RESUMO
We studied sows (Landrace × Yorkshire line, DanBred Hybrid) to evaluate the possible changes in progesterone receptor (PR) expression in the uterus and ovary caused by different non-hypophyseal gonadotropins treatments: equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG). Varying concentrations of eCG and hCG were evaluated (Groups 1, 2, 3, 4). PR expression was determined by immunohistochemistry, and labelling intensity was determined by the HScore method. In the ovary, PR expression in the granulosa cells of follicles did not differ significantly between Groups 1 and 2 (p < 0.05) but differed significantly from that in Groups 3 and 4 (p < 0.05), which in turn did not differ from each other. This PR expression pattern was similar across groups in the internal and external theca cells. Conversely, in the uterus, PR expression in the lining epithelium was lower in Group 4 than that in Group 1 (p < 0.05). Increased expression was observed in the endometrial lamina propria in all groups 2 and 4 compared to that in the control group (p < 0.05). Decreased expression was observed in the glandular epithelium and myometrium in Group 4 compared to that in Group 1 (p < 0.05). In the ovary, PR expression in the granulosa and outer and inner theca of the follicles was not significantly different (p < 0.05) between Groups 1 and 2 or Groups 3 and 4; however, the expression in these pairs of groups differed from each other. Thus, changes in PR expression may depend on the concentrations and proportions of exogenous hormones used in the treatments, indicating an alteration in the reproductive process.
Assuntos
Gonadotropina Coriônica , Ovário , Receptores de Progesterona , Útero , Animais , Feminino , Receptores de Progesterona/metabolismo , Gonadotropina Coriônica/farmacologia , Útero/metabolismo , Útero/efeitos dos fármacos , Ovário/metabolismo , Ovário/efeitos dos fármacos , Suínos , Cavalos , Humanos , Imuno-Histoquímica/veterinária , Gonadotropinas Equinas/farmacologia , Células da Granulosa/metabolismo , Células da Granulosa/efeitos dos fármacos , Endométrio/metabolismo , Endométrio/efeitos dos fármacosRESUMO
Fungal keratitis, which is caused primarily by Neocosmospora and Fusarium species, is a significant global health issue that affects more than a million people annually in tropical and subtropical regions. Neocosmospora solani (formerly Fusarium solani) is a leading cause of corneal infections, along with members of the Fusarium oxysporum species complex (FOSC). This study provides new insights by reporting a series of ocular fusariosis cases caused by FOSC members and presenting molecular evidence linking specific haplotypes within FOSC to human infections. We describe three cases of Fusarium keratitis selected from a comprehensive review of clinicopathological data in our institution's archives. These cases were chosen for their distinctive clinical presentations and the involvement of less common Fusarium species. Two of these patients were diagnosed with keratitis and anterior endophthalmitis, and the third patient had a corneal ulcer previously treated with topical antivirals and antibiotics. All patients were successfully treated with topical amphotericin B. The Fusarium isolates from these patients were subjected to detailed molecular characterization, including DNA sequencing (tef1α, rpb2, CaM, tub2, and LSU), amplified fragment length polymorphism (AFLP) marker analysis, and MALDI-TOF MS analysis. Remarkably, our study reports the first case of human infection by F. veterinarium, alongside cases involving F. contaminatum and F. curvatum. Furthermore, a molecular survey using haplotypic networks based on tef1α sequences identified genotypes associated with human infections and revealed the emergence of F. veterinarium clade VII. Our findings emphasize the need for vigilance regarding emerging species within the FOSC, particularly F. veterinarium. This highlights the need for improved diagnostic tools and targeted research to combat fusarioid-related infections effectively.
Assuntos
Fusariose , Fusarium , Ceratite , Fusarium/genética , Fusarium/isolamento & purificação , Fusarium/classificação , Humanos , Ceratite/microbiologia , Ceratite/diagnóstico , Fusariose/microbiologia , Fusariose/diagnóstico , Masculino , Feminino , Análise de Sequência de DNA , Pessoa de Meia-Idade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Adulto , Haplótipos , Antifúngicos/uso terapêutico , Antifúngicos/farmacologia , Idoso , DNA Fúngico/genéticaRESUMO
Obesity, a global epidemic, is linked to adverse reproductive outcomes, including infertility and ovulation dysfunction. The cafeteria diet (CAF) serves as an animal model mirroring Western diet habit. Coenzyme Q10 (CoQ10), known for enhancing reproductive outcomes in various pathologies, is not fully understood for its effects on obesity treatment. Here, obesity was modeled using CAF-fed rats to assess CoQ10's impact on metabolic and ovarian disruptions caused by obesity. Wistar rats were divided into control (standard diet) and obese (CAF diet) groups. After 75 days, half of each group received oral CoQ10 (5 mg/kg) for 13 days, while the rest received a vehicle. Animals were euthanized during the estrus phase, and blood and ovaries were collected for analysis. CAF caused increased body weight gain (p < 0.01) associated with hyperglycemia, hypertriglyceridemia, and hypercholesterolemia (p < 0.05). Moreover, it caused a reduction in the number of AMH + follicles (p < 0.001), increasing follicular atresia (p < 0.05) and serum estradiol levels (p < 0.05). Obesity also altered the estrous cycle and reduced the ovulation rate (p < 0.05). CoQ10 administration showed beneficial effects on all ovarian disruptions but had no effect on the metabolic alterations induced by obesity. In summary, CoQ10 could be an additional treatment for obesity-related infertility in patients with normal metabolic profiles. While CoQ10 does not affect metabolic parameters influenced by obesity, crucial for reproductive issues and offspring health, it is recommended as part of a treatment plan that includes a balanced diet and increased physical activity for obese individuals with metabolic alterations seeking pregnancy.
