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1.
Cell ; 152(5): 1065-76, 2013 Feb 28.
Artículo en Inglés | MEDLINE | ID: mdl-23452854

RESUMEN

Medulloblastoma is the most common pediatric malignant brain tumor. Although current therapies improve survival, these regimens are highly toxic and are associated with significant morbidity. Here, we report that placental growth factor (PlGF) is expressed in the majority of medulloblastomas, independent of their subtype. Moreover, high expression of PlGF receptor neuropilin 1 (Nrp1) correlates with poor overall survival in patients. We demonstrate that PlGF and Nrp1 are required for the growth and spread of medulloblastoma: PlGF/Nrp1 blockade results in direct antitumor effects in vivo, resulting in medulloblastoma regression, decreased metastasis, and increased mouse survival. We reveal that PlGF is produced in the cerebellar stroma via tumor-derived Sonic hedgehog (Shh) and show that PlGF acts through Nrp1-and not vascular endothelial growth factor receptor 1-to promote tumor cell survival. This critical tumor-stroma interaction-mediated by Shh, PlGF, and Nrp1 across medulloblastoma subtypes-supports the development of therapies targeting PlGF/Nrp1 pathway.


Asunto(s)
Neoplasias Cerebelosas/patología , Cerebelo/metabolismo , Meduloblastoma/patología , Neuropilina-1/metabolismo , Proteínas Gestacionales/metabolismo , Transducción de Señal , Animales , Células Cultivadas , Neoplasias Cerebelosas/metabolismo , Humanos , Meduloblastoma/metabolismo , Ratones , Ratones Noqueados , Trasplante de Neoplasias , Comunicación Paracrina , Factor de Crecimiento Placentario , Trasplante Heterólogo , Receptor 1 de Factores de Crecimiento Endotelial Vascular/metabolismo
2.
J Pharmacol Exp Ther ; 390(2): 250-259, 2024 Jul 18.
Artículo en Inglés | MEDLINE | ID: mdl-38866563

RESUMEN

Mutations in the GNAO1 gene, which encodes the abundant brain G-protein Gα o, result in neurologic disorders characterized by developmental delay, epilepsy, and movement abnormalities. There are over 50 mutant alleles associated with GNAO1 disorders; the R209H mutation results in dystonia, choreoathetosis, and developmental delay without seizures. Mice heterozygous for the human mutant allele (Gnao1 +/R209H) exhibit hyperactivity in open field tests but no seizures. We developed self-complementary adeno-associated virus serotype 9 (scAAV9) vectors expressing two splice variants of human GNAO1 Gα o isoforms 1 (GoA, GNAO1.1) and 2 (GoB, GNAO1.2). Bilateral intrastriatal injections of either scAAV9-GNAO1.1 or scAAV9-GNAO1.2 significantly reversed mutation-associated hyperactivity in open field tests. GNAO1 overexpression did not increase seizure susceptibility, a potential side effect of GNAO1 vector treatment. This represents the first report of successful preclinical gene therapy for GNAO1 encephalopathy applied in vivo. Further studies are needed to uncover the molecular mechanism that results in behavior improvements after scAAV9-mediated Gα o expression and to refine the vector design. SIGNIFICANCE STATEMENT: GNAO1 mutations cause a spectrum of developmental, epilepsy, and movement disorders. Here we show that intrastriatal delivery of scAAV9-GNAO1 to express the wild-type Gα o protein reduces the hyperactivity of the Gnao1 +/R209H mouse model, which carries one of the most common movement disorder-associated mutations. This is the first report of a gene therapy for GNAO1 encephalopathy applied in vivo on a patient-allele model.


Asunto(s)
Dependovirus , Subunidades alfa de la Proteína de Unión al GTP Gi-Go , Heterocigoto , Animales , Subunidades alfa de la Proteína de Unión al GTP Gi-Go/genética , Ratones , Dependovirus/genética , Humanos , Masculino , Vectores Genéticos/administración & dosificación , Vectores Genéticos/genética , Hipercinesia/genética , Mutación , Terapia Genética/métodos , Ratones Endogámicos C57BL , Locomoción/genética
3.
Adv Exp Med Biol ; 1443: 33-61, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38409415

