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1.
Emerg Infect Dis ; 30(5): 1022-1025, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38666647

RESUMO

We investigated molecular evolution and spatiotemporal dynamics of atypical Legionella pneumophila serogroup 1 sequence type 1905 and determined its long-term persistence and linkage to human disease in dispersed locations, far beyond the large 2014 outbreak epicenter in Portugal. Our finding highlights the need for public health interventions to prevent further disease spread.


Assuntos
Surtos de Doenças , Evolução Molecular , Legionella pneumophila , Doença dos Legionários , Análise Espaço-Temporal , Legionella pneumophila/genética , Legionella pneumophila/classificação , Portugal/epidemiologia , Humanos , Doença dos Legionários/epidemiologia , Doença dos Legionários/microbiologia , História do Século XXI , Recidiva , Filogenia , Sorogrupo
2.
Artigo em Inglês | MEDLINE | ID: mdl-38696083

RESUMO

BACKGROUD: Although not fully investigated, studies show that Legionella pneumophila can develop antibiotic resistance. As there is limited data available for Portugal, we determined the antibiotic susceptibility profile of Portuguese L. pneumophila serogroup 1 (LpnSg1) isolates against antibiotics used in the clinical practice in Portugal. METHODS: Minimum inhibitory concentrations (MICs) were determined for LpnSg1 clinical (n = 100) and related environmental (n = 7) isolates, collected between 2006-2022 in the context of the National Legionnaire´s Disease Surveillance Programme, against azithromycin, clarithromycin, erythromycin, levofloxacin, ciprofloxacin, moxifloxacin, rifampicin, doxycycline, tigecycline, and amoxicillin/clavulanic acid, using three different assays. Isolates were also PCR-screened for the presence of the lpeAB gene. RESULTS: Twelve isolates had azithromycin MICs above the EUCAST tentative highest WT MIC, 9 of which were lpeAB negative; for erythromycin and clarithromycin, all isolates tested within the susceptible range. The number of isolates with MICs above the tentative highest WT MIC for the remaining antibiotics was: ciprofloxacin: 7; levofloxacin: 17; moxifloxacin: 8; rifampicin: 11; doxycycline: 82; tigecycline: 4. EUCAST breakpoints are not available for amoxicillin/clavulanic acid. We estimated the ECOFFs and one isolate had a MIC eightfold higher than the E-test ECOFF. Additionally, a clinical isolate generated three colonies growing on the E-test inhibition zone that resulted in MICs fourfold higher than for the parental isolate. CONCLUSIONS: We report, for the first time, elevated MICs against first-line and other antibiotics (including azithromycin, fluoroquinolones and amoxicillin/clavulanic acid commonly used to treat pneumonia patients in Portugal) in Portuguese L. pneumophila strains. Results point towards decreased susceptibility in circulating strains, justifying further investigation.

3.
Phys Chem Chem Phys ; 26(16): 12799-12805, 2024 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-38619871

RESUMO

Isothiazolinones represent a class of heterocyclic compounds widely used in various applications, including as biocides in cosmetics, detergents, and paints, as well as in industrial wastewater treatment. Indeed, the presence of isothiazolinones in the environment and their associated potential health hazards have raised significant concerns. In this study, a non-adiabatic dynamics investigation was conducted using state-of-the-art methodologies to explore the photochemistry of isothiazolinones. A simplified model, isothiazol-3(2H)-one (ISO), was employed to represent this compound class. The study validated the model and demonstrated that ISO can return to its ground state through the cleavage of the S-N or S-C bonds, with no significant energy barrier observed. Non-adiabatic dynamics simulations provided insights into the time scales and detailed processes of isothiazolinone photodissociation. The preferred route for deactivation was found to be the cleavage of the S-N bond. This research enhances our understanding of the photodeactivation processes of isothiazolinones and their potential environmental impact.

