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1.
J Biol Chem ; 300(9): 107692, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39159809

RESUMO

Monoxenous trypanosomatid Strigomonas culicis harbors an endosymbiotic bacterium, which enables the protozoa to survive without heme supplementation. The impact of H2O2 resistance and symbiont elimination on intracellular heme and Fe2+ availability was analyzed through a comparison of WT strain with both WT H2O2-resistant (WTR) and aposymbiotic (Apo) protozoa. The relative quantification of the heme biosynthetic pathway through label-free parallel reaction monitoring targeted mass spectrometry revealed that H2O2 resistance does not influence the abundance of tryptic peptides. However, the Apo strain showed increased coproporphyrinogen III oxidase and ferrochelatase levels. A putative ferrous iron transporter, homologous to LIT1 and TcIT from Leishmania major and Trypanosoma cruzi, was identified for the first time. Label-free parallel reaction monitoring targeted mass spectrometry also showed that S. culicis Iron Transporter (ScIT) increased 1.6- and 16.4-fold in WTR and Apo strains compared to WT. Accordingly, antibody-mediated blockage of ScIT decreased by 28.0% and 40.0% intracellular Fe2+concentration in both WTR and Apo strains, whereas no effect was detected in WT. In a heme-depleted medium, adding 10 µM hemin decreased ScIT transcript levels in Apo, whereas 10 µM PPIX, the substrate of ferrochelatase, increased intracellular Fe2+ concentration and ferric iron reduction. Overall, the data suggest mechanisms dependent on de novo heme synthesis (and its substrates) in the Apo strain to overcome reduced heme availability. Given the importance of heme and Fe2+ as cofactors in metabolic pathways, including oxidative phosphorylation and antioxidant systems, this study provides novel mechanistic insights associated with H2O2 resistance in S. culicis.


Assuntos
Heme , Peróxido de Hidrogênio , Simbiose , Heme/metabolismo , Peróxido de Hidrogênio/metabolismo , Trypanosomatina/metabolismo , Trypanosomatina/genética , Ferro/metabolismo , Resistência a Medicamentos , Proteínas de Protozoários/metabolismo , Proteínas de Protozoários/genética
3.
Reprod Domest Anim ; 59(8): e14701, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39109447

RESUMO

This study aimed to evaluate the effect of chemical gasification and HEPES as alternative systems to pH control during in vitro maturation on bovine oocytes competence. Groups of 20 bovine cumulus oocytes complexes (COCs) were randomly distributed and cultured for 24 h in one of the following experimental groups: (i) chemical reaction (ChRG) system: CO2 generated from sodium bicarbonate and citric acid reaction (ii) culture media TCM-HEPES (HEPES-G); and (iii) control group (CNTG) in conventional incubator. After in vitro maturation (IVM), the COCs were in vitro fertilized (IVF), and in vitro cultivated (IVC) in a conventional incubator. We evaluated oocyte nuclear maturation, cleavage and blastocyst rates, in addition to the relative mRNA expression of BAX, BMP-15, AREG and EREG genes in oocytes and cumulus cells. The proportion of oocytes in metaphase II was higher in CNTG and ChRG (77.57% and 77.06%) than in the HEPES-G (65.32%; p = .0408 and .0492, respectively). The blastocyst production was similar between CNTG and ChRG (26.20% and 28.47%; p = .4232) and lower (p = .001) in the HEPES-G (18.71%). The relative mRNA expression of BAX gene in cumulus cells was significantly higher (p = .0190) in the HEPES-G compared to the CNTG. Additionally, the relative mRNA expression of BMP-15 gene was lower (p = .03) in oocytes from HEPES-G compared to the CNTG. In conclusion, inadequate atmosphere control has a detrimental effect on oocyte maturation. Yet, the use of chemical gasification can be an efficient alternative to bovine COCs cultivation.


