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Background: Recent analysis of the human proteome via proteogenomics and ribosome profiling of the transcriptome revealed the existence of thousands of previously unannotated microprotein-coding small open reading frames (smORFs). Most functional microproteins were chosen for characterization because of their evolutionary conservation. However, one example of a non-conserved immunomodulatory microprotein in mice suggests that strict sequence conservation misses some intriguing microproteins., Results: We examine the ability of gene regulation to identify human microproteins with potential roles in inflammation or fibrosis of the intestine. To do this, we collected ribosome profiling data of intestinal cell lines and peripheral blood mononuclear cells and used gene expression of microprotein-encoding transcripts to identify strongly regulated microproteins, including several examples of microproteins that are only conserved with primates., Conclusion: This approach reveals a number of new microproteins worthy of additional functional characterization and provides a dataset that can be queried in different ways to find additional gut microproteins of interest., (© 2024. The Author(s).)

Parabens are preservatives widely used in personal care products, pharmaceuticals, and foodstuffs. However, they are still unregulated chemical compounds. Given their extensive use and presence in different environmental compartments, parabens can adversely affect animal health. Thus, the current study aimed to summarize and critically analyze the bioaccumulation and ecotoxicity of parabens in aquatic species. Studies have been mostly conducted in laboratory conditions (75%), using mainly fish and crustaceans. Field studies were carried out across 128 sampling sites in six countries. Paraben bioaccumulation was predominantly detected in fish muscle, liver, brain, gills, ovary, and testes. Among the parent parabens, methylparaben (MeP), ethylparaben (EtP), and propylparaben (PrP) have been detected frequently and more abundantly in tissues of marine and freshwater specimens, as well as the metabolite 4-hydroxybenzoic acid (4-HB). Parabens can induce lethal and sublethal effects on aquatic organisms, such as oxidative stress, endocrine disruption, neurotoxicity, behavioral changes, reproductive impairment, and developmental abnormalities. The toxicity of parabens varied according to species, taxonomic group, developmental stage, exposure time, and concentrations tested. This study highlights the potential bioaccumulation and ecotoxicological impacts of parabens and their metabolites on aquatic invertebrates and vertebrates. Additionally, future research recommendations are provided to evaluate the environmental risks posed by paraben contamination more effectively., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Neotropical primates rarely exhibit active tuberculosis. A brown howler monkey was found injured in an urban area. Histopathology revealed granulomatous inflammation in the lungs, lymph nodes, and liver. Immunohistochemistry and molecular analysis confirmed the presence of Mycobacterium tuberculosis complex. The findings highlight the importance of TB surveillance in nonhuman primates., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Genome annotation has historically ignored small open reading frames (smORFs), which encode a class of proteins shorter than 100 amino acids, collectively referred to as microproteins. This cutoff was established to avoid thousands of false positives due to limitations of pure genomics pipelines. Proteogenomics, a computational approach that combines genomics, transcriptomics, and proteomics, makes it possible to accurately identify these short sequences by overlaying different levels of omics evidence. In this chapter, we showcase the use of μProteInS, a bioinformatics pipeline developed for the identification of unannotated microproteins encoded by smORFs in bacteria. The workflow covers all the steps from quality control and transcriptome assembly to the scoring and post-processing of mass spectrometry data. Additionally, we provide an example on how to apply the pipeline's machine learning method to identify high-confidence spectra and pinpoint the most reliable identifications from large datasets., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Tuberculosis (TB) is one of the main causes of death from a single pathological agent, Mycobacterium tuberculosis (Mtb). In addition, the emergence of drug-resistant TB strains has exacerbated even further the treatment outcome of TB patients. It is thus needed the search for new therapeutic strategies to improve the current treatment and to circumvent the resistance mechanisms of Mtb. The shikimate kinase (SK) is the fifth enzyme of the shikimate pathway, which is essential for the survival of Mtb. The shikimate pathway is absent in humans, thereby indicating SK as an attractive target for the development of anti-TB drugs. In this work, a combination of in silico and in vitro techniques was used to identify potential inhibitors for SK from Mtb (MtSK). All compounds of our in-house database (Centro de Pesquisas em Biologia Molecular e Funcional, CPBMF) were submitted to in silico toxicity analysis to evaluate the risk of hepatotoxicity. Docking experiments were performed to identify the potential inhibitors of MtSK according to the predicted binding energy. In vitro inhibitory activity of MtSK-catalyzed chemical reaction at a single compound concentration was assessed. Minimum inhibitory concentration values for in vitro growth of pan-sensitive Mtb H37Rv strain were also determined. The mixed approach implemented in this work was able to identify five compounds that inhibit both MtSK and the in vitro growth of Mtb., (© 2022. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Microproteins (MPs) are a potentially rich source of uncharacterized metabolic regulators. Here, we use ribosome profiling (Ribo-seq) to curate 3,877 unannotated MP-encoding small ORFs (smORFs) in primary brown, white, and beige mouse adipocytes. Of these, we validated 85 MPs by proteomics, including 33 circulating MPs in mouse plasma. Analyses of MP-encoding mRNAs under different physiological conditions (high-fat diet) revealed that numerous MPs are regulated in adipose tissue in vivo and are co-expressed with established metabolic genes. Furthermore, Ribo-seq provided evidence for the translation of Gm8773, which encodes a secreted MP that is homologous to human and chicken FAM237B. Gm8773 is highly expressed in the arcuate nucleus of the hypothalamus, and intracerebroventricular administration of recombinant mFAM237B showed orexigenic activity in obese mice. Together, these data highlight the value of this adipocyte MP database in identifying MPs with roles in fundamental metabolic and physiological processes such as feeding., Competing Interests: Declaration of interests All authors affiliated with the Novo Nordisk Research Center Seattle, Inc. have worked for a for-profit commercial pharmaceuticals company that produces and sells medicines for the treatment of obesity and diabetes. B.C.S. is a founder and shareholder of Proteome Software. M.J.M. has a sponsored research agreement with and is a paid consultant for Thermo Fisher Scientific. A. Saghatelian is a paid consultant and shareholder for and cofounder of Exo Therapeutics and Velia Therapeutics. T.F.M. is a paid consultant and shareholder of Velia Therapeutics. C.A.B. is a current employee of Velia Therapeutics., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)