Assuntos
Suplementos Nutricionais , Obesidade , Ratos Wistar , Reprodução , Ubiquinona , Animais , Ubiquinona/análogos & derivados , Ubiquinona/farmacologia , Ubiquinona/uso terapêutico , Feminino , Obesidade/tratamento farmacológico , Obesidade/metabolismo , Ratos , Reprodução/efeitos dos fármacos , Ovário/efeitos dos fármacos , Ovário/metabolismoRESUMO
Rhodococcus equi is an opportunistic soil-borne bacterium that is eliminated in feces of multi-host animals. An increase in multidrug-resistant R. equi isolates has been reported in humans and domestic animals, and it has been hypothesized that the treatment of R. equi in foals could increase the selective pressure on multidrug-resistant isolates and favor human infections by resistant isolates. We investigated the in vitro antimicrobial susceptibility/resistance of 41 R. equi strains from humans, which were isolated from patients with pulmonary signs, using 19 antimicrobials from 10 distinct classes, recommended exclusively to humans, recommended exclusively to domestic animals and used in both. All isolates were subjected to mass spectrometry and identified as R. equi. Among the antimicrobials used exclusively in humans, tigecycline and vancomycin showed 100% efficacy. Amikacin, amoxicillin/clavulanic acid, imipenem, levofloxacin, clarithromycin, rifampin, ciprofloxacin, and gentamicin, used in both humans and animals, revealed high efficacy (97-100%). Conversely, a higher frequency of isolates was resistant to penicillin (87.8%) and trimethoprim/sulfamethoxazole (43.9%), which are used in both humans and animals. Among the antimicrobials used only in animals, isolates were resistant to florfenicol (46.4%), ceftiofur (17.1%), and enrofloxacin (2.5%). Multidrug resistance was observed in 34% of isolates. The identification of drug-resistant R. equi isolated from humans used exclusively in animals is circumstantial evidence of the pathogen transmission from domestic animals to humans. This study contributes to the molecular identification of Rhodococcus species from humans and to the epidemiological vigilance of the multidrug-resistant isolates.
Assuntos
Infecções por Actinomycetales , Antibacterianos , Testes de Sensibilidade Microbiana , Rhodococcus equi , Rhodococcus equi/efeitos dos fármacos , Rhodococcus equi/isolamento & purificação , Animais , Antibacterianos/farmacologia , Humanos , Infecções por Actinomycetales/microbiologia , Infecções por Actinomycetales/tratamento farmacológico , Infecções por Actinomycetales/veterinária , Farmacorresistência Bacteriana Múltipla , Animais Domésticos/microbiologia , Cavalos/microbiologiaRESUMO
Hepatic ischemia reperfusion injury (HIRI) is a pathophysiological and complex systemic process involving multiple tissues and organs. Gastrodin (GSTD), a natural compound from Gastrodia elata, displays a variety of interesting pharmacological activities. Heme oxygenase-1 (HO-1), a stress-responsive protein, has a cytoprotective defense response against oxidative and inflammatory injuries. The aim of this investigation was to elucidate whether GSTD plays a protective role against HIRI by regulating HO-1 expression. GSTD (100 mg/kg) or zinc protoporphyrin (15 mg/kg; an HO-1 inhibitor) was administered to HIRI C57 male mice. GSTD decreased glutamic pyruvic transaminase and glutamic oxaloacetic transaminase levels in HIRI mice. Inflammatory (TNF-α and IL-6) and oxidative-stress (malondialdehyde, MDA) markers of HIRI mice were decreased by GSTD. GSTD up-regulated HO-1 protein and mRNA expression in HIRI mice but decreased caspase-3 and -9 protein expression. GSTD lowered mRNA expression of apoptosis-related genes (caspase-3, -9, -12, and Bax) in the liver of HIRI mice but enhanced mRNA level of the anti-apoptotic Bcl-2 gene. Consistent with in vivo results, GSTD displayed a similar regulatory effect on the expression of mRNA (HO-1, caspase-3, -9, -12, Bax, and Bcl-2) and protein (HO-1, caspase-3 and -9) as well as inflammatory (TNF-α and IL-6) and on oxidative stress factors (superoxide dismutase and MDA) in BRL-3A cells transfected with small interfering HO-1 RNA in a hypoxia-reperfusion model. In conclusion, GSTD up-regulated HO-1 expression to play a protective role in HIRI by anti-apoptotic, anti-inflammatory, and antioxidant effects. GSTD is a promising natural compound that alleviated HIRI in liver surgery.
