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1.
Data Brief ; 53: 110124, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38357455

RESUMO

This paper presents a comprehensive augmented lexicon sentiment analysis dataset for the Hausa language. The dataset was created by adopting words and phrases from a Hausa Language dictionary and then using the data augmentation method to expand the quantity of the dataset. The researchers manually annotated each phrase/sentence with positive, negative, or neutral polarity. The dataset consists of 14,663 rows, with 4,154 positives, 4,310 negatives, and 6,199 neutrals. The dataset is valuable because it contributes to the available resources for sentiment analysis, especially for Hausa, which is a low-resource language. The dataset will benefit researchers in sentiment analysis who want to develop a model to analyze Hausa posts on social media or product reviews in the Hausa language.

2.
J Water Health ; 22(1): 21-35, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38295070

RESUMO

Cholera is a leading cause of mortality in Nigeria. The two most significant predictors of cholera are a lack of access to clean water and poor sanitary conditions. Other factors such as natural disasters, illiteracy, and internal conflicts that drive people to seek sanctuary in refugee camps may contribute to the spread of cholera in Nigeria. The aim of this research is to develop a cholera outbreak risk prediction (CORP) model using machine learning tools and data science. In this study, we developed a CORP model using design science perspectives and machine learning to detect cholera outbreaks in Nigeria. Nonnegative matrix factorization (NMF) was used for dimensionality reduction, and synthetic minority oversampling technique (SMOTE) was used for data balancing. Outliers were detected using density-based spatial clustering of applications with noise (DBSCAN) were removed improving the overall performance of the model, and the extreme-gradient boost algorithm was used for prediction. The findings revealed that the CORP model outcomes resulted in the best accuracy of 99.62%, Matthews's correlation coefficient of 0.976, and area under the curve of 99.2%, which were improved compared with the previous findings. The developed model can be helpful to healthcare providers in predicting possible cholera outbreaks.


Assuntos
Cólera , Humanos , Cólera/epidemiologia , Aprendizado de Máquina , Surtos de Doenças , Algoritmos , Pesquisa sobre Serviços de Saúde
3.
MethodsX ; 11: 102460, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38023300

RESUMO

Natural Language Processing (NLP) has transformed machine translation, sentiment analysis, information retrieval, and conversation systems. NLP applications rely on complete linguistic resources, which might be difficult for low-resource languages. NLP solutions for every language require a language-specific dataset. Dataset in a language is essential for NLP solution creation. Over 7000 languages are spoken worldwide. Only around 20 languages have text corpora for NLP applications. English has the most datasets, then Chinese and Spanish. Japanese has several Western European language datasets. For an accurate NLP system, most Asian and African languages lack training datasets. To address this challenge, we propose a methodology for building a lexicon-based sentiment analysis model for languages with limited resources. The Hausa language was used as training and evaluation language. The methodology combines lexicon creation; augmentation, annotation, and fine-tuning model, and has been tested on a corpus of Hausa tweets achieving an accuracy of 98 %. The results suggest that our proposed model is a promising tool for sentiment analysis in a variety of applications, such as social media monitoring, customer service, and market research. Our methodology can be used for any low-resource language. The outline of the work done in this paper can be shown as follows:•We propose a methodology for building a lexicon-based sentiment analysis model for languages with limited resources, using the Hausa language as a case study.•The methodology combines lexicon creation, augmentation, annotation, and fine-tuning model, and achieves an accuracy of 98 % on a corpus of Hausa tweets.•The results suggest that the proposed model is a promising tool for sentiment analysis in a variety of applications for low-resource languages.

4.
Heliyon ; 9(2): e13483, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36776910

RESUMO

Purpose: The COVID-19 pandemic has affected more than 192 countries. The condition results in a respiratory illness (e.g., influenza) with signs and symptoms such as cold, cough, fever, and breathing difficulties. Predicting new instances of COVID-19 is always a challenging task. Methods: This study improved the autoregressive integrated moving average (ARIMA)-based time series prediction model by incorporating statistical significance for feature selection and k-means clustering for outlier detection. The accuracy of the improved model (ARIMAI) was examined using World Health Organization's official data on the COVID-19 pandemic worldwide and compared with that of many modern, cutting-edge algorithms. Results: The ARIMAI model (RSS score = 0.279, accuracy = 97.75%) outperformed the current ARIMA model (RSS score = 0.659, accuracy = 93%). Conclusions: The ARIMAI model is not only an efficient but also a rapid and simple technique to forecast COVID-19 trends. The usage of this model enables the prediction of any disease that will affect patients in the future pandemics.