4.
Reprod Domest Anim ; 59(1): e14510, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38013657

RESUMO

While sperm migrate within the reproductive tract of cows experiencing negative energy balance (NEB), they come into contact with elevated concentrations of non-esterified fatty acids (NEFA). For this reason, this study aimed to investigate the effects of three different NEFA - palmitic acid (PA), stearic acid (SA), and oleic acid (OA) - on bovine sperm motility, kinetic parameters, oxidative status, and morphology. Frozen thawed semen samples from Bos taurus bulls were incubated with varying concentrations of each fatty acid, and the sperm's characteristics were analysed at different time points. Computer-Assisted Sperm Analysis (CASA) was employed to assess sperm motility and kinetic parameters. Concurrently, the production of the reactive oxygen species (ROS) and total antioxidant capacity were measured to determine the oxidative status. Additionally, sperm morphology was evaluated. In Experiment 1, different concentrations of PA did not show significant effects on total motility, progressive motility, or any kinetic parameters analysed. Similarly, PA did not have a significant impact on the oxidative status or sperm morphology. In Experiment 2, SA at various concentrations did not lead to significant changes in total motility, progressive motility, or any kinetic parameters evaluated. Furthermore, SA did not affect oxidative status or sperm morphology. In Experiment 3, the concentrations of OA used did not result in significant changes in total motility, progressive motility, or any kinetic parameters studied. Likewise, OA did not induce any alterations in oxidative status or sperm morphology. Overall, the results from all three experiments indicate that PA, SA and OA, at the in vitro conditions and tested concentrations, do not exert detrimental effects on bovine sperm function and morphology. These results provide insights that contribute to our understanding of how fatty acids can impact the reduction of fertility rates in cows facing NEB. This, in turn, lays the foundation for additional critical investigations in this area. Further studies are necessary to validate these findings in vivo.


Assuntos
Preservação do Sêmen , Sêmen , Feminino , Bovinos , Masculino , Animais , Ácidos Graxos , Motilidade dos Espermatozoides , Ácidos Graxos não Esterificados , Espermatozoides , Preservação do Sêmen/veterinária , Estresse Oxidativo
5.
Eur Arch Otorhinolaryngol ; 281(3): 1105-1114, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-37864748

RESUMO

PURPOSE: Our study goal is to review the efficacy of tranexamic acid in reducing blood loss and operative time in nasal surgeries. METHODS: We included randomized clinical trials using oral or intravenous tranexamic acid, excluded non-randomized studies, topic administration, coagulopathy, and using other drugs interfering in the coagulation cascade. Online databases, National Library of Medicine (MEDLINE-PubMED), Latin American and Caribbean Literature on Health Sciences (Lilacs), Cochrane Library, Embase and Google Scholar were used to perform the search. The review was registered in PROSPERO by no CRD42022310977. Two authors, independently, selected the articles meeting the inclusion criteria. They extracted the data and used RevMan 5 software to perform the meta-analysis. RESULTS: Our search resulted in 16 RCTs that were included in the meta-analysis totalizing 1108 patients. Studies were evaluated resulting in a low risk of bias for the five domains. The use of tranexamic acid resulted in significant reduction in duration of surgery (DOS) and intraoperative blood loss (IBL) had significant reduction. The level of evidence according to GRADE System was high in all studies and variables. CONCLUSION: Tranexamic acid has an important role in reducing intraoperative blood loss and duration of surgery. Our study has some limitations due to the low number of RCTs available in the literature.


Assuntos
Antifibrinolíticos , Procedimentos Cirúrgicos Nasais , Ácido Tranexâmico , Humanos , Ácido Tranexâmico/uso terapêutico , Perda Sanguínea Cirúrgica/prevenção & controle , Antifibrinolíticos/uso terapêutico , Duração da Cirurgia , Procedimentos Cirúrgicos Nasais/efeitos adversos
6.
J Therm Biol ; 121: 103833, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38527387

RESUMO

Dairy cows in pasture-based systems are more susceptible to heat stress. Holstein cows have the black or red phenotypes, the latter having lower absorbance of solar radiation. Therefore, the study's objective was to evaluate whether cows with red (R) coats are more resistant than black (B) cows to hot weather in a subtropical climate. R and B lactating Holstein cows were evaluated during the cold and hot seasons for internal and surface temperature and sweating rate. In the cold season, body temperature (n = 9/group) did not differ between groups, but the average superficial temperature (n = 13/group) was lower in R cows (B: 30.9 ± 0.3 °C; RW: 29.6 ± 0.3 °C; p = 0.02). In the hot season, under mild to moderate heat stress, mean body temperature (n = 9/group) of R cows was lower (B: 38.75 ± 0.01 °C; R: 38.62 ± 0.1 °C; p=<0.0001), whereas no difference was observed in superficial temperature (n = 17/group). The maximum internal temperature and sweating rate (n = 11/group), measured in the hot season, and the number of evaluations in hyperthermia in both seasons did not differ. Therefore, there were differences in thermoregulation between phenotypes under mild to moderate heat stress conditions. However, considering that only discrete differences were observed, the red and white coat is unlikely to benefit the Holstein cow's welfare under mild to moderate thermal stress.