Assuntos
Álcoois Benzílicos , Glucosídeos , Heme Oxigenase-1 , Camundongos Endogâmicos C57BL , Traumatismo por Reperfusão , Animais , Traumatismo por Reperfusão/metabolismo , Traumatismo por Reperfusão/prevenção & controle , Masculino , Glucosídeos/farmacologia , Álcoois Benzílicos/farmacologia , Heme Oxigenase-1/metabolismo , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Fígado/efeitos dos fármacosRESUMO
Alpinia zerumbet, a plant native to East Asia, is widely found on the Brazilian coast, where it is used in folk medicine as an antihypertensive, diuretic, and anxiolytic. This study investigated the effects of the hydroalcoholic extract obtained from Alpinia zerumbet leaves (AZE) on cardiovascular changes and oxidative status in spontaneously hypertensive rats (SHR). SHR and Wistar-Kyoto male rats, 90 days old, treated or not with AZE (50 mg/kg/day in drinking water) for six weeks, were used in this study. Blood pressure (BP) was assessed weekly by tail plethysmography. At the end of treatment, the animals were anesthetized with thiopental (70 mg/kg, ip), blood was collected through abdominal aorta puncture, the thoracic aorta and left ventricle were isolated for morphometric analysis and immunostaining of NOX-4, SOD-2, 8-isoprostane, and angiotensin II AT1 receptors (AT1R), and the mesenteric arterial bed (MAB) was isolated for the assessment of vascular function. Oxidative damage in lipids and proteins and the enzymatic antioxidant activity were evaluated in plasma samples by spectrophotometry. AZE normalized BP in SHR. Although the treatment did not improve the MAB vascular dysfunction, it reversed the cardiovascular remodeling in the aorta and left ventricle. In addition, AZE improved antioxidant activity in plasma and SOD-2 immunostaining in the thoracic aorta and left ventricle, decreased protein carbonylation in plasma, and reduced 8-isoprostane, NOX-4, and AT1R immunostaining in the cardiovascular system. The results suggested that AZE reversed hypertension and cardiovascular remodeling in SHR, which was associated with lower oxidative stress and AT1R.
Assuntos
Alpinia , Anti-Hipertensivos , Hipertensão , Extratos Vegetais , Folhas de Planta , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Animais , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Hipertensão/tratamento farmacológico , Hipertensão/fisiopatologia , Masculino , Alpinia/química , Ratos , Folhas de Planta/química , Anti-Hipertensivos/farmacologia , Pressão Sanguínea/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Ventrículos do Coração/efeitos dos fármacos , Ventrículos do Coração/fisiopatologia , Remodelação Ventricular/efeitos dos fármacosRESUMO
Osteosarcoma (OS) remains the most common bone tumor and the prognosis for many patients remains stagnant due to the unsatisfactory therapeutic effect of conventional treatment regimens. This research explored the effect and mechanism of a novel natural product, Eriocalyxin B (EB), in pathogenesis and immunotherapy in OS. Cell Count Kit 8 assay, colony formation assay, and wound healing assay were employed to detect the proliferative, colony-forming, and migratory abilities of human OS cells following EB treatment. Moreover, xenograft growth assay was performed to assess the effect of EB on OS in vivo. Subcutaneous OS models constructed in immunocompetent mice were employed to evaluate the effect of EB treatment in combination with immune checkpoint blockades (ICBs) PD1ab and CTLA4ab. Immunohistochemistry (IHC) staining was utilized to detect the level of CD8+ T cells infiltration and Ki67 expression. TARGET database, RNA interference technology, and qPCR assay were employed to explore the mechanism of EB on OS. EB inhibited the proliferative, colony-forming, and migratory abilities of the human OS cells MG63 and U2OS both in vitro and in vivo. TARGET data analysis demonstrated that up-regulation of TCEA3 was significantly negatively correlated with overall survival in OS patients. EB exerted anti-tumor activity via downregulation of TCEA3. EB, in conjunction with ICBs, synergistically optimized anti-tumorigenic activity against OS in immunocompetent mice. EB may promote infiltration of CD8+ T cells and down-regulate Ki67 expression. These results signaled that EB may have a role as a candidate therapeutic or preventive agent for the treatment of OS.
Assuntos
Neoplasias Ósseas , Regulação para Baixo , Inibidores de Checkpoint Imunológico , Osteossarcoma , Osteossarcoma/tratamento farmacológico , Animais , Humanos , Regulação para Baixo/efeitos dos fármacos , Camundongos , Inibidores de Checkpoint Imunológico/farmacologia , Inibidores de Checkpoint Imunológico/uso terapêutico , Neoplasias Ósseas/tratamento farmacológico , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto , Antígenos de Neoplasias , DiterpenosRESUMO
Necrotizing enterocolitis (NEC) is a severe intestinal disease of multifactorial origin that primarily affects premature infants. Approximately 27% of NEC babies develop short gut (SG) secondary to extensive intestinal resection, and 10% will have chronic dependence on total parenteral nutrition. We evaluated the Botox treatment in SG model rats. Twenty-day-old weanling male rats (weight range 38-70 g, n=72) were divided into four groups (n=18 each): 1) Control (fed a regular liquid diet); 2) Botox (Control submitted to laparotomy and intestinal injection of Botox®); 3) SG (short gut); and 4) SG and Botox (SG+Botox®). After seven post-operative days, samples were collected for biometrics [body weight (BW), intestine weight (IW) and IW/BW ratio (IBR), and intestine length (IL) and height (IH)], histometric analysis [villous height (VH), crypt depth (CD), muscular thickness (MT), and PCNA index)], and intestinal transit time (ITT). BW, IW, and IL decreased in SG (P<0.05). IH, VH, and PCNA index increased in Botox groups [Control = SG < Botox and SG+Botox (P<0.05)], CD increased in Botox, SG, and SG+Botox (P<0.005), and MT was higher in SG and SG+Botox. Botox groups had lower ITT (P<0.05). Botox provided dilatation and histological changes in SG. These findings suggested that Botox improved adaptation and might be applied in SG with promising results.
