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1.
Cell ; 186(18): 3862-3881.e28, 2023 08 31.
Artículo en Inglés | MEDLINE | ID: mdl-37572660

RESUMEN

Male sexual behavior is innate and rewarding. Despite its centrality to reproduction, a molecularly specified neural circuit governing innate male sexual behavior and reward remains to be characterized. We have discovered a developmentally wired neural circuit necessary and sufficient for male mating. This circuit connects chemosensory input to BNSTprTac1 neurons, which innervate POATacr1 neurons that project to centers regulating motor output and reward. Epistasis studies demonstrate that BNSTprTac1 neurons are upstream of POATacr1 neurons, and BNSTprTac1-released substance P following mate recognition potentiates activation of POATacr1 neurons through Tacr1 to initiate mating. Experimental activation of POATacr1 neurons triggers mating, even in sexually satiated males, and it is rewarding, eliciting dopamine release and self-stimulation of these cells. Together, we have uncovered a neural circuit that governs the key aspects of innate male sexual behavior: motor displays, drive, and reward.


Asunto(s)
Vías Nerviosas , Conducta Sexual Animal , Animales , Masculino , Neuronas/fisiología , Recompensa , Conducta Sexual Animal/fisiología , Ratones
2.
Cell ; 185(23): 4298-4316.e21, 2022 11 10.
Artículo en Inglés | MEDLINE | ID: mdl-36323317

RESUMEN

After ingestion of toxin-contaminated food, the brain initiates a series of defensive responses (e.g., nausea, retching, and vomiting). How the brain detects ingested toxin and coordinates diverse defensive responses remains poorly understood. Here, we developed a mouse-based paradigm to study defensive responses induced by bacterial toxins. Using this paradigm, we identified a set of molecularly defined gut-to-brain and brain circuits that jointly mediate toxin-induced defensive responses. The gut-to-brain circuit consists of a subset of Htr3a+ vagal sensory neurons that transmit toxin-related signals from intestinal enterochromaffin cells to Tac1+ neurons in the dorsal vagal complex (DVC). Tac1+ DVC neurons drive retching-like behavior and conditioned flavor avoidance via divergent projections to the rostral ventral respiratory group and lateral parabrachial nucleus, respectively. Manipulating these circuits also interferes with defensive responses induced by the chemotherapeutic drug doxorubicin. These results suggest that food poisoning and chemotherapy recruit similar circuit modules to initiate defensive responses.


Asunto(s)
Eje Cerebro-Intestino , Núcleos Parabraquiales , Nervio Vago , Animales , Ratones , Neuronas/fisiología , Neuronas Aferentes/fisiología , Nervio Vago/fisiología
3.
Cell ; 173(5): 1265-1279.e19, 2018 05 17.
Artículo en Inglés | MEDLINE | ID: mdl-29775595

RESUMEN

Chronic social isolation causes severe psychological effects in humans, but their neural bases remain poorly understood. 2 weeks (but not 24 hr) of social isolation stress (SIS) caused multiple behavioral changes in mice and induced brain-wide upregulation of the neuropeptide tachykinin 2 (Tac2)/neurokinin B (NkB). Systemic administration of an Nk3R antagonist prevented virtually all of the behavioral effects of chronic SIS. Conversely, enhancing NkB expression and release phenocopied SIS in group-housed mice, promoting aggression and converting stimulus-locked defensive behaviors to persistent responses. Multiplexed analysis of Tac2/NkB function in multiple brain areas revealed dissociable, region-specific requirements for both the peptide and its receptor in different SIS-induced behavioral changes. Thus, Tac2 coordinates a pleiotropic brain state caused by SIS via a distributed mode of action. These data reveal the profound effects of prolonged social isolation on brain chemistry and function and suggest potential new therapeutic applications for Nk3R antagonists.


Asunto(s)
Encéfalo/metabolismo , Neuroquinina B/metabolismo , Precursores de Proteínas/metabolismo , Aislamiento Social , Estrés Psicológico , Taquicininas/metabolismo , Animales , Antipsicóticos/farmacología , Conducta Animal/efectos de los fármacos , Encéfalo/patología , Femenino , Vectores Genéticos/administración & dosificación , Vectores Genéticos/genética , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Neuroquinina B/genética , Neuronas/citología , Neuronas/metabolismo , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Precursores de Proteínas/antagonistas & inhibidores , Precursores de Proteínas/genética , Interferencia de ARN , ARN Interferente Pequeño/genética , Receptores de Taquicininas/antagonistas & inhibidores , Receptores de Taquicininas/metabolismo , Taquicininas/antagonistas & inhibidores , Taquicininas/genética , Regulación hacia Arriba/efectos de los fármacos
4.
Proc Natl Acad Sci U S A ; 120(22): e2300282120, 2023 05 30.
Artículo en Inglés | MEDLINE | ID: mdl-37216560