Assuntos
Fertilização in vitro , Técnicas de Maturação in Vitro de Oócitos , Oócitos , Animais , Bovinos , Técnicas de Maturação in Vitro de Oócitos/veterinária , Técnicas de Maturação in Vitro de Oócitos/métodos , Oócitos/efeitos dos fármacos , Fertilização in vitro/veterinária , Feminino , Meios de Cultura , Blastocisto/efeitos dos fármacos , Células do Cúmulo/efeitos dos fármacos , Dióxido de Carbono/farmacologia , Bicarbonato de Sódio/farmacologia , Ácido Cítrico/farmacologia , Técnicas de Cultura Embrionária/veterinária
4.
J Phys Condens Matter ; 36(46)2024 Aug 23.
Artigo em Inglês | MEDLINE | ID: mdl-39111342

RESUMO

The final structure and properties of layers grown by epitaxy techniques are determined in the very early stage of the process. This review describes one-dimensional models for epitaxial growth, emphasizing the basic theoretical concepts employed to analyze nucleation and aggregation phenomena in the submonolayer regime. The main findings regarding the evolution of quantities that define the properties of the system, such as monomer and island densities, and the associated island size, gap length, and capture zone distributions are discussed, as well as the analytical tools used to evaluate them. This review provides a concise overview of the most widely used algorithms for simulating growth processes, discusses relevant experimental results, and establishes connections with existing theoretical studies.

5.
Biomedica ; 44(2): 248-257, 2024 05 30.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-39088534

RESUMO

Introduction. El Alférez, a village in Los Montes de María (Bolívar, Colombia) and a macro-focus of leishmaniasis, recorded its first case in 2018, evidencing changes in the distribution and eco-epidemiology of the disease, although interactions between vectors and local fauna remain unknown. Objective. To evaluate the diversity of sandflies and their blood meal sources in the community of El Alférez in the municipality of El Carmen de Bolívar (Bolívar, Colombia). Materials and methods. In 2018, sandflies were collected using LED-based light traps in domestic, peridomestic, and sylvatic ecotopes and identified at the species level. Multiplex polymerase chain reaction targeting the mitochondrial cytochrome B gene was used to analyze blood from the digestive tract. Results. Lutzomyia evansi was the most abundant species (71.85%; n = 485/675), followed by Lu. panamensis, Lu. gomezi, Lu. trinidadensis, Lu. dubitans, Lu. abonnenci, and Lu.aclydifera. Twenty-five percent of the species had blood meals from Canis familiaris (36.00%; n = 9/25), Ovis aries (36.00%; n=9:/25), Bos taurus (24.00%; n = 6/25), Sus scrofa (20.00%; n = 5/25), and Homo sapiens (8.00%; n = 2/25). Lutzomyia evansi registered the highest feeding frequency (68.00%; n = 17/25), predominantly on a single (44.00%; n = 11/25) or multiple species (24.00%; n = 6/25). Conclusion. Results indicate a eclectic feeding behavior in Lu. evansi, implying potential reservoir hosts for Leishmania spp. and increasing transmission risk. This study is a first step towards understanding the diversity of mammalian blood sources used by sandflies, that may be crucial for vector identification and formulation of effective control measures.


Introducción. En 2018, en la vereda El Alférez de Los Montes de María (Bolívar, Colombia), un macrofoco de leishmaniasis, se reportó el primer caso y se evidenciaron cambios en la distribución y ecoepidemiología de la enfermedad. No obstante, las interacciones entre vectores y fauna local aún son desconocidas. Objetivo. Evaluar la diversidad de flebotomíneos y sus fuentes de alimentación sanguínea en la comunidad de El Alférez del municipio de El Carmen de Bolívar (Bolívar, Colombia). Materiales y métodos. En el 2018, se recolectaron flebotomíneos mediante trampas de luz led ubicadas en el domicilio, el peridomicilio y en el área silvestre, y se identificaron a nivel de especie. Se utilizó la reacción en cadena de la polimerasa múltiple dirigida al gen mitocondrial citocromo B para analizar la sangre del aparato digestivo. Resultados. Lutzomyia evansi fue la especie más abundante (71,85 %; n = 485/675), seguida por Lu. panamensis, Lu. gomezi, Lu. trinidadensis, Lu. dubitans, Lu. abonnenci y Lu. aclydifera. El 25 % (n = 25/100) de las especies analizadas tuvieron como fuentes de ingesta sanguínea a Canis familiaris (36 %; n = 9/25), Ovis aries (36 %; n = 9/25), Bos taurus (24 %; n = 6/25), Sus scrofa (20 %; n = 5/25) y Homo sapiens (8 %; n = 2/25). Lutzomyia evansi fue la especie con la mayor frecuencia de alimentación (68 %; n = 17/25), predominantemente de una sola especie (44 %; n = 11/25) o de varias (24 %; n = 6/25).