Assuntos
Adaptação Fisiológica , Síndrome do Intestino Curto , Animais , Masculino , Síndrome do Intestino Curto/tratamento farmacológico , Adaptação Fisiológica/efeitos dos fármacos , Ratos , Modelos Animais de Doenças , Ratos Wistar , Toxinas Botulínicas Tipo A/farmacologia , Toxinas Botulínicas Tipo A/administração & dosagem , Enterocolite Necrosante/tratamento farmacológico , Animais Recém-Nascidos , Desmame , Intestinos/efeitos dos fármacosRESUMO
Nasopharyngeal carcinoma (NPC) is a malignant tumor predominantly influenced by Epstein-Barr virus infection and genetic factors. The transforming growth factor-beta (TGF-ß) superfamily is implicated in various cellular processes, including tumorigenesis. This study aimed to detect the role of one TGF-ß superfamily member activin receptor type IIB (ACTRIIB) in NPC. This study analyzed NPC datasets, including GSE12452, GSE102349, and GSE53819. ACTRIIB expression and N-glycosylation levels were assessed by western blot, real-time PCR, immunofluorescence, and immunohistochemistry in NPC cells and tissues. As indicated by the datasets, ACTRIIB was significantly upregulated in NPC tissues, and the up-regulation was associated with poor prognosis. This study confirmed the N-glycosylation of ACTRIIB primarily at the forty-second amino acid, an asparagine. The N-glycosylation of ACTRIIB promoted the localization of ACTRIIB to the cell membrane and prevented the degradation of the protein by lysosomes, through which ACTRIIB activated the downstream Smard1/2 to promote tumor cell proliferation and invasion. Inhibition of N-glycosylation or knockdown of ACTRIIB resulted in reduced cell proliferation and invasion and increased the cell sensitivity to docetaxel. In conclusion, N-glycosylation of ACTRIIB was a critical post-translational modification that enhanced protein stability and induced membrane localization, which facilitates the functions of ACTRIIB in cell proliferation and invasion in NPC. Inhibition of ACTRIIB N-glycosylation could potentially serve as a therapeutic strategy to improve the efficacy of chemotherapy in NPC.
Assuntos
Antineoplásicos , Proliferação de Células , Docetaxel , Resistencia a Medicamentos Antineoplásicos , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas , Humanos , Docetaxel/farmacologia , Docetaxel/uso terapêutico , Carcinoma Nasofaríngeo/metabolismo , Carcinoma Nasofaríngeo/tratamento farmacológico , Carcinoma Nasofaríngeo/patologia , Carcinoma Nasofaríngeo/genética , Proliferação de Células/efeitos dos fármacos , Glicosilação/efeitos dos fármacos , Neoplasias Nasofaríngeas/metabolismo , Neoplasias Nasofaríngeas/tratamento farmacológico , Neoplasias Nasofaríngeas/patologia , Neoplasias Nasofaríngeas/genética , Antineoplásicos/farmacologia , Resistencia a Medicamentos Antineoplásicos/genética , Receptores de Activinas Tipo II/metabolismo , Receptores de Activinas Tipo II/genética , Estabilidade Proteica/efeitos dos fármacos , Linhagem Celular Tumoral , Imuno-Histoquímica , Reação em Cadeia da Polimerase em Tempo Real , Western Blotting , Regulação para CimaRESUMO
Cancer is the second leading cause of death worldwide. Cancer cachexia is a multifactorial catabolic syndrome responsible for almost one third of cancer-related deaths. Drug repurposing has been used in oncological research and drugs like clenbuterol and metformin seem to be reasonable candidates in the context of cancer cachexia, because the former is a ß2-agonist that stimulates muscle gain and the latter has anti-inflammatory properties. The aim of this study was to assess the effects of a short-term treatment with metformin and clenbuterol, isolated or combined, on tumor growth and cancer cachexia parameters in Walker 256 tumor-bearing rats, a model of cancer cachexia. To this end, Wistar rats were separated into 8 groups and 4 of them were injected with Walker 256 tumor cells (W groups). Control (C) and W groups received the following treatments: metformin (M), clenbuterol (Cb), or metformin combined with clenbuterol (MCb). Body and tumor weight, metabolic parameters, and oxidative damage in the tumor were assessed. Compared to the C group, the W group showed body weight loss, hypoglycemia, hyperlactatemia, and hypertriacylglycerolemia. None of the treatments could reverse body weight loss, although they reversed the alterations of the assessed plasma metabolic parameters. Surprisingly, only clenbuterol alone reduced tumor weight. Hydrogen peroxide production and lipid peroxidation in tumor tissue was increased in this group. In conclusion, metformin and clenbuterol ameliorated metabolic cachexia parameters in Walker tumor-bearing rats, but only clenbuterol reduced the tumor weight, probably, through a lipid peroxidation-dependent cell death.
