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1.
Proc Natl Acad Sci U S A ; 121(25): e2401159121, 2024 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-38865261

RESUMO

Trichomonas vaginalis, a common sexually transmitted parasite that colonizes the human urogenital tract, secretes extracellular vesicles (TvEVs) that are taken up by human cells and are speculated to be taken up by parasites as well. While the crosstalk between TvEVs and human cells has led to insight into host:parasite interactions, roles for TvEVs in infection have largely been one-sided, with little known about the effect of TvEV uptake by T. vaginalis. Approximately 11% of infections are found to be coinfections of multiple T. vaginalis strains. Clinical isolates often differ in their adherence to and cytolysis of host cells, underscoring the importance of understanding the effects of TvEV uptake within the parasite population. To address this question, our lab tested the ability of a less adherent strain of T. vaginalis, G3, to take up fluorescently labeled TvEVs derived from both itself (G3-EVs) and TvEVs from a more adherent strain of the parasite (B7RC2-EVs). Here, we showed that TvEVs generated from the more adherent strain are internalized more efficiently compared to the less adherent strain. Additionally, preincubation of G3 parasites with B7RC2-EVs increases parasite aggregation and adherence to host cells. Transcriptomics revealed that TvEVs up-regulate expression of predicted parasite membrane proteins and identified an adherence factor, heteropolysaccharide binding protein (HPB2). Finally, using comparative proteomics and superresolution microscopy, we demonstrated direct transfer of an adherence factor, cadherin-like protein, from TvEVs to the recipient parasite's surface. This work identifies TvEVs as a mediator of parasite:parasite communication that may impact pathogenesis during mixed infections.


Assuntos
Vesículas Extracelulares , Trichomonas vaginalis , Vesículas Extracelulares/metabolismo , Trichomonas vaginalis/metabolismo , Trichomonas vaginalis/genética , Humanos , Interações Hospedeiro-Parasita , Regulação para Cima , Adesão Celular , Feminino , Proteínas de Protozoários/metabolismo , Proteínas de Protozoários/genética
2.
Proc Natl Acad Sci U S A ; 119(38): e2123117119, 2022 09 20.
Artigo em Inglês | MEDLINE | ID: mdl-36099298

RESUMO

Acinetobacter baumannii is a clinically important, predominantly health care-associated gram-negative bacterium with high rates of emerging resistance worldwide. Given the urgent need for novel antibacterial therapies against A. baumannii, we focused on inhibiting lipoprotein biosynthesis, a pathway that is essential for envelope biogenesis in gram-negative bacteria. The natural product globomycin, which inhibits the essential type II signal peptidase prolipoprotein signal peptidase (LspA), is ineffective against wild-type A. baumannii clinical isolates due to its poor penetration through the outer membrane. Here, we describe a globomycin analog, G5132, that is more potent against wild-type and clinical A. baumannii isolates. Mutations leading to G5132 resistance in A. baumannii map to the signal peptide of a single hypothetical gene, which we confirm encodes an alanine-rich lipoprotein and have renamed lirL (prolipoprotein signal peptidase inhibitor resistance lipoprotein). LirL is a highly abundant lipoprotein primarily localized to the inner membrane. Deletion of lirL leads to G5132 resistance, inefficient cell division, increased sensitivity to serum, and attenuated virulence. Signal peptide mutations that confer resistance to G5132 lead to the accumulation of diacylglyceryl-modified LirL prolipoprotein in untreated cells without significant loss in cell viability, suggesting that these mutations overcome a block in lipoprotein biosynthetic flux by decreasing LirL prolipoprotein substrate sensitivity to processing by LspA. This study characterizes a lipoprotein that plays a critical role in resistance to LspA inhibitors and validates lipoprotein biosynthesis as a antibacterial target in A. baumannii.