RESUMEN

In actinobacteria, an OmpR/PhoB subfamily protein called GlnR acts as an orphan response regulator and globally coordinates the expression of genes responsible for nitrogen, carbon, and phosphate metabolism in actinobacteria. Although many researchers have attempted to elucidate the mechanisms of GlnR-dependent transcription activation, progress is impeded by lacking of an overall structure of GlnR-dependent transcription activation complex (GlnR-TAC). Here, we report a co-crystal structure of the C-terminal DNA-binding domain of GlnR (GlnR_DBD) in complex with its regulatory cis-element DNA and a cryo-EM structure of GlnR-TAC which comprises Mycobacterium tuberculosis RNA polymerase, GlnR, and a promoter containing four well-characterized conserved GlnR binding sites. These structures illustrate how four GlnR protomers coordinate to engage promoter DNA in a head-to-tail manner, with four N-terminal receiver domains of GlnR (GlnR-RECs) bridging GlnR_DBDs and the RNAP core enzyme. Structural analysis also unravels that GlnR-TAC is stabilized by complex protein-protein interactions between GlnR and the conserved ß flap, σAR4, αCTD, and αNTD domains of RNAP, which are further confirmed by our biochemical assays. Taken together, these results reveal a global transcription activation mechanism for the master regulator GlnR and other OmpR/PhoB subfamily proteins and present a unique mode of bacterial transcription regulation.


Asunto(s)
Actinobacteria , Actinobacteria/genética , Actinobacteria/metabolismo , Activación Transcripcional/genética , Proteínas Bacterianas/metabolismo , Transactivadores/metabolismo , Regiones Promotoras Genéticas/genética , Regulación Bacteriana de la Expresión Génica
5.
Infect Immun ; 92(7): e0007224, 2024 Jul 11.
Artículo en Inglés | MEDLINE | ID: mdl-38899880

RESUMEN

Invasive fungal infections impose an enormous clinical, social, and economic burden on humankind. One of the most common species responsible for invasive fungal infections is Candida albicans. More than 30% of patients with disseminated candidiasis fail therapy with existing antifungal drugs, including the widely used azole class. We previously identified a collection of 13 medications that antagonize the activity of the azoles on C. albicans. Although gain-of-function mutations responsible for antifungal resistance are often associated with reduced fitness and virulence, it is currently unknown how exposure to azole antagonistic drugs impacts C. albicans physiology, fitness, or virulence. In this study, we examined how exposure to seven azole antagonists affects C. albicans phenotype and capacity to cause disease. Most of the azole antagonists appear to have little impact on fungal growth, morphology, stress tolerance, or gene transcription. However, aripiprazole had a modest impact on C. albicans hyphal growth and increased cell wall chitin content. It also aggravated the disseminated C. albicans infections in mice. This effect was abrogated in immunosuppressed mice, indicating that it is at least in part dependent upon host immune responses. Collectively, these data provide proof of principle that unanticipated drug-fungus interactions have the potential to influence the incidence and outcomes of invasive fungal disease.


Asunto(s)
Antifúngicos , Aripiprazol , Candida albicans , Candidiasis , Candida albicans/efectos de los fármacos , Candida albicans/genética , Animales , Ratones , Antifúngicos/farmacología , Candidiasis/tratamiento farmacológico , Candidiasis/microbiología , Aripiprazol/farmacología , Aripiprazol/uso terapéutico , Antipsicóticos/farmacología , Antipsicóticos/uso terapéutico , Virulencia , Femenino , Azoles/farmacología , Modelos Animales de Enfermedad
6.
Antimicrob Agents Chemother ; 68(9): e0077924, 2024 Sep 04.
Artículo en Inglés | MEDLINE | ID: mdl-39136466

RESUMEN

Despite the overall decline in malaria cases in Thailand, continuous surveillance in endemic areas remains crucial. This retrospective analysis examined Plasmodium falciparum samples from Tak province, Thailand, collected in 1998, 1999, and 2001, to investigate the prevalence and evolution of antimalarial genotypic drug resistance. The study revealed a high prevalence of drug-resistant P. falciparum, particularly to mefloquine and sulfadoxine/pyrimethamine, with significant mutations in genes associated with resistance. Notably, mutations indicative of artemisinin resistance, such as those in the kelch13 gene, were detected at low frequencies, suggesting an evolving resistance pattern. The underlying cause of these resistance mutations appears to be the historical and widespread use of these antimalarial drugs, which exerted selective pressure on the parasite population. These findings underscore the necessity of ongoing surveillance and adaptive control strategies to manage drug resistance, guide treatment policies, and prevent potential outbreaks, even as malaria cases decrease. Continuous monitoring and research are imperative to sustain malaria elimination efforts and address the dynamic challenges posed by evolving drug-resistant strains.