Assuntos
Insetos Vetores , Leishmaniose , Psychodidae , Animais , Colômbia/epidemiologia , Psychodidae/parasitologia , Insetos Vetores/parasitologia , Humanos , Leishmaniose/epidemiologia , Leishmaniose/transmissão , Comportamento Alimentar , Cães , Bovinos , Citocromos b/genética , Feminino , Masculino
6.
Arch Argent Pediatr ; 122(6): e202410317, 2024 12 01.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-39024496

RESUMO

Childhood leprosy, which affects children up to 14 years old, is characterized by a delay in diagnosis since it is usually confused with other dermatoses. Its presence in a child is a relevant epidemiological indicator since it signals active disease transmission. We present 4 patients between 5 and 14 years old who attended a public hospital in Buenos Aires -two patients with borderline tuberculoid leprosy, one with lepromatous leprosy, and one with indeterminate leprosy.The World Health Organization provides therapy for people between 10 and 14 but does not consider children under 10. This difficulty implies adapting the dosage and pharmaceutical form to each patient under this age. Finally, it should be noted that the diagnosis of the patients led to the diagnosis and treatment of the disease in adult cohabitants.


La lepra infantil, aquella que afecta a niños de hasta 14 años, se caracteriza por la demora en su diagnóstico, ya que habitualmente es confundida con otras dermatosis. Su presencia en un niño es un indicador epidemiológico relevante, dado que señala la transmisión activa de la enfermedad. Presentamos 4 pacientes de entre 5 y 14 años asistidos en un hospital público de la Ciudad de Buenos Aires. Dos pacientes presentaron lepra borderline tuberculoide; uno, lepra lepromatosa; y otra, lepra indeterminada. La Organización Mundial de la Salud facilita la terapéutica para la población entre 10 y 14 años, pero no contempla a los menores de 10 años. Esta dificultad implica adecuar la dosis y la forma farmacéutica a cada paciente menor de la edad referida. Finalmente, es de destacar que el diagnóstico de los pacientes llevó al diagnóstico y tratamiento de la enfermedad en convivientes adultos.


Assuntos
Hanseníase , Humanos , Argentina , Adolescente , Criança , Pré-Escolar , Masculino , Feminino , Hanseníase/diagnóstico , Saúde da População Urbana
7.
J Sci Food Agric ; 104(14): 9035-9045, 2024 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-38989963

RESUMO

BACKGROUND: Extrusion cooking of cereal-legume flour mixture is an innovative strategy to introduce nutrient-enriched ready-to-eat snacks to the market. However, this thermal process triggers the formation of compounds that could impact safety aspects of these products. Maillard reaction markers and the end products known as melanoidins were evaluated to assess the toxicological and bioactive profiles of extruded snacks from corn-plus-common-bean-flour combinations. Different molecular weight fractions were isolated and purified to analyze their antioxidant activity and to investigate the role of melanoidins. RESULTS: The snack formulated with an 84:16 ratio of corn:common bean flours exhibited an enhanced toxicological profile. It displayed the lowest levels of acrylamide and furanic compounds, along with reduced blockage of lysine residues in the protein. Extrusion increased the antioxidant activity of uncooked flours (30 to 64%) and total phenolic compounds (26 to 50%), and decreased the available lysine (-72.7 to -79.5%). During the fractionation process, it was established that compounds within the range of 3-10 kDa made the greatest contribution to antioxidant activity. The fraction greater than 10 kDa, which included melanoidins, displayed 7 to 33% lower antioxidant activity. The purification of the fraction greater than 10 kDa revealed that pure melanoidins represented approximately one-third of the antioxidant activity in that fraction. Non-covalent adducts linked to the melanoidin core therefore had a relevant role in the antioxidant action of formulated snacks. CONCLUSION: This investigation illustrates the importance of considering both potential risks and associated benefits of compounds formed during the Maillard reaction while developing new extruded snacks. © 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Assuntos
Antioxidantes , Farinha , Reação de Maillard , Polímeros , Lanches , Zea mays , Antioxidantes/química , Antioxidantes/análise , Zea mays/química , Farinha/análise , Polímeros/química , Culinária , Fabaceae/química , Fenóis/química , Fenóis/análise , Acrilamida/química
8.
Biomolecules ; 14(7)2024 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-39062466