Assuntos
Regulação da Temperatura Corporal , Lactação , Estações do Ano , Animais , Bovinos/fisiologia , Feminino , Brasil , Resposta ao Choque Térmico , Temperatura Alta , Temperatura Corporal , Temperatura Baixa , Sudorese
7.
Surg Radiol Anat ; 46(3): 353-362, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38329522

RESUMO

PURPOSE: The aim of this systematic review and meta-analysis was to systematically review and perform a meta-analysis on the anatomical variations of the RLN. METHODS: We performed online research for studies that addressed anatomical variations of the RLN and laterality, published between 2015 and 2021. We found 230 articles, and nine were included. RESULTS: Eight variations were found, with Type I prevailing (41.17%; 95% CI 19.44-64.88), extra laryngeal divergence of the RLN. The other types were: II-fan shape; III-distance greater than 5 mm to the cricothyroid joint; IV-thickening and adipopexy in the elderly; V-non-recurrent laryngeal nerve; VI-intracranial branch; VII-tortuous ascending RLN; and VIII-combination between the inferior branch of the NV and the ascending trunk of the RLN. Types I (p = 0) and III (p < 0.01) prevailed on the left and types II (p < 0.01) and V (p < 0.01) on the right. CONCLUSIONS: It was observed that variations occurred due to the path of the RLN to the entrance to the larynx, its shape, and the age of the evaluated individual. The most frequent variation and side were, respectively, Type I, extra laryngeal divergence and left.


Assuntos
Laringe , Traumatismos do Nervo Laríngeo Recorrente , Humanos , Idoso , Nervo Laríngeo Recorrente/fisiologia , Cartilagem Cricoide , Tireoidectomia
8.
Reproduction ; 166(6): 401-410, 2023 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-37855431

RESUMO

In brief: Ubiquitination plays a pivotal role in a multitude of cellular functions; however, the precise contributions of various ubiquitin ligases in governing early developmental processes remain largely unexplored. This study revealed that the E3 ubiquitin ligases DCAF13 and RNF114 are both necessary for the normal regulation of early porcine embryo development. Abstract: Ubiquitylation is required for normal regulation of many biological functions by modulating several protein facets such as structure, stability, interaction, localization, and degradation. In this study, we explored the roles of two E3 ubiquitin ligases (E3s), the DDB1- and CUL4-associated factor 13 (DCAF13) and the Ring finger protein 114 (RNF114), in the regulation of porcine embryo development. Attenuation of DCAF13 mRNA decreased embryo development at the blastocyst stage, while the development of RNF114-attenuated embryos was not significantly different than that of control embryos. The average number of cells per blastocyst was decreased in DCAF13-attenuated embryos and increased in RNF114-attenuated embryos compared to controls. The relative mRNA abundance of the histone methyltransferase SUV39H1, which regulates histone H3 lysine 9 trimethylation (H3K9me3), was increased in both DCAF13- and RNF114-attenuated embryos, but nuclear immunofluorescence signal for H3K9me3 on day 3 embryos was not significantly altered between attenuated and control embryos. Nuclear immunofluorescence signal for H3K4m3 was decreased in DCAF13-attenuated embryos, but it was increased in RNF114-attenuated embryos compared to controls. Attenuation of DCAF13 and RNF114 mRNAs increased transcript levels for the DNA recombinase RAD51 and decreased expression of phosphorylated histone H2A.X (γH2AX), which suggests an impact on DNA damage repair. In addition, lower mRNA expression of the lysine demethylases 5B (KDM5B) and 5C (KDM5C), both involved in embryo genome activation and DNA repair, was detected in DCAF13-attenuated embryos. These findings indicated that both DCAF13 and RNF114 have important roles in the regulation of the early development of porcine embryos.