RESUMEN

Mass spectrometry (MS) is a powerful analytical technique that plays a central role in modern protein analysis and the study of proteostasis. In the field of advanced molecular technologies, MS-based proteomics has become a cornerstone that is making a significant impact in the post-genomic era and as precision medicine moves from the research laboratory to clinical practice. The global dissemination of COVID-19 has spurred collective efforts to develop effective diagnostics, vaccines, and therapeutic interventions. This chapter highlights how MS seamlessly integrates with established methods such as RT-PCR and ELISA to improve viral identification and disease progression assessment. In particular, matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) takes the center stage, unraveling intricate details of SARS-CoV-2 proteins, revealing modifications such as glycosylation, and providing insights critical to formulating therapies and assessing prognosis. However, high-throughput analysis of MALDI data presents challenges in manual interpretation, which has driven the development of programmatic pipelines and specialized packages such as MALDIquant. As we move forward, it becomes clear that integrating proteomic data with various omic findings is an effective strategy to gain a comprehensive understanding of the intricate biology of COVID-19 and ultimately develop targeted therapeutic paradigms.


Asunto(s)
COVID-19 , Proteómica , Humanos , Proteómica/métodos , COVID-19/diagnóstico , SARS-CoV-2 , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Proteínas , Prueba de COVID-19
4.
Reprod Domest Anim ; 59(3): e14543, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38459831

RESUMEN

This study aims to investigate the effects of melatonin on follicular growth, viability and ultrastructure, as well as on the levels of mRNA for antioxidant enzymes, reactive oxygen species (ROS) and meiotic progression in oocytes from in vitro cultured bovine early antral follicles. To this end, isolated early antral follicles (500-600 µm) were cultured in TCM-199+ alone or supplemented with 10-6 , 10-7 or 10-8 M melatonin at 38.5°C with 5% CO2 for 8 days. Follicle diameters were evaluated at days 0, 4 and 8 of culture. At the end of culture, ultrastructure, chromatin configuration, viability (calcein-AM and ethidium homodimer-1 staining), and the levels of ROS and mRNA for catalase (CAT), superoxide dismutase (SOD) and peroxiredoxin 6 (PRDX6) and glutathione peroxidase (GPx) were investigated in oocyte-granulosa cell complexes (OGCs). The results showed that early antral follicles cultured with 10-6 and 10-8 M melatonin had a progressive and significant increase in their diameters throughout the culture period (p < .05). Additionally, oocytes from follicles cultured with 10-7 or 10-8 M melatonin had increased fluorescence for calcein-AM, while those cultured with 10-6 or 10-7 M had reduced fluorescence for ethidium homodimer-1. Different from follicles cultured in other treatments, those cultured with 10-8 M melatonin had well-preserved ultrastructure of oocyte and granulosa cells. Melatonin, however, did not influence the levels of ROS, the mitochondrial activity, oocyte meiotic resumption and expression mRNA for SOD, CAT, GPX1 and PRDX6. In conclusion, the presence of 10-8 M melatonin in culture medium improves viability and preserves the ultrastructure of oocyte and granulosa cells of early antral follicles cultured in vitro.


Asunto(s)
Fluoresceínas , Melatonina , Femenino , Animales , Bovinos , Melatonina/farmacología , Melatonina/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Oocitos , Superóxido Dismutasa , ARN Mensajero/metabolismo
5.
J Sci Food Agric ; 104(5): 2660-2668, 2024 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-37985208

RESUMEN

BACKGROUND: Coffee cultivation and agroindustry generate residues that are rich in several metabolites. These compounds, such as phenolic compounds and alkaloids, are known for their antioxidant activity and are usually consumed as nutraceuticals. The purpose of this study was to evaluate the occurrence of chemical and antioxidant components of low-pruned coffee stems under different fertilizer regimes. Extractives and lignin composition, histochemical, chromatographic, and antioxidant analyses were performed. RESULTS: Multiple compounds were found to accumulate in the stems of coffee trees. Furthermore, the presence of phenolic compounds such as chlorogenic acid, vanillin, resveratrol, and the alkaloids caffeine and trigonelline varied depending on the type of fertilization. In all samples examined, optimal performance was observed at the highest tested concentration (500 µg mL-1 ). All samples analyzed presented a great performance at the highest concentration tested (500 µg mL-1 ), with the dose 70% and the dose 100%, which is the recommended for the culture, showing the highest values for most of the concentrations and the best half-maximal inhibitory concentration (IC50 ) when compared with the other samples tested. CONCLUSION: As shown in the results, the reuse of stem residues as antioxidant material, with the potential to be profitable, and has the added benefit of providing a sustainable destination for material that until now has been underutilized. © 2023 Society of Chemical Industry.