5.
Adv Pharm Bull ; 11(4): 746-754, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34888222

RESUMO

Purpose: Cisplatin is a cancer chemotherapeutic drug that has been extensively used in the treatment of a variety of cancers. However, the full usage of cisplatin is limited due to its treatment associated development of multiple side effects in the host. In the present study, the modulatory effect of rutin, a type of flavonoid, on the cisplatin mediated antitumor activity and allied genotoxicity in ascites Dalton's lymphoma (DL)-bearing mice were investigated. Methods: The antitumor activity was determined by calculating the percent increase in the life span of mice, cell viability and scanning electron microscopy (SEM) of DL cells. Further, the modulatory effect of rutin on the cisplatin-induced genotoxic effects in the same DL-bearing mice was assessed by the analysis of micronuclei, chromosomal aberration and sperm abnormality. Results: The combination treatment of mice with rutin and cisplatin showed a considerable increase in the life span of the DL-bearing mice depicting better antitumor efficacy. SEM of these DL cells showed severe membrane deformities in DL cells such as fusion of cell membrane, membrane blebbing, cell shrinkage, membrane folding and loss in microvilli from the tumor cell surface which may lead to cell death. Cisplatin alone treatment caused an increase in the frequency of chromosomal aberrations, micronuclei and sperms abnormality. However, the combination treatment of DL-bearing mice with rutin and cisplatin comparatively reduced these genotoxic effects. Conclusion: The overall findings suggest that rutin enhances the cisplatin-mediated antitumor activity and cytotoxicity against DL cells and at the same time diminishes the genotoxic effects induced by cisplatin in the DL-bearing mice.

6.
Biol Trace Elem Res ; 193(2): 536-547, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31044358

RESUMO

In this study, the pattern of metals concentration in water, sediment, plants, and three edible fish species (Channa striata, Labeo rohita, and Catla catla) of different trophic levels, captured from Jajmau (Kanpur), an important fishery area of river Ganga in Uttar Pradesh, India was examined. The heavy metals, Ni, Pb, Fe, Cu, Zn, Cd, Cr, and Co, were estimated in the liver, kidney, muscles, and gill tissues of abovesaid species of fish. The highest metal concentration was reported in the bottom feeder fish as compared with the column and surface feeders. The result obtained after analysis of water sample reflects the order of occurrence of heavy metals as Fe > Cr > Pb > Ni > Cd > Zn > Cu > Co. Sediments analysis indicates high concentration of Fe and Cr, making the entire environment from top to bottom, stressful. Target hazard quotient (THQ) and hazard index (HI) of the three species suggest a potential risk to the health of consumers, the humans. Thus, it is inevitable that the river Ganga should be closely monitored to safeguard human health. Graphical Abstract.


Assuntos
Monitoramento Ambiental/métodos , Peixes/metabolismo , Água Doce/análise , Sedimentos Geológicos/análise , Metais Pesados/análise , Plantas/metabolismo , Animais , Peixes/classificação , Cadeia Alimentar , Contaminação de Alimentos/análise , Brânquias/química , Humanos , Índia , Rim/química , Fígado/química , Músculos/química , Rios
7.
Environ Toxicol Pharmacol ; 71: 103209, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31207396

RESUMO

Ameliorative potential of curcumin against Cr6+-induced eco-toxicological manifestations was assessed in liver of exposed Channa punctatus (Actinopterygii) in six groups for 45 d; Group I as control. Group II with 3 mg/L of curcumin; group III with 7.89 mg/L of Cr6+. Groups IV, V and VI were simultaneously co-exposed with 7.89 mg/L of Cr6+ and three different curcumin concentrations, 1, 2, and 3 mg/L, respectively. In group III, SOD-CAT, GR significantly (p < 0.05) increased; decreased GSH level; elevated MN and AC frequencies; and a significant (p < 0.05) up-regulation of cat (2.72-fold), p53 (1.73-fold), bax (1.33-fold) and apaf-1 (2.13-fold) together with a significant (p < 0.05) down-regulation of bcl-2 (0.51-fold). Co-exposure significantly (p < 0.05) brought down activities of SOD-CAT, GR, raised GSH, decreased micronuclei and apoptotic frequencies along with recovery of histopathological anomalies in liver. This study establishes the protective role of curcumin against Cr6+-induced hepatotoxicity in fish.