Assuntos
Acinetobacter baumannii , Antibacterianos , Ácido Aspártico Endopeptidases , Proteínas de Bactérias , Farmacorresistência Bacteriana , Furanos , Deleção de Genes , Lipoproteínas , Inibidores de Proteases , Piridinas , Acinetobacter baumannii/efeitos dos fármacos , Acinetobacter baumannii/enzimologia , Acinetobacter baumannii/genética , Antibacterianos/farmacologia , Ácido Aspártico Endopeptidases/genética , Proteínas de Bactérias/genética , Farmacorresistência Bacteriana/genética , Furanos/farmacologia , Lipoproteínas/biossíntese , Lipoproteínas/genética , Peptídeos/farmacologia , Inibidores de Proteases/farmacologia , Sinais Direcionadores de Proteínas/genética , Piridinas/farmacologia
3.
J Am Chem Soc ; 146(7): 4346-4350, 2024 02 21.
Artigo em Inglês | MEDLINE | ID: mdl-38346011

RESUMO

In situ modification of glycans requires extraordinary molecular recognition of highly complex and subtly different carbohydrates, followed by reactions at precise locations on the substrate. We here report synthetic catalysts that under physiological conditions cleave a predetermined oligosaccharide block such as a branched trimannose or the entire N-glycan of a glycoprotein, while nontargeted glycoproteins stay intact. The method also allows α2-6-sialylated galactosides to be removed preferentially over the α2-3-linked ones from cell surfaces, highlighting the potential of these synthetic glycosidases for glycan editing.


Assuntos
Glicoproteínas , Polissacarídeos , Glicoproteínas/química , Polissacarídeos/química , Oligossacarídeos , Glicosídeo Hidrolases
4.
Langmuir ; 40(10): 5106-5120, 2024 03 12.
Artigo em Inglês | MEDLINE | ID: mdl-38427698

RESUMO

The present study focuses on exploring the physical properties of lipid membranes based on the polyhydroxy oxanorbornane (PH-ONB) headgroup, designed as synthetic analogues of naturally occurring archaeal lipid membranes. Specifically, we study two variants of PH-ONB headgroup-based lipids differing in the number of hydroxy groups present in the headgroup, with one having two hydroxy groups (ONB-2OH) and the other having three (ONB-3OH). These lipids form stable bilayer membranes. The study begins with a comprehensive analysis of the fluorescence characteristics of nitrobenzoxadiazole (NBD)-tagged ONB-based lipids in different solvent environments and within a model lipid membrane 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). Subsequently, the physical properties of the ONB-based membranes were examined by using an NBD-tagged ONB-based probe and a commonly used extrinsic 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescent probe. The steady-state and time-resolved fluorescence properties of the NBD-tagged ONB-based probe and DPH were used to compare the physical properties of the ONB-based membranes, including polarity, fluidity, phase transition, order, hydration, location, heterogeneity, and rotational diffusion. The solid gel to liquid crystalline phase transition temperatures of ONB-2OH and ONB-3OH lipid membranes are found to be (68 ± 1) °C and (74 ± 1) °C, respectively. The variation in organization (size), fluidity, and phase transition temperature of ONB-based lipid membranes is explained by the extent of hydrogen bonding interactions between lipid head groups. ONB-based membranes exhibit characteristics similar to those of phospholipid membranes and possess a notably high phase transition temperature. These properties make them a promising and cost-effective synthetic alternative to archaeal lipid membranes with a wide range of potential applications.


Assuntos
Corantes Fluorescentes , Fosfolipídeos , Corantes Fluorescentes/química , Fosfolipídeos/química , Fenômenos Químicos , Temperatura , Transição de Fase , Bicamadas Lipídicas/química , Fosfatidilcolinas/química
5.
Inflamm Res ; 73(5): 877-896, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38555555