RESUMO

Dihydroxyacetone phosphate (DHAP)-dependent aldolases catalyze the aldol addition of DHAP to a variety of aldehydes and generate compounds with two stereocenters. This reaction is useful to synthesize chiral acyclic nucleosides, which constitute a well-known class of antiviral drugs currently used. In such compounds, the chirality of the aliphatic chain, which mimics the open pentose residue, is crucial for activity. In this work, three DHAP-dependent aldolases: fructose-1,6-biphosphate aldolase from rabbit muscle, rhanmulose-1-phosphate aldolase from Thermotoga maritima, and fuculose-1-phosphate aldolase from Escherichia coli, were used as biocatalysts. Aldehyde derivatives of thymine and cytosine were used as acceptor substrates, generating new acyclic nucleoside analogues containing two new stereocenters with conversion yields between 70% and 90%. Moreover, structural analyses by molecular docking were carried out to gain insights into the diasteromeric excess observed.


Assuntos
Aldeído Liases , Escherichia coli , Frutose-Bifosfato Aldolase , Simulação de Acoplamento Molecular , Nucleosídeos de Pirimidina , Thermotoga maritima , Animais , Escherichia coli/enzimologia , Nucleosídeos de Pirimidina/química , Nucleosídeos de Pirimidina/síntese química , Aldeído Liases/metabolismo , Aldeído Liases/química , Coelhos , Frutose-Bifosfato Aldolase/química , Frutose-Bifosfato Aldolase/metabolismo , Thermotoga maritima/enzimologia , Fosfato de Di-Hidroxiacetona/metabolismo , Fosfato de Di-Hidroxiacetona/química , Estereoisomerismo
9.
J Anim Sci ; 1022024 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-39028536

RESUMO

With global warming, there are growing challenges for raising taurine and composite beef cattle populations in tropical regions, including elevated temperatures, limited forage availability, parasite infestation, and infectious diseases. These environmental factors can trigger specific physiological responses in the developing fetus, which may have long-term implications on its performance. Therefore, the main objective of this study was to assess the influence of naturally induced thermal stress during the gestation period on the subsequent performance of tropical composite beef cattle progeny. Furthermore, we aimed to investigate the impact of genotype-by-gestational thermal environment interaction (G×Eg) on traits under selection pressure in the breeding population. A total of 157,414 animals from 58 farms located in various Brazilian states were recorded for birth weight (BW), preweaning weight gain (PWG), yearling weight (YW), hip height (HH), scrotal circumference (SC), and days to first calving (DFC). We first applied a linear regression model to the BW data, which revealed that the last 40 d of gestation were suitable for calculating the mean temperature humidity index (THIg). Subsequent regression analyses revealed that for every 10-unit increase in THIg, detrimental effects of approximately 1.13% to 16.34% are expected for all traits evaluated. Genetic parameters were estimated through a reaction norm model using THIg as the environmental descriptor. The posterior means of heritability estimates (SD) were 0.35 (0.07), 0.25 (0.03), 0.31 (0.03), 0.37 (0.01), 0.29 (0.07), and 0.20 (0.09) for the direct effect of BW, PWG, YW, HH, SC, and DFC, respectively. These estimates varied along the range of THIg values, suggesting a variable response to selection depending on the thermal environment during gestation. Genetic correlation estimates between more divergent THIg values were low or negative for YW, PWG, and DFC, indicating that the best-performing individuals at low THIg values may not perform as well at high THIg values and vice versa. Overall, thermal stress during gestation impacts the future performance of beef cattle offspring. Our results indicate the need for developing effective breeding strategies that take into account G×Eg effects and the re-ranking of breeding animals along the THIg scale, particularly for traits such as DFC that are highly sensitive to thermal stress.