Assuntos
Apoptose/efeitos dos fármacos , Cromo/toxicidade , Curcumina/farmacologia , Peixes , Fígado/efeitos dos fármacos , Micronúcleos com Defeito Cromossômico/induzido quimicamente , Estresse Oxidativo/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Transcriptoma/efeitos dos fármacos , Animais , Apoptose/genética , Fígado/metabolismo , Fígado/patologia , Estresse Oxidativo/genética , Espécies Reativas de Oxigênio/sangue , Toxicogenética
8.
Chemosphere ; 231: 588-606, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31154237

RESUMO

Tremendous explosion of population has led to about 200% increment of total energy consumptions in last twenty-five years. Apart from conventional fossil fuel as limited energy source, alternative non-conventional sources are being explored worldwide to cater the energy requirement. Lignocellulosic biomass conversion for biofuel production is an important alternative energy source due to its abundance in nature and creating less harmful impacts on the environment in comparison to the coal or petroleum-based sources. However, lignocellulose biopolymer, the building block of plants, is a recalcitrant substance and difficult to break into desirable products. Commonly used chemical and physical methods for pretreating the substrate are having several limitations. Whereas, utilizing microbial potential to hydrolyse the biomass is an interesting area of research. Because of the complexity of substrate, several enzymes are required that can act synergistically to hydrolyse the biopolymer producing components like bioethanol or other energy substances. Exploring a range of microorganisms, like bacteria, fungi, yeast etc. that utilizes lignocelluloses for their energy through enzymatic breaking down the biomass, is one of the options. Scientists are working upon designing organisms through genetic engineering tools to integrate desired enzymes into a single organism (like bacterial cell). Studies on designer cellulosomes and bacteria consortia development relating consolidated bioprocessing are exciting to overcome the issue of appropriate lignocellulose digestions. This review encompasses up to date information on recent developments for effective microbial degradation processes of lignocelluloses for improved utilization to produce biofuel (bioethanol in particular) from the most plentiful substances of our planet.


Assuntos
Biodegradação Ambiental , Biocombustíveis/microbiologia , Fontes Geradoras de Energia , Etanol/metabolismo , Bactérias , Biomassa , Fungos , Engenharia Genética , Hidrólise , Lignina , Leveduras
9.
Mol Biochem Parasitol ; 222: 70-80, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29753659

RESUMO

Plasmodium falciparum DJ1 (PfDJ1) belongs to the DJ-1/ThiJ/PfpI superfamily whose members are present in all the kingdoms of life and exhibit diverse cellular functions and biochemical activities. The common feature of the superfamily is the class I glutamine amidotransferase domain with a conserved redox-active cysteine residue, which mediates various activities of the superfamily members, including anti-oxidative activity in PfDJ1 and human DJ1 (hDJ1). As the superfamily members represent diverse functional classes, to investigate if there is any sequence feature unique to hDJ1-like proteins, sequences of the representative proteins of different functional classes were compared and analysed. A novel motif unique to PfDJ1 and several other hDJ1-like proteins, with the consensus sequence of TSXGPX5FXLX5L, was identified that we designated as the hDJ1-subfamily motif (DJSM). Several mutations that have been associated with Parkinson's disease are also present in DJSM, suggesting its functional importance in hDJ1-like proteins. Mutations of the conserved residues of DJSM of PfDJ1 did not significantly affect overall secondary structure, but caused both a significant loss (S151A and P154A) and gain (L168A) of anti-oxidative activity. We also report that PfDJ1 has deglycase activity, which was significantly decreased in its mutants of the catalytic cysteine (C106A) and DJSM (S151A and P154A). Episomal expression of the catalytic cysteine (C106A) or DJSM (P154A) mutant decreased growth rates of parasites as compared to that of wild type parasites or parasites expressing wild type PfDJ1. S151 appears to properly position the nucleophilic elbow containing C106 and P154 forms a hydrogen bond with C106, which could be a reason for the loss of activities of PfDJ1 upon their mutations. Taken together, DJSM delineates PfDJ1 and other hDJ1-subfamily proteins from the remaining superfamily, and is critical for anti-oxidative and deglycase activities of PfDJ1.