RESUMO

BACKGROUND: Urticaria is a clinical condition characterized by the appearance of wheals (hives), angioedema, or both. Over the last several decades, a better understanding of the mechanisms at play in the immunopathogenesis of urticaria has underscored the existence of numerous urticaria subtypes. Separating the different kinds of urticaria explicitly helps find the best detection method for the management of this skin disorder. Subtypes of urticaria also include both spontaneous and physical types. The conventional ones include spontaneous urticaria, constituting both acute and chronic urticaria. Therefore, a broad and effective therapy is essential for the diagnosis and treatment of urticaria. METHODS: To understand the immunopathogenesis of urticaria, various databases, including PubMed, Scopus, and Web of Science, were used to retrieve original articles and reviews related to urticaria. While information on several clinical trials were obtained from clinicaltrials.gov database. RESULTS: This article highlights the immunopathogenesis involved in the intricate interaction between cellular infiltration, immune reactions, coagulation cascades, and autoantibodies that underlie urticaria's pathophysiology. CONCLUSION: The recent progress in understanding urticaria can help to understand the intricate characteristics in the immunopathogenesis of urticaria and could play a beneficial role in the management of urticaria.


Assuntos
Citocinas , Histamina , Urticária , Humanos , Urticária/imunologia , Histamina/imunologia , Citocinas/imunologia , Animais
6.
Mol Biol Rep ; 51(1): 199, 2024 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-38270712

RESUMO

BACKGROUND: Brassica species is the second most important edible oilseed crop in India. Albugo candida (Pers.) Kuntze, a major oomycete disease of oilseed brassica causing white rust, leads to 60% yield loss globally. The prevalence of A. candida race 2 (Ac2V) that specifically infects B. juncea, coupled with limitations of conventional methods has resulted in a dearth of white rust resistance resources in cultivated varieties. METHODS AND RESULTS: In an effort to develop resistant plants, Agrobacterium mediated genetic transformation of three B. juncea genotypes viz., susceptible host var. Varuna, along with its doubled haploid mutant lines C66 and C69 (showing moderate tolerance to field isolates of A. candida) was initiated to transfer resistance genes (WRR8Sf-2 and WRR9Hi-0) identified in Arabidopsis thaliana against race Ac2V, that encode for Toll-like/interleukin-1 receptor-nucleotide binding-leucine-rich repeat proteins that recognize effectors of the pathogen races. CONCLUSIONS: Our results demonstrate that introduction of resistance genes from a tertiary gene pool by genetic transformation enhances disease resistance in B. juncea genotypes to a highly virulent Ac2V isolate.


Assuntos
Arabidopsis , Oomicetos , Mostardeira/genética , Genótipo , Agrobacterium , Arabidopsis/genética , Candida
7.
Phys Chem Chem Phys ; 26(35): 23363-23371, 2024 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-39212031

RESUMO

The atmospheric oxidation of 1,1-dichloro-1,3,3,3-tetrafluoropropane, HCFC-234fb (DTP), leads to the formation of harmful radicals, contributing to stratospheric ozone depletion. Thus, a detailed study of the gas-phase oxidation of the first-generation chlorofluorocarbon alternative HCFC-234fb by a reaction with hydroxyl radicals and chlorine atoms is important to understand its harmful effects on the environment. In this work, we have performed quantum chemical calculations to investigate the thermodynamic and kinetic aspects of the titled reaction. The potential energy profile calculated at the CCSD(T)/aug-cc-pVTZ//MP2/cc-pVTZ level of theory shows that the major reaction pathway involves the abstraction of the H-atom from the central carbon atom, C2, giving rise to the product radical CF3C˙HCCl2F. The calculated rate-coefficients for the reaction with ˙OH and Cl-atoms are 3.89 × 10-15 and 2.54 × 10-17 cm3 molecule-1 s-1, respectively, at 298 K and are in accordance with the experimental rate coefficients. The results suggest that the rate-coefficient for the reaction of DTP with ˙OH is two orders of magnitude higher than that with Cl-atoms, indicating greater significance of the former reaction in the atmosphere. With the rate-coefficient values, the lifetime and the radiative efficiency were calculated to be 8.2 years and 0.246 W m-2 ppb-1, respectively. A 100-year global warming potential (GWP) of 712 was also estimated using the lifetime corrected radiative efficiency value.