Assuntos
Caquexia , Carcinoma 256 de Walker , Clembuterol , Peroxidação de Lipídeos , Metformina , Ratos Wistar , Animais , Caquexia/tratamento farmacológico , Caquexia/etiologia , Metformina/farmacologia , Metformina/uso terapêutico , Clembuterol/farmacologia , Clembuterol/uso terapêutico , Carcinoma 256 de Walker/tratamento farmacológico , Carcinoma 256 de Walker/complicações , Masculino , Peroxidação de Lipídeos/efeitos dos fármacos , Ratos , Hipoglicemiantes/farmacologia , Hipoglicemiantes/uso terapêutico , Modelos Animais de Doenças , Agonistas Adrenérgicos beta/farmacologiaRESUMO
Tau is a neuronal protein that confers stability to microtubules; however, its hyperphosphorylation and accumulation can lead to an impairment of protein degradation pathways, such as autophagy. Autophagy is a lysosomal catabolic process responsible for degrading cytosolic components, being essential for cellular homeostasis and survival. In this context, autophagy modulation has been postulated as a possible therapeutic target for the treatment of neurodegenerative diseases. Studies point to the modulatory and neuroprotective role of the cannabinoid system in neurodegenerative models and here it was investigated the effects of cannabidiol (CBD) on autophagy in a human neuroblastoma strain (SH-SY5Y) that overexpresses the EGFP-Tau WT (Wild Type) protein in an inducible Tet-On system way. The results demonstrated that CBD (100 nM and 10 µM) decreased the expression of AT8 and total tau proteins, activating autophagy, evidenced by increased expression of light chain 3-II (LC3-II) protein and formation of autophagosomes. Furthermore, the cannabinoid compounds CBD, ACEA (CB1 agonist) and GW-405,833 (CB2 agonist) decreased the fluorescence intensity of EGFP-Tau WT; and when chloroquine, an autophagic blocker, was used, there was a reversal in the fluorescence intensity of EGFP-Tau WT with CBD (1 and 10 µM) and GW-405,833 (2 µM), demonstrating the possible participation of autophagy in these groups. Thus, it was possible to conclude that CBD induced autophagy in EGFP-Tau WT cells which increased tau degradation, showing its possible neuroprotective role. Hence, this study may contribute to a better understanding of how cannabinoids can modulate autophagy and present a potential therapeutic target in a neurodegeneration model.
Assuntos
Autofagia , Canabidiol , Proteínas tau , Canabidiol/farmacologia , Autofagia/efeitos dos fármacos , Humanos , Proteínas tau/metabolismo , Linhagem Celular Tumoral , Proteínas de Fluorescência Verde/metabolismo , Proteínas de Fluorescência Verde/genéticaRESUMO
Parkinson's disease (PD) is a complex neurodegenerative disorder that affects multiple neurotransmitters, and its exact cause is still unknown. Developing new drugs for PD is a lengthy and expensive process, making it difficult to find new treatments. This study aims to create a detailed dataset to build strong predictive models with various machine learning algorithms. An ensemble modeling approach was employed to screen the DrugBank database, aiming to repurpose approved medications as potential treatments for Parkinson's disease (PD). The dataset was constructed using pIC50 values of various compounds targeting the inhibition of leucine-rich repeat kinase 2 (LRRK2). The best ensemble model showed exceptional predictive performance, with five-fold cross-validation and external validation metrics exceeding 0.8 (Q2cv = 0.864 and Q2ext = 0.873). The DrugBank screening resulted in three promising drugs-triamterene, phenazopyridine, and CRA_1801-with predicted pIC50 values greater than 7, warranting further investigation as novel PD treatments. Molecular docking and molecular dynamics simulations were performed to provide a comprehensive understanding of the interactions between LRRK2 and the inhibitors in the data set and best molecules of the screening. Free energy of binding calculation along with hydrogen bond occupancy analysis and RMSD of the ligand in the pocket show CRA_1801 as the best candidate to be repurposed as LRRK2 inhibitor.
Assuntos
Serina-Treonina Proteína Quinase-2 com Repetições Ricas em Leucina , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Doença de Parkinson , Inibidores de Proteínas Quinases , Serina-Treonina Proteína Quinase-2 com Repetições Ricas em Leucina/antagonistas & inibidores , Serina-Treonina Proteína Quinase-2 com Repetições Ricas em Leucina/metabolismo , Doença de Parkinson/tratamento farmacológico , Humanos , Inibidores de Proteínas Quinases/química , Inibidores de Proteínas Quinases/farmacologia , Inibidores de Proteínas Quinases/uso terapêutico , Simulação por Computador , Aprendizado de MáquinaRESUMO
The crisis of bacterial resistance is an emerging One Health challenge, driven by the overuse of antimicrobials in medical and agricultural settings. This study aimed to investigate extended-spectrum ß-lactamase (ESBL), Ampicillinase (AmpC), and carbapenemase production, and the presence of genes encoding these enzymes in Escherichia coli, Klebsiella spp., and Proteus spp., major contributors to infections and resistance isolates from animals. From 2016 to 2021, 130 multidrug-resistant (MDR) or extensively drug-resistant (XDR) isolates were recovered from the secretions, excretions, and organs of companion and production animals with active infections. Antibacterial sensitivity tests, along with phenotypic and genotypic detection of resistance enzymes, were performed. To the best of our knowledge, this is the first study in Brazil to estimate the prevalence of XDR Enterobacteriales isolated from companion and production animals, which accounted for 13.8% of the strains. Statistically significant differences (P < 0.05) in resistant bacteria between different classes and within the same class of antibacterial bacteria were found. The statistical probability between genotypic detection of ESBL (OR = 3.1) and phenotypic tests for AmpC (OR = 2.3) was also established. Approximately 32.3%, 17.6%, and 16.8% of the strains had positive phenotypic tests for ESBL, AmpC, and carbapenemases, respectively. Genetic analysis revealed the presence of blaCTX-M (60.0%), blaAmpC (9.18%), blaKPC-2 (0.76%), and blaNDM (1.52%). AmpC genes were identified in 8.46% of the samples, with blaCMY being the most frequent (6.92%), followed by blaDHA (0.77%), and blaFOX (0.77%). The sequenced amplicons were deposited in NCBI. This study reveals critical data on Enterobacteriaceae with antibacterial resistance genes isolated from animals and may pose a significant threat to One health.