With global warming posing increasing challenges in tropical regions, this study aimed to assess the impact of thermal stress during gestation on the performance of composite beef cattle offspring. Environmental factors such as high temperatures, humidity, limited forage availability, and parasite infestation can elicit physiological responses in the developing fetus, affecting its long-term performance and welfare. Using the temperature humidity index (THIg) of the late gestation as a measure of thermal environment, a reaction norm model was applied to analyze the birth weight, preweaning weight gain, yearling weight, hip height, scrotal circumference, and days to first calving (DFC). Results revealed that increasing THIg values were associated with a detrimental effect in these traits. Genotype-by-environment interaction was found to significantly influence trait variability, with DFC showing the strongest effect. Negative genetic correlations were observed between divergent THIg values, suggesting that individuals performing well in mild thermal environments may not excel in high thermal stress conditions. The heritability estimates varied along the THIg scale, indicating that selection response may vary depending on the thermal environment during gestation. These findings emphasize the need for breeding strategies that account for genotype-by-environment effects and consider the impact of thermal stress on cattle performance.


Assuntos
Genótipo , Animais , Bovinos/genética , Bovinos/fisiologia , Feminino , Gravidez , Brasil , Masculino , Clima Tropical , Peso ao Nascer , Cruzamento , Aumento de Peso , Temperatura
10.
Sci Rep ; 14(1): 17001, 2024 07 24.
Artigo em Inglês | MEDLINE | ID: mdl-39043835

RESUMO

The Continuous Visual Attention Test (CVAT) is a test that detects visuomotor reaction time (RT, alertness), variability of reaction time (VRT, sustained attention), omission errors (OE, focused attention), and commission errors (CE, response inhibition). The standard test takes 15 min, while the ultrafast version only 90 s. Besides overall task length, the two versions differ by target probability (20% and 80% in the 15-min vs. only 80% in the 90-s test) and stimulus-onset asynchrony (SOA) (1, 2, and 4 s in the 15-min vs. only 1 s in the 90-s test. We aimed to analyze the effect of target probability, SOA, and time length on the CVAT variables across the 15-min task and to verify correlations and agreements between the 15-min and the 90-s CVATs. 205 healthy participants performed the two CVATs on the same day. Considering the 15-min task, RT and CE were strongly affected by target probability. Conversely, VRT was not affected. When the 15-min task was compared to the 90-s task, we found no significant difference in the VRT variable. Additionally, a significant agreement between the two tasks was found for the VRT variable. We concluded that sustained attention can be measured with the 90-s CVAT.


Assuntos
Atenção , Tempo de Reação , Humanos , Atenção/fisiologia , Tempo de Reação/fisiologia , Masculino , Feminino , Adulto , Adulto Jovem , Desempenho Psicomotor/fisiologia , Testes Neuropsicológicos , Adolescente , Pessoa de Meia-Idade , Percepção Visual/fisiologia , Estimulação Luminosa
11.
J Mol Model ; 30(8): 278, 2024 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-39034330

RESUMO

CONTEXT: This study investigates the ethylene insertion reaction mechanism for polymerization catalysis, aiming to discern differences between Ni-α-imine ketone-type catalyst and their SiO2-supported counterpart. The reaction force analysis unveils a more intricate mechanism with SiO2 support, shedding light on unexplored factors and elucidating the observed lower catalytic activity. Furthermore, reactivity indexes suggest earlier ethylene activation in the supported catalyst, potentially enhancing overall selectivity. Finally, the reaction electronic flux analysis provides detailed insights into the electronic activity at each step of the reaction mechanism. In sum, this study offers a comprehensive understanding of the ethylene insertion reaction mechanism in both molecular and supported catalysts, underscoring the pivotal role of structural and electronic factors in catalytic processes. METHODS: Density functional theory (DFT) calculations were conducted using the ωB97XD functional and the 6-31 + G(d,p) basis sets with Gaussian16 software. Computational techniques utilized in this study encompassed the IRC method, reaction force analysis, and evaluation of electronic descriptors such as electronic chemical potential, molecular hardness, and electrophilicity reactivity indexes. Additionally, reaction electronic flux analysis was employed to investigate electronic activity along the reaction coordinate.