8.
Childs Nerv Syst ; 40(1): 253-256, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37470835

RESUMO

BACKGROUND: The prevalence of encephalocele is estimated to be 0.8-5.0 per 10,000 live births. The most frequent encephalocele is the occipital encephalocele. It is a congenital neural tube defect characterized by the protrusion or herniation of intracranial contents through a cranial defect. The term "giant/massive/large encephalocele" is used to describe an encephalocele that is significantly larger than the size of the head. CASE DESCRIPTION: A 2-month-old male infant presented in the neurosurgery outpatient department with one of the largest head masses over the posterior aspect since birth. The swelling was gradually progressive and developed ulceration over the swelling with intermittent cerebrospinal fluid (CSF) discharge but no associated weakness in limbs. Magnetic Resonance Imaging (MRI) brain showed a large occipital meningoencephalocele containing predominantly cyst with part of the cerebellar and occipital lobe. The surgery was planned. The sac contained CSF with the gliotic occipital lobe. The sac and gliotic brain tissue was excised. He had an uneventful postoperative course. CONCLUSION: Surgery serves several functions, including reducing the torque and weight of the head to allow for more normal motor development, removal of the thin, leaking scalp and dural closure to prevent CSF leak and subsequent infection, and improving the cosmetic and social issues that the child and family may have to endure.


Assuntos
Encefalocele , Defeitos do Tubo Neural , Lactente , Criança , Humanos , Masculino , Encefalocele/complicações , Encefalocele/diagnóstico por imagem , Encefalocele/cirurgia , Imageamento por Ressonância Magnética , Cerebelo , Neuroimagem , Gliose
9.
Pediatr Cardiol ; 2024 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-38777858

RESUMO

Right ventricular (RV) afterload due to elevated pulmonary arterial (PA) pressure in pulmonary hypertension (PH) causes long-term right atrial (RA) remodeling and dysfunction. RA function has been shown to correlate with PA pressures and outcome in both adult and pediatric patients with PH. We studied the role of RA strain in estimating PA pressures in congenital heart disease (CHD)-associated PH. Children below 12 years undergoing elective repair of CHD with left-to-right shunts and echocardiographic evidence of PH were included. RA reservoir, conduit and contractile strain along with conventional measures of RV function and PA pressure were measured using transthoracic echocardiography after induction of anaesthesia. Pre-and post-repair invasive PA pressures were measured after surgical exposure. 51 children with a median age of 24 months (range 4-144 months) were included, most of whom were undergoing VSD closure. Contractile RA strain showed good correlation with pre-repair systolic PA pressure in mmHg (r = 0.59, 95%CI 0.37-0.75) or expressed as a percentage of SBP (r = 0.67, 95%CI 0.49-0.80). It also predicted persistent postoperative PH as well as pre-repair pulmonary artery acceleration time and right ventricular systolic pressure measured from tricuspid regurgitation jet. The trends of correlation observed suggest a possible prognostic role of RA strain in ACHD with PH and potential utility in its echocardiographic assessment. The observed findings merit deeper evaluation in larger cohorts.

10.
Int J Biometeorol ; 68(7): 1397-1409, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38602552

RESUMO

Selection of the most stably expressed reference genes is key to monitoring accurate target gene expression across any tissue or cell type. The mRNA in spermatozoa stores valuable information related to changes in spermatogenesis due to variations in environmental conditions, especially during heat stress, which affects various sperm functions. Semen quality in buffalo bulls is significantly influenced by the seasons. In the study, a panel of nine genes was evaluated to identify the most stably expressed internal control gene (ICG) for the normalization of real-time gene expression data generated across various seasons for Murrah buffalo bulls' spermatozoa. Sperm cells were purified from the semen samples collected during different seasons, with temperature-humidity index (THI) ranging from 80.80 ± 1.47 (hot summer) to 55.88 ± 1.98 (winter), using the BoviPure™ gradient purification method. The RNA isolated from the purified spermatozoa fraction was quality checked prior to reverse transcription and subjected to qPCR (quantitative real-time PCR) based expression analysis. An automated 'endoGene' pipeline was employed to apply the geNorm, NormFinder, and BestKeeper algorithms for data analysis. The result indicated that GAPDH and PP1A were the most stably expressed among the gene panel, whereas ATPSF1 and ACTB were the two least stable expressed reference genes. Further, the most suitable ICGs identified were validated by normalization of real time expression data of heat stress and sperm quality genes, HSFY2 and AKAP4, respectively. The genes identified would help in generating the most reliable results for the expression profiling of the genes dictating sperm quality and heat stress cope-up mechanism in buffalo spermatozoa, collected during different seasons.