Assuntos
Estresse Oxidativo , Plasmodium falciparum/enzimologia , Plasmodium falciparum/metabolismo , Proteína Desglicase DJ-1/química , Proteína Desglicase DJ-1/metabolismo , Proteínas de Protozoários/química , Proteínas de Protozoários/metabolismo , Motivos de Aminoácidos , Sequência de Aminoácidos , Catálise , Sequência Conservada , Humanos , Plasmodium falciparum/química , Plasmodium falciparum/genética , Proteína Desglicase DJ-1/genética , Proteínas de Protozoários/genética , Alinhamento de Sequência
10.
Aquat Toxicol ; 200: 158-167, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29753883

RESUMO

Present study was designed to assess the hexavalent chromium (Cr6+) mediated oxidative stress that induces DNA damage and apoptosis in adult fish, Channa punctatus (35 ±â€¯3.0 g; 14.5 ±â€¯1.0 cm; Actinopterygii). Fishes were maintained in three groups for 15, 30 and 45 d of exposure periods. They were treated with 5% (Group T1) and 10% (Group T2) of 96 h-LC50 of chromium trioxide (Cr6+). Controls were run for the similar duration. A significant (p < 0.05) increment in the activities of antioxidant enzymes, SOD and CAT in liver tissues of the exposed fish evinces the persistence of oxidative stress. A significant (p < 0.05) increase in induction of micronuclei (MN) coupled with transcriptional responses of target genes related to antioxidant enzymes, DNA damage and apoptosis (sod, cat, gsr, nox-1, p53, bax, bcl-2, apaf-1 and casp3a) establishes the impact of oxidative stress due to in vivo, Cr6+ accumulation in liver as compared to control (0 mg/L), in a dose and exposure-dependent manner. Initially, the increased level of reactive oxygen species (ROS) in liver coincided with that of enhanced mRNA expression of antioxidant enzymes, sod, cat, gsr and nox-1 but, later, the overproduction of ROS, after 45 d of exposure of Cr6+, resulted in a significant (p < 0.05) up-regulation of p53. Our findings also unveil that the up-regulation of bax, apaf-1 and casp3a and down-regulation of bcl-2 are associated with Cr6+-induced oxidative stress mediated-apoptosis in liver of test fish. Aforesaid molecular markers can, thus, be efficiently utilized for bio-monitoring of aquatic regimes and conservation of fish biodiversity.


Assuntos
Apoptose/genética , Cromo/toxicidade , Dano ao DNA/genética , Peixes/genética , Perfilação da Expressão Gênica , Fígado/metabolismo , Mutagênicos/toxicidade , Estresse Oxidativo/genética , Animais , Antioxidantes/metabolismo , Apoptose/efeitos dos fármacos , Catalase/metabolismo , Fígado/efeitos dos fármacos , Testes para Micronúcleos , Estresse Oxidativo/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismo , Transcrição Gênica/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade
11.
Ecotoxicol Environ Saf ; 151: 10-20, 2018 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-29304413

RESUMO

Zn2+ is essential for normal physiological functioning of all organisms in small quantities, but when its concentration enhances in surrounding environment it acts as a toxicant to organisms. Common sources of Zn2+ pollution are electroplating, alloying, mining, and allied industrial operations. The present study aims to assess the biochemical, histopathological and genotoxicological implications under Zn2+ intoxication along with its accumulation patterns in prime biotransformation sites-liver and kidney, of a bottom feeder fish, Channa punctatus. Fish were chronically exposed to two different concentrations of Zn2+i.e., 5mg/L (permissible limit, T1) and 10mg/L (twice the permissible limit, T2). Simultaneous control was maintained. A significant (p<0.05) increment in Zn2+ bioaccumulation, antioxidant enzymes activities of SOD, CAT and GR and induction in micronuclei frequencies along with the significant (p<0.05) decrement in total protein and GSH were observed in all the exposed groups after 28 d. Altered biochemical parameters coupled with enhanced induction in micronuclei and accumulation of Zn2+ in liver and kidney of fish can be regarded as sensitive biomarkers of Zn2+ induced toxicological manifestations and thus, they may be effectively utilized for reliable ecotoxicological biomonitoring of aquatic regimes polluted with Zn2+.