Assuntos
Desenvolvimento Embrionário , Fator XIII , Suínos , Ubiquitina-Proteína Ligases , Animais , Blastocisto , Desenvolvimento Embrionário/genética , Fator XIII/metabolismo , Lisina/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Suínos/embriologia , Proteínas de Ligação a RNA , Ubiquitina-Proteína Ligases/metabolismo
9.
Reprod Domest Anim ; 58(12): 1662-1671, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37743826

RESUMO

In the postpartum period, there is an increase in non-esterified fatty acids (NEFA) in both serum and follicular fluid (FF) of cattle. The increase in fatty acid concentration results in increased production of reactive oxygen species (ROS) that can compromise bovine fertility. The objectives of this study were to characterize the lipid profile found in the FF of cows experiencing induced negative energy balance (NEB) and to evaluate the effect of α-tocopherol in the prevention of oxidative stress in the serum and FF of cows. Twenty-nine beef cows were divided into groups: (1) control; (2) Fasting for 24 days; and (3) Fasting + VitE. Between D0 and D4 blood samples were taken to assess concentrations of NEFA, ROS production, total antioxidant capacity (FRAP), lipid peroxidation, and α-tocopherol (vitamin E). On D4, follicular aspiration was performed for analysis of FF from the dominant follicle. Our results demonstrate that fasting was effective in causing increased fat mobilization in animals. The increase in serum concentration of C18:1c9 was reflected in the FF of fasting cows. Serum α-tocopherol concentration was higher in the control and Fasting + VitE groups compared to the Fasting group. In FF, there was an increase of α-tocopherol in the Fasting + VitE group in comparison to Fasting cows. There was an increase in ROS production in the serum of fasting cows. ROS production in FF was higher in the Fasting compared to the Fasting + VitE group. Vitamin E has beneficial effects in reducing ROS production in the dominant follicle of cows in NEB.


Assuntos
Ácidos Graxos não Esterificados , Vitamina E , Feminino , Bovinos , Animais , Espécies Reativas de Oxigênio , Vitamina E/farmacologia , Lactação/metabolismo , alfa-Tocoferol/farmacologia
10.
Sensors (Basel) ; 23(12)2023 Jun 09.
Artigo em Inglês | MEDLINE | ID: mdl-37420626

RESUMO

Cyber-Physical Systems (CPS) are prone to many security exploitations due to a greater attack surface being introduced by their cyber component by the nature of their remote accessibility or non-isolated capability. Security exploitations, on the other hand, rise in complexities, aiming for more powerful attacks and evasion from detections. The real-world applicability of CPS thus poses a question mark due to security infringements. Researchers have been developing new and robust techniques to enhance the security of these systems. Many techniques and security aspects are being considered to build robust security systems; these include attack prevention, attack detection, and attack mitigation as security development techniques with consideration of confidentiality, integrity, and availability as some of the important security aspects. In this paper, we have proposed machine learning-based intelligent attack detection strategies which have evolved as a result of failures in traditional signature-based techniques to detect zero-day attacks and attacks of a complex nature. Many researchers have evaluated the feasibility of learning models in the security domain and pointed out their capability to detect known as well as unknown attacks (zero-day attacks). However, these learning models are also vulnerable to adversarial attacks like poisoning attacks, evasion attacks, and exploration attacks. To make use of a robust-cum-intelligent security mechanism, we have proposed an adversarial learning-based defense strategy for the security of CPS to ensure CPS security and invoke resilience against adversarial attacks. We have evaluated the proposed strategy through the implementation of Random Forest (RF), Artificial Neural Network (ANN), and Long Short-Term Memory (LSTM) on the ToN_IoT Network dataset and an adversarial dataset generated through the Generative Adversarial Network (GAN) model.


Assuntos
Inteligência Artificial , Segurança Computacional , Inteligência , Memória de Longo Prazo , Redes Neurais de Computação
11.
Reprod Fertil Dev ; 34(16): 1023-1033, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36116787

RESUMO

CONTEXT: The establishment of pregnancy in cows requires uterine activity regulation of the main Hippo signalling effector yes-associated protein 1 (YAP). It remains unknown (1) how YAP activity at the corpus luteum (CL) correlates with early pregnancy-related events in ruminants; and (2) if YAP activity in the uterus and CL can be affected by metabolic disorders that may lead to pregnancy failure in ruminants. AIMS AND METHODS: To determine the effect of early pregnancy on total and phospho-YAP expression and its transcriptional activity in the CL, we compared non-pregnant vs pregnant ewes. To understand the YAP activity dysregulation with disorders that may result in pregnancy loss, we induced negative energy balance in pregnant ewes. KEY RESULTS AND CONCLUSIONS: Our main results indicate that early pregnancy alters the expression and activity patterns of YAP in the ovine CL but not in the endometrium. In addition, while our NEB-induced model fails to alter YAP activity at the endometrium level, we found that fasting during the first but not second week of pregnancy affects YAP activity in the CL of pregnant ewes. IMPLICATIONS: The data presented herein provide considerable insight into the activity of a signalling pathway that may be a key player in pregnancy recognition and establishment in ewes.


Assuntos
Prenhez , Proteínas de Sinalização YAP , Animais , Bovinos , Corpo Lúteo/metabolismo , Endométrio/metabolismo , Feminino , Gravidez , Ovinos , Útero/metabolismo
12.
Zygote ; 30(2): 239-243, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-34405788

RESUMO

The Hippo pathway is involved in the proliferation of intrafollicular cells and in early embryonic development, mainly because effectors of this pathway are key transcription regulators of genes such as CTGF and CYR61, which are involved in cell proliferation. Recent studies by our group found that fibroblast growth factor 18 (FGF18) is present in the fallopian tube during early embryonic development, leading to the hypothesis that FGF18 may have a role during embryonic development. Therefore, the aim of the following study was to determine whether FGF18 modulates the expression of Hippo pathway target genes, CTGF and CYR61, during oocyte maturation and early embryonic development. Three experiments were carried out, with in vitro maturation (IVM) of cumulus-oocyte complexes (COCs) and embryo culture. In experiment one, FGF18 (100 ng/ml) induced an increase (P < 0.05) in CTGF gene expression at 12 h post-exposure. In experiment two, FGF18 (100 ng/ml) induced a reduction (P < 0.05) in CTGF expression at 3 h post-exposure. In the third experiment, day 7 embryos exposed to FGF18 during oocyte IVM expressed greater CTGF mRNA abundance, whereas FGF18 exposure during embryo in vitro embryo culture did not alter CTGF expression in comparison with untreated controls. The preliminary data presented here show that FGF18 modulates CTGF expression in critical periods of oocyte nuclear maturation, cumulus expansion and early embryonic development in cattle.


Assuntos
Células do Cúmulo , Técnicas de Maturação in Vitro de Oócitos , Animais , Blastocisto , Bovinos , Desenvolvimento Embrionário , Feminino , Fatores de Crescimento de Fibroblastos , Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos , Gravidez , Dados Preliminares , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
13.
J Assist Reprod Genet ; 39(2): 481-492, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-35091965

RESUMO

PURPOSE: To determine if the inhibition of the interaction between the Hippo effector YAP or its transcriptional co-activator TAZ with the TEAD family of transcription factors is critical for the cumulus expansion-related events induced by the EGF network in cumulus-oocyte complexes (COCs). METHODS: We performed a series of experiments using immature bovine COCs subjected to an IVM protocol for up 24 h in which cumulus expansion was stimulated with EGF recombinant protein or FSH. RESULTS: The main results indicated that EGFR activity stimulation in bovine cumulus cells (CC) increases mRNA levels encoding the classic YAP/TAZ-TEAD target gene CTGF. To determine if important genes for cumulus expansion are transcriptional targets of YAP/TAZ-TEAD interaction in CC, COCs were then subjected to IVM in the presence of FSH with or without distinct concentrations of Verteporfin (VP; a small molecule inhibitor that interferes with YAP/TAZ binding to TEADs). COCs were then collected at 6, 12, 18, and 24 h for total RNA extraction and RT-qPCR analyses. This experiment indicated that VP inhibits in a time- and concentration-dependent manner distinct cumulus expansion and oocyte maturation-related genes, by regulating EGFR and CTGF expression in CC. CONCLUSIONS: Taken together, the results presented herein represent considerable insight into the functional relevance of a completely novel signaling pathway underlying cumulus expansion and oocyte maturation in monovulatory species. YAP/TAZ or CTGF may represent potential targets to improve the efficiency of IVM systems, not only for monovulatory species of agricultural importance as the cow, but for human embryo production.