Asunto(s)
Antimaláricos , Resistencia a Medicamentos , Malaria Falciparum , Mutación , Plasmodium falciparum , Plasmodium falciparum/efectos de los fármacos , Plasmodium falciparum/genética , Antimaláricos/farmacología , Antimaláricos/uso terapéutico , Tailandia/epidemiología , Resistencia a Medicamentos/genética , Malaria Falciparum/parasitología , Malaria Falciparum/epidemiología , Malaria Falciparum/tratamiento farmacológico , Humanos , Estudios Retrospectivos , Prevalencia , Mefloquina/farmacología , Mefloquina/uso terapéutico , Animales , Artemisininas/farmacología , Artemisininas/uso terapéutico , Pirimetamina/farmacología , Pirimetamina/uso terapéutico , Sulfadoxina/farmacología , Sulfadoxina/uso terapéutico , Combinación de Medicamentos
7.
Antimicrob Agents Chemother ; : e0002224, 2024 Apr 16.
Artículo en Inglés | MEDLINE | ID: mdl-38624217

RESUMEN

Candida parapsilosis has recently emerged as a major threat due to the worldwide emergence of fluconazole-resistant strains causing clonal outbreaks in hospitals and poses a therapeutic challenge due to the limited antifungal armamentarium. Here, we used precise genome editing using CRISPR-Cas9 to gain further insights into the contribution of mutations in ERG11, ERG3, MRR1, and TAC1 genes and the influence of allelic dosage to antifungal resistance in C. parapsilosis. Seven of the most common amino acid substitutions previously reported in fluconazole-resistant clinical isolates (including Y132F in ERG11) were engineered in two fluconazole-susceptible C. parapsilosis lineages (ATCC 22019 and STZ5). Each mutant was then challenged in vitro against a large array of antifungals, with a focus on azoles. Any possible change in virulence was also assessed in a Galleria mellonella model. We successfully generated a total of 19 different mutants, using CRISPR-Cas9. Except for R398I (ERG11), all remaining amino acid substitutions conferred reduced susceptibility to fluconazole. However, the impact on fluconazole in vitro susceptibility varied greatly according to the engineered mutation, the stronger impact being noted for G583R acting as a gain-of-function mutation in MRR1. Cross-resistance with newer azoles, non-medical azoles, but also non-azole antifungals such as flucytosine, was occasionally noted. Posaconazole and isavuconazole remained the most active in vitro. Except for G583R, no fitness cost was associated with the acquisition of fluconazole resistance. We highlight the distinct contributions of amino acid substitutions in ERG11, ERG3, MRR1, and TAC1 genes to antifungal resistance in C. parapsilosis.

8.
Exp Cell Res ; 433(2): 113806, 2023 12 15.
Artículo en Inglés | MEDLINE | ID: mdl-37844792

RESUMEN

Heart failure (HF) is a complex clinical syndrome associated with significant morbidity and mortality. Dysregulation of long non-coding RNA (lncRNA) has been implicated in the pathogenesis of HF. The present study aims to investigate the role of lncRNA HOX transcript antisense RNA (HOTAIR) in cardiomyocyte pyroptosis in a murine HF model. A murine HF model was established through transverse aortic contraction surgery, and an in vitro HF cell model was developed by treating HL-1 cells with H2O2. HOTAIR was overexpressed in TAC mice and HL-1 cells via pcDNA3.1-HOTAIR transfection. Cardiac function was assessed in TAC mice, and myocardial changes were evaluated using HE staining. The expression of NLRP3 was examined by immunohistochemistry. Myocardial injury markers and pyroptosis-related inflammatory cytokines were quantified using ELISA. Protein levels of NLRP3, cleaved-caspase-1, and GSDMD-N were analyzed by Western blot. Dual-luciferase assays and RNA immunoprecipitation were employed to confirm the binding interactions between HOTAIR and miR-17-5p, miR-17-5p and RORA. Functional rescue experiments were conducted by overexpressing miR-17-5p or silencing RORA in HL-1 cells. HOTAIR exhibited reduced expression in TAC mice and H2O2-induced cardiomyocytes. Overexpression of HOTAIR ameliorated cardiac dysfunction, reduced myocardial pathological injury, enhanced cardiomyocyte viability, and decreased myocardial injury and pyroptosis. HOTAIR interacted with miR-17-5p to repress RORA transcription. Overexpression of miR-17-5p or silencing of RORA abolished the inhibitory effect of HOTAIR overexpression on cardiomyocyte pyroptosis. In conclusion, HOTAIR competitively bound to miR-17-5p, relieving its inhibition of RORA transcription and leading to increased RORA expression and suppressed cardiomyocyte pyroptosis in HF models.