Assuntos
Dano ao DNA , Peixes/metabolismo , Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Sulfato de Zinco/toxicidade , Animais , Biomarcadores/metabolismo , Ecotoxicologia , Monitoramento Ambiental , Eritrócitos/efeitos dos fármacos , Peixes/sangue , Peixes/genética , Rim/metabolismo , Rim/patologia , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia
12.
Folia Microbiol (Praha) ; 62(1): 17-55, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27718043

RESUMO

Bacteriophages (or phages), the most abundant viral entity of the planet, are omni-present in all the ecosystems. On the basis of their unique characteristics and anti-bacterial property, phages are being freshly evaluated taxonomically. Phages replicate inside the host either by lytic or lysogenic mode after infecting and using the cellular machinery of a bacterium. Since their discovery by Twort and d'Herelle in the early 1900s, phage became an important agent for combating pathogenic bacteria in clinical treatments and its related research gained momentum. However, due to recent emergence of bacterial resistance on antibiotics, applications of phage (phage therapy) become an inevitable option of research. Phage particles become popular as a biotechnological tool and treatment of pathogenic bacteria in a range of applied areas. However, there are few concerns over the application of phage-based solutions. This review deals with the updated phage taxonomy (ICTV 2015 Release and subsequent revision) and phage biology and the recent development of its application in the areas of biotechnology, biosensor, therapeutic medicine, food preservation, aquaculture diseases, pollution remediation, and wastewater treatment and issues related with limitations of phage-based remedy.


Assuntos
Infecções Bacterianas/terapia , Bacteriófagos/classificação , Bacteriófagos/fisiologia , Terapia por Fagos/métodos , Pesquisa Biomédica/tendências , Biotecnologia/métodos , Humanos
13.
Bioorg Med Chem ; 25(1): 221-232, 2017 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-27816268

RESUMO

Novel series of naphthyl bearing 1,2,3-triazoles (4a-t) were synthesized and evaluated for their in vitro antiplasmodial activity against pyrimethamine (Pyr)-sensitive and resistant strains of Plasmodium falciparum. The synthesized compounds were assessed for their cytotoxicity employing human embryonic kidney cell line (HEK-293), and none of them was found to be toxic. Among them 4j, 4k, 4l, 4m, 4n, 4t exhibited significant antiplasmodial activity in both strains, of which compounds 4m, 4n and 4t (∼3.0-fold) displayed superior activity to Pyr against resistant strain. Pyr and selected compounds (4n, 4p and 4t) that repressed parasite development also inhibited PfDHFR activity of the soluble parasite extract, suggesting that anti-parasitic activity of these compounds is a result of inhibition of the parasite DHFR. In silico studies suggest that activity of these compounds might be enhanced due to π-π stacking.


Assuntos
Antiprotozoários/farmacologia , Antagonistas do Ácido Fólico/farmacologia , Naftalenos/farmacologia , Triazóis/farmacologia , Antiprotozoários/síntese química , Antiprotozoários/química , Antiprotozoários/toxicidade , Cristalografia por Raios X , Ensaios Enzimáticos , Antagonistas do Ácido Fólico/síntese química , Antagonistas do Ácido Fólico/química , Antagonistas do Ácido Fólico/toxicidade , Células HEK293 , Humanos , Simulação de Acoplamento Molecular , NADP/metabolismo , Naftalenos/síntese química , Naftalenos/química , Naftalenos/toxicidade , Plasmodium falciparum/efeitos dos fármacos , Pirimetamina/química , Triazóis/síntese química , Triazóis/química , Triazóis/toxicidade
14.
Mol Biochem Parasitol ; 202(2): 11-22, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26439304