Assuntos
Células do Cúmulo , Fator de Crescimento Epidérmico , Proteínas com Motivo de Ligação a PDZ com Coativador Transcricional , Proteínas de Sinalização YAP , Animais , Bovinos , Células do Cúmulo/metabolismo , Fator de Crescimento Epidérmico/farmacologia , Feminino , Via de Sinalização Hippo , Técnicas de Maturação in Vitro de Oócitos , Oócitos/metabolismo , Transdução de Sinais , Proteínas com Motivo de Ligação a PDZ com Coativador Transcricional/metabolismo , Proteínas de Sinalização YAP/metabolismo
14.
Int J Mol Sci ; 23(17)2022 Aug 25.
Artigo em Inglês | MEDLINE | ID: mdl-36077022

RESUMO

DNA damage in early-stage embryos impacts development and is a risk factor for segregation of altered genomes. DNA damage response (DDR) encompasses a sophisticated network of proteins involved in sensing, signaling, and repairing damage. DDR is regulated by reversible post-translational modifications including acetylation, methylation, phosphorylation, ubiquitylation, and SUMOylation. While important regulators of these processes have been characterized in somatic cells, their roles in early-stage embryos remain broadly unknown. The objective of this study was to explore how ubiquitylation and SUMOylation are involved in the regulation of early development in porcine embryos by assessing the mRNA profile of genes encoding ubiquitination (UBs), deubiquitination (DUBs), SUMOylation (SUMOs) or deSUMOylation (deSUMOs) enzymes in oocyte and embryos at different stages of development, and to evaluate if the induction of DNA damage at different stages of embryo development would alter the mRNA abundance of these genes. Pig embryos were produced by in vitro fertilization and DNA damage was induced by ultraviolet (UV) light exposure for 10 s on days 2, 4 or 7 of development. The relative mRNA abundance of most UBs, DUBs, SUMOs, and deSUMOs was higher in oocytes and early-stage embryos than in blastocysts. Transcript levels for UBs (RNF20, RNF40, RNF114, RNF169, CUL5, DCAF2, DECAF13, and DDB1), DUBs (USP16), and SUMOs (CBX4, UBA2 and UBC9), were upregulated in early-stage embryos (D2 and/or D4) compared to oocytes and blastocysts. In response to UV-induced DNA damage, transcript levels of several UBs, DUBs, SUMOs, and deSUMOs decreased in D2 and D4 embryos, but increased in blastocysts. These findings revealed that transcript levels of genes encoding for important UBs, DUBs, SUMOs, and deSUMOs are regulated during early embryo development and are modulated in response to induced DNA damage. This study has also identified candidate genes controlling post-translational modifications that may have relevant roles in the regulation of normal embryo development, repair of damaged DNA, and preservation of genome stability in the pig embryo.


Assuntos
Blastocisto , Ubiquitina , Animais , Blastocisto/metabolismo , Dano ao DNA , Desenvolvimento Embrionário/genética , Oócitos/fisiologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Suínos , Ubiquitina/metabolismo
15.
Int J Mol Sci ; 23(22)2022 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-36430640

RESUMO

The molecular mechanisms that drive the granulosa cells' (GC) differentiation into a more estrogenic phenotype during follicular divergence and establishment of follicle dominance have not been completely elucidated. The main Hippo signaling effector, YAP, has, however, emerged as a potential key player to explain such complex processes. Studies using rat and bovine GC demonstrate that, in conditions where the expression of the classic YAP-TEAD target gene tissue growth factor (CTGF) is augmented, CYP19A1 expression and activity and, consequently, estradiol (E2) secretion are reduced. These findings led us to hypothesize that, during ovarian follicular divergence in cattle, FSH downregulates YAP-TEAD-dependent transcriptional activity in GC to allow the future dominant follicle to exert its augmented estrogenic capacity. To address this, we performed a series of experiments employing distinct bovine models. Our in vitro and ex vivo experiments indicated that indeed FSH downregulates, in a concentration-dependent manner, mRNA levels not only for CTGF but also for the other classic YAP-TEAD transcriptional target genes ANKRD1 and CYR61 by a mechanism that involves increased YAP phosphorylation. To better elucidate the functional importance of such FSH-induced YAP activity regulation, we then cultured GC in the presence of verteporfin (VP) or peptide 17 (P17), two pharmacological inhibitors known to interfere with YAP binding to TEADs. The results showed that both VP and P17 increased CYP19A1 basal mRNA levels in a concentration-dependent manner. Most interestingly, by using GC samples obtained in vivo from dominant vs. subordinate follicles, we found that mRNA levels for CTGF, CYR61, and ANKRD1 are higher in subordinate follicles following the follicular divergence. Taken together, our novel results demonstrate that YAP transcriptional activity is regulated in bovine granulosa cells to allow the increased estrogenic capacity of the selected dominant follicle.