Asunto(s)
Insuficiencia Cardíaca , MicroARNs , ARN Largo no Codificante , Animales , Ratones , Insuficiencia Cardíaca/genética , Peróxido de Hidrógeno , MicroARNs/genética , Miocitos Cardíacos , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Piroptosis/genética , ARN Largo no Codificante/genética , Miembro 1 del Grupo F de la Subfamilia 1 de Receptores Nucleares/genética , Miembro 1 del Grupo F de la Subfamilia 1 de Receptores Nucleares/metabolismo
9.
Nutr J ; 23(1): 87, 2024 Jul 31.
Artículo en Inglés | MEDLINE | ID: mdl-39085886

RESUMEN

BACKGROUND: No study has investigated the relationship between dietary total antioxidant capacity and sarcopenia so far. OBJECTIVE: This study aimed to investigate the association between dietary Total Antioxidant Capacity (d-TAC) and sarcopenia in elderly adults. METHODS: In this cross-sectional study we enrolled 300 elderly people (150 men and 150 women) aged ≥ 55 years using cluster random sampling method. Sarcopenia was defined based on European Working Group on Sarcopenia (EWGSOP) definition. A DXA scanner, a squeeze bulb dynamometer and a 4-Meter walk gait speed test was used to measure Appendicular Skeletal Muscle (ASM), muscle strength and muscle performance respectively. We also used a Block-format 117-item food frequency questionnaire (FFQ) to assess dietary intakes of participants. Multivariable logistic regression models were applied to examine the association between d-TAC and sarcopenia. RESULTS: Mean ± SD age of study participants and their BMI was 66.8 ± 7.72 year and 27.3 ± 4.2 kg/m2, respectively. People in the highest tertile of d-TAC had the greatest hand grip strength (11.9 ± 3.63 vs. 10.4 ± 3.55 psi, p = 0.009) and had lower odds of sarcopenia compared with those in the lowest tertile, either before (OR = 0.39; 95% CI: 0.17, 0.88) or after adjustment for potential confounders (OR = 0.33; 95% CI: 0.11, 0.95). No other significant association was seen between d-TAC and components of sarcopenia. CONCLUSION: We found an inverse association between dietary total antioxidant capacity and odds of sarcopenia. No significant association was seen between d-TAC and individual components of sarcopenia. Further studies are needed to confirm our findings.


Asunto(s)
Antioxidantes , Dieta , Fuerza de la Mano , Sarcopenia , Humanos , Masculino , Estudios Transversales , Femenino , Antioxidantes/administración & dosificación , Antioxidantes/análisis , Anciano , Persona de Mediana Edad , Dieta/métodos , Dieta/estadística & datos numéricos , Fuerza de la Mano/fisiología , Fuerza Muscular/fisiología , Músculo Esquelético/fisiopatología , Índice de Masa Corporal
10.
Cryobiology ; 114: 104836, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38092234

RESUMEN

Geographically distributed ovarian tissue cryobanks remain limited due to the high facility and staff costs, and cold transportation to centers is associated with ischemia-induced tissue damage that increases with transport distance. It is ideal to perform the cryopreservation procedure at a tissue removal site or local hospital before shipment to cost-effective centralized cryobanks. However, conventional liquid nitrogen-based freezers are not portable and require expensive infrastructure. To study the possibility of an ovarian tissue cryopreservation network not dependent on liquid nitrogen, we cryopreserved bovine ovarian tissue using three cooling techniques: a controlled rate freezer using liquid nitrogen, a liquid nitrogen-free controlled rate freezer, and liquid nitrogen-free passive cooling. Upon thawing, we evaluated a panel of viability metrics in frozen and fresh groups to examine the potency of the portable liquid nitrogen-free controlled and uncontrolled rate freezers in preserving the ovarian tissue compared to the non-portable conventional controlled rate freezer. We found similar outcomes for reactive oxygen species (ROS), total antioxidant capacity (TAC), follicular morphology, tissue viability, and fibrosis in the controlled rate freezer groups. However, passive slow cooling was associated with the lowest tissue viability, follicle morphology, and TAC, and the highest tissue fibrosis and ROS levels compared to all other groups. A stronger correlation was found between follicle morphology, ovarian tissue viability, and fibrosis with the TAC/ROS ratio compared to ROS and TAC alone. The current study undergirds the possibility of centralized cryobanks using a controlled rate liquid nitrogen-free freezer to prevent ischemia-induced damage during ovarian tissue shipment.