Background: The indiscriminate use of antimicrobials is considered a major contributing factor to the increase in antimicrobial resistance. Community pharmacies are the main source of access to antimicrobials, and pharmacists are in a strategic position to promote rational use of these medicines. Therefore, it is important to know dispensing service quality., Objective: To evaluate the behavior of pharmacists in dispensing antimicrobials in community pharmacies in northeast Brazil., Methods: This cross-sectional pilot study was conducted from August to October 2021 in a private community pharmacy chain in Sergipe. Dispensing was evaluated using the simulated patient (SP) technique. Two SP asked the pharmacists for the antimicrobials (case clinic 1: upper respiratory infection; case clinic 2: urinary tract infection) and recorded the service through audio. Dispensing practices were independently analyzed by two researchers based on the tools available in the literature. Data were presented using descriptive statistics., Results: A total of 54 simulated patient visits were conducted. Based on the 12 steps recommended by the research team for good dispensing, pharmacists asked an average of 1 (±1.17) question for upper respiratory infections and 0.3 (±0.54) for urinary tract infections, as well as provided counseling (mean number of recommendations, 2.6 (±1.44) and 4.5 (±2.35), respectively). As for communication skills, pharmacists had a regular score (3.07 ± 0.34). Furthermore, there was no significant difference in the number of steps and counseling recommendations by pharmacists in dispensing clinical cases 1 and 2 (p = 0.0674)., Conclusion: The quality of antimicrobial dispensing was evaluated as suboptimal, requiring improvements in practice and multifaceted strategies to promote continuing education of these professionals. In addition, awareness actions for the population must be implemented to promote the rational use of antimicrobials and reduce microbial resistance., (© 2022. The Author(s).)