Assuntos
Búfalos , Estações do Ano , Espermatozoides , Animais , Búfalos/genética , Masculino , Espermatozoides/metabolismo , Expressão Gênica , Temperatura , Reação em Cadeia da Polimerase em Tempo Real , Perfilação da Expressão Gênica , Análise do Sêmen/veterinária , Umidade , Padrões de Referência
11.
Cardiol Young ; 34(2): 268-271, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37318004

RESUMO

BACKGROUND/AIM: Double-chambered right ventricle is a rare and progressive condition that is characterised by obstruction of the right ventricular tract. Double-chambered right ventricle is usually associated with ventricular septal defect. Early surgical intervention is recommended in patients with these defects. Based on this background, the present study aimed to review early and midterm outcomes of primary repair after double-chambered right ventricle. METHODS: Between January 2014 and June 2021, 64 patients with a mean age of 13.42 ± 12.31 years underwent surgical repair for double-chambered right ventricle. The clinical outcomes of these patients were reviewed and assessed retrospectively. RESULTS: An associated ventricular septal defect was present in all the recruited patients; 48 (75%) patients of sub-arterial type, 15 (23.4%) of perimembranous, and 1 (1.6%) patient of muscular type. The patients were followed up for a mean period of 46.73 ± 27.37 months. During their follow-up, a significant decrease in the mean pressure gradient from 62.33 ± 5.52 mmHg preoperatively to 15.73 ± 2.94 mmHg postoperatively was observed (p < 0.001). Notably, there were no hospital deaths. CONCLUSIONS: The development of double-chambered right ventricle in association with ventricular septal defect results in an increased pressure gradient within the right ventricle. The defect needs correction in a timely manner. In our experience, the surgical correction of double-chambered right ventricle is safe and shows excellent early and mid-term results.


Assuntos
Comunicação Interventricular , Ventrículos do Coração , Humanos , Lactente , Pré-Escolar , Criança , Adolescente , Adulto Jovem , Adulto , Ventrículos do Coração/diagnóstico por imagem , Ventrículos do Coração/cirurgia , Estudos Retrospectivos , Comunicação Interventricular/cirurgia , Fatores de Tempo , Artérias
12.
Ann Pharm Fr ; 2024 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-39089366

RESUMO

OBJECTIVE: Regardless of having desired therapeutic properties many of the recently approved drugs are removed from the developmental pipeline for their clinical use due to low solubility and permeability. Conventional dosage forms are found relatively unsuitable for achieving desired pharmacokinetic and pharmacodynamics profiles. Cilnidipine is 1,4 dihydropyridine derivative calcium channel blocker used for the treatment of hypertension. METHOD: The aim and objective of this study was to develop a precise and significant method in LC-MS/MS for quantification of pharmacokinetic parameters of a cilnidipine-loaded self-micro-emulsifying drug delivery system in rat plasma and simultaneously assessed pharmacodynamic characters in comparison with the marketed cilnidipine tablet. Another potential aim of this study is to reduce the dose of the drug in order to counter the dose-dependent toxicities related to chronic use. In the present study, the parent and product ion of cilnidipine was m/z 491.3\237.1. RESULT: The plasma was extracted by protein precipitation technique. The calibration standard concentrations were 1.875, 3.75, 7.50, 15.00, 30.00, 60.00ng/mL and LLOQ, low-quality control, middle-quality control and high-quality control were 1.87, 5.62, 22.50, 45.00ng/mL, respectively. The mobile phase composition was 0.1% formic acid in Milli Q water with 10mM Ammonium acetate as an aqueous solvent and 0.1% formic acid in methanol as an organic solvent. Following oral administration of optimized formulation Cmax (peak plasma concentration) was achieved 21.02±3.17ng/mL at 0.866±0.11h (Tmax), whereas in the case of marketed tablet Cmax (peak plasma concentration) was achieved 10.16±0.89ng/mL at 0.93±0.11h (Tmax). DISCUSSION: The in-vivo characterizations of the optimized SMEDDS showed significantly better pharmacokinetic parameters in Wistar rats and showed almost 2.4 times enhanced relative bioavailability as compared to the marketed tablet of cilnidipine which was observed to be correlating to our findings with noninvasive blood pressure parameter of Wistar rats.