Asunto(s)
Criopreservación , Nitrógeno , Humanos , Femenino , Animales , Bovinos , Congelación , Criopreservación/métodos , Especies Reactivas de Oxígeno , Supervivencia Celular , Isquemia , Fibrosis
11.
Mikrochim Acta ; 191(8): 465, 2024 07 16.
Artículo en Inglés | MEDLINE | ID: mdl-39012354

RESUMEN

A novel Fe-MoOx nanozyme, engineered with enhanced peroxidase (POD)-like activity through strategic doping and the creation of oxygen vacancies, is introduced to catalyze the oxidation of TMB with high efficiency. Furthermore, Fe-MoOx is responsive to single electron transfer (SET) and hydrogen atom transfer (HAT) mechanisms related to antioxidants and can serve as a desirable nanozyme for total antioxidant capacity (TAC) determination. The TAC colorimetric platform can reach a low LOD of 0.512 µM in solution and 24.316 µM in the smartphone-mediated RGB hydrogel (AA as the standard). As proof of concept, the practical application in real samples was explored. The work paves a promising avenue to design diverse nanozymes for visual on-site inspection of food quality.


Asunto(s)
Antioxidantes , Colorimetría , Oxidación-Reducción , Antioxidantes/química , Antioxidantes/análisis , Antioxidantes/metabolismo , Colorimetría/métodos , Catálisis , Molibdeno/química , Límite de Detección , Hierro/química , Bencidinas/química , Teléfono Inteligente , Hidrogeles/química , Transporte de Electrón , Técnicas Biosensibles/métodos , Óxidos/química
12.
J Environ Manage ; 351: 119845, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38109825

RESUMEN

Pyrogallol promotes free radicals leading to oxidative stress and toxicity. There are however a lack of studies on oxidative stress and the antioxidant system of fish following exposure to pyrogallol. This study measured oxidative stress markers, antioxidant responses, and histological changes in catfish exposed to pyrogallol. Fish were divided into one of four experimental groups: control only, or 1, 5 or 10 mg/L pyrogallol. After 15 days, glutathione-S-transferase in the serum was decreased in fish exposed to either 5 or 10 mg/L pyrogallol relative to controls while superoxide dismutase and total antioxidant capacity were decreased significantly in fish exposed to 1, 5, or 10 mg/L pyrogallol. Conversely, catalase was increased in serum of fish exposed to 1, 5, or 10 mg/L pyrogallol compared to controls. The liver of fish treated with 1, 5, or 10 mg/L pyrogallol had significantly higher levels of oxidative stress markers (malondialdehyde, lipid peroxidation, hydroperoxide content, oxidised protein content, and DNA fragmentation %) that varied with concentration. Catfish exposed to either 1, 5, or 10 mg/L pyrogallol presented with notable histological alterations in the intestine, kidney, and muscles with prominent fibrosis, as intense deposition of collagen fibre was observed by Masson's trichrome staining. Overall, endpoints related to oxidative stress and antioxidant defence enzymes in fish may be early biomarkers of pyrogallol exposure and contamination in aquatic ecosystems. Additional studies should characterize oxidative stress indicators for their utility as biomarkers of effect.


Asunto(s)
Bagres , Contaminantes Químicos del Agua , Animales , Antioxidantes/metabolismo , Pirogalol/toxicidad , Pirogalol/metabolismo , Ecosistema , Estrés Oxidativo , Bagres/metabolismo , Biomarcadores/metabolismo , Peroxidación de Lípido , Contaminantes Químicos del Agua/toxicidad , Contaminantes Químicos del Agua/metabolismo
13.
BMC Oral Health ; 24(1): 960, 2024 Aug 17.
Artículo en Inglés | MEDLINE | ID: mdl-39153972