12.
J Mol Model ; 30(8): 294, 2024 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-39080111

RESUMO

CONTEXT: Hydrazones have been studied for a myriad of chemical and physiochemical properties, such as sensors, chelators and numerous biological activities. Experimental data indicates that hydrazones are unstable under cathodic potentials irrespective of the solvent. The single electron reduction of hydrazones to produce radical anions result in unstable species that cleaves at the N-N bond in a heterolytic manner. The literature has proposed a mechanism favouring the radical on the imine moiety, however in this study DFT calculations suggest the radical on the amine product is more likely upon bond cleavage. This has implications on electrochemical mechanisms, and the active molecule in biological studies viz the method of delivery to target areas. METHODS: Density functional theory calculations were carried out using the GAMESS software package. The structures were optimized in the gas phase (B3LYP/6-31G(d,p)) as indicated by the absence of imaginary frequencies in the Hessian, and in CH3CN (B3LYP/6-31G(d,p)/SMD) with the Pople polarization functions. As a comparison, selected pathways were fully optimized using PBE0/6-31G(d,p) and PBE0/6-31G(d,p)/SMD for gas phase and CH3CN, respectively with the Pople polarization functions. The values were not significantly different (< 5% difference). As such only the B3LYP is evaluation is discussed.

13.
Cardiol Ther ; 13(3): 593-602, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39008026

RESUMO

INTRODUCTION: This prospective, single-arm, crossover pharmacodynamic study assessed the effect of Bayer® low-dose enteric-coated aspirin 81 mg tablets (LD EC-ASA) (Bayer AG, Leverkusen, North Rhine-Westphalia, Germany) compared to Vazalore® low-dose phospholipid-aspirin liquid-filled 81 mg capsules (LD PL-ASA) (PLx Pharma Inc., Sparta, NJ, USA) on platelet reactivity with respect to aspirin reaction units (ARU). METHODS: Forty-seven healthy volunteers were recruited. Platelet function was evaluated with the VerifyNow™ ARU assay (Werfen, Bedford, MA, USA) and assessed post-initiation of Bayer® LD EC-ASA daily for 14 days, with a washout period of 28 days, followed by Vazalore® LD PL-ASA daily for 14 days, again followed by ARU testing. RESULTS: Participants on LD EC-ASA had a mean ARU score of 426, with 19.1% of participants having an ARU > 550; patients on LD PL-ASA derived a mean ARU score of 435, with 14.9% achieving an ARU > 550. There were no significant differences in aspirin resistance (ARU > 550) according to the formulation (Bayer® LD EC-ASA vs. Vazalore® LD PL-ASA) used. Aspirin resistance was independent of ethnicity regardless of the formulation used. In addition, there were no significant associations between body surface area (BSA) and Bayer® LD EC-ASA ARU value (p value 0.788) or Vazalore® LD PL-ASA ARU value (p value 0.477). No patients experienced any serious adverse events or treatment-emergent adverse events. CONCLUSIONS: There were no significant differences in aspirin resistance between Bayer® LD EC-ASA and Vazalore® LD PL-ASA. This dedicated pharmacodynamic study could potentially be informative and applicable for Trinidadian patients on dual antiplatelet therapy (DAPT). Further studies are required to confirm these exploratory findings. TRIAL REGISTRATION: ClinicalTrials.gov identifier, NCT06228820, prospectively registered 1/18/2024.

14.
J Fungi (Basel) ; 10(7)2024 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-39057394

RESUMO

INTRODUCTION: The study of Candida glabrata genes associated with fluconazole resistance, from a molecular perspective, increases the understanding of the phenomenon with a view to its clinical applicability. OBJECTIVE: We sought to establish the predictive molecular profile of fluconazole resistance in Candida glabrata by analyzing the ERG11, ERG3, CgCDR1, and CgSNQ2 genes. METHOD: Expression was quantified using RT-qPCR. Metrics were obtained through molecular docking and Fisher discriminant functions. Additionally, a predictive classification was made against the susceptibility of C. glabrata to fluconazole. RESULTS: The relative expression of the ERG3, CgCDR1, and CgSNQ2 genes was higher in the fluconazole-resistant strains than in the fluconazole-susceptible, dose-dependent strains. The gene with the highest relative expression in the fluconazole-exposed strains was CgCDR1, and in both the resistant and susceptible, dose-dependent strains exposed to fluconazole, this was also the case. The molecular docking model generated a median number of contacts between fluconazole and ERG11 that was lower than the median number of contacts between fluconazole and ERG3, -CgCDR1, and -CgSNQ2. The predicted classification through the multivariate model for fluconazole susceptibility achieved an accuracy of 73.5%. CONCLUSION: The resistant strains had significant expression levels of genes encoding efflux pumps and the ERG3 gene. Molecular analysis makes the identification of a low affinity between fluconazole and its pharmacological target possible, which may explain the lower intrinsic susceptibility of the fungus to fluconazole.