Background: Antimicrobial resistance remains a major global public health concern, and antimicrobial dispensing in community pharmacies is an important factor in preventing this damage. However, the current literature focuses on the technical and attitudinal aspects related to antimicrobial dispensing, with little emphasis on the interventions provided in this service. Thus, this study aimed to determine the antimicrobial dispensing process in community pharmacies., Methods: A scoping review was performed in September 2020 using the PubMed, EMBASE, LILACS, Web of Science, and Cochrane databases. The search terms included words related to dispensing, antibacterial agents, and pharmacies in various combinations. Two reviewers screened the titles, abstracts, and full-text articles according to the eligibility criteria, and extracted the data. The findings were presented in a descriptive form., Results: Of the 7713 studies screened, 35 were included, of which 22 (63%) were published in Asia. Most studies followed a cross-sectional design (n = 27), and the simulated patient was the most often used method to assess the antimicrobial dispensing process (n = 22). Moreover, 31 (89%) studies investigated antimicrobial dispensing without prescription, and only four (11%) studies evaluated antimicrobial dispensing with prescription. In the 35 studies, the most frequently asked questions were about drug allergies (n = 19) and patient symptoms (n = 18), and counseling mainly focused on the side effects (n = 14), precautions (n = 14), how to take the medication (n = 12), and duration of medication use (n = 11). Another common intervention was referral (n = 15). Among clinical cases, counseling on medication use occurred often in cases of urinary tract infection (51%) and otitis media (50%)., Conclusions: Antimicrobial dispensing processes have been primarily investigated in low- and middle-income countries, with a focus on dispensing antimicrobials without prescriptions. During the dispensing process, pharmacists mostly posed minimal questions and counseling, highlighting the deficiencies that persist in this practice. Our results indicate the need for multifaceted strategies, such as implementing educational, regulatory or administrative strategies and changes in cultural background, especially in low- and middle-income countries, that aim to reduce indiscriminate use of antimicrobials. Therefore, qualifying the antimicrobial dispensing process is a fundamental factor for improving the rational use of antimicrobials and reducing microbial resistance., (© 2022. The Author(s).)

Tuberculosis (TB) remains one of the leading causes of death due to a single pathogen. The emergence and proliferation of multidrug-resistant (MDR-TB) and extensively drug-resistant strains (XDR-TB) represent compelling reasons to invest in the pursuit of new anti-TB agents. The shikimate pathway, responsible for chorismate biosynthesis, which is a precursor of important aromatic compounds, is required for Mycobacterium tuberculosis growth. The enzyme 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase ( Mtb DAHPS) catalyzes the first step in the shikimate pathway and it is an attractive target for anti-tubercular agents. Here, we used a CRISPRi system to evaluate the DAHPS as a vulnerable target in M. tuberculosis. The silencing of aroG significantly reduces the M. tuberculosis growth in both rich medium and, especially, in infected murine macrophages. The supplementation with amino acids was only able to partially rescue the growth of bacilli, whereas the Aro supplement (aromix) was enough to sustain the bacterial growth at lower rates. This study shows that Mtb DAHPS protein is vulnerable and, therefore, an attractive target to develop new anti-TB agents. In addition, the study contributes to a better understanding of the biosynthesis of aromatic compounds and the bacillus physiology. IMPORTANCE Determining the vulnerability of a potential target allows us to assess whether its partial inhibition will impact bacterial growth. Here, we evaluated the vulnerability of the enzyme 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase (DAHPS) from M. tuberculosis by silencing the DAHPS-coding aroG gene in different contexts. These results could lead to the development of novel and potent anti-tubercular agents in the near future.

Enoyl-ACP reductases (ENRs) are enzymes that catalyze the last step of the elongation cycle during fatty acid synthesis. In recent years, new bacterial ENR types were discovered, some of them with structures and mechanisms that differ from the canonical bacterial FabI enzymes. Here, we briefly review the diversity of structural and catalytic properties of the canonical FabI and the new FabK, FabV, FabL, and novel ENRs identified in a soil metagenome study. We also highlight recent efforts to use the newly discovered Fabs as targets for drug development and consider the complex evolutionary history of this diverse set of bacterial ENRs., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Hopf, Roth, de Souza, Galina, Czeczot, Machado, Basso and Bizarro.)

Summary: Genome annotation pipelines traditionally exclude open reading frames (ORFs) shorter than 100 codons to avoid false identifications. However, studies have been showing that these may encode functional microproteins with meaningful biological roles. We developed µProteInS, a proteogenomics pipeline that combines genomics, transcriptomics and proteomics to identify novel microproteins in bacteria. Our pipeline employs a model to filter out low confidence spectra, to avoid the need for manually inspecting Mass Spectrometry data. It also overcomes the shortcomings of traditional approaches that usually exclude overlapping genes, leaderless transcripts and non-conserved sequences, characteristics that are common among small ORFs (smORFs) and hamper their identification., Availability and Implementation: µProteInS is implemented in Python 3.8 within an Ubuntu 20.04 environment. It is an open-source software distributed under the GNU General Public License v3, available as a command-line tool. It can be downloaded at https://github.com/Eduardo-vsouza/uproteins and either installed from source or executed as a Docker image., Supplementary Information: Supplementary data are available at Bioinformatics online., (© The Author(s) 2022. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Drug dispensing aims to promote rational medicine use. However, in many countries, the work processes are still not well defined. In this sense, the perception of pharmacists about dispensing practices presents an overview of how the service is being performed in the country and its main challenges. Thus, the purpose of this study was to determine the self-reported work process of Brazilian community pharmacists in relation to drug dispensing, challenges, and strategies for carrying out the service., Method: A cross-sectional survey was conducted between May and July 2021, with community pharmacists from all regions of Brazil. Pharmacists were invited to answer a validated, self-administered questionnaire, implemented through Google Forms, containing 33 questions related to the steps of drug dispensing (questions and counseling) and the main challenges and strategies to perform the service. The data were exported to Microsoft Office Excel and SPSS®. Multiple linear regression analysis was used to assess the association between responses and demographic information, with a significance level of less than 5% (p < 0.05). This study was approved by the Research Ethics Committee (number: 4.295.171)., Results: A total of 625 community pharmacists responded to the survey. Most pharmacists reported always or frequently performing 17 (54%) of the 31 steps described in the instrument. The steps that pharmacists reported performing more frequently were forming the medication name (n = 569, 91.04%), verifying the completeness and adequacy of the prescription according to current legislation (n = 567, 90.72%) and providing counseling on dosage (n = 549, 87.84%). Documentation was the main step in which pharmacists reported never or rarely performing (n = 424, 67.84%). The results showed that there was a significant influence of the variables of public education institution, age, and postgraduate education on the frequency of dispensing steps (F(3, 621) = 14.884, p < 0.001; R 2 ajdusted  = 0,063)., Conclusion: This study showed that most pharmacists reported always or frequently asking most of the questions and performing counseling contained in the instrument during drug dispensing. These results can contribute to an understanding of current dispensing practices and generate insights for developing strategies to qualify the service., (© 2022. The Author(s).)

The epidemiological importance of mycobacterial species is indisputable, and the necessity to find new molecules that can inhibit their growth is urgent. The shikimate pathway, required for the synthesis of important bacterial metabolites, represents a set of targets for inhibitors of Mycobacterium tuberculosis growth. The aroA -encoded 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) enzyme catalyzes the sixth step of the shikimate pathway. In this study, we combined gene disruption, gene knockdown, point mutations (D61W, R134A, E321N), and kinetic analysis to evaluate aroA gene essentiality and vulnerability of its protein product, EPSPS, from Mycolicibacterium ( Mycobacterium ) smegmatis ( Ms EPSPS). We demonstrate that aroA -deficient cells are auxotrophic for aromatic amino acids (AroAAs) and that the growth impairment observed for aroA -knockdown cells grown on defined medium can be rescued by AroAA supplementation. We also evaluated the essentiality of selected Ms EPSPS residues in bacterial cells grown without AroAA supplementation. We found that the catalytic residues R134 and E321 are essential, while D61, presumably important for protein dynamics and suggested to have an indirect role in catalysis, is not essential under the growth conditions evaluated. We have also determined the catalytic efficiencies ( K cat / K m ) of recombinant wild-type (WT) and mutated versions of Ms EPSPS (D61W, R134A, E321N). Our results suggest that drug development efforts toward EPSPS inhibition may be ineffective if bacilli have access to external sources of AroAAs in the context of infection, which should be evaluated further. In the absence of AroAA supplementation, aroA from M. smegmatis is essential, its essentiality is dependent on Ms EPSPS activity, and Ms EPSPS is vulnerable. IMPORTANCE We found that cells from Mycobacterium smegmatis, a model organism safer and easier to study than the disease-causing mycobacterial species, when depleted of an enzyme from the shikimate pathway, are auxotrophic for the three aromatic amino acids (AroAAs) that serve as building blocks of cellular proteins: l-tryptophan, l-phenylalanine, and l-tyrosine. That supplementation with only AroAAs is sufficient to rescue viable cells with the shikimate pathway inactivated was unexpected, since this pathway produces an end product, chorismate, that is the starting compound of essential pathways other than the ones that produce AroAAs. The depleted enzyme, the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), catalyzes the sixth step of shikimate pathway. Depletion of this enzyme inside cells was performed by disrupting or silencing the EPSPS-encoding aroA gene. Finally, we evaluated the essentiality of specific residues from EPSPS that are important for its catalytic activity, determined with experiments of enzyme kinetics using recombinant EPSPS mutants.

This study combined conventional epidemiology of human astroviruses. From 2010 to 2016, 232 stool samples from children under 5 years of age were screened using NGS and conventional RT-PCR followed by genetic analysis in order to investigate the genotypic diversity of classical human astrovirus (HAstV) circulating in Tocantins State, Brazil. HAstV was detected in 16 cases (6.9%). Seven specimens (43.7%; 7/16) were positive according RT-PCR and next-generation sequencing (NGS) to investigate the molecular to both NGS and RT-PCR. NGS and RT-PCR individually revealed six (37.5%; 6/16) and three (18.8%; 3/16) additional positive samples, respectively. Sequencing of the HAstV-positive samples revealed HAstV-1a (9/16), HAstV-4c (3/16), and HAstV-5c (4/16) lineages., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Objectives : This study investigated the relationship between single-nucleotide polymorphisms (SNPs) in cytokine genes and the susceptibility to Squamous Intraepithelial Lesions (SIL), cervical cancer and HPV infection through a systematic review with meta-analysis. To verify the effect of SNPs, we also analyzed the transcription factor binding affinity using bioinformatics tools. Methods : Seven electronic databases (MEDLINE, Scielo, BIREME, PubMed, Scopus, Web of Science and Science Direct) were searched for case-control studies. Results : A total of 35 relevant case-control studies were meta-analyzed, including 7 cytokine genes and 15 SNPs. SNPs in IL-17A (rs2275913, rs3748067); IL-17 F (rs763780); IL-12A (rs568408); IL-12B (rs3212227); TNFA (rs1800629, rs361525); IL-1B (rs16944); IL-6 (rs1800795); IL-10 (rs1800896) genes were associated with increased risk for cervical cancer. No association was observed between meta-analyzed polymorphisms and SIL. Additional bioinformatics analysis suggested a possible transcriptional regulation pathway of the TNFA and IL-10 genes through the MZF1 ( TNFA -308 G > A and IL-10  - 1082A>G) and ZNF263 ( TNFA -238 G > A) transcription factors binding. Conclusion : Overall, 10 SNPs in cytokine genes were associated with increased risk for cervical cancer. Therefore, in our meta-analysis, these SNPs demonstrated to be potential biomarkers for predicting or identifying cases of high risk for SIL and cervical cancer.

Objective and Design: Biochanin A (BCA), a phytoestrogen, has various pharmacological properties. This study was conducted to compare BCA's therapeutic property against 17-β estradiol replacement therapy in zymosan-induced arthritis (ZIA) in mice. Additionally, we further investigated in vitro the anti-inflammatory action on neutrophils., Treatment: Ovariectomized (OVX) and non-OVX mice were pretreated with BCA (1, 3 and 9 mg/kg) or estrogen (50 µg/kg) for 14 days prior to ZIA. Neutrophils were pretreated with BCA (1, 10 and 100 μM) for 1 h prior to phorbol 12-myristate 13-acetate., Methods: Anti-inflammatory effects of BCA were evaluated by cellular infiltrate, paw edema and cytokine measurement. In vitro, apoptosis was assessed by morphology and flow cytometry. Neutrophil extracellular traps (NET) were determined by fluorescent microscopy and DNA release. Statistical differences were determined by one- or two-way ANOVA., Results: BCA inhibited neutrophil accumulation, paw edema and proinflammatory cytokine (TNF-α and IFN-γ) and increased anti-inflammatory cytokines (IL-4 and IL-10) in OVX and non-OVX mice, similar to 17-β estradiol replacement therapy. In vitro, BCA increased apoptosis and consequently reduced NETs., Conclusion: BCA has a notable anti-inflammatory effect, similar to 17-β estradiol, and is especially effective for treatment of ZIA. These results suggest that BCA may be promising for the treatment of postmenopausal arthritis.

The global epidemic of tuberculosis (TB) imposes a sustained epidemiologic vigilance and investments in research by governments. Mycobacterium tuberculosis , the main causative agent of TB in human beings, is a very successful pathogen, being the main cause of death in the population among infectious agents. In 2018, ~10 million individuals were contaminated with this bacillus and became ill with TB, and about 1.2 million succumbed to the disease. Most of the success of the M. tuberculosis to linger in the population comes from its ability to persist in an asymptomatic latent state into the host and, in fact, the majority of the individuals are unaware of being contaminated. Even though TB is a treatable disease and is curable in most cases, the treatment is lengthy and laborious. In addition, the rise of resistance to first-line anti-TB drugs elicits a response from TB research groups to discover new chemical entities, preferably with novel mechanisms of action. The pathway to find a new TB drug, however, is arduous and has many barriers that are difficult to overcome. Fortunately, several approaches are available today to be pursued by scientists interested in anti-TB drug development, which goes from massively testing chemical compounds against mycobacteria, to discovering new molecular targets by genetic manipulation. This review presents some difficulties found along the TB drug development process and illustrates different approaches that might be used to try to identify new molecules or targets that are able to impair M. tuberculosis survival., (Copyright © 2020 Dalberto, de Souza, Abbadi, Neves, Rambo, Ramos, Macchi, Machado, Bizarro and Basso.)

Background: Inflammation could be a risk factor for the development of depression and change the outcome of this common chronic-recurrent mental disorder., Aims: This study aimed to investigate if bone marrow mononuclear cell (BMMC) transplantation is effective in restoring sucrose preference in rats subjected to chronic stress (CS), if it has an anti-inflammatory effect and is able to restore damaged DNA., Methods: The effect of BMMC transplantation was studied in a controlled protocol (compared with a control group and a selective serotonin reuptake inhibitor escitalopram group) involving sucrose preference in CS in rats. Measurements were taken of the amygdala, hippocampus, frontal cortex, and other brain areas, the spleen and blood pro-inflammatory cytokines, namely interleukin-1β, interleukin-6, tumor necrosis factor-alpha, and interferon-gamma, as well as anti-inflammatory cytokine interleukin-10. Finally, 8-hydroxy-2'-deoxyguanosine (a DNA damage marker) was determined., Results: BMMC transplantation was as effective as escitalopram in restoring sucrose preference. It also had an anti-inflammatory effect and slightly improved damaged DNA after one week., Conclusions: These findings suggest administration of BMMC in rats subjected to CS restores sucrose preference, resolves inflammation in both the peripheral and central nervous system, as well as diminishes DNA damage. This effect was similar to that of escitalopram, which is effective in the treatment of depressive patients.

The human umbilical cord blood (HUCB) is an excellent source of adult stem cells, having the benefit of being younger than the bone marrow stem cells. The role of stem cells in the lesion repair mechanism is still being studied. We evaluated the capability of HUCB to interfere into the fibroblast dedifferentiation plasticity through cocultivation. Direct and indirect cocultures were maintained for 24, 48, and 72 hours. Coculture viability was evaluated by MTT assay. The messenger RNA was extracted, and the expression of p16 and p21 genes was estimated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). The direct or indirect contact did not interfere with fibroblast cell viability. However, these direct and indirect contacts reduced the expression of p16 and p21 genes. A sigmoidal curve was applied to adjust gene expression against time, and a mathematical function was established for gene expression according to cell culture type. These results suggest that the differentiated cells were influenced by immature cells (HUCB) either by the direct contact or by signaling molecules, which alter their behavior and plasticity. Therefore our data may contribute to paracrine effects other than the commonly known to be responsible for the repair of lesions in stem cell therapy.

Background: Ice-pack is widely used for alleviating postpartum perineal pain sustained after birth related perineal trauma. However, it lacks robust evidence on timing and frequency of applications, to ensure the effective and safe use of this therapy., Aims: To evaluate if a 10min ice-pack application relieved postpartum perineal pain and if the analgesic effect was maintained for up to 2h., Methods: A randomised controlled trial conducted from December 2012 to February 2013 with 69 primiparous women ≥18 years old, 6-24h postpartum, with perineal pain ≥3, who had not received anti-inflammatory medication or analgesics after childbirth, who were randomised to a single ice-pack application on the perineum for 10min or standard care. The primary and secondary outcomes were a reduction ≥30% in perineal pain intensity, immediately after the application and the maintenance of the analgesic effect for up to 2h, respectively., Findings: Immediately post-intervention, the proportion of women whose perineal pain decreased ≥30% was significantly higher in the experimental group. Within 2h, there was no significant difference in the pain levels in both groups. Within 2h, for 61.9% and 89.3% of women in the experimental and control group, respectively, the perineal pain levels remained unchanged. For the remaining participants, perineal pain was increasing after an average time of 1h 45min and 1h 56min for the experimental and control groups, respectively., Conclusion: By applying an ice-pack for 10min to the perineum, effective pain relief is achieved, that is maintained for between 1h 45min and 2h., (Copyright © 2018 Australian College of Midwives. Published by Elsevier Ltd. All rights reserved.)

Wound healing is a complex process that involves several biological events, and a delay in this process may cause economic and social problems for the patient. The search continues for new alternative treatments to aid healing, including the use of herbal medicines. Members of the genus Caesalpinia are used in traditional medicine to treat wounds. The related species Poincianella pluviosa (DC.) L.P. Queiroz increases the cell viability of keratinocytes and fibroblasts and stimulates the proliferation of keratinocytes in vitro. The crude extract (CE) from bark of P. pluviosa was evaluated in the wound-healing process in vivo, to validate the traditional use and the in vitro activity. Standardized CE was incorporated into a gel and applied on cutaneous wounds (TCEG) and compared with the formulation without CE (Control) for 4, 7, 10, or 14 days of treatment. The effects of the CE on wound re-epithelialization; cell proliferation; permeation, using photoacoustic spectroscopy (PAS); and proteins, including vascular endothelial growth factor (VEGF), superoxide dismutase 2 (SOD-2) and cyclooxygenase 2 (COX-2) were evaluated. The TCEG stimulated the migration of keratinocytes at day 4 and proliferation on the following days, with a high concentration of cells in metaphase at 7 days. Type I collagen formed more rapidly in the TCEG. PAS showed that the CE had permeated through the skin. TCEG stimulated VEGF at day 4 and SOD-2 and COX-2 at day 7. The results suggest that the CE promoted the regulation of proteins and helped to accelerate the processes involved in healing, promoting early angiogenesis. This led to an increase in the re-epithelialized surface, with significant mitotic activity. Maturation of collagen fibers was also enhanced, which may affect the resistance of the extracellular matrix. PAS indicated a correlation between the rate of diffusion and biological events during the healing process. The CE from P. pluviosa appears promising as an aid in healing.