RESUMEN

BACKGROUND: Pattern of dental anomalies encountered in cleft patients shows subtle signs of genetic involvement. This study aimed to evaluate the prevalence and pattern of tooth agenesis and supernumerary teeth in Thai cleft population according to the cleft type. METHODS: Data collected from patients with cleft lip and palate, who had been treated at Tawanchai Cleft Center, Khon Kaen University, Thailand, available during year 2012-2022, were investigated. Records from 194 patients with non-syndromic clefts met the inclusion criteria. Standard dental records, and at least either orthopantomogram (OPG) or cone beam computed tomography (CBCT), were examined. Statistical analysis was performed using chi-square and binominal test (p ≤ 0.05). RESULTS: Prevalence of tooth agenesis was higher (77.3%) than that of supernumerary teeth (5.7%) and was more common in bilateral cleft lip and palate (BCLP) (88.1%) than in unilateral cleft lip and palate (UCLP) (72.6%) (p = 0.017). The upper lateral incisor was more frequently affected (46.4%), followed by the upper second premolar. The number of missing teeth observed on the left side was significantly higher. Patients with left UCLP (ULCLP) had the highest prevalence of tooth agenesis. A total of 41 tooth agenesis code (TAC) patterns was found. The prevalence of supernumerary teeth was comparable with 6.6% of ULCLP, 5.1% of BCLP, and 4.5% of URCLP. Tooth-number anomalies were observed more often in the BCLP and were most likely to occur on the left side of the maxilla. Both types of anomalies could be featured in a small proportion of cleft patients. CONCLUSIONS: More than half of the patients with non-syndromic cleft lip and palate in this study, presented with tooth-number anomalies. Tooth agenesis was approximately 10-time more prevalent than supernumerary teeth. Tooth agenesis was likely to appear on the left-side of the maxilla regardless of the laterality of the cleft.


Asunto(s)
Anodoncia , Labio Leporino , Fisura del Paladar , Tomografía Computarizada de Haz Cónico , Diente Supernumerario , Humanos , Labio Leporino/epidemiología , Fisura del Paladar/epidemiología , Diente Supernumerario/epidemiología , Diente Supernumerario/diagnóstico por imagen , Tailandia/epidemiología , Prevalencia , Masculino , Femenino , Anodoncia/epidemiología , Anodoncia/diagnóstico por imagen , Adolescente , Niño , Radiografía Panorámica , Adulto Joven , Pueblos del Sudeste Asiático
14.
J Cell Sci ; 134(5)2021 03 08.
Artículo en Inglés | MEDLINE | ID: mdl-33589495

RESUMEN

Proper mitochondrial genome inheritance is important for eukaryotic cell survival. Trypanosoma brucei, a protozoan parasite, contains a singular mitochondrial genome, the kinetoplast (k)DNA. The kDNA is anchored to the basal body via the tripartite attachment complex (TAC) to ensure proper segregation. Several components of the TAC have been described; however, the connection of the TAC to the kDNA remains elusive. Here, we characterize the TAC-associated protein TAP110. We find that both depletion and overexpression of TAP110 leads to a delay in the separation of the replicated kDNA networks. Proteome analysis after TAP110 overexpression identified several kDNA-associated proteins that changed in abundance, including a TEX-like protein that dually localizes to the nucleus and the kDNA, potentially linking replication and segregation in the two compartments. The assembly of TAP110 into the TAC region seems to require the TAC but not the kDNA itself; however, once TAP110 has been assembled, it also interacts with the kDNA. Finally, we use ultrastructure expansion microscopy in trypanosomes for the first time, and reveal the precise position of TAP110 between TAC102 and the kDNA, showcasing the potential of this approach.This article has an associated First Person interview with the first author of the paper.


Asunto(s)
Genoma Mitocondrial , Trypanosoma brucei brucei , ADN de Cinetoplasto/genética , Genoma Mitocondrial/genética , Mitocondrias , Proteínas Protozoarias/genética , Trypanosoma brucei brucei/genética
15.
Biochem Biophys Res Commun ; 642: 175-184, 2023 01 29.
Artículo en Inglés | MEDLINE | ID: mdl-36584481

RESUMEN

Dantrolene (DAN) directly binds to cardiac ryanodine receptor 2 (RyR2) through Leu601-Cys620 in the N-terminal domain and subsequently inhibits diastolic Ca2+ leakage through RyR2. We previously reported that therapy using RyR2 V3599K mutation, which inhibits diastolic Ca2+ leakage by enhancing calmodulin (CaM) binding ability to RyR2, prevents left ventricular (LV) remodeling in transverse aortic constriction (TAC) heart failure. Here, we examined whether chronic administration of DAN prevents LV remodeling in TAC heart failure via the same mechanism as genetic therapy. A pressure-overloaded hypertrophy mouse model was developed using TAC. Wild-type (WT) mice were divided into three groups: sham-operated mice (Sham group), TAC mice (TAC group), and TAC mice treated with DAN (TAC-DAN group, 20 mg/kg/day, i.p.). They were then followed up for 8 weeks. The survival rate was higher in the TAC-DAN group (83%) than in the TAC group (49%), and serial echocardiography studies and pathological tissue analysis showed that LV remodeling was significantly prevented in the TAC-DAN group compared to the TAC group. An increase in the diastolic Ca2+ spark frequency and a decrease in the binding affinity of CaM to RyR2 were observed at 8 weeks in the TAC group but not in the TAC-DAN group. Stabilization of RyR2 with DAN prevented LV remodeling and improved survival after TAC by enhancing CaM binding to RyR2 and inhibiting RyR2-mediated diastolic Ca2+ leakage.