Assuntos
Hormônio Foliculoestimulante , Folículo Ovariano , Animais , Bovinos/genética , Bovinos/metabolismo , Feminino , Ratos , Estrona/metabolismo , Hormônio Foliculoestimulante/farmacologia , Hormônio Foliculoestimulante/metabolismo , Células da Granulosa/metabolismo , Folículo Ovariano/metabolismo , RNA Mensageiro/metabolismo , Verteporfina , Fatores de Transcrição de Domínio TEA/metabolismo , Proteínas de Sinalização YAP/metabolismo
16.
Int J Environ Health Res ; 32(5): 963-971, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-32814444

RESUMO

Association between selection pressure caused by the use of azole fungicides in sawmills and the development of fungal resistance has been described. The aim of this study was to implement an algorithm to assess the presence of Aspergillus section Fumigati resistant strains in sawmills.Eighty-six full-shift inhalable dust samples were collected from eleven industrial sawmills in Norway. Different culture media were used and molecular identification to species level in Aspergillus section Fumigati was done by calmodulin sequencing and TR34/L98H and TR46/Y121F/T289A mutations were screened by real-time PCR assay and confirmed by cyp51A sequencing. Six Fumigati isolates were identified as A. fumigatus sensu stricto and two of these grew on azole-supplemented media and were further analyzed by real-time PCR. One was confirmed to be a TR34/L98H mutant.The obtained results reinforce the need to assess the presence of A. fumigatus sensu stricto resistant isolates at other workplaces with fungicide pressure.


Assuntos
Aspergillus fumigatus , Azóis , Algoritmos , Antifúngicos/farmacologia , Aspergillus fumigatus/genética , Farmacorresistência Fúngica/genética , Proteínas Fúngicas/genética , Testes de Sensibilidade Microbiana
17.
Chembiochem ; 22(5): 865-875, 2021 03 02.
Artigo em Inglês | MEDLINE | ID: mdl-33084150

RESUMO

The SARS-CoV-2 pandemic is the biggest health concern today, but until now there is no treatment. One possible drug target is the receptor binding domain (RBD) of the coronavirus' spike protein, which recognizes the human angiotensin-converting enzyme 2 (hACE2). Our in silico study discusses crucial structural and thermodynamic aspects of the interactions involving RBDs from the SARS-CoV and SARS-CoV-2 with the hACE2. Molecular docking and molecular dynamics simulations explain why the chemical affinity of the new SARS-CoV-2 for hACE2 is much higher than in the case of SARS-CoV, revealing an intricate pattern of hydrogen bonds and hydrophobic interactions and estimating a free energy of binding, which is consistently much more negative in the case of SARS-CoV-2. This work presents a chemical reason for the difficulty in treating the SARS-CoV-2 virus with drugs targeting its spike protein and helps to explain its infectiousness.


Assuntos
Enzima de Conversão de Angiotensina 2/química , COVID-19 , SARS-CoV-2/química , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave/química , Glicoproteína da Espícula de Coronavírus/química , Enzima de Conversão de Angiotensina 2/metabolismo , Sítios de Ligação , Cristalografia por Raios X , Humanos , Ligação de Hidrogênio , Simulação de Dinâmica Molecular , Ligação Proteica , Domínios e Motivos de Interação entre Proteínas , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave/metabolismo , SARS-CoV-2/metabolismo , Glicoproteína da Espícula de Coronavírus/metabolismo
18.
Andrologia ; 53(8): e14148, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34171138

