Invivo and invitro evaluation of antitumor effects of iron oxide and folate core shell-iron oxide nanoparticles

Abstract Nanoparticles are considered viable options in the treatment of cancer. This study was conducted to investigate the effect of magnetite nanoparticles (MNPs) and magnetite folate core shell (MFCS) on leukemic and hepatocarcinoma cell cultures as well as their effect on the animal model of acute myelocytic leukemia (AML). Through current study nanoparticles were synthesized, characterized by various techniques, and their properties were studied to confirm their nanostructure. Invivo study, nanoparticles were evaluated to inspect their cytotoxic activity against SNU-182 (human hepatocellular carcinoma), K562 (human leukemia), and THLE2 (human normal epithelial liver) cells via MTT test. Apoptotic signaling proteins Bcl-2 and Caspase-3 expression were inspected through RT-PCR method. A cytotoxic effect of MNPs and MFCS was detected in previous cell cultures. Moreover, the apoptosis was identified through significant up-regulation of caspase-3, with Bcl-2 down-regulation. Invitro study, AML was induced in rats by N-methyl-N-nitrosourea followed by oral treatment with MNPS and MFCS. Biochemical indices such as aspartate and alanine amino transferases, and lactate dehydrogenase activities, uric acid, complete blood count, and Beta -2-microglubulin were assessed in serum. Immunophenotyping for CD34 and CD38 detection was performed. Liver, kidney, and bone marrow were microscopically examined. Bcl-2 promoter methylation, and mRNA levels were examined. Although, both MNPs and MFCS depict amelioration in biochemical parameters, MFCS alleviated them toward normal control. Anticancer activity of MNPs and MFCS was approved especially for AML. Whenever, administration of MFCS was more effective than MNPs. The present work is one of few studies used MFCS as anticancer agent.

Resumo Nanopartículas são consideradas opções viáveis no tratamento do câncer. Este estudo foi conduzido para investigar o efeito de nanopartículas de magnetita (MNPs) e núcleo de folato de magnetita (MFCS) em culturas de células leucêmicas e de hepatocarcinoma, bem como seu efeito no modelo animal de leucemia mielocítica aguda (LMA). Através do atual estudo, nanopartículas foram sintetizadas, caracterizadas por várias técnicas, e suas propriedades foram estudadas para confirmar sua nanoestrutura. No estudo in vivo, as nanopartículas foram avaliadas para inspecionar sua atividade citotóxica contra células SNU-182 (carcinoma hepatocelular humano), K562 (leucemia humana) e THLE2 (fígado epitelial humano normal) por meio do teste MTT. A expressão das proteínas sinalizadoras apoptóticas Bcl-2 e Caspase-3 foram inspecionadas através do método RT-PCR. Um efeito citotóxico de MNPs e MFCS foi detectado em culturas de células anteriores. Além disso, a apoptose foi identificada por meio de regulação positiva significativa da Caspase-3, com regulação negativa de Bcl-2. No estudo in vitro, a AML foi induzida em ratos por N-metil-N-nitrosoureia seguida por tratamento oral com MNPS e MFCS. Índices bioquímicos como aspartato e alanina aminotransferases e atividades de lactato desidrogenase, ácido úrico, hemograma completo e Beta-2-microglubulina foram avaliados no soro. A imunofenotipagem para detecção de CD34 e CD38 foi realizada. Fígado, rim e medula óssea foram examinados microscopicamente. A metilação do promotor Bcl-2 e os níveis de mRNA foram examinados. Embora tanto os MNPs quanto os MFCS representem uma melhora nos parâmetros bioquímicos, o MFCS os aliviou em direção ao controle normal. A atividade anticâncer de MNPs e MFCS foi aprovada especialmente para AML. Sempre, a administração de MFCS foi mais eficaz do que MNPs. O presente trabalho é um dos poucos estudos que utilizou o MFCS como agente anticâncer.

Invivo and invitro evaluation of antitumor effects of iron oxide and folate core shell-iron oxide nanoparticles / Avaliação in vivo e in vitro dos efeitos antitumorais de nanopartículas de óxido de ferro e óxido de ferro com núcleo de folato