15.
Methods Mol Biol ; 2827: 417-433, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38985286

RESUMO

In situ RT-PCR presents advantages over other expression analysis methods due to its rapid processing and low-cost equipment. However, this technique is not without its challenges. A protocol based on a capsule made from centrifuge tubes that offers advantages over slides is presented. This capsule protects histological sections from drying out, and its easy assembly reduces time pauses between incubations. In addition, the container size where the sample is deposited allows the addition and withdrawal of the different solutions. The capsule does not need previous sealing after each incubation, and, above all, it is a low-cost and accessible material. A guideline for tissue sectioning using a cryostat that offers advantages over other sectioning methods is also described.


Assuntos
Centrifugação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Centrifugação/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Plantas/genética , RNA de Plantas/genética
16.
Vet Anim Sci ; 25: 100372, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39022766

RESUMO

Enzootic Bovine Leukosis (EBL), caused by the bovine leukosis virus (BLV), is a global infectious disease affecting livestock. This study focuses on studying the frequency and genetic traits of BLV in three Creole breeds including Chino Santandereano (Chino), Casanareño (CAS), and Sanmartinero (SM) in Eastern Colombia. We implemented a cross-sectional survey between 2019 and 2020 across four departments (Arauca, Casanare, Santander and Meta) in Eastern Colombia to assess the molecular characteristics of BLV infection in these breeds. A total of 253 cattle were analyzed, of which 42.6 %, 28.8 %, and 28.4 % belonged to the Chino, CAS, and SM breeds, respectively. BLV provirus was detected using nested polymerase chain reaction (n-PCR) targeting the conserved region of the env viral gene. Subsequently, the obtained amplicons were sequenced and subjected to phylogenetic analyses. The overall BLV infection frequency was 26.48 % (95 % CI: 21.01 - 31.98 %), with Chino exhibiting the highest frequency (35.1 %) following by SAM and CAS, respectively (P < 0.05). Other epidemiological variables associated with the infection included age, department, and season (P < 0.05). BLV-positive animals exhibited elevated levels of total serum proteins (P < 0.05), while molecular characterization revealed the exclusive circulation of BLV genotype 1 within these breeds. This study provides an updated assessment of BLV infection in Creole breeds from the eastern of Colombia, underscoring their lower infection frequency compared to introduced breeds and their reduced susceptibility to developing clinical signs. The epidemiological and molecular characteristics observed should be considered in developing control programs aimed at improving genetic resistance to BLV in Colombian cattle.

17.
New Phytol ; 2024 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-39014516

RESUMO

Through enviromics, precision breeding leverages innovative geotechnologies to customize crop varieties to specific environments, potentially improving both crop yield and genetic selection gains. In Brazil's four southernmost states, data from 183 distinct geographic field trials (also accounting for 2017-2021) covered information on 164 genotypes: 79 phenotyped maize hybrid genotypes for grain yield and their 85 nonphenotyped parents. Additionally, 1342 envirotypic covariates from weather, soil, sensor-based, and satellite sources were collected to engineer 10 K synthetic enviromic markers via machine learning. Soil, radiation light, and surface temperature variations remarkably affect differential genotype yield, hinting at ecophysiological adjustments including evapotranspiration and photosynthesis. The enviromic ensemble-based random regression model showcases superior predictive performance and efficiency compared to the baseline and kernel models, matching the best genotypes to specific geographic coordinates. Clustering analysis has identified regions that minimize genotype-environment (G × E) interactions. These findings underscore the potential of enviromics in crafting specific parental combinations to breed new, higher-yielding hybrid crops. The adequate use of envirotypic information can enhance the precision and efficiency of maize breeding by providing important inputs about the environmental factors that affect the average crop performance. Generating enviromic markers associated with grain yield can enable a better selection of hybrids for specific environments.