RESUMO

Postpartum cows, mainly with metabolic diseases, such as ketosis, usually experience an increased number of services per conception. During ketosis, high concentrations of ß-hydroxybutyrate (BHBA) in follicular, uterine and oviductal fluid have been considered to cause subfertility in cows. However, the effect of sperm exposure to an environment with high BHBA concentration is not known. This study investigated the influence of high levels of BHBA on kinetics, oxidative status and morphology of bovine spermatozoa. To assess the effect of BHBA after sperm selection, bovine spermatozoa were incubated (180 min) with different BHBA concentrations: 0 (Control), 0.8, 2.4 or 5 mM. Sperm kinetics was evaluated after 30, 60, 120 and 180 min, and oxidative status and morphology were analysed at 180 min. Oxidative status was evaluated through the production of reactive oxidative species (ROS), total antioxidant capacity and lipid peroxidation. High concentrations of BHBA decreased the curvilinear velocity, straight line velocity, mean path velocity, linearity, straightness and hyperactivity of spermatozoa. However, there was no effect of BHBA on oxidative and antioxidant capacity as well as on sperm morphology. In conclusion, exposure of bovine spermatozoa to high levels of BHBA impairs sperm kinetics without altering oxidative and antioxidant mechanisms.


Assuntos
Doenças dos Bovinos , Cetose , Ácido 3-Hidroxibutírico , Animais , Bovinos , Feminino , Cinética , Masculino , Espermatozoides
19.
J Environ Manage ; 279: 111800, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-33340962

RESUMO

Several ethnobotanical studies have attempted to understand the criteria for the differential use of plant resources. However, we need more effort to understand the interaction between local uses: how using a species for a given purpose may affect its use for another purpose. Thus, we hypothesize that high importance species in a more specialized category of use may have their use reduced for other categories with a more generalist nature. We have conducted the study in three rural communities in northeastern Brazil, set in seasonally dry tropical forest areas. We applied the free-list technique to identify woody species used for medicinal and/or wood purposes (fuelwood, construction and technology). Respondents rated the species according to their efficiency for wood purposes and their local availability. We performed a multiple regression to assess the effects of medicinal popularity, perceived availability, and perceived efficiency for wood uses over the species popularity for wood uses. Our results showed that medicinal use has a significant protective effect against wood uses. Perceived availability and efficiency were significant explanatory variables for wood use. Maintaining the medicinal importance of certain species can be a powerful tool in protecting their populations against more harmful uses.


Assuntos
Conservação dos Recursos Naturais , Madeira , Brasil , Etnobotânica , Florestas , Humanos
20.
Mol Reprod Dev ; 87(8): 899-909, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32761819

RESUMO

Interferon tau (IFNT) is the cytokine responsible for the maternal recognition of pregnancy in ruminants and plays a role modulating embryo-maternal communication in the oviduct inducing a local response from immune cells. We aimed to investigate IFNT production, reactive oxygen species, and oxidative stress under the influence of heat stress (HS) during different stages of bovine in vitro embryo production. HS was established when the temperature was gradually raised from 38.5°C to 40.5°C in laboratory incubator, sustained for 6 hr, and decreased back to 38.5°C. To address the HS effects on IFNT production, reactive oxygen species, and oxidative stress, ovaries from a slaughterhouse were used according to treatments: control group (38.5°C); oocytes matured under HS; oocytes fertilized under HS; zygotes cultured in the first day under HS; and cells submitted to HS at oocyte maturation, fertilization, and the first day of zygote culture. The HS negatively affected cleavage and blastocyst rates, in all HS groups. On Day 7, all HS-treated embryos showed decrease IFNT gene and protein expressions, whereas reactive oxygen species were increased in comparison to the control. In conclusion, the compromised early embryo development due to higher temperatures during in vitro oocyte maturation, fertilization, and/or zygote stage have diminished IFNT expression and increased reactive oxygen species in bovine.


Assuntos
Bovinos/embriologia , Desenvolvimento Embrionário/fisiologia , Resposta ao Choque Térmico/fisiologia , Oócitos/fisiologia , Estresse Oxidativo/fisiologia , Zigoto/fisiologia , Animais , Células Cultivadas , Embrião de Mamíferos , Feminino , Fertilização in vitro/veterinária , Transtornos de Estresse por Calor/embriologia , Transtornos de Estresse por Calor/metabolismo , Transtornos de Estresse por Calor/fisiopatologia , Temperatura Alta , Técnicas de Maturação in Vitro de Oócitos , Interferon Tipo I/metabolismo , Oócitos/citologia , Oogênese/fisiologia , Proteínas da Gravidez/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Zigoto/citologia
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