RESUMO

Falcipain-3 (FP3) is an essential and drug target cysteine protease of the most lethal human malaria parasite Plasmodium falciparum. FP3 and its majority of homologs in malaria parasites prefer Leu at the P2 position in substrates and inhibitors, whereas its major host homolog cathepsin L prefers Phe. However, FP3 is much less active on peptide substrates and has negligible activity against a P2 Arg-containing substrate (Z-RR-AMC) compared to its paralog falcipain-2A (FP2A). To identify the specificity determinants, the S2/3 pocket residues of FP3 were substituted with the corresponding residues in FP2 or cathepsin L, and the wild type and mutant proteases were assessed for hydrolysis of peptide and protein substrates. Our results indicate that the S2 pocket residues I94 and P181 of FP3 are chiefly responsible for its P2 Leu preference and negligible activity for Z-RR-AMC, respectively. E243 in FP3 and the corresponding residue D234 in FP2 have a key role in Z-RR-AMC hydrolysing activity, possibly through stabilization of side chain interactions, as their substitution with Ala abolished the activity. Several FP3 mutants, which retained P2 Leu preference and showed similar or more activity than wild type FP3 on peptide substrates, degraded haemoglobin less efficiently than wild type FP3, suggesting that multiple residues contribute to haemoglobinase activity. Furthermore, P181 and E243 appear to contribute to the optimum activity of FP3 in the food vacuole milieu (≈pH 5.5). The identification of residues determining specificity of FP3 could aid in developing specific inhibitors of FP3 and its homologs in malaria parasites.


Assuntos
Cumarínicos/metabolismo , Cisteína Endopeptidases/química , Dipeptídeos/metabolismo , Plasmodium falciparum/enzimologia , Proteínas de Protozoários/química , Sequência de Aminoácidos , Domínio Catalítico , Catepsina L/química , Cisteína Endopeptidases/genética , Hemoglobinas/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Hidrólise , Leucina/química , Leucina/genética , Malária Falciparum/parasitologia , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Peptídeos/química , Proteínas de Protozoários/genética , Especificidade por Substrato
15.
PLoS One ; 8(9): e73530, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24023882

RESUMO

Among key potential drug target proteolytic systems in the malaria parasite Plasmodium falciparum are falcipains, a family of hemoglobin-degrading cysteine proteases, and the ubiquitin proteasomal system (UPS), which has fundamental importance in cellular protein turnover. Inhibition of falcipains blocks parasite development, primarily due to inhibition of hemoglobin degradation that serves as a source of amino acids for parasite growth. Falcipains prefer P2 leucine in substrates and peptides, and their peptidyl inhibitors with leucine at the P2 position show potent antimalarial activity. The peptidyl inhibitor MG132 (Z-Leu-Leu-Leu-CHO) is a widely used proteasome inhibitor, which also has P2 leucine, and has also been shown to inhibit parasite development. However, the antimalarial targets of MG132 are unclear. We investigated whether MG132 blocks malaria parasite development by inhibiting hemoglobin degradation and/or by targeting the UPS. P. falciparum was cultured with inhibitors of the UPS (MG132, epoxomicin, and lactacystin) or falcipains (E64), and parasites were assessed for morphologies, extent of hemoglobin degradation, and accumulation of ubiquitinated proteins. MG132, like E64 and unlike epoxomicin or lactacystin, blocked parasite development, with enlargement of the food vacuole and accumulation of undegraded hemoglobin, indicating inhibition of hemoglobin degradation by MG132, most likely due to inhibition of hemoglobin-degrading falcipain cysteine proteases. Parasites cultured with epoxomicin or MG132 accumulated ubiquitinated proteins to a significantly greater extent than untreated or E64-treated parasites, indicating that MG132 inhibits the parasite UPS as well. Consistent with these findings, MG132 inhibited both cysteine protease and UPS activities present in soluble parasite extracts, and it strongly inhibited recombinant falcipains. MG132 was highly selective for inhibition of P. falciparum (IC50 0.0476 µM) compared to human peripheral blood mononuclear cells (IC50 10.8 µM). Thus, MG132 inhibits two distinct proteolytic systems in P. falciparum, and it may serve as a lead molecule for development of dual-target inhibitors of malaria parasites.