Assuntos
Antibacterianos , Proteínas de Bactérias , Infecções por Enterobacteriaceae , Enterobacteriaceae , Testes de Sensibilidade Microbiana , beta-Lactamases , beta-Lactamases/genética , beta-Lactamases/metabolismo , Animais , Brasil , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Antibacterianos/farmacologia , Infecções por Enterobacteriaceae/microbiologia , Infecções por Enterobacteriaceae/veterinária , Enterobacteriaceae/genética , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/enzimologia , Enterobacteriaceae/isolamento & purificação , Plasmídeos/genética , Farmacorresistência Bacteriana Múltipla/genética , Animais de Estimação/microbiologia , Fenótipo , Genótipo , Enterobacteriáceas Resistentes a Carbapenêmicos/genética , Enterobacteriáceas Resistentes a Carbapenêmicos/isolamento & purificação , Enterobacteriáceas Resistentes a Carbapenêmicos/efeitos dos fármacos , Enterobacteriáceas Resistentes a Carbapenêmicos/enzimologiaRESUMO
OBJECTIVES: To carry out physicomechanical characterization of the HA/DCPA/TiO2 and to evaluate the tissue repair in rat calvaria. METHODS: Two bone defects were made in the calvaria of 36 Wistar rats, divided into groups: HA/DCPA, HA/DCPA/TiO2 and sham (blood clot). The animals were euthanized at 30, 60 and 90 days and calvaria slides were processed with hematoxylin/eosin. The newly formed bone, connective tissue, biomaterial remnant, and total tissue repair percentages were calculated in relation to the total defect area. The HA/DCPA/TiO2 was characterized structurally by scanning electron microscopy (SEM), and chemically by energy-dispersive X-ray spectroscopy (EDS) and X-ray diffraction (XRD). It was submitted to apparent density (AD), apparent porosity (AP), water absorption (WA) and compressive strength (CS) physical tests. The ANOVA test was applied, followed by Turkey's test and Student's t-test (p ≤ 0,05). RESULTS: The SEM showed biomaterials inside the bone defects and newly formed bone. EDS identified oxygen, calcium, phosphorus, and titanium in the sample. The HA/DCPA/TiO2 and HA/DCPA groups presented a total tissue repair area that was larger than the sham group (p < 0.001). CONCLUSIONS: The physical-mechanical assays showed that HA/DCPA/TiO2 has AD and CS properties within the limits of trabecular bone and with values higher than HA/DCPA.HA/DCPA/TiO2 presented higher densification and compressive strength rates than HA/DCPA. CLINICAL RELEVANCE: Potential as a scaffold for bone.
Assuntos
Microscopia Eletrônica de Varredura , Ratos Wistar , Crânio , Titânio , Animais , Titânio/química , Ratos , Crânio/cirurgia , Difração de Raios X , Casca de Ovo/química , Espectrometria por Raios X , Regeneração Óssea/efeitos dos fármacos , Teste de Materiais , Materiais Biocompatíveis/farmacologia , Materiais Biocompatíveis/química , Masculino , Força Compressiva , Bioengenharia , Fosfatos de Cálcio/química , Fosfatos de Cálcio/farmacologia , Propriedades de Superfície , Porosidade , Durapatita/farmacologia , Durapatita/químicaRESUMO
Agricultural waste, such as rice straw, has become increasingly valuable as biocomposites in various industries. For cosmetic and pharmaceutical sectors, these biocomposites have improved active substance incorporation and waste reduction, which is pivotal for mitigating environmental impact. This study reports the encapsulation of a protein derivative derived from rice straw within a nanoemulsion for skin care applications, emphasizing stability and efficacy. Protein hydrolysates were produced by extracting proteins in an alkaline medium, followed by precipitation at the isoelectric point. The hydrolysates were enzymatically treated with Alcalase® at 80 °C and pH 10 for 45 min to generate antioxidant-rich formulations. Utilizing Hydrophilic-Lipophilic Deviation (HLD) theory, oil-in-water (O/W) emulsions were formulated by adjusting variables to achieve an HLD near zero. Sunflower oil and surfactants were combined, stirred at 70 °C, and homogenized using a rotor-stator. The final formulation's stability and permeability were evaluated through fluorescence microscopy, particle size analysis, zeta potential measurements, and accelerated stability assays. Nanoemulsion ENE37 showed high stability with 47.25 nm size, PDI 0.21, and excellent dispersion, maintaining integrity without phase separation. Hydrolyzed protein into ENE37 (NE37-HP) improved stability, increasing zeta potential and preventing aggregation while maintaining structure without phase inversion. NE37-HP exhibited shear-thinning behavior and good diffusion capacity, achieving 20.14 µg/cm2.h. The HLD theory and ternary diagrams are valuable methodological tools for formulating stable nanoscale emulsions. Additionally, this dosage form, containing protein hydrolysates derived from rice straw, demonstrated potential for adequate dermal absorption in humans.