Abstract Nanoparticles are considered viable options in the treatment of cancer. This study was conducted to investigate the effect of magnetite nanoparticles (MNPs) and magnetite folate core shell (MFCS) on leukemic and hepatocarcinoma cell cultures as well as their effect on the animal model of acute myelocytic leukemia (AML). Through current study nanoparticles were synthesized, characterized by various techniques, and their properties were studied to confirm their nanostructure. Invivo study, nanoparticles were evaluated to inspect their cytotoxic activity against SNU-182 (human hepatocellular carcinoma), K562 (human leukemia), and THLE2 (human normal epithelial liver) cells via MTT test. Apoptotic signaling proteins Bcl-2 and Caspase-3 expression were inspected through RT-PCR method. A cytotoxic effect of MNPs and MFCS was detected in previous cell cultures. Moreover, the apoptosis was identified through significant up-regulation of caspase-3, with Bcl-2 down-regulation. Invitro study, AML was induced in rats by N-methyl-N-nitrosourea followed by oral treatment with MNPS and MFCS. Biochemical indices such as aspartate and alanine amino transferases, and lactate dehydrogenase activities, uric acid, complete blood count, and Beta -2-microglubulin were assessed in serum. Immunophenotyping for CD34 and CD38 detection was performed. Liver, kidney, and bone marrow were microscopically examined. Bcl-2 promoter methylation, and mRNA levels were examined. Although, both MNPs and MFCS depict amelioration in biochemical parameters, MFCS alleviated them toward normal control. Anticancer activity of MNPs and MFCS was approved especially for AML. Whenever, administration of MFCS was more effective than MNPs. The present work is one of few studies used MFCS as anticancer agent.

Resumo Nanopartículas são consideradas opções viáveis ​​no tratamento do câncer. Este estudo foi conduzido para investigar o efeito de nanopartículas de magnetita (MNPs) e núcleo de folato de magnetita (MFCS) em culturas de células leucêmicas e de hepatocarcinoma, bem como seu efeito no modelo animal de leucemia mielocítica aguda (LMA). Através do atual estudo, nanopartículas foram sintetizadas, caracterizadas por várias técnicas, e suas propriedades foram estudadas para confirmar sua nanoestrutura. No estudo in vivo, as nanopartículas foram avaliadas para inspecionar sua atividade citotóxica contra células SNU-182 (carcinoma hepatocelular humano), K562 (leucemia humana) e THLE2 (fígado epitelial humano normal) por meio do teste MTT. A expressão das proteínas sinalizadoras apoptóticas Bcl-2 e Caspase-3 foram inspecionadas através do método RT-PCR. Um efeito citotóxico de MNPs e MFCS foi detectado em culturas de células anteriores. Além disso, a apoptose foi identificada por meio de regulação positiva significativa da Caspase-3, com regulação negativa de Bcl-2. No estudo in vitro, a AML foi induzida em ratos por N-metil-N-nitrosoureia seguida por tratamento oral com MNPS e MFCS. Índices bioquímicos como aspartato e alanina aminotransferases e atividades de lactato desidrogenase, ácido úrico, hemograma completo e Beta-2-microglubulina foram avaliados no soro. A imunofenotipagem para detecção de CD34 e CD38 foi realizada. Fígado, rim e medula óssea foram examinados microscopicamente. A metilação do promotor Bcl-2 e os níveis de mRNA foram examinados. Embora tanto os MNPs quanto os MFCS representem uma melhora nos parâmetros bioquímicos, o MFCS os aliviou em direção ao controle normal. A atividade anticâncer de MNPs e MFCS foi aprovada especialmente para AML. Sempre, a administração de MFCS foi mais eficaz do que MNPs. O presente trabalho é um dos poucos estudos que utilizou o MFCS como agente anticâncer.

Antifungal efficacy of Moringa oleifera leaf and seed extracts against Botrytis cinerea causing gray mold disease of tomato (Solanum lycopersicum L. ) / Eficácia antifúngica de extratos de folha e de semente de Moringa oleifera contra Botrytis cinerea causador da doença do molde cinza de tomate (Solanum lycopersicum L. )

Abstract Drawbacks associated with the use of chemical fungicides to control plant pathogenic fungi such as Botrytis cinerea stimulate the need for alternatives. Therefore, the present study was carried out to determine the antifungal potentials of Moringa oleifera extracts against B. cinerea. Phytochemical analysis using qualitative chemical tests revealed the presence of huge amount of crucial phytochemicals compounds like phenolic compounds, alkaloids and saponins in the M. oleifera leaf extract. Antifungal bioassay of the crude extracts indicated better mycelial growth inhibition by methanol leaf extract (99%). The minimum inhibitory concentration (MIC) was 5 mg/ml with 100% spore germination inhibition and minimum fungicidal concentration (MFC) was 10 mg/ml with 98.10% mycelial growth inhibition using broth micro dilution and poisoned food techniques. Gas chromatography-mass spectrometry (GC-MS) analysis led to the identification of 67 volatile chemical compounds in the leaf extract with 6-decenoic acid (Z)- (19.87%) was the predominant compound. Further chemical elucidation of the crude extracts performed by liquid chromatography with tandem mass spectrometry (LC-MS/MS) showed the presence of non-volatile chemical compounds, mostly flavones, flavonoids and phenolic acids (i.e. quercetin and kaempferol). Scanning electron microscopy and transmission electron microscopy analysis showed positive effect of M. oleifera leaf extract on the treated conidia and mycelium of B. cinerea. Findings revealed that irreversible surface and ultra-structural changes with severe detrimental effects on conidia and mycelium morphology compared to control treatment. Overall findings suggested that M. oleifera leaf extract is a promising candidate for biological control of fungal pathogens, thus limiting overdependence on chemical fungicides.