13.
Microvasc Res ; 145: 104454, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36347299

RESUMO

OBJECTIVE: Subclinical life style disease can cause endothelial dysfunction associated with perfusion abnormalities and reduced vascular compliance. Subclinical elevated beta type natriuretic peptide (BNP) has been associated with altered fluid shift from extra to intracellular space during acute hypoxia. Therefore we measured vascular response and BNP levels during acute hypoxia to study endothelial functions among healthy individuals. METHODS: Individuals were exposed to acute normobaric hypoxia of FiO2 = 0.15 for one hour in supine position and their pulmonary and systemic vascular response to hypoxia was compared. Individuals were divided into two groups based on either no response (Group 1) or rise in systolic pulmonary artery pressure to hypoxia (Group 2) and their BNP levels were compared. RESULTS: BNP was raised after hypoxia exposure in group 2 only from 18.52 ± 7 to 21.56 ± 10.82 picogram/ml, p < 0.05. Group 2 also showed an increase in mean arterial pressure and no fall in total body water in response to acute hypoxia indicating decreased endothelial function compared to Group 1. CONCLUSION: Rise in pulmonary artery pressure and BNP level in response to acute normobaric hypoxia indicates reduced endothelial function and can be used to screen subclinical lifestyle disease among healthy population.


Assuntos
Hipóxia , Peptídeo Natriurético Encefálico , Humanos , Hipóxia/diagnóstico , Pulmão/irrigação sanguínea , Vasodilatadores , Estilo de Vida , Artéria Pulmonar
14.
Mol Pharm ; 20(6): 2966-2977, 2023 06 05.
Artigo em Inglês | MEDLINE | ID: mdl-37216314

RESUMO

Coenzyme Q10 is a potent antioxidant that plays an important role in the maintenance of various biochemical pathways of the body and has a wide range of therapeutic applications. However, it has low aqueous solubility and oral bioavailability. Mesoporous silica nanoparticles (MCM-41 and SBA-15 types) exhibiting varying pore sizes and modified with phosphonate and amino groups were used to study the influence of pore structure and surface chemistry on the solubility, in vitro release profile, and intracellular ROS inhibition activity of coenzyme Q10. The particles were thoroughly characterized to confirm the morphology, size, pore profile, functionalization, and drug loading. Surface modification with phosphonate functional groups was found to have the strongest impact on the solubility enhancement of coenzyme Q10 when compared to that of pristine and amino-modified particles. Phosphonate-modified MCM-41 nanoparticles (i.e., MCM-41-PO3) induced significantly higher coenzyme Q10 solubility than the other particles studied. Furthermore, MCM-41-PO3 led to a twofold decrease in ROS generation in human chondrocyte cells (C28/I2), compared to the free drug in a DMSO/DMEM mixture. The results confirmed the significant contribution of small pore size and negative surface charge of MSNs that enable coenzyme Q10 confinement to allow enhanced drug solubility and antioxidant activity.