Assuntos
Antioxidantes , Emulsões , Interações Hidrofóbicas e Hidrofílicas , Oryza , Tamanho da Partícula , Emulsões/química , Antioxidantes/química , Antioxidantes/administração & dosagem , Antioxidantes/farmacologia , Oryza/química , Tensoativos/química , Química Farmacêutica/métodos , Nanopartículas/química , Absorção Cutânea/fisiologia , Absorção Cutânea/efeitos dos fármacos , Permeabilidade , Hidrolisados de Proteína/química , Hidrolisados de Proteína/administração & dosagem , Óleo de Girassol/química , Administração Cutânea , Estabilidade de MedicamentosRESUMO
Peptides binding overexpressed breast and cervical cancer cell surface proteins can be isolated by phage display technology, and their affinity to their potential receptors can be assessed by molecular docking. We isolated 44 phage clones displaying dodecapeptides with high affinity to HeLa cervical cancer and MDA-MB-231 (MDA) breast cancer cells by repeated biopanning of an MK13 phage library and explored their affinity to specific proteins by molecular docking. Six peptides appeared repeatedly during biopanning: two with affinity to HeLa (H5/H21), and four with affinity to MDA cells (M3/M7/M15/M17). Peptide pairs M3/H5 and H1/M17 had affinity to both cell lines. A systematic review identified Annexin A2, EGFR, CD44, CD146, and Integrin alpha V as potential protein targets in HeLa cells, and Vimentin, Galectin-1, and Annexins A1 and A5 in MDA cells. Via virtual screening, we selected six peptides with the highest total docking scores: H1 (-916.32), H6 (-979.21), H19 (-1093.24), M6 (-732.21), M16 (-745.5), and M19 (-739.64), and identified that docking scores were strengthened by the protein type, the interacting amino acid side chains, and the polarity of peptides. This approach facilitates the selection of relevant peptides that could be further explored for active targeting in cancer diagnosis and treatment.
Assuntos
Simulação de Acoplamento Molecular , Biblioteca de Peptídeos , Peptídeos , Humanos , Peptídeos/química , Peptídeos/metabolismo , Peptídeos/farmacologia , Células HeLa , Linhagem Celular Tumoral , Neoplasias do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/patologia , Proteínas de Neoplasias/metabolismo , Proteínas de Neoplasias/química , Proteínas de Membrana/química , Proteínas de Membrana/metabolismoRESUMO
Dysregulated renin-angiotensin system (RAS) signaling contributes to elevated blood pressure (BP), inflammation, and organ damage in systemic arterial hypertension (HTN). We have demonstrated that hypertensive humans and rats exhibit higher expression of classic RAS components and lower expression of counterregulatory RAS components in the lungs compared with normotensive counterparts. Here, we investigated whether BP control could restore the balance between classic [angiotensin I-converting enzyme 2 (ACE)/angiotensin II (Ang II)] and counterregulatory [angiotensin I-converting enzyme 2 (ACE2)/Ang (1-7)] RAS, thereby mitigating lung inflammation. Male spontaneously hypertensive rats (SHRs) were treated with either losartan or amlodipine, both of which effectively reduced BP. These interventions up-regulated lung Ace2 and down-regulated Ace gene expression. Pulmonary membrane ACE2 abundance and activity were higher in losartan- and amlodipine-treated SHRs than in vehicle-treated SHRs, whereas ACE protein and function remained unchanged. Drug-treated SHRs exhibited lower levels of lung Ang II and higher levels of Ang (1-7) than vehicle-treated SHRs. Rebalancing the pulmonary RAS remarkably reduced macrophage number and down-regulated pro-inflammatory genes in SHR lungs, with lower expression of lung pro-inflammatory genes correlating with lower circulating levels of ACE2. Serum analysis in healthy and hypertensive individuals supported these findings, showing higher ACE2 levels in uncontrolled compared with controlled hypertension and normotension. Collectively, these findings suggest that high blood pressure may induce lung inflammation via an ACE/ACE2 imbalance. BP control with either an RAS inhibitor or a calcium channel blocker rebalances RAS in SHR lungs and alleviates inflammation. Furthermore, this study provides a mechanistic link between inflammatory lung diseases (such as COVID-19) and hypertension as a major risk factor.
Assuntos
Enzima de Conversão de Angiotensina 2 , Anti-Hipertensivos , Pressão Sanguínea , Hipertensão , Losartan , Pulmão , Ratos Endogâmicos SHR , Sistema Renina-Angiotensina , Animais , Sistema Renina-Angiotensina/efeitos dos fármacos , Masculino , Pressão Sanguínea/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/efeitos dos fármacos , Pulmão/fisiopatologia , Enzima de Conversão de Angiotensina 2/metabolismo , Enzima de Conversão de Angiotensina 2/genética , Ratos , Hipertensão/fisiopatologia , Hipertensão/tratamento farmacológico , Hipertensão/metabolismo , Losartan/farmacologia , Anti-Hipertensivos/farmacologia , Anti-Hipertensivos/uso terapêutico , Anlodipino/farmacologia , Anlodipino/uso terapêutico , Angiotensina I/metabolismo , Angiotensina II/metabolismo , Humanos , Fragmentos de Peptídeos/metabolismo , COVID-19/complicações , COVID-19/metabolismo , Peptidil Dipeptidase A/metabolismo , Peptidil Dipeptidase A/genética , SARS-CoV-2RESUMO
BACKGROUND: Recurrent vulvovaginal candidosis (RVVC) has been associated with increased antifungal resistance. Recently, we reported that Candida isolates from Colombian patients with RVVC did not show an increase in antifungal resistance. OBJECTIVE: The aim of this study was to evaluate the virulence of Candida isolates from patients with RVVC. METHODS: A total of 40 Candida isolates were evaluated (37 C. albicans and 3 C. lusitaniae ). C. albicans isolates were divided into two groups based on the number of VVC episodes in patients per year: Group 1 (four to seven episodes; n = 26) and Group 2 (≥ eight episodes; n = 11). The XTT assay was used to assess biofilm formation. Galleria mellonella larvae were used for survival analysis and fungal load assessment, and the qPCR technique to determine the expression of the PRA1 gene. RESULTS: It was observed that C. lusitaniae and C. albicans isolates from patients with ≥ eight VVC episodes per year exhibited a greater capacity to form biofilms compared to those from patients with four to seven VVC episodes. Moreover, in the G. mellonella model, larvae inoculated with isolates from RVVC patients exhibited approximately 80% mortality. Similarly, larvae infected with C. albicans from patients who experienced ≥ eight VVC episodes showed a significantly higher fungal load compared to the other evaluated groups; likewise, the expression of the PRA1 gene was significantly higher in isolates from patients with ≥ eight VVC episodes. CONCLUSION: These results indicate that Candida isolates from patients with RVVC exhibit a high degree of virulence and suggest that virulence may be one of the mechanisms explaining recurrence rather than antifungal resistance itself.