18.
Braz J Cardiovasc Surg ; 39(4): e20230236, 2024 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-39038115

RESUMO

INTRODUCTION: Perfusion safety in cardiac surgery is vital, and this survey explores perfusion practices, perspectives, and challenges related to it. Specifically, it examines the readiness of on-call and emergency operation rooms for perfusion-related procedures during urgent situations. The aim is to identify gaps and enhance perfusion safety protocols, ultimately improving patient care. METHODS: This was a preliminary survey conducted as an initial exploration before committing to a comprehensive study. The sample size was primarily determined based on a one-month time frame. The survey collected data from 236 healthcare professionals, including cardiac surgeons, perfusionists, and anesthetists, using an online platform. Ethical considerations ensured participant anonymity and voluntary participation. The survey comprised multiple-choice and open-ended questions to gather quantitative and qualitative data. RESULTS: The survey found that 53% preferred a dry circuit ready for emergencies, 19.9% preferred primed circuits, and 19.1% chose not to have a ready pump at all. Various reasons influenced these choices, including caseload variations, response times, historical practices, surgeon preferences, and backup perfusionist availability. Infection risk, concerns about error, and team dynamics were additional factors affecting circuit readiness. CONCLUSION: This survey sheds light on current perfusion practices and challenges, emphasizing the importance of standardized protocols in regards to readiness of on-call and emergency operation rooms. It provides valuable insights for advancing perfusion safety and patient care while contributing to the existing literature on the subject.


Assuntos
Salas Cirúrgicas , Humanos , Inquéritos e Questionários , Perfusão/métodos , Procedimentos Cirúrgicos Cardíacos , Segurança do Paciente , Serviço Hospitalar de Emergência/organização & administração
19.
Clinics (Sao Paulo) ; 79: 100405, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38968666

RESUMO

CONTEXT: Vehicle driving depends on the integration of motor, visual, and cognitive skills to respond appropriately to different situations that occur in traffic. OBJECTIVES: To analyze a model of performance predictor for braking time in the driving simulator, using a battery of tests divided by gender. METHODS: Selected were 100 male drivers with a mean age of 72.6 ± 5.7 years. Sociodemographic variables, braking time in the driving simulator, and motor, visual, and cognitive skills were evaluated. RESULTS: Comparing genders, men were older than women (p = 0.002) and had longer driving times (p = 0.001). Men had more strength in hand grip (p ≤ 0.001). In the linear regression analysis, the model explained 68 % of the braking time in men and 50.8 % in women. In the stepwise multiple linear regression analysis, the variable that remained in the model was the strength of the right plantar flexors, which explained 13 % of the braking time in women and men, and the cognitive variables explained 38.9 %. CONCLUSION: Sociodemographic, motor, visual, and cognitive variables, explained a substantial portion of the variability in braking time for both older women and men, the specific variables driving this performance differed between the sexes. For older women, factors such as muscle strength emerged as critical determinants of braking ability, highlighting the importance of physical health in maintaining driving skills. On the other hand, cognitive conditions emerged as the primary predictor of braking performance in older men, underscoring the role of mental acuity and decision-making processes in safe driving.


Assuntos
Condução de Veículo , Humanos , Masculino , Idoso , Condução de Veículo/psicologia , Feminino , Fatores Sexuais , Fatores de Tempo , Fatores Etários , Cognição/fisiologia , Força da Mão/fisiologia , Fatores Socioeconômicos , Idoso de 80 Anos ou mais , Desempenho Psicomotor/fisiologia , Modelos Lineares , Simulação por Computador , Tempo de Reação/fisiologia , Destreza Motora/fisiologia , Estudos Transversais
20.
Heliyon ; 10(11): e32573, 2024 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-38961942

RESUMO

In this study, a novel 3,3'-bipyrazolo [3,4-b]pyridine-type structure was synthesized from 5-acetylamino-3-methyl-1-phenylpyrazole using the Vilsmeier-Haack reaction as a key step. The spectroscopic properties and structural elucidation of the compound were determined with the use of FT-IR, HRMS, 1H NMR, and 13C NMR. Likewise, the theoretical analysis of the IR and NMR spectra allowed peaks to be assigned and a solid correlation was demonstrated between the experimental and theoretical results. Finally, ab initio calculations based on the density functional theory method at the B3LYP/6-311G (d,p) level of theory were used to determine the conformational energy barrier, facilitating the identification of the most probable conformers of the synthesized compound. Overall, our findings contribute to the understanding of bipyrazolo [3,4-b]pyridine derivatives.

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