Assuntos
Hemoglobinas/metabolismo , Leupeptinas/farmacologia , Plasmodium falciparum/efeitos dos fármacos , Plasmodium falciparum/crescimento & desenvolvimento , Complexo de Endopeptidases do Proteassoma/metabolismo , Proteólise/efeitos dos fármacos , Ubiquitina/metabolismo , Cisteína Endopeptidases/metabolismo , Sinergismo Farmacológico , Eritrócitos/parasitologia , Humanos , Plasmodium falciparum/metabolismo , Plasmodium falciparum/fisiologia , Inibidores de Proteassoma/farmacologia
16.
PLoS One ; 7(12): e51619, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23251596

RESUMO

Papain-like cysteine proteases of malaria parasites degrade haemoglobin in an acidic food vacuole to provide amino acids for intraerythrocytic parasites. These proteases are potential drug targets because their inhibitors block parasite development, and efforts are underway to develop chemotherapeutic inhibitors of these proteases as the treatments for malaria. Plasmodium knowlesi has recently been shown to be an important human pathogen in parts of Asia. We report expression and characterization of three P. knowlesi papain-like proteases, termed knowpains (KP2-4). Recombinant knowpains were produced using a bacterial expression system, and tested for various biochemical properties. Antibodies against recombinant knowpains were generated and used to determine their cellular localization in parasites. Inhibitory effects of the cysteine protease inhibitor E64 were assessed on P. knowlesi culture to validate drug target potential of knowpains. All three knowpains were present in the food vacuole, active in acidic pH, and capable of degrading haemoglobin at the food vacuolar pH (≈5.5), suggesting roles in haemoglobin degradation. The proteases showed absolute (KP2 and KP3) to moderate (KP4) preference for peptide substrates containing leucine at the P2 position; KP4 preferred arginine at the P2 position. While the three knowpains appear to have redundant roles in haemoglobin degradation, KP4 may also have a role in degradation of erythrocyte cytoskeleton during merozoite egress, as it displayed broad substrate specificity and was primarily localized at the parasite periphery. Importantly, E64 blocked erythrocytic development of P. knowlesi, with enlargement of food vacuoles, indicating inhibition of haemoglobin hydrolysis and supporting the potential for inhibition of knowpains as a strategy for the treatment of malaria. Functional expression and characterization of knowpains should enable simultaneous screening of available cysteine protease inhibitor libraries against knowpains for developing broadly effective compounds active against multiple human malaria parasites.


Assuntos
Malária/parasitologia , Papaína/metabolismo , Parasitos/enzimologia , Plasmodium knowlesi/citologia , Plasmodium knowlesi/enzimologia , Proteínas de Protozoários/metabolismo , Sequência de Aminoácidos , Animais , Inibidores de Cisteína Proteinase/farmacologia , Proteínas do Citoesqueleto/metabolismo , Estabilidade Enzimática/efeitos dos fármacos , Eritrócitos/efeitos dos fármacos , Eritrócitos/metabolismo , Eritrócitos/parasitologia , Haplorrinos/parasitologia , Hemoglobinas/metabolismo , Humanos , Concentração de Íons de Hidrogênio/efeitos dos fármacos , Hidrólise/efeitos dos fármacos , Cinética , Leucina/análogos & derivados , Leucina/farmacologia , Dados de Sequência Molecular , Papaína/química , Parasitos/efeitos dos fármacos , Parasitos/crescimento & desenvolvimento , Plasmodium knowlesi/efeitos dos fármacos , Plasmodium knowlesi/crescimento & desenvolvimento , Transporte Proteico/efeitos dos fármacos , Proteólise/efeitos dos fármacos , Proteínas de Protozoários/química , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Alinhamento de Sequência , Especificidade por Substrato/efeitos dos fármacos
17.
Int J Biol Macromol ; 50(2): 369-75, 2012 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-22210527

RESUMO

This paper reports a novel microwave initiated method for synthesis of polyacrylamide grafted Psyllium (Psy-g-PAM). Psyllium was modified through grafting of polyacrylamide (PAM) chains on it using microwave radiations only, in absence of any other free radical initiator. The grafting was confirmed by intrinsic viscosity study and characterization techniques like FTIR spectroscopy, elemental analysis (C, H, N, O and S) and SEM morphology study. Further, the flocculation efficacy of the synthesized graft copolymers was studied in kaolin and coal fine suspension through standard 'Jar test' procedure.