Assuntos
Antioxidantes , Nanopartículas , Humanos , Solubilidade , Antioxidantes/farmacologia , Espécies Reativas de Oxigênio , Nanopartículas/química , Dióxido de Silício/química , Porosidade , Portadores de Fármacos/química
15.
Langmuir ; 39(44): 15572-15586, 2023 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-37882047

RESUMO

Evaporation is a ubiquitous phenomenon. Rapid evaporation of the continuous phase from micrometric colloidal droplets can be used to realize nanostructured microgranules, constituting the assembled nanoparticles. One of the important aspects of such nonequilibrium assembly is the nature of the packing of nanoparticles in the microgranules. The present work demonstrates the evidence of size stratification of the nanoparticles in such far-from-equilibrium configurations. Small-angle X-ray scattering, in combination with particle packing simulation, reveals the "large on top"-type stratification in such assembled microgranules, where the larger particles get concentrated at the outer shell of the granules while the smaller particles reside in the core region. It also reveals the presence of local clusters in such a rapid evaporative assembly in aerosolized colloidal droplets.

16.
J Org Chem ; 88(1): 475-482, 2023 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-36520416

RESUMO

A facile and sustainable protocol for the thiolation of hydrazones with sodium sulfinates has been developed in the presence of CuBr2 and DBU in DMF to afford diverse benzylic thioethers. Control experiments reveal a radical pathway involving a thiyl radical as a key intermediate.


Assuntos
Cobre , Sulfetos , Sódio , Hidrazonas , Catálise
17.
Mol Biol Rep ; 50(8): 6717-6727, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37378747

RESUMO

BACKGROUND: Being highly fragmented and low in concentration, isolation of good quality RNA from sperm cells is a big challenge. Attempts have been made to evaluate various sperm RNA isolation methods from purified buffalo bull sperm cells. METHODS: Both, non-membrane and membrane-based methods have been evaluated for isolating RNA from Murrah buffalo sperms and compared for their respective efficacies. The traditional TRIzol, TRIzol-heat lysed (H-TRIzol) and cocktail of TCEP-RLT lysis buffer (Qiagen RNeasy mini kit)-TRIzol (C-TRIzol) based isopropanol isolation methods have been evaluated. RESULTS: H-TRIzol yielded best results among conventional methods. The combined T-RLT RNA isolation protocol yielded best quality and quantity compared to other membrane-based methods, due to high lytic property of cocktail of lysis reagents, necessary for complete breakdown of sperm membrane and RNA binding membrane for RNA isolation. Combined lysis performed by treatment with RLT-T and T-RLT differing in order of reagents used were also evaluated. T-RLT combination giving better results compared to RLT-T due to high gDNA contamination and membrane clogging in later protocol steps. CONCLUSION: Overall, in terms of total RNA quantity and quality per million spermatozoa, the heat-lysed TRIzol method (H-TRIzol) performs best among RNA separation techniques employed and is also quite easy to perform. This comparative evaluation of sperm RNA isolation protocols can be useful in deciding the best protocol for isolation of good quality and high concentration sperm RNA from buffalo semen, for transcriptome and other downstream studies.


Assuntos
RNA , Preservação do Sêmen , Animais , Masculino , RNA/metabolismo , Búfalos/genética , Búfalos/metabolismo , Sêmen/metabolismo , Espermatozoides/metabolismo , Preservação do Sêmen/métodos , Criopreservação/métodos
18.
Environ Res ; 221: 115271, 2023 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-36640933

RESUMO

The residual imidacloprid, a widely used insecticide is causing serious environmental concerns. Knowledge of its biodegradation will help in assessing its residual mass in soil. In view of this, a soil microcosm-based study was performed to test the biodegradation potential of Agrobacterium sp. InxBP2. It achieved ∼88% degradation in 20 days and followed the pseudo-first-order kinetics (k = 0.0511 day-1 and t1/2=7 days). Whole genome sequencing of Agrobacterium sp. InxBP2 revealed a genome size of 5.44 Mbp with 5179 genes. Imidacloprid degrading genes at loci K7A42_07110 (ABC transporter substrate-binding protein), K7A42_07270 (amidohydrolase family protein), K7A42_07385 (ABC transporter ATP-binding protein), K7A42_16,845 (nitronate monooxygenase family protein), and K7A42_20,660 (FAD-dependent monooxygenase) having sequence and functional similarity with known counterparts were identified. Molecular docking of proteins encoded by identified genes with their respective degradation pathway intermediates exhibited significant binding energies (-6.56 to -4.14 kcal/mol). Molecular dynamic simulation discovered consistent interactions and binding depicting high stability of docked complexes. Proteome analysis revealed differential protein expression in imidacloprid treated versus untreated samples which corroborated with the in-silico findings. Further, the detection of metabolites proved the bacterial degradation of imidacloprid. Thus, results provided a mechanistic link between imidacloprid and associated degradative genes/enzymes of Agrobacterium sp. InxBP2. These findings will be of immense significance in carrying out the lifecycle analysis and formulating strategies for the bioremediation of soils contaminated with insecticides like imidacloprid.