Assuntos
Biofilmes , Candida , Candidíase Vulvovaginal , Candidíase Vulvovaginal/microbiologia , Humanos , Feminino , Virulência , Biofilmes/crescimento & desenvolvimento , Animais , Candida/patogenicidade , Candida/genética , Candida/isolamento & purificação , Candida/classificação , Candida/efeitos dos fármacos , Candida/fisiologia , Recidiva , Larva/microbiologia , Candida albicans/patogenicidade , Candida albicans/genética , Candida albicans/isolamento & purificação , Candida albicans/fisiologia , Farmacorresistência Fúngica , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Mariposas/microbiologiaRESUMO
The lignocellulosic materials of some plant species are rich in chemical compounds that can serve as a source of natural and environmentally less aggressive biocides for treating wood. The aim of this study was to verify the potential of the lignocellulosic materials (wood and bark) of Ateleia glazioviana and Hovenia dulcis as a natural wood preservative. The extracts were prepared by heating the materials to 100 °C at a concentration of 10%. The wood samples were treated in a laboratory autoclave using the empty cell method. Control samples (untreated) and samples treated with chromated copper borate - CCB (2.5%) were used as a comparison. The accelerated rot test in the laboratory was carried out using a sample of the colony of the white rot fungus, Trametes versicolor. Mass loss, solubility in 1% NaOH, scanning electron spectroscopy (SEM) and attenuated total reflection infrared spectroscopy (ATR-FTIR) were evaluated. A rotting field test was also carried out in a forest fragment for 180 days and the mass loss, apparent specific mass, ATR-FTIR, and dynamic bending of the wood samples were evaluated. In laboratory tests, natural extractive solutions from the bark and wood of Hovenia dulcis and only from the bark of Ateleia glazioviana have fungitoxic potential against the white rot fungus, when compared to material without preservative impregnations. The resistance of the wood in the field to rot did not obtain significant results with the application of the natural preservatives, and future studies will need to increase the concentration of the extracts in an attempt to improve their performance as natural biocides.
Assuntos
Lignina , Casca de Planta , Madeira , Madeira/química , Madeira/microbiologia , Lignina/farmacologia , Lignina/química , Lignina/análise , Casca de Planta/química , Microscopia Eletrônica de Varredura , Espectroscopia de Infravermelho com Transformada de Fourier , Extratos Vegetais/farmacologia , Extratos Vegetais/químicaRESUMO
Stryphnodendron adstringens (Mart.) Coville, commonly known as "barbatimão," is native to the Cerrado biome in Brazil and belongs to the botanical family Fabaceae. The objective of this study was to evaluate the biological activity of crude hydroethanolic extracts formulated from the bark, leaves, and stems of S. adstringens. Soluble solids were determined using the incubation drying methodology. Colorimetric methods of complexation with ferric chloride were employed as a qualitative assay to identify the presence of tannins, while phenolics and flavonoids were quantified by the Folin-Ciocalteu method and aluminum chloride complexation, respectively. Antioxidant activity was assessed by the capture of DPPH free radicals. Antibacterial and antifungal analyses in vitro were conducted using the disk diffusion method against Escherichia coli, Mycobacterium tuberculosis, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, and Cryptococcus neoformans var. grubii. The MTT assay was used to determine antiparasitic activity against Leishmania amazonensis and to assess cytotoxicity using differentiated THP-1 macrophages. The extracts demonstrated efficacy against yeasts, especially the stem extract against C. albicans (7.62 mm), and against bacteria, with emphasis on the stem and leaf extracts against M. tuberculosis (both 9 mm). All extracts exhibited high antioxidant capacity, particularly the leaf and stem extracts (both over 92%) and low cytotoxicity (Cytotoxic Concentration - CC50 > 300 µg/mL). No extract was effective against L. amazonensis (Inhibitory Concentration - IC50 > 100 µg/mL). In conclusion, S. adstringens is a potential source of compounds with antibacterial properties (particularly against Gram-positive bacteria) and antifungal activity, with low cytotoxicity and high antioxidant activity. This work emphasizes the use of this plant as a source of molecules for the development of drugs against bacterial and fungal infectious diseases, as well as for combating diseases, such as cancer and neurodegenerative disorders, that are linked to cellular and DNA damage due to oxidative stress.