Asunto(s)
Estenosis de la Válvula Aórtica , Insuficiencia Cardíaca , Ratones , Animales , Canal Liberador de Calcio Receptor de Rianodina/genética , Canal Liberador de Calcio Receptor de Rianodina/metabolismo , Dantroleno/farmacología , Dantroleno/uso terapéutico , Remodelación Ventricular/genética , Insuficiencia Cardíaca/metabolismo , Señalización del Calcio
16.
J Mol Recognit ; 36(7): e3016, 2023 07.
Artículo en Inglés | MEDLINE | ID: mdl-37042157

RESUMEN

The present work determines efficiency of domestic food waste like tea waste in removing pharmaceutical waste such as ceftriaxone (CEF) from synthetic wastewater. Carbonaceous material; Tea waste activated carbon (TAC) has been employed and it showed high removal capacity of 787.5 mg/g. TAC was characterized using; XPS, XRD, SEM, FT-IR, and BET as well as it approved that the adsorbent a has high surface area of .6 m2 /g. Various experimental parameters are evaluated for the removal efficiency of the synthesized adsorbent under the present study. During the adsorption study through batch experiments, it approved that the adsorption isotherm was fitted to Langmuir, while kinetically fitted to pseudo-second-order; the adsorption process was chemisorption process as the adsorption energy was 23.7 kJ mol-1 . From evaluation thermodynamic parameters the adsorption reaction was endothermic and spontaneous reaction. The different real samples spiked with CEF and studies the efficiency of TAC to remove it. On the other hand, investigated the regeneration efficiency of the TAC and exhibit high regeneration efficiency as it will be used after four cycles with good efficiency of about 84.2%.


Asunto(s)
Eliminación de Residuos , Contaminantes Químicos del Agua , Antibacterianos , Ceftriaxona , Carbón Orgánico , Adsorción , Agua , Alimentos , Espectroscopía Infrarroja por Transformada de Fourier , Termodinámica , , Agricultura , Contaminantes Químicos del Agua/análisis , Cinética , Concentración de Iones de Hidrógeno
17.
Eur J Neurol ; 30(10): 2955-2979, 2023 10.
Artículo en Inglés | MEDLINE | ID: mdl-37515405

RESUMEN

BACKGROUND AND PURPOSE: Cluster headache is a relatively rare, disabling primary headache disorder with a major impact on patients' quality of life. This work presents evidence-based recommendations for the treatment of cluster headache derived from a systematic review of the literature and consensus among a panel of experts. METHODS: The databases PubMed (Medline), Science Citation Index, and Cochrane Library were screened for studies on the efficacy of interventions (last access July 2022). The findings in these studies were evaluated according to the recommendations of the European Academy of Neurology, and the level of evidence was established using GRADE (Grading of Recommendations Assessment, Development, and Evaluation). RECOMMENDATIONS: For the acute treatment of cluster headache attacks, there is a strong recommendation for oxygen (100%) with a flow of at least 12 L/min over 15 min and 6 mg subcutaneous sumatriptan. Prophylaxis of cluster headache attacks with verapamil at a daily dose of at least 240 mg (maximum dose depends on efficacy and tolerability) is recommended. Corticosteroids are efficacious in cluster headache. To reach an effect, the use of at least 100 mg prednisone (or equivalent corticosteroid) given orally or at up to 500 mg iv per day over 5 days is recommended. Lithium, topiramate, and galcanezumab (only for episodic cluster headache) are recommended as alternative treatments. Noninvasive vagus nerve stimulation is efficacious in episodic but not chronic cluster headache. Greater occipital nerve block is recommended, but electrical stimulation of the greater occipital nerve is not recommended due to the side effect profile.


Asunto(s)
Cefalalgia Histamínica , Humanos , Cefalalgia Histamínica/terapia , Calidad de Vida , Sumatriptán/uso terapéutico , Oxígeno/uso terapéutico
18.
Appl Microbiol Biotechnol ; 107(7-8): 2469-2481, 2023 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-36912903