Assuntos
Resinas Acrílicas/química , Resinas Acrílicas/síntese química , Cinza de Carvão/química , Micro-Ondas , Psyllium/química , Floculação , Testes de Floculação , Caulim/química , Suspensões , Viscosidade
18.
Biochem Biophys Res Commun ; 389(2): 333-7, 2009 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-19723504

RESUMO

Work from our laboratory has previously demonstrated the requirement of membrane cholesterol in the function of the serotonin(1A) receptor, a member of the G-protein coupled receptor (GPCR) superfamily. In order to monitor the effect of cellular organization on the function of human serotonin(1A) receptors, we explored receptor function following cholesterol depletion in live cells and membranes isolated from cholesterol-depleted cells. We report here the novel observation that while ligand binding of serotonin(1A) receptors displays an increase in membranes isolated from cholesterol-depleted cells, such trend is absent when binding is performed on cholesterol-depleted intact cells. Importantly, we show here, for the first time, that G-protein coupling of the serotonin(1A) receptor is enhanced in membranes isolated from cholesterol-depleted cells. These results assume pharmacological relevance in view of the recently described structural evidence of specific cholesterol binding sites in GPCRs, and may help in designing better therapeutic strategies for diseases related to GPCRs.


Assuntos
Membrana Celular/metabolismo , Colesterol/metabolismo , Receptor 5-HT1A de Serotonina/metabolismo , Animais , Células CHO , Cricetinae , Cricetulus , Humanos , Ligantes , Receptor 5-HT1A de Serotonina/genética
19.
Biochem Biophys Res Commun ; 390(1): 93-6, 2009 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-19781522

RESUMO

Membrane lipid composition of cells in the nervous system is unique and displays remarkable diversity. Cholesterol metabolism and homeostasis in the central nervous system and their role in neuronal function represent important determinants in neuropathogenesis. The serotonin(1A) receptor is an important member of the G-protein coupled receptor superfamily, and is involved in a variety of cognitive, behavioral, and developmental functions. We report here, for the first time, that the ligand binding function of human serotonin(1A) receptors exhibits an increase in membranes isolated from cholesterol-depleted neuronal cells. Our results gain pharmacological significance in view of the recently described structural evidence of specific cholesterol binding site(s) in GPCRs, and could be useful in designing better therapeutic strategies for neurodegenerative diseases associated with GPCRs.


Assuntos
Membrana Celular/metabolismo , Colesterol/metabolismo , Neurônios/metabolismo , Receptor 5-HT1A de Serotonina/metabolismo , Humanos , Ligantes
20.
Glycoconj J ; 26(6): 733-8, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18802747

RESUMO

The serotonin(1A) receptor is an important member of the G-protein coupled receptor family, and is involved in the generation and modulation of a variety of cognitive, behavioral, and developmental functions. In order to examine the role of membrane material properties in ligand binding activity of the hippocampal serotonin(1A) receptor, we monitored the function of the receptor in presence of capsaicin. Capsaicin has been previously shown to increase the elasticity of membrane bilayers. Our results show that the ligand binding activity of the hippocampal serotonin(1A) receptor is reduced in the presence of capsaicin in a linear concentration-dependent manner. This is accompanied by no appreciable change in G-protein coupling of the receptor and overall membrane order. We conclude that material properties of membrane bilayers could play an important role in the function of the serotonin(1A) receptor in particular, and membrane proteins in general.


Assuntos
8-Hidroxi-2-(di-n-propilamino)tetralina/metabolismo , Antipruriginosos/farmacologia , Capsaicina/farmacologia , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Receptor 5-HT1A de Serotonina/metabolismo , Animais , Bovinos , Polarização de Fluorescência , Ligação Proteica/efeitos dos fármacos
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