanofi, and Amgen. João Eduardo Nunes Salles: has received speaker’s fees and sat on boards for Abbott Nutrition, Abbott ADC, AstraZeneca, Bayer, Boeringher Ingelheim, Eli Lilly, Merck Serono, MSD, and Novo Nordisk. José Silva-Nunes: has received research support or honoraria for consultancy or training activities from Abbott, AstraZeneca, Bial, Boehringer Ingelheim, Eli Lilly & Company, Janssen Pharmaceuticals, Medinfar, Merck SA, Merck Sharp & Dohme, Mundipharma, Novartis Pharmaceuticals, Novo Nordisk, Roche, Sanofi, Servier, and Tecnimede. Hermelinda Cordeiro Pedrosa: sits on an advisory board for Roche and has received support for participation in scientific conferences from AstraZeneca, Novo Nordisk, and Servier. Rodrigo Oliveira Moreira: AstraZeneca, Boehringer Ingelheim, Novo Nordisk, Merck Serono, Servier, Takeda, Libbs, Sanofi-Aventis. Rui Manuel Calado da Silva Duarte: has received research grants and/or honoraria as a consultant, member of advisory board(s), or speaker from Abbott, AstraZeneca, Boehringer Ingelheim, Lilly, Medinfar, MSD, Novartis, Novo-Nordisk, Tecnimede, and Sanofi. Davide Mauricio da Costa Carvalho: has sat on advisory boards and is speaker for AstraZeneca, Boehringer Ingelheim, Eli Lilly, Mundipharma, Novo Nordisk, Novartis, and Sanofi. Fábio Rogério Trujilho: has received speaker’s fees from AstraZeneca, Boehringer-Lilly, Eurofarma, NovoNordisk, Sanofi, Servier, and Takeda. João Filipe Cancela dos Santos Raposo: has received speaker’s fees from or sat on advisory boards for Novo Nordisk, AstraZeneca, Eli Lilly, Abbott, and Boehringer Ingelheim. Erika Bezerra Parente: has received speaker’s fees from Sanofi, Abbott, and Eli Lilly; sits on the advisory board of Sanofi. Fernando Valente: NovoNordisk, AstraZeneca. Fábio Ferreira de Moura: has received speaker’s fees and/or support for participation in scientific conferences from Novo Nordisk, AstraZeneca, and Boehringer Ingelheim. Alexandre Hohl: has served as a scientific consultant for AstraZeneca, Novo Nordisk, and Sanofi. Miguel Melo: has consulting relationships with Bial and has participated in advisory panels and presented lectures for: Abbott, AstraZeneca, Bial, Boehringer Ingelheim, Johnson & Johnson, Lilly, NovoNordisk, Sanofi. Francisco Garcia Pestana Araujo: has received speaker’s fees from and/or served as an investigator in trials sponsored by Astra, Lilly, Medinfar, MSD, Tecninfar. Rosa Maria Monteiro Castro de Araújo Principe: has served as a clinical investigator in trials sponsored by Novo Nordisk, Bayer, and Sanofi. Adriana Costa e Forti: has received clinical research grants and/or support for participation in scientific conferences from AstraZeneca, Boehringer, Lilly, MSD, Novo Nordisk, Sanofi, Takeda. Cynthia Melissa Valerio: has received clinical research grants, speaker’s fees, and/or support for participation in scientific conferences from Novo Nordisk, Takeda, and Novartis. João Manuel Sequeira Duarte: has received grants and fees as consultant, speaker and investigator in clinical trials from Sanofi, Lilly, AstraZeneca, Novo Nordisk, Boehringer Ingelheim, Mundipharma, MSD, Tecnimed, and Bial. Melanie Rodacki: has received event sponsorship and speaker’s fees from Novo Nordisk, Abbott, and Sanofi. Pedro Manuel Patricio de Matos: has sat on advisory boards for MSD and AstraZeneca, and has received honoraria and research support from MSD, AstraZeneca, Boehringer-Lilly, Novo Nordisk, Abbott, and Bayer AG. Pedro Carneiro de Melo Pereira de Magalhães: has served as principal investigator in clinical trials sponsored by Bayer Pharma AG and Novo Nordisk; sits on advisory or expert boards for AstraZeneca; and has received support for participation in scientific conferences from Bial, Medinfar, Novo Nordisk, Sanofi. Rosângela Roginski Réa: has received clinical research grants, speaker’s fees, and/or support for participation in scientific conferences from AstraZeneca, Boehringer-Ingelheim, Eli Lilly, Janssen, Merck, MSD, Novo Nordisk, Sanofi, and Takeda. Roberto Tadeu Barcellos Betti, Patrícia Quadros Branco, Rosane Kupfer, Thaisa Dourado Guedes Trujilho, Maria Helane Costa Gurgel Castelo, Cristiane Bauermann Leitão, Hélder José Ferreira, José Francisco Kerr Saraiva, Mariana Pereira Monteiro, Lana Catani Ferreira Pinto declare no conflicts of interest., (© The Author(s) 2020.)