RESUMEN

The Antarctic bacterium Pseudoalteromonas haloplanktis TAC125 (PhTAC125) is considered an interesting alternative host for the recombinant protein production, that can be explored when the conventional bacterial expression systems fail. Indeed, the manufacture of all the difficult-to-express proteins produced so far in this bacterial platform gave back soluble and active products. Despite these promising results, the low yield of recombinant protein production achieved is hampering the wider and industrial exploitation of this psychrophilic cell factory. All the expression plasmids developed so far in PhTAC125 are based on the origin of replication of the endogenous pMtBL plasmid and are maintained at a very low copy number. In this work, we set up an experimental strategy to select mutated OriR sequences endowed with the ability to establish recombinant plasmids at higher multiplicity per cell. The solution to this major production bottleneck was achieved by the construction of a library of psychrophilic vectors, each containing a randomly mutated version of pMtBL OriR, and its screening by fluorescence-activated cell sorting (FACS). The selected clones allowed the identification of mutated OriR sequences effective in enhancing the plasmid copy number of approximately two orders of magnitude, and the production of the recombinant green fluorescent protein was increased up to twenty times approximately. Moreover, the molecular characterization of the different mutant OriR sequences allowed us to suggest some preliminary clues on the pMtBL replication mechanism that deserve to be further investigated in the future. KEY POINTS: • Setup of an electroporation procedure for Pseudoalteromonas haloplanktis TAC125. • Two order of magnitude improvement of OriR-derived psychrophilic expression systems. • Almost twenty times enhancement in Green fluorescent protein production.


Asunto(s)
Variaciones en el Número de Copia de ADN , Pseudoalteromonas , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Proteínas Recombinantes/metabolismo , Plásmidos/genética , Pseudoalteromonas/genética , Pseudoalteromonas/metabolismo
19.
Appl Microbiol Biotechnol ; 107(13): 4301-4309, 2023 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-37256328

RESUMEN

Total antioxidant capacity (TAC) is currently considered as a vital indicator of food quality in antioxidant ability and attracts much attention for human healthcare. It is thus of great significance to realize the accurate and rapid detection of TAC in foods. Herein, we have constructed a preferable hybrid nanozyme based on the mesoporous silica-stabilized CuO composited Fe3O4 nanoparticles (Fe3O4@MSNs@CuO, FMC NPs), which possess the enhanced peroxidase (POD)-like activity via cascade response for specific and sensitive determination of TAC in fruit foods. The results showed the hybrid nanozyme displayed a remarkable POD-like activity, excellent selectivity and sensitivity, and the limit of detection (LOD) of the colorimetric sensor was 6.13 mM with the concentration range from 10 to 45 mM. Therefore, the fabricated hybrid nanozyme can be regarded as an effective biosensor for the evaluation of antioxidant quality in fruit foods in future. KEY POINTS: • The stabilized bimetallic nanozyme was constructed for TAC analysis in fruits. • The hybrid nanozyme possessed the enhanced POD-like activity by cascading effects. • The nanozyme was an effective biosensor for antioxidant quality evaluation in fruits.


Asunto(s)
Antioxidantes , Frutas , Humanos , Antioxidantes/análisis , Frutas/química , Dióxido de Silicio , Cobre , Colorimetría/métodos , Peróxido de Hidrógeno , Peroxidasa
20.
BMC Vet Res ; 19(1): 148, 2023 Sep 07.
Artículo en Inglés | MEDLINE | ID: mdl-37679743

RESUMEN

BACKGROUND: Leptospirosis is a neglected but widespread zoonotic disease throughout the world. Most mammals are hosts of Leptospira spp., including domestic cats, species in which no consensus has been reached on the clinical presentation or diagnosis of the disease. The study of acute-phase proteins (APPs) and biomarkers of oxidative status would contribute to knowledge about the disease in cats. This report evaluated four APPs: Serum amyloid A-SAA, Haptoglobin-Hp, albumin and Paraoxonase 1-PON1 and the antioxidant response through Total Antioxidant Capacity-TAC, in 32 free-roaming cats. Cats were classified as seroreactive for anti-leptospiral antibodies (group 1, n = 8), infected with Leptospira spp (group 2, n = 5) and leptospires-free cats (group 3, n = 19). RESULTS: SAA differences were observed between groups 1 and 2 (p-value = 0.01) and between groups 2 and 3 (p-value = 0.0001). Hp concentration differences were only detected between groups 2 and 3 (p-value = 0.001). Albumin concentrations only differed between groups 1 and 3 (p-value = 0.017) and 2 and 3 (p-value < 0.005). Cats in groups 1 (p-value < 0.005) and 2 (p-value < 0.005) had lower PON1 concentrations than group 3. No statistically significant differences between pairs of groups were detected for TAC concentrations. The principal component analysis (PCA) retained two principal components, (PC1 and PC2), explaining 60.1% of the observed variability of the inflammatory proteins and the antioxidant TAC. CONCLUSIONS: Increases in Serum SAA, Hp, and decreases in PON1 activity may indicate an active inflammatory state in infected cats (currently or recently infected).


Asunto(s)
Proteínas de Fase Aguda , Leptospira , Gatos , Animales , Antioxidantes , Proteína Amiloide A Sérica , Haptoglobinas , Albúminas , Mamíferos
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