Asunto(s)
Alcaloides , Coffea , Antioxidantes/química , Alcaloides/análisis , Cafeína/análisis , Dieta , Suplementos Dietéticos/análisis , Ácido Clorogénico/análisis , Fenoles/análisis , Coffea/química
6.
Brain ; 145(2): 655-669, 2022 04 18.
Artículo en Inglés | MEDLINE | ID: mdl-34410345

RESUMEN

GM1 gangliosidosis is a fatal neurodegenerative disease caused by a deficiency of lysosomal ß-galactosidase. In its most severe form, GM1 gangliosidosis causes death by 4 years of age, and no effective treatments exist. Previous work has shown that injection of the brain parenchyma with an adeno-associated viral (AAV) vector provides pronounced therapeutic benefit in a feline GM1 model. To develop a less invasive treatment for the brain and increase systemic biodistribution, intravenous injection of AAV9 was evaluated. AAV9 expressing feline ß-galactosidase was intravenously administered at 1.5×1013 vector genomes/kg body weight to six GM1 cats at ∼1 month of age. The animals were divided into two cohorts: (i) a long-term group, which was followed to humane end point; and (ii) a short-term group, which was analysed 16 weeks post-treatment. Clinical assessments included neurological exams, CSF and urine biomarkers, and 7 T MRI and magentic resonance spectroscopy (MRS). Post-mortem analysis included ß-galactosidase and virus distribution, histological analysis and ganglioside content. Untreated GM1 animals survived 8.0 ± 0.6 months while intravenous treatment increased survival to an average of 3.5 years (n = 2) with substantial improvements in quality of life and neurological function. Neurological abnormalities, which in untreated animals progress to the inability to stand and debilitating neurological disease by 8 months of age, were mild in all treated animals. CSF biomarkers were normalized, indicating decreased CNS cell damage in the treated animals. Urinary glycosaminoglycans decreased to normal levels in the long-term cohort. MRI and MRS showed partial preservation of the brain in treated animals, which was supported by post-mortem histological evaluation. ß-Galactosidase activity was increased throughout the CNS, reaching carrier levels in much of the cerebrum and normal levels in the cerebellum, spinal cord and CSF. Ganglioside accumulation was significantly reduced by treatment. Peripheral tissues such as heart, skeletal muscle, and sciatic nerve also had normal ß-galactosidase activity in treated GM1 cats. GM1 histopathology was largely corrected with treatment. There was no evidence of tumorigenesis or toxicity. Restoration of ß-galactosidase activity in the CNS and peripheral organs by intravenous gene therapy led to profound increases in lifespan and quality of life in GM1 cats. These data support the promise of intravenous gene therapy as a safe, effective treatment for GM1 gangliosidosis.


Asunto(s)
Gangliosidosis GM1 , Enfermedades Neurodegenerativas , Animales , Biomarcadores , Gatos , Dependovirus/genética , Gangliósido G(M1)/uso terapéutico , Gangliósidos , Gangliosidosis GM1/genética , Gangliosidosis GM1/patología , Gangliosidosis GM1/terapia , Terapia Genética/métodos , Humanos , Calidad de Vida , Distribución Tisular , beta-Galactosidasa/genética , beta-Galactosidasa/metabolismo
7.
Sensors (Basel) ; 23(8)2023 Apr 13.
Artículo en Inglés | MEDLINE | ID: mdl-37112314

RESUMEN

Robust methods to compute tissue displacements in optical coherence elastography (OCE) data are paramount, as they play a significant role in the accuracy of tissue elastic properties estimation. In this study, the accuracy of different phase estimators was evaluated on simulated OCE data, where the displacements can be accurately set, and on real data. Displacement (∆d) estimates were computed from (i) the original interferogram data (Δφori) and two phase-invariant mathematical manipulations of the interferogram: (ii) its first-order derivative (Δφd) and (iii) its integral (Δφint). We observed a dependence of the phase difference estimation accuracy on the initial depth location of the scatterer and the magnitude of the tissue displacement. However, by combining the three phase-difference estimates (Δdav), the error in phase difference estimation could be minimized. By using Δdav, the median root-mean-square error associated with displacement prediction in simulated OCE data was reduced by 85% and 70% in data with and without noise, respectively, in relation to the traditional estimate. Furthermore, a modest improvement in the minimum detectable displacement in real OCE data was also observed, particularly in data with low signal-to-noise ratios. The feasibility of using Δdav to estimate agarose phantoms' Young's modulus is illustrated.


Asunto(s)
Diagnóstico por Imagen de Elasticidad , Tomografía de Coherencia Óptica , Tomografía de Coherencia Óptica/métodos , Diagnóstico por Imagen de Elasticidad/métodos , Módulo de Elasticidad , Fantasmas de Imagen
8.
Trop Anim Health Prod ; 55(3): 217, 2023 May 22.
Artículo en Inglés | MEDLINE | ID: mdl-37212890

RESUMEN

This study aimed to evaluate the thermoregulatory capacity and performance of Saanen goat kids from birth to weaning in a hot climate. Twelve newborn males and female goat kids with an initial body weight of 4.17 ± 0.81 kg were used. Physiological responses, climatic variables, and biometric traits data were collected. Univariate and multivariate analysis techniques were used. Heart rate (HR) was high up to the 6th week of life, with a reduction from the 7th week on (P < 0.001). Rectal temperature (RT) was lower in the first 2 weeks (P < 0.001), with an increase and stabilization occurring in the 7th and 8th weeks. Coat surface temperature (ST) was more activated from the 5th week onwards (P < 0.001). Body weight (BW) and withers height (WH) were higher in later weeks of the calving phase with a linear effect (P < 0.001). The first principal component demonstrated the relationship of sensible heat dissipation × body area of the goat kids; the second component shows the relationship of meteorological data with RT, having a positive relationship between RT with RH and negative with AT, and the third component points to the association of RR and HR. Of the animals, 81.3% were correctly classified in their group of origin in discriminant canonical analysis, with emphasis on the classification of the kids in the 1st-2nd and 3rd-4th weeks (classification percentage Æ© = 95.8%). It is concluded that (i) newborn kids activate latent mechanisms to maintain their homoeothermic during the first 2 weeks of life, and as they grow, they use sensitive heat loss processes, especially from the 5th week of life onwards and (ii) male and female goats do not show sexual dimorphism effect on body performance and body morphometric measurements up to 60 days of life.


Asunto(s)
Cabras , Parto , Embarazo , Masculino , Femenino , Animales , Destete , Cabras/fisiología , Peso Corporal , Clima
9.
J Phys Chem A ; 126(13): 2082-2090, 2022 Apr 07.
Artículo en Inglés | MEDLINE | ID: mdl-35345882

RESUMEN

Computational modeling at the DLPNO-CCSD(T)/CBS//M06-L/def2-TZVP level of theory was used to propose four different iron catalysts whose structures were inspired on the [Fe]-hydrogenase active site: [Fe(MePNNHNP)(acmp)] (C(1), MePNNHNP = 2,6-bis(dimethylphosphine), acmp = acylmethylpyridine), [Fe(CNNHNC)(acmp)] (C(2), CNNHNC = 2,6-bis(methylimidazol-2-ylidene)), [Fe(MePNNNP)(acmp)] (D(1), MePNNNP = 2,6-bis((dimethylphosphine)pyridine)), and [Fe(CNNNC)(acmp)] (D(2), CNNNC = 2,6-bis((methylimidazol-2-ylidene) pyridine)). Through these electronic structure calculations, the catalytic mechanism of the reaction was explored. The intermediates and transition states present along the reaction coordinate were identified and described as to their equilibrium geometries, vibrational frequencies, and energies. Quasi-harmonic corrections were performed considering conditions analogous to those used experimentally. To compare the catalytic activities of the studied catalysts, turnover frequencies (TOFs) were calculated. Based on the explored catalytic cycles and TOF values (D(1) > C(1) > D(2) > C(2)), the most suitable iron catalysts are those with tridentate phosphine pincer-type ligands coordinated to the metal center. These systems are new promising iron catalysts to promote the CO2 hydrogenation to formic acid without any use of bases or additives.


Asunto(s)
Materiales Biomiméticos , Dióxido de Carbono , Hidrogenasas , Materiales Biomiméticos/química , Dióxido de Carbono/química , Catálisis , Hidrogenasas/química , Hidrogenación , Proteínas Hierro-Azufre/química
10.
J Water Health ; 20(12): 1733-1747, 2022 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-36573676

RESUMEN

This study assessed the bacterial community composition of a drinking water system (DWS) serving a mid-sized city (120,000 inhabitants) in Brazil. Water samples, including raw and treated water, were collected at seven points throughout the DWS. DNA was extracted and analysed using high-throughput sequencing (Ion Torrent). Free chlorine and turbidity were measured in situ. Results showed that the highest relative abundance of 16S rRNA genes was from phyla Proteobacteria, followed by Bacteroidetes and Actinobacteria. The next most abundant phylum was Cyanobacteria, represented by Arthronema, Calothrix, and Synechococcus. An interesting observation was that the DNA-based analysis suggested a bacterial community change in the distribution network, with treated reservoir water being very different from the network samples. This suggests active microbiology within the distribution network and a tendency for bacterial diversity to decrease after chlorine disinfection but increase after pipeline distribution. In raw water, a predominance of Proteobacteria was observed with reduced Cyanobacteria, showing a negative correlation. In treated water, Proteobacteria were negatively correlated with Bacteroidetes. Finally, 16S rRNA genes from Firmicutes (especially Staphylococcus) had a high abundance in the chlorinated water, which may indicate the phylum's resistance to chlorine residuals. Opportunistic pathogens, e.g., Mycobacteria, Legionella, and Staphylococcus, were also observed.


Asunto(s)
Cianobacterias , Agua Potable , Agua Potable/microbiología , Cloro/farmacología , ARN Ribosómico 16S/genética , Brasil , Proteobacteria/genética , Cianobacterias/genética , Bacteroidetes/genética , Abastecimiento de Agua
11.
Zygote ; 30(3): 365-372, 2022 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-34851249

RESUMEN

In vitro culture of ovarian tissue containing primordial follicles is an important tool to study the initiation of follicular populations and to develop efficient culture systems to support in vitro follicle growth. Considering that in vitro culture favours oxidative stress, it is very important to supplement culture medium with antioxidant substances such as Aloe vera extract. This study aims to evaluate the effects of different concentrations of Aloe vera on the distribution of collagen fibres in the extracellular matrix, follicular activation, development and survival in bovine ovarian cortical tissues cultured in vitro, as well as on expression of mRNAs for antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT), peroxiredoxin 6 (PRDX6) and glutathione peroxidase 1 (GPX1)]. To this end, ovarian cortical tissues were cultured for 6 days in α-MEM alone or supplemented with different concentrations of Aloe vera extract (1.0, 5.0, 10.0 or 50.0%). After culture, fragments were fixed and processed histologically to evaluate follicular morphology and activation, as well as the extracellular matrix by staining with picrosirius red. The levels of mRNA for SOD, CAT, PRDX6 and GPX1 in cultured ovarian tissues were evaluated by real-time polymerase chain reaction (PCR). Ovarian tissues cultured with 10.0 or 50.0% Aloe vera had higher percentages of collagen fibres than tissues cultured in control medium. A significant increase in developing follicles was observed in ovarian tissues cultured in α-MEM alone or supplemented with 10% Aloe vera when compared with fresh control or tissues cultured with 1.0% Aloe vera. Presence of Aloe vera did not influence the percentage of morphologically normal follicles when compared with control medium. Ovarian tissues cultured with 50.0% Aloe vera had higher percentages of morphologically normal follicles than those cultured with 10.0% Aloe vera. Furthermore, 10% Aloe vera significantly increased mRNA levels for PRDX6. In conclusion, 10.0% Aloe vera improves extracellular matrix distribution in cultured tissues and increases the expression of mRNA for PRDX6 after 6 days in vitro.


Asunto(s)
Aloe , Aloe/genética , Animales , Antioxidantes , Bovinos , Colágeno/genética , Matriz Extracelular , Peroxiredoxina VI , Extractos Vegetales , ARN Mensajero/genética , Superóxido Dismutasa , Técnicas de Cultivo de Tejidos
12.
Zygote ; 30(6): 882-890, 2022 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-36148786

RESUMEN

This study aims to evaluate the effects of N-acetylcysteine (NAC) on bovine oocyte maturation, mitochondrial activity and transzonal projections (TZP), as well as on the levels of reactive oxygen species (ROS) and messenger RNA (mRNA) for catalase (CAT) superoxide dismutase (SOD), periredoxin-6 (Prdx6), glutathione peroxidase (GPx), growth and differentiation factor-9 (GDF9), histone H1Foo, cyclin B1 (CCNB1) and c-Mos. Bovine cumulus-oocyte complexes (COC) of medium-sized antral follicles (3.0-6.0 mm) were prematured in TCM-199 for 8 h at 38.5°C in 5% CO2. After prematuration in the presence of forskolin and C-type natriuretic peptide, COCs were matured in TCM-199 alone or with 0.1, 0.5 or 2.5 mM NAC. Then, oocytes were classified according to the stage of chromatin. Furthermore, mitochondrial activity and intracellular levels of ROS and TZP were also evaluated. The levels of mRNAs for CAT, SOD, Prdx6, GPx, GDF9, H1Foo, CCNB1 and c-Mos were evaluated using real-time polymerase chain reaction (RT-PCR). The results showed that NAC significantly increased the percentages of oocytes with resumption of meiosis when compared with those oocytes matured in control medium. Oocytes had homogeneous mitochondrial distribution, and those cultured with 0.1 and 0.5 mM NAC had lower levels of ROS when compared with the control. In addition, 0.5 mM NAC reduced TZP and the levels of mRNA for CCNB1. In contrast, NAC did not influence the expression of CAT, GPx, Prdx6, SOD, GDF9, H1Foo, and c-Mos. In conclusion, 0.5 mM NAC reduced the levels of ROS, TZP and mRNA for CCNB1, and improved in vitro resumption of meiosis in oocytes from medium-sized bovine antral follicles.


Asunto(s)
Acetilcisteína , Técnicas de Maduración In Vitro de los Oocitos , Bovinos , Animales , Técnicas de Maduración In Vitro de los Oocitos/métodos , Especies Reactivas de Oxígeno/metabolismo , Acetilcisteína/farmacología , Acetilcisteína/metabolismo , Oocitos , Meiosis , Superóxido Dismutasa/metabolismo , Glutatión Peroxidasa/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo
13.
Sensors (Basel) ; 22(24)2022 Dec 11.
Artículo en Inglés | MEDLINE | ID: mdl-36560071

RESUMEN

Two-photon imaging (TPI) microscopy, namely, two-photon excited fluorescence (TPEF), fluorescence lifetime imaging (FLIM), and second-harmonic generation (SHG) modalities, has emerged in the past years as a powerful tool for the examination of biological tissues. These modalities rely on different contrast mechanisms and are often used simultaneously to provide complementary information on morphology, metabolism, and structural properties of the imaged tissue. The cornea, being a transparent tissue, rich in collagen and with several cellular layers, is well-suited to be imaged by TPI microscopy. In this review, we discuss the physical principles behind TPI as well as its instrumentation. We also provide an overview of the current advances in TPI instrumentation and image analysis. We describe how TPI can be leveraged to retrieve unique information on the cornea and to complement the information provided by current clinical devices. The present state of corneal TPI is outlined. Finally, we discuss the obstacles that must be overcome and offer perspectives and outlooks to make clinical TPI of the human cornea a reality.


Asunto(s)
Colágeno , Córnea , Humanos , Córnea/diagnóstico por imagen , Microscopía , Procesamiento de Imagen Asistido por Computador , Imagen Óptica , Microscopía de Fluorescencia por Excitación Multifotónica/métodos
14.
Phys Chem Chem Phys ; 23(16): 9980-9990, 2021 Apr 28.
Artículo en Inglés | MEDLINE | ID: mdl-33870397

RESUMEN

In this work the H2 adsorption at a Cu(i)-SSZ-13 exchanged zeolite was theoretically investigated. A systematic cluster approach was used and different density functionals (B3LYP, B3LYP-D3(BJ), M06L, PBE, PBE-D3(BJ) and ωB97XD) and a def2-SVP basis set were benchmarked. In order to select the best approach to the H2 adsorption over a Cu(i)-SSZ-13 cluster with 78 atoms (16 T-sites), two main tasks were performed: (1) a comparison between theoretical and experimental structures and (2) a comparison between theoretical and experimental adsorption enthalpies. By employing the most suitable functional - the ωB97X-D - the H2 interaction with the zeolite structure was studied by means of NBO, NCI, AIM and DLPNO-CCSD(T)/LED analyses.

15.
Mol Ther ; 28(10): 2150-2160, 2020 10 07.
Artículo en Inglés | MEDLINE | ID: mdl-32592687

RESUMEN

The GM2 gangliosidoses, Tay-Sachs disease (TSD) and Sandhoff disease (SD), are fatal lysosomal storage disorders caused by mutations in the HEXA and HEXB genes, respectively. These mutations cause dysfunction of the lysosomal enzyme ß-N-acetylhexosaminidase A (HexA) and accumulation of GM2 ganglioside (GM2) with ensuing neurodegeneration, and death by 5 years of age. Until recently, the most successful therapy was achieved by intracranial co-delivery of monocistronic adeno-associated viral (AAV) vectors encoding Hex alpha and beta-subunits in animal models of SD. The blood-brain barrier crossing properties of AAV9 enables systemic gene therapy; however, the requirement of co-delivery of two monocistronic AAV vectors to overexpress the heterodimeric HexA protein has prevented the use of this approach. To address this need, we developed multiple AAV constructs encoding simultaneously HEXA and HEXB using AAV9 and AAV-PHP.B and tested their therapeutic efficacy in 4- to 6-week-old SD mice after systemic administration. Survival and biochemical outcomes revealed superiority of the AAV vector design using a bidirectional CBA promoter with equivalent dose-dependent outcomes for both capsids. AAV-treated mice performed normally in tests of motor function, CNS GM2 ganglioside levels were significantly reduced, and survival increased by >4-fold with some animals surviving past 2 years of age.


Asunto(s)
Dependovirus/genética , Terapia Genética , Vectores Genéticos/genética , Enfermedad de Sandhoff/terapia , Animales , Manejo de la Enfermedad , Modelos Animales de Enfermedad , Gangliósido G(M2)/metabolismo , Expresión Génica , Predisposición Genética a la Enfermedad , Terapia Genética/métodos , Vectores Genéticos/administración & dosificación , Ratones , Mutación , Enfermedad de Sandhoff/genética , Enfermedad de Tay-Sachs/genética , Enfermedad de Tay-Sachs/metabolismo , Enfermedad de Tay-Sachs/terapia , Transgenes , beta-N-Acetilhexosaminidasas/genética , beta-N-Acetilhexosaminidasas/metabolismo
16.
Mol Ther ; 28(2): 411-421, 2020 02 05.
Artículo en Inglés | MEDLINE | ID: mdl-31813800

RESUMEN

Global gene delivery to the CNS has therapeutic importance for the treatment of neurological disorders that affect the entire CNS. Due to direct contact with the CNS, cerebrospinal fluid (CSF) is an attractive route for CNS gene delivery. A safe and effective route to achieve global gene distribution in the CNS is needed, and administration of genes through the cisterna magna (CM) via a suboccipital puncture results in broad distribution in the brain and spinal cord. However, translation of this technique to clinical practice is challenging due to the risk of serious and potentially fatal complications in patients. Herein, we report development of a gene therapy delivery method to the CM through adaptation of an intravascular microcatheter, which can be safely navigated intrathecally under fluoroscopic guidance. We examined the safety, reproducibility, and distribution/transduction of this method in sheep using a self-complementary adeno-associated virus 9 (scAAV9)-GFP vector. This technique was used to treat two Tay-Sachs disease patients (30 months old and 7 months old) with AAV gene therapy. No adverse effects were observed during infusion or post-treatment. This delivery technique is a safe and minimally invasive alternative to direct infusion into the CM, achieving broad distribution of AAV gene transfer to the CNS.


Asunto(s)
Cisterna Magna/metabolismo , Dependovirus/genética , Expresión Génica , Técnicas de Transferencia de Gen , Vectores Genéticos/genética , Transducción Genética , Animales , Catéteres , Sistema Nervioso Central/metabolismo , Genes Reporteros , Terapia Genética , Vectores Genéticos/administración & dosificación , Humanos , Inyecciones Espinales , Imagen por Resonancia Magnética , Modelos Animales , Ovinos , Cirugía Asistida por Computador , Tomografía Computarizada por Rayos X , Transgenes , Grabación en Video
17.
J Hum Nutr Diet ; 34(4): 758-767, 2021 08.
Artículo en Inglés | MEDLINE | ID: mdl-33955072

RESUMEN

INTRODUCTION: Patients undergoing haemodialysis are at great cardiovascular risk. Neck circumference (NC) is a simple and low-cost measure for estimating this risk precociously. The present study aimed to evaluate whether the cardiovascular risk obtained by NC is associated with the main cardiometabolic risk factors in patients on haemodialysis treatment who are on a waiting list for transplantation. METHODS: A cross-sectional study was conducted including 96 patients in a single transplantation centre. Socio-demographic, clinical-laboratory and anthropometric data were collected. NC was considered as a dependent variable and the independent variables were body mass index (BMI), fasting glycaemia and lipid profile, triglyceride and high-density lipoprotein-cholesterol (TGL/HDL-C) ratio, and triglyceride and glycaemia (TyG) index. For the comparison of averages, we used Mann-Whitney and Student's t tests, as well as one-way analysis of variance and Kruskal-Wallis tests. Bivariate and multivariate logistic regression was performed for the association between NC and cardiometabolic risk factors. p < 0.05 was considered statistically significant. RESULTS: There were higher BMI averages, blood glucose, triglycerides (TGL), TGL/HDL-C ratio and TyG index in the tertile 3 of the NC, whereas the HDL-C decreased as the tertile increased. There was a statistically significant risk of cardiovascular disease that was asscoiated, according to NC, with being overweight, high levels of TGL, TGL/HDL-C, TyG index and low HDL-C. CONCLUSIONS: NC is shown to be associated with cardiometabolic risk factors in kidney patients undergoing haemodialysis who are on a transplant waiting list.


Asunto(s)
Pesos y Medidas Corporales , Factores de Riesgo Cardiometabólico , Factores de Riesgo de Enfermedad Cardiaca , Cuello/anatomía & histología , Diálisis Renal , Insuficiencia Renal Crónica/terapia , Adiposidad , Adulto , Anciano , Índice de Masa Corporal , Brasil/epidemiología , Estudios Transversales , Femenino , Hospitales Universitarios , Humanos , Trasplante de Riñón , Masculino , Persona de Mediana Edad , Listas de Espera
18.
An Acad Bras Cienc ; 93(1): e20181091, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33759949

RESUMEN

The objective of this study was to evaluate the subjective, chemical and sensorial meat characteristics of ½ Santa Inês (SI) x ½ No Defined Racial Standard (NDRS) and ½ Brazilian Somalis (BS) x ½ No Defined Racial Standard (NDRS) crossbred lambs, finished in confinement. Sixteen uncastrated male lambs with initial weight of 19.7 ± 2.03 kg and approximately 90 days of age. A randomized block design was used, with blocks represented by the initial weight of each genetic group, with eight animals per group. There was a higher degree and distribution of marbling, percentage of lipids and meat color for ½ BS x ½ NDRS lambs. The conjugated linoleic acid profile was higher for ½ SI x ½ NDRS lambs. Considering the meat quality of the evaluated genetic groups, Santa Inês crossbred lambs have a better nutritional value for meat, especially taking into account the production of foods that are beneficial to human health.


Asunto(s)
Composición Corporal , Oveja Doméstica , Alimentación Animal/análisis , Animales , Brasil , Humanos , Masculino , Carne/análisis , Ovinos/genética , Oveja Doméstica/genética , Somalia
19.
Water Sci Technol ; 84(10-11): 2903-2912, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34850702

RESUMEN

The effects of depth and climate seasonality on zooplankton, algal biomass, coliforms and Escherichia coli in a small full-scale shallow maturation pond receiving pre-treated domestic wastewater were evaluated during a tropical climatic seasonal cycle. The experiment revealed that the zooplankton community was dominated by rotifers and protozoans, and concentrations were influenced by seasonality. A negative correlation between zooplankton, and pH, dissolved oxygen, temperature and ultraviolet radiation, and chlorophyll-a and Escherichia coli were observed at all depths. The major driving forces influencing Escherichia coli were pH, dissolved oxygen, ultraviolet radiation and the zooplankton. A significant difference between Escherichia coli removal throughout the three different depths were observed. Both bacterial and zooplankton concentrations were greater closer to the bottom of the pond, therefore reinforcing the integral role of solar radiation on bacterial removal. These results give an insight on the dynamics of these groups in pond systems treating domestic wastewater, by correlating the variation of zooplankton with biotic and abiotic variables and seasonal changes in a tropical climate, where few studies have been performed on this topic.


Asunto(s)
Escherichia coli , Estanques , Animales , Biomasa , Clima Tropical , Rayos Ultravioleta , Zooplancton
20.
Phys Chem Chem Phys ; 22(29): 16943-16948, 2020 Aug 07.
Artículo en Inglés | MEDLINE | ID: mdl-32672262

RESUMEN

We report a computational study of the mechanism and determination of the rate constants of the Fe + CO2→ FeO + CO reaction, in the 1000-3000 K temperature range, at the CCSD(T)/CBS//B3LYP/def2-TZVP level of theory. The overall rate constant was obtained by a Kinetic Monte Carlo simulation. The calculated rate constant, at 2000 K, is 9.72 × 10-13 cm3 molecule-1 s-1, in agreement with experimental measurements: 2.97 × 10-13 cm3 molecule-1 s-1 [A. Giesen et al., Phys. Chem. Chem. Phys., 2002, 4, 3665] and 1.13 × 10-13 cm3 molecule-1 s-1 [V. N. Smirnov, Kinet. Catal., 2008, 49, 607]. Our study shows that this reaction follows a complex mechanism, with multiple reaction paths contributing to the overall rate, and that CCSD(T) accurately describes this transition metal reaction.

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