Background: Serum 25-hydroxyvitamin D [25(OH)D] and parathormone (PTH) have an inverse relation. The 25(OH)D threshold required to maximally suppress PTH has been used as a marker of optimal vitamin D status. Obesity is associated with lower serum levels of 25(OH)D and higher levels of PTH; however, the relation between these hormones in this setting is not well established. We aimed to assess the influence of excessive weight and weight loss after bariatric surgery on the relation between serum 25(OH)D and PTH., Methods: Anthropometric and analytical parameters of phospho-calcium metabolism from 290 patients undergoing bariatric surgery were analyzed. The association between 25(OH)D and PTH before and 1 year after surgery was evaluated through Student's t test and simple and multivariable linear regression., Results: At baseline, there was an inverse association between 25(OH)D and PTH plasma levels (β = - 0.571; p = 0.001). The threshold for a significant increase of PTH at baseline was a 25(OH)D level of < 10 ng/mL (p < 0.001). At 1 year after surgery (n = 194), the association between the two variables was weaker (β = - 0.291; p = 0.014). The 25(OH)D threshold for a significant increase in serum PTH levels was lower in patients who maintained obesity (n = 77) [25(OH)D 5 vs 10 ng/mL]., Conclusions: PTH suppression threshold observed in patients with morbid obesity before and after bariatric surgery is lower than that described for the general population, suggesting that the 25(OH)D plasma levels may not be representative of the body's vitamin D reserves. Other studies are required to demonstrate which optimal vitamin D levels need to be aimed for in obesity, in order to improve obese patients' bone health.

The impact of various degrees of acetylation on improving the thermal stability of xylan isolated from different botanical source has been studied; methylglucuronoxylan from birch and eucalyptus, arabinoglucuronoxylan from spruce and glucuronoarabinoxylan from sugarcane bagasse and straw. The lower molecular weight of non- acetylated methylglucuronoxylan (17.7-23.7 kDa) and arabinoglucuronoxylan (16.8 kDa) meant that they were more soluble in water than glucuronoarabinoxylan (43.0-47.0 kDa). The temperature at the onset of degradation increased by 17-61 °C and by 75-145 °C for low and high acetylated xylans respectively, as a result of acetylation. A glass transition temperature in the range of 121-132 °C was observed for the samples non-acetylated and acetylated at low degree of acetylation (0.0-0.6). The acetylation to higher degrees (1.4-1.8) increased the glass transition temperature of the samples to 189-206 °C. Acetylation proved to be an efficient method for functionalization of the xylan to increase the thermal stability., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Visceral leishmaniasis (VL) is considered a major public health concern in Brazil and several regions of the world. A recent advance in the diagnosis of infectious diseases was the development of loop-mediated isothermal amplification (LAMP). The aim of this study was to develop and evaluate a new LAMP assay for detection of K26 antigen-coding gene of L. donovani complex. A total of 219 blood samples of immunocompetent patients, including 114 VL cases and 105 non-VL cases, were analyzed for the diagnosis of VL in the present study. Diagnostic accuracy was calculated against a combination of parasitological and/or serological tests as a reference standard. The results were compared to those of kDNA Leishmania- PCR. The detection limit for the K26-Lamp assay was 1fg L. infantum purified DNA and 100 parasites/mL within 60 min of amplification time with visual detection for turbidity. The assay was specific for L. donovani complex. Sensitivity, specificity, and accuracy were 98.2%, 98.1%, and 98.2%, respectively, for K26-LAMP and 100%, 100%, and 100%, respectively, for kDNA Leishmania -PCR. Excellent agreement was observed between K26-LAMP and kDNA Leishmania- PCR assays (K = 0.96). A highly sensitive and specific LAMP assay targeting K26 antigen-coding gene of L. donovani complex was developed for diagnosis in peripheral blood samples of VL patients., Competing Interests: The authors declare no conflicts of interest.

Aims: To evaluate (1) the prevalence of diabetic distal symmetrical sensory-motor polyneuropathy (DSPN) and painful DSPN among patients with type 1 diabetes mellitus (DM1) aged over 18 years and (2) the determinant factors of neuropathy and pain in those patients. Materials and Methods: An epidemiological, cross-sectional, observational study was performed; 330,386 people were included, and a total of 444 people were diagnosed with DM1. After exclusion of possible confounders, 360 patients were assessed for painless and painful DSPNs using neurological examination and questionnaires for neuropathy and pain. Odds ratio (OR) and confidence intervals (95% CI) were estimated using multinomial logistic regression models. The analysis was based on a framework with four conceptual levels that consider feasible pathways between several risk factors: (1) socio-demographic factors and diabetes duration, (2) patient habits, (3) co-morbidities, and (4) metabolic factors and disease complications. Results: The prevalence of DSPN and painful DSPN were 42.8 and 18.9%, respectively. Diabetes duration was positively associated with painful (OR = 1.107, 95% CI: 1.107-1.139) and painless DSPN (OR = 1.069, 95% CI: 1.043-1.096). Education level was negatively associated with painful DSPN (OR = 0.889, 95% CI: 0.826-0.957). Sex (female) was positively associated only with painless DSPN (OR = 1.769, 95% CI: 1.007-3.107). Being a current or former smoker was positively associated only with painless DSPN (OR = 1.940, 95% CI: 1.069-3.518). Hypertension was positively associated with painful DSPN (OR = 2.474, 95% CI: 1.110-5.512) and painless DSPN (OR = 2.565, 95% CI: 1.252-5.256). Glycated hemoglobin (HbA1c) was positively associated only with painless DSPN (OR = 1.193, 95% CI: 1.018-1.399). Conclusions: Diabetes duration and hypertension have a direct impact on the development of painful and painless DSPN. However, female sex and HbA1c have a direct effect only on the development of painless DSPN, and education level has an indirect effect on the development of painful DSPN. Therefore, it can be concluded that different etiological factors have different contributions to the development of neuropathy and pain.

Considering that fluoxetine (FLX) is used to treat depressive states during pregnancy and that it is a cytochrome P450 (CYP)2D6 inhibitor, which is involved in the metabolism of both of its enantiomers, this study aims to describe the enantioselective distribution and metabolism of FLX and of its metabolite norfluoxetine (NorFLX) following a single oral dose. Nine healthy pregnant women received 20 mg FLX at 32 weeks of gestation and later at the day of delivery. The apparent clearance of (S)-(+)-FLX (1.45 vs. 0.66 L/hour/kg) and the area under the plasma concentration vs. time curve (AUC) of the (S)-(+)-NorFLX (AUC 0-∞ 942.7 vs. 498.6 ng hour/mL) were higher (P < 0.05) than those of the respective (R)-(-) enantiomers, indicating that the (S)-(+)-FLX enantiomer is preferentially metabolized to (S)-(+)-NorFLX. The placental transfer (umbilical vein/maternal vein) of FLX and NorFLX is low (30-40%), with the predominant transfer of (S)-(+)-FLX (44 vs. 33%). The distribution of the enantiomers of FLX and NorFLX to amniotic fluid is low (< 10%)., (© 2018 The Authors Clinical Pharmacology & Therapeutics © 2018 American Society for Clinical Pharmacology and Therapeutics.)

Structure-activity relationship and crystallographic data revealed that quinazolinone-containing fragments flip between two distinct modes of binding to activin receptor-like kinase-2 (ALK2). We explored both binding modes to discover potent inhibitors and characterized the chemical modifications that triggered the flip in binding mode. We report kinase selectivity and demonstrate that compounds of this series modulate ALK2 in cancer cells. These inhibitors are attractive starting points for the discovery of more advanced ALK2 inhibitors.

A two-dimensional liquid chromatography system coupled to triple quadrupole tandem mass spectrometer (2D LC-MS/MS) was employed for the determination of fluoxetine (FLU) and norfluoxetine (N-FLU) in colostrum and mature milk by direct sample injection. With a run time of 12 min representing a gain in throughput analysis, the validated methods furnished selectivity, extraction efficiency, accuracy, and precision in accordance with the criteria preconized by the European Medicines Agency guidelines. With a linear range of 3.00-150 ng/mL for FLU and 4.00-200 ng/mL for N-FLU they were applied to the analysis of colostrum and mature milk samples from nursing mothers. The paper discusses the differences and similarity of sample preparation for this two sample matrices. The herein reported methods are an advance in sample preparation procedures providing waste reduction and a sustainable approach., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The article discusses competition between midwives and doctors offering birth-related services in the city of Rio de Janeiro from 1835 to 1900. The research analyzed minutes from meetings, textbooks on births, and terms from qualification examinations for physicians, surgeons, apothecaries, and midwives at the Rio de Janeiro Medical School (Faculdade de Medicina do Rio de Janeiro), as well as announcements by and advertisements for doctors and midwives in columns featuring advertisements and useful recommendations in the Jornal do Commercio newspaper. An increase in the number of delivery physicians, and their scientific discourses led midwives to feel an obligation to diversify their clientele, consequently establishing themselves and working in lower-class and inhospitable areas.

Aims: Diabetes mellitus can inhibit cytochrome P450 3A4, an enzyme responsible for the metabolism of nifedipine, used for the treatment of hypertension in pregnant women. We aimed to assess the effect of type 2 diabetes mellitus (T2DM) on the pharmacokinetics, placental transfer and distribution of nifedipine in amniotic fluid in hypertensive pregnant women., Methods: The study was conducted in 12 hypertensive pregnant women [control group (CG)] and 10 hypertensive pregnant women with T2DM taking slow-release nifedipine (20 mg, 12/12 h). On the 34th week of gestation, serial blood samples were collected (0-12 h) after administration of the medication. At delivery, samples of maternal and fetal blood and amniotic fluid were collected for determination of nifedipine distribution in these compartments., Results: The median pharmacokinetic parameters of CG were: peak plasma concentration (C max ) 26.41 ng ml -1 , time to reach C max (t max ) 1.79 h, area under the plasma concentration vs. time curve from 0-12 h (AUC 0-12 ) 235.99 ng.h ml -1 , half-life (t½) 4.34 h, volume of distribution divided by bioavailability (Vd/F) 560.96 l, and Cl T /F 84.77 l h -1 . The parameters for T2DM group were: C max 23.52 ng ml -1 , t max 1.48 h, AUC 0-12 202.23 ng.h ml -1 , t½ 5.00 h, Vd/F 609.40 l, and apparent total clearance (Cl T /F) 98.94 l h -1 . The ratios of plasma concentrations of nifedipine in the umbilical vein, intervillous space and amniotic fluid to those in the maternal vein for CG and T2DM were 0.53 and 0.44, 0.78 and 0.87, respectively, with an amniotic fluid/maternal plasma ratio of 0.05 for both groups. The ratios of plasma concentrations in the umbilical artery to those in the umbilical vein were 0.82 for CG and 0.88 for T2DM., Conclusions: There was no influence of T2DM on the pharmacokinetics or placental transfer of nifedipine in hypertensive women with controlled diabetes., (© 2017 The British Pharmacological Society.)

Aim: This was to compare the periodontal status of children with primary and mixed dentition at the time of their first consultation., Methods: Children (200), aged 0-12 years (156 with mixed and 44 with primary dentition), were examined by assessing their simplified plaque index (PI) and simplified periodontal record (PSR). Statistical analysis (Chi-square test) was performed with appropriate software to find any significant associations between sex, type of dentition and PI with the PSR codes (0, 1, 2, 3 and 4)., Results: There was no statistically significant difference with regard to gender (p = 0.82). Generalised PI was associated more significantly with mixed dentition (p = 0.025 and higher PSR scores (p < 0.001). There was no significant relationship between sex and PSR codes (p = 0.82)., Conclusion: Children presenting with a mixed dentition had worse PI values and PSR scores. It is important to perform periodontal examination in children to diagnose and prevent future periodontal disease and maintain their dentition as well as to identify any associated systemic conditions.

Objectives: The capacity of a human cell line to secrete recombinant factor VIII with a F309S point mutation was investigated, as was the effect of the addition of chemical chaperones (betaine and sodium-4-phenylbutyrate) on the secretion of factor VIII., Methods: This work used a vector with a F309S mutation in the A1 domain to investigate FVIII production in the HEK 293 human cell line. Factor VIII activity was measured by chromogenic assay. Furthermore, the effects of chemical drugs on the culture were evaluated., Results: The addition of the F309S mutation to a previously described FVIII variant increased FVIII secretion by 4.5 fold. Moreover, the addition of betaine or sodium-4-phenylbutyrate increased the secretion rate of FVIIIΔB proteins in HEK 293 cells, but the same effect was not seen for FVIIIΔB-F309S indicating that all the recombinant protein produced had been efficiently secreted., Conclusion: Bioengineering factor VIII expressed in human cells may lead to an efficient production of recombinant factor VIII and contribute toward low-cost coagulation factor replacement therapy for hemophilia A. FVIII-F309S produced in human cells can be effective in vivo., (Copyright © 2016 Associação Brasileira de Hematologia, Hemoterapia e Terapia Celular. Published by Elsevier Editora Ltda. All rights reserved.)

Primitive neuroectodermal tumors of the central nervous system (CNS-PNETs) are highly aggressive, poorly differentiated embryonal tumors occurring predominantly in young children but also affecting adolescents and adults. Herein, we demonstrate that a significant proportion of institutionally diagnosed CNS-PNETs display molecular profiles indistinguishable from those of various other well-defined CNS tumor entities, facilitating diagnosis and appropriate therapy for patients with these tumors. From the remaining fraction of CNS-PNETs, we identify four new CNS tumor entities, each associated with a recurrent genetic alteration and distinct histopathological and clinical features. These new molecular entities, designated "CNS neuroblastoma with FOXR2 activation (CNS NB-FOXR2)," "CNS Ewing sarcoma family tumor with CIC alteration (CNS EFT-CIC)," "CNS high-grade neuroepithelial tumor with MN1 alteration (CNS HGNET-MN1)," and "CNS high-grade neuroepithelial tumor with BCOR alteration (CNS HGNET-BCOR)," will enable meaningful clinical trials and the development of therapeutic strategies for patients affected by poorly differentiated CNS tumors., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Two experiments were conducted to evaluate the inclusion of 0, 14, 27.5 and 41 g/kg roasted soybean (RSB) in protein-energy supplements for the finishing of beef cattle on pastures of Brachiaria brizantha cv. Marandu in the rainy period. In experiment 1, for the evaluation of the nutritional parameters, five rumen-cannulated steers with an initial average body weight of 474.5 kg were utilised. In experiment 2, for the evaluation of performance, 25 intact male zebu cattle with an initial average body weight of 418 kg were utilised. Supply of RSB led to an increase in the intake of crude protein and non-fibre carbohydrates (P < 0.05). The concentrations of rumen ammonia nitrogen increased significantly (P < 0.05) 4 h after supplementation. The animals that received protein-energy supplements had superior (52 %) average daily gain in relation to those receiving mineral mixture. The supply of protein-energy supplements formulated with roasted soybean provided an increase in average daily weight gain and an improvement in the nutritional parameters of finishing beef cattle on pasture in the rainy period.

Background: Neonatal effects of drugs administered to mothers before delivery depend on the quantity that crosses the placental barrier, which is determined by the pharmacokinetics of the drug in the mother, fetus, and placenta. Diabetes mellitus can alter the kinetic disposition and the metabolism of drugs. This study investigated the placental transfer of lidocaine and its metabolite monoethylglycinexylidide (MEGX) in pregnant women with gestational diabetes mellitus (GDM) submitted to peridural anesthesia., Patients and Methods: A total of 10 normal pregnant women (group 1) and 6 pregnant women with GDM (group 2) were studied, all at term. The patients received 200 mg 2% lidocaine hydrochloride by the peridural locoregional route. Maternal blood samples were collected at the time of delivery and, after placental expulsion, blood samples were collected from the intervillous space, umbilical artery, and vein for determination of lidocaine and MEGX concentrations and analysis of the placental transfer of the drug., Results: The following respective lidocaine ratios between the maternal and the fetal compartments were obtained for groups 1 and 2: umbilical vein/maternal peripheral blood, 0.60 and 0.46; intervillous space/maternal blood, 1.01 and 0.88; umbilical artery/umbilical vein, 0.77 and 0.91; and umbilical vein/intervillous space, 0.53 and 0.51. The following MEGX ratios for groups 1 and 2 were, respectively, fetal/maternal, 0.43 and 0.97; intervillous space/maternal blood, 0.64 and 0.90; umbilical artery/umbilical vein, 1.09 and 0.99; and umbilical vein/intervillous space, 0.55 and 0.78., Conclusion: Gestational diabetes mellitus did not affect the transplacental transfer of lidocaine but interfered with the transfer of MEGX, acting as a mechanism facilitating the transport of the metabolite., (© The Author(s) 2015.)

Nifedipine is a dihydropyridine calcium channel blocker used for the treatment of hypertension in pregnant women. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for analysis of nifedipine in human plasma and amniotic fluid. Separation of nifedipine and nitrendipine (IS) was performed using a LiChroCART(®) RP-Select B column and a mixture of water:acetonitrile:glacial acetic acid (30:70:0.5 v/v) as the mobile phase. Aliquots of 500μL of biological samples were extracted at pH 13 using dichloromethane:n-pentane (3:7 v/v). The validated method was applied to a study of the pharmacokinetics of nifedipine in human plasma and amniotic fluid samples collected up to 12h after administration of the last slow-release nifedipine (20mg/12h) dose to 12 hypertensive pregnant women. The estimated pharmacokinetic parameters of nifedipine showed a mean AUC(0-12) of 250.2ngh/mL, ClT/F of 89.2L/h, Vd/F of 600.0L and t1/2 5.1h. The mean amniotic fluid/plasma concentration ratio was 0.05. The methods proved to be highly sensitive by showing a lower quantification limit of 0.1ng/mL for both matrices. And this study reports for the first time the complete development and validation of the method to quantify nifedipine in amniotic fluid using LC-MS-MS., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Background: Familial partial lipodystrophy, Dunnigan variety, is a recognised autosomal dominant disorder which is caused by heterozygous missense mutations in the lamin A/C gene. Dunnigan lipodystrophy is characterised by a variable loss of fat from the extremities and trunk, as well as an excess of subcutaneous fat in the chin and supraclavicular area. The associated metabolic abnormalities include: insulin resistance, diabetes, dyslipidaemia and low leptin levels., Case Presentation: The authors studied the case of a 24-year-old caucasian pregnant woman, with a past medical history of acute pancreatitis, combined dyslipidaemia and diabetes mellitus. At 7 weeks of pregnancy she was referred to the outpatient endocrinology and obstetrics clinic for diabetes care. A physical examination revealed that she presented a loss of fat from the extremities and trunk and also had an excess of subcutaneous fat in the chin. Triglyceride levels were persistently high, and glycaemic control was only achieved through the administration of high doses of insulin (1.8 U/Kg/day). Dunnigan lipodystrophy was suspected and thus a genetic study was requested, which revealed the presence of c.1444C > T (p.Arg482Trp) heterozygote mutation in the lamin A/C gene., Conclusion: This case is used to illustrate the importance of being able to recognise the clinical signs of this rare lipodystrophic syndrome, which may cause potentially severe consequences, and also the difficulties in treating it during pregnancy.

Different resource use strategies manifest as differences in the realized niches of species. Niche segregation may involve several dimensions of the niche, such as diet, space, and time. We measured the level of redundancy and complementarity of a bee-plant interaction network in an agricultural system. Because flower resource diversity is high and resource abundance associated with flowering phenology varies throughout the year, we hypothesized that trophic overlap in the community would be low (i.e., high niche complementarity). In contrast, we expected a combination of physiological constraints and exploitation competition to create high temporal overlap, leading to high redundancy in the time of use of floral resources. Dietary overlap was low (NOih = 0.18): niches of 88% of species pairs had less than 30% overlap. In contrast, temporal overlap was intermediate (NOih = 0.49): niches of 65% of species pairs had 30% to 60% overlap. Network analysis showed that bees separated their dietary niches and had intermediate complementary specialization (H2' = 0.46). In terms of their temporal niches (H2' = 0.12), bees were generalists, with high temporal redundancy. Temperature was not a key factor in the determination of niche overlap, suggesting that environmental factors do not likely have a primary role in determining high redundancy in the temporal use of floral resources. Rather, temporal overlap is likely associated with the timing of nectar production by flowers. Our results suggest that bees partition a wide variety of available floral resources, resulting in low dietary overlap and intermediate temporal overlap.

Reference genes presenting stable expression profiles over a wide variety of conditions are required in relative expression studies of specific bacterial genes by quantitative reverse transcription PCR (RT-qPCR). High-throughput sequencing of bacterial transcriptomes using the RNA-seq methodology now provides a wealth of data that may be searched for identification of the most stably expressed genes of a given bacterium. Herein, we searched a RNA-seq dataset from various experiments with the pathogenic bacterium Corynebacterium pseudotuberculosis, grown under different stress conditions, in order to select appropriate candidate reference genes for this species. Nineteen genes involved in maintenance of basic cellular functions, so-called housekeeping genes, were chosen for study and their expression profiles in C. pseudotuberculosis were evaluated throughout all growth conditions. Eight of these genes (atpA, dnaG, efp, fusA, gyrA, gyrB, rpoB, and rpoC), mostly participating in DNA replication and transcription, matched the defined criteria to be included as candidate reference genes. Transcriptional levels of these genes were quantified by RT-qPCR assays after growth of C. pseudotuberculosis under two additional conditions. Expression stability analysis by NormFinder indicated the combination of genes encoding DNA gyrase subunit A (gyrA) and elongation factor P (fusA) as the most suitable for normalization of RT-qPCR studies in C. pseudotuberculosis.

Background. During dengue virus (DV) infection, monocytes produce tumor necrosis factor alpha (TNF-α) and nitric oxide (NO) which might be critical to immunopathogenesis. Since intensity of DV replication may determine clinical outcomes, it is important to know the effects of viral nonstructural protein 1 (NS1) on innate immune parameters of infected patients. The present study investigates the relationships between dengue virus nonstructural protein 1 (NS1) serum levels and innate immune response (TLR4 expression and TNF-α/NO production) of DV infected patients presenting different clinical outcomes. Methodology/Principal Findings. We evaluated NO, NS1 serum levels (ELISA), TNF-α production by peripheral blood mononuclear cells (PBMCs), and TLR4 expression on CD14(+) cells from 37 dengue patients and 20 healthy controls. Early in infection, increased expression of TLR4 in monocytes of patients with dengue fever (DF) was detected compared to patients with dengue hemorrhagic fever (DHF). Moreover, PBMCs of DHF patients showed higher NS1 and lower NO serum levels during the acute febrile phase and a reduced response to TLR4 stimulation by LPS (with a reduced TNF-α production) when compared to DF patients. Conclusions/Significance. During DV infection in humans, some innate immune parameters change, depending on the NS1 serum levels, and phase and severity of the disease which may contribute to development of different clinical outcomes.

Unlabelled: Sound evidence on the effectiveness of fluoride varnishes (FV) to reduce caries incidence in preschool children is lacking., Objective: To assess whether the application of FV in preschool children at 6-month intervals decreases the incidence of caries and produces any adverse effects., Methods: A randomized, examiner- and patient-blind, placebo-controlled, parallel-group design, clinical trial, comprising 1- to 4-year-old children, 100 in each group (FV or placebo varnish, PV), was conducted in Rio de Janeiro, Brazil. Two trained pediatric dentists performed the clinical examinations (kappa = 0.85). Dental caries was recorded at the d2 (cavitated enamel) and d3 (dentine) levels using the International Caries Diagnosis and Assessment System., Results: At baseline, the mean age of the participants was 2.4 years (SD 0.9) and the mean d3mfs was 0.8 (SD 1.9). Most of the children brushed their teeth with fluoride toothpaste and consumed fluoridated tap water. After 24 months, 89 and 92 children of the test and the control groups were analyzed, respectively. A total of 32 (35.9%) children in the FV group and 43 (46.7%) in the PV group presented new dentine caries lesions (χ(2) test; p = 0.14), showing relative and absolute risk reductions of 23% (95% CI: -9.5 to 45.9) and 11% (95% CI: -3.5 to 25.0). The mean caries increment differences between the test and control groups were -0.8 (95% CI: -2.0 to 0.4) at the d2 level and -0.7 (95% CI: -1.9 to 0.4) at the d3 level. Only 2 minor complaints regarding the intervention were reported., Conclusion: Although safe and well accepted, twice-yearly professional FV application, during 2 years, did not result in a significant decrease in caries incidence.

This article describes the experience of in-service training for pedagogical coordinators of the Brazilian School Nutrition Program (PNAE) developed by the Cooperation Center for Student Food and Nutrition of the Federal University of Bahia (CECANE-UFBA). Participant observation and analysis of data documented in films, group discussion summaries, procedural and final evaluations and field diaries are used. The formative experiences were in 2011, in two municipalities in Bahia and Sergipe states, involving 118 pedagogical coordinators from 79 municipalities in both states. It was revealed that the objective of raising awareness and guiding pedagogical coordinators for mainstreaming the theme of food and nutrition has been achieved. The educational proposal was evaluated by the coordinators as "dynamic," "productive" and "interactive," promoting the exchange of experiences among participants. Despite considering the theme relevant, the pedagogical coordinators do not have sufficient training about food and nutrition to back their teaching practices. It is considered that their training has some limitations on the impact in the fieldwork, as the knowledge acquired night be dissipated when the coordinators return to their workplace due to the lack of ongoing training.

Viral hepatitis A, B, C, D and E--systemic hepatotropic viral infections--present as acute hepatitis that, depending on the etiological agent, viral load and host conditions, may evolve into chronic hepatitis, cirrhosis, liver cancer and acute fulminant disease. The ecological versatility of these viruses, their spectrum of transmission in time and space, potentialized by the sub-clinical course of a large proportion of infections, comprise an epidemiological challenge. This essay describes scenarios and tendencies in the socioepidemiologic profile, based on the history of these infections, and indicates the need to overcome patterns, models, and protocols and instead investigate each particular situation. In other words, it highlights the need to explore singularities in order to be able to develop new proposals for general actions tailored to local specificities.

An inventory of the theses on hepatitis of the Faculdade de Medicina of the Universidade Federal do Rio de Janeiro between 1837 and 2000 is presented. The analysis indicates the potential and limits for discussion of the evolutionary framework of scientific knowledge on these health problems in Brazil. The theories are also discussed in light of their scientific reference points and the technological and social changes that influenced them. The landmarks in medical education and knowledge about hepatitis are identified and categorized, considering that the theses reveal at the very least the state of the art on the subject. The study makes it possible to explore the foundations upon which the scientific knowledge on hepatitis were built and indicate possibilities for research in the reconstruction of scientific knowledge of other health problems.