Assuntos
Inseticidas , Poluentes do Solo , Biodegradação Ambiental , Simulação de Acoplamento Molecular , Multiômica , Neonicotinoides/análise , Inseticidas/análise , Nitrocompostos/análise , Nitrocompostos/química , Nitrocompostos/metabolismo , Bactérias/metabolismo , Poluentes do Solo/análise , Solo
19.
World J Surg ; 47(12): 3222-3228, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37787777

RESUMO

BACKGROUND: Tracheal airflow limitation is frequently reported in patients with goiter but is severely underestimated, and studies on how goiter and its treatment affect trachea are scarce. Moreover, the choice of the optimal treatment for individual patient with asymptomatic goiter is not straightforward. Therefore, in this study we aim to investigate the effect of goiter and subsequent thyroidectomy on tracheal anatomy and change in airflow in asymptomatic patient with goiter. METHODS: Seventy patients undergoing total/hemithyroidectomy (TT/HT) from Feb 2020 to Feb 2021 satisfying inclusion criteria were enrolled in the study. Neck radiograph (NR) and forced spirometry (FS) were performed preoperatively and on postoperative day 10 and 6 weeks and 3 months. RESULTS: Out of 70 patients, 84.3% patients were female, and mean duration and weight of goiter were 54.7 months and 72.21 gm, respectively. Of 70 patients, 57 were of benign pathology. Significant improvement in tracheal compression with moderate improvement in deviation was observed after surgery. Preoperative spirometry showed significant reduction in almost all parameters. After surgery, a weak improvement was observed at postoperative day 10 and 6 weeks; however, significant improvement in FEV1, PEFR, FEV1/FEV0.5, and FEF50%/FIF50% was observed at postoperative 3 months. Patient with right sided and those with ≥ 8 mm deviation were associated with poorer pulmonary function. Weak correlation was observed between neck NR and spirometry parameters. Weight of the thyroid gland significantly correlated with ratio of MVV/FEV1. CONCLUSION: Patients with asymptomatic goiter can have significant abnormal changes in airflow as evidenced by FS and NR. Thyroidectomy is followed by gradual restoration of tracheal deviation and compression with significant improvement in pulmonary airflow.


Assuntos
Bócio , Traqueia , Humanos , Feminino , Masculino , Traqueia/diagnóstico por imagem , Traqueia/cirurgia , Tireoidectomia , Estudos Prospectivos , Bócio/complicações , Bócio/cirurgia , Pulmão
20.
Clin Radiol ; 78(5): 333-339, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-36710122

RESUMO

The aim of this review is to discuss end-stage renal disease (ESRD) and renal transplant complications and present the nuclear medicine imaging findings. The conditions discussed are renal osteodystrophy, metastatic calcification, and renal transplant complications, such as vascular occlusion and acute tubular necrosis. A total of eight nuclear medicine imaging scintigraphy primarily of bone and renal scintigraphy were selected and the imaging features of the complications are discussed. This article highlights the role of nuclear medicine imaging in diagnosis, quantitative and qualitative assessment of renal function, and monitoring of complications associated with ESRD and renal transplant.


Assuntos
Falência Renal Crônica , Transplante de Rim , Cintilografia , Humanos , Falência Renal Crônica/complicações , Falência Renal Crônica/diagnóstico por imagem , Falência Renal Crônica/cirurgia , Transplante de Rim/efeitos adversos , Complicações Pós-Operatórias/diagnóstico por imagem , Complicações Pós-Operatórias/etiologia
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