Diabetes mellitus is a serious public health problem in which a major complication is impaired wound healing. Among the strategies developed to foster tissue repair is the use of medicinal plants. The bark of Stryphnodendron adstringens, which is popularly used as an aid in wound healing, has a documented effect on wound repair in normal rats. This study evaluated the healing action of the crude extract of S. adstringens in diabetic rats, and its chemical content. Compounds present in the crude extract were characterized by mass spectrometry. In diabetic rats (streptozotocin 35 mg/kg), two wounds made in the skin were treated daily for 4, 7, 10, and 14 days with gel containing 1 % crude extract or with base gel. Histological analyses involved the measurement of the length and thickness of the re-epithelialized surface, quantification of the number of cells in mitosis, and types I and III collagen fibers. Also, cutaneous permeation by photoacoustic spectroscopy, and the expression of cyclooxygenase-2 and vascular endothelial growth factor by Western blot were assessed. The crude extract fingerprint showed masses indicating proanthocyanidins. The crude extract mainly stimulated cell migration and proliferation of keratinocytes at the beginning of the treatment in addition to stimulating the replacement of type III collagen fibers by type I collagen fibers at 10 and 14 days. The photoacoustic spectroscopy technique showed that the gel containing 1 % of crude extract permeated through the skin to the dermis, where the crude extract was found. Vascular endothelial growth factor was stimulated after 7 days of treatment with the crude extract and cyclooxygenase-2 at 4, 7, and 10 days. The crude extract of S. adstringens acted in tissue repair in wounds in diabetic rats by stimulating the production of collagen fibers at the wound site. The crude extract favored the formation of a more organized extracellular matrix and filled the entire extent of the wound, and also fostered the upregulation of cyclooxygenase-2 and vascular endothelial growth factor, which are essential to this process. These crude extract actions in diabetic wounds are probably due to the presence of proanthocyanidins., (Georg Thieme Verlag KG Stuttgart · New York.)

The application of synthetic aminopolycarboxylic acids to soil increases metal solubility, and therefore enhances phytoextraction. However, synthetic chelants degrade poorly in soil, and metal leaching threatens human and animal health. The aim of this study is to assess the use of a biodegradable chelant (NTA) for Pb phytoextraction from a soil contaminated by battery-casing disposal. EDTA was also included in the experiment to assess the behavior of a non-degradable chelant. Each synthetic chelant was applied to soil pots cultivated with maize plants at rates of 0, 2, 5, 10, and 20 mmol kg(-1). Soil samples were extracted with CaCl(2) and by sequential extraction for Pb. In addition, a soil column experiment was set up to study the leaching of Pb from the chelant-amended soil. The results showed that both NTA and EDTA were highly effective in solubilizing Pb from soil. The Pb distribution into soil fractions after chelant addition followed the sequence: Ex (exchangeable)>OM (organic matter)>AFeOx (amorphous iron oxides)>CFeOx (crystalline iron oxides). The 5 mmol kg(-1) dose of EDTA increased the Pb concentration in maize shoots to 1.1%, but it promoted unacceptable Pb leaching rates. On the other hand, the results showed that phytoremediating the site using 5 mmol kg(-1) NTA could be feasible with no environmental effects due to Pb leaching over a five-year period.