Resumo As desvantagens associadas ao uso de fungicidas químicos para controlar fungos fitopatogênicos, como Botrytis cinerea, estimulam a necessidade de alternativas. Portanto, o presente estudo foi realizado para determinar o potencial antifúngico de extratos de Moringa oleifera contra B. cinerea. A análise fitoquímica usando testes químicos qualitativos revelou a presença de uma grande quantidade de compostos fitoquímicos cruciais, como compostos fenólicos, alcaloides e saponinas no extrato da folha de M. oleifera. O bioensaio antifúngico dos extratos brutos indicou melhor inibição do crescimento micelial pelo extrato de folhas de metanol (99%). A concentração inibitória mínima (MIC) foi de 5 mg/mL com 100% de inibição da germinação de esporos e a concentração fungicida mínima (MFC) foi de 10 mg/mL com 98,10% de inibição do crescimento micelial usando microdiluição em caldo e técnicas de alimentos envenenados. A análise por cromatografia gasosa-espectrometria de massa (GC-MS) levou à identificação de 67 compostos químicos voláteis no extrato da folha, sendo o ácido 6-decenoico (Z) (19,87%) o composto predominante. Elucidação química adicional dos extratos brutos realizada por cromatografia líquida com espectrometria de massa em tandem (LC-MS/MS) mostrou a presença de compostos químicos não voláteis, principalmente flavonas, flavonoides e ácidos fenólicos (ou seja, quercetina e kaempferol). As análises de microscopia eletrônica de varredura e microscopia eletrônica de transmissão mostraram efeito positivo do extrato de folhas de M. oleifera sobre os conídios e micélios tratados de B. cinerea. Os resultados revelaram a superfície irreversível e alterações ultraestruturais com graves efeitos prejudiciais sobre os conídios e a morfologia micelial, em comparação com o tratamento de controle. Os resultados gerais sugeriram que o extrato da folha de M. oleifera é um candidato promissor para o controle biológico de patógenos fúngicos, limitando assim a dependência excessiva de fungicidas químicos.

Suppression of Xanthomonas oryzae pv. oryzae biofilm formation by Acacia mangium methanol leaf extract / Supressão de Xanthomonas oryzae pv. oryzae: formação do biofilme oryzae pelo extrato de folhas de metanol de Acacia mangium

Abstract Xanthomonas oryzae pv. oryzae (Xoo), a pathogen responsible for rice bacterial leaf blight, produces biofilm to protect viable Xoo cells from antimicrobial agents. A study was conducted to determine the potency of Acacia mangium methanol (AMMH) leaf extract as a Xoo biofilm inhibitor. Four concentrations (3.13, 6.25, 9.38, and 12.5 mg/mL) of AMMH leaf extract were tested for their ability to inhibit Xoo biofilm formation on a 96-well microtiter plate. The results showed that the negative controls had the highest O.D. values from other treatments, indicating the intense formation of biofilm. This was followed by the positive control (Streptomycin sulfate, 0.2 mg/mL) and AMMH leaf extract at concentration 3.13 mg/mL, which showed no significant differences in their O.D. values (1.96 and 1.57, respectively). All other treatments at concentrations of 6.25, 9.38, and 12.5 mg/mL showed no significant differences in their O.D. values (0.91, 0.79, and 0.53, respectively). For inhibition percentages, treatment with concentration 12.5 mg/mL gave the highest result (81.25%) followed by treatment at concentrations 6.25 and 9.38 mg/mL that showed no significant differences in their inhibition percentage (67.75% and 72.23%, respectively). Concentration 3.13 mg/mL resulted in 44.49% of biofilm inhibition and the positive control resulted in 30.75% of biofilm inhibition. Confocal laser scanning microscopy (CLSM) analysis of Xoo biofilm inhibition and breakdown showed the presence of non-viable Xoo cells and changes in aggregation size due to increase in AMMH leaf extract concentration. Control slides showed the absence of Xoo dead cells.

Resumo Xanthomonas oryzae pv. oryzae (Xoo), um patogênico responsável pela influência bacteriana na folha do arroz, produz biofilme para proteger células Xoo viáveis de agentes antimicrobianos. Foi conduzido um estudo para determinar a potência do extrato de folha de Acacia mangium methanol (AMMH) como um inibidor de biofilme Xoo. Quatro concentrações (3,13, 6,25, 9,38 e 12,5 mg/mL) de extrato de folha de AMMH foram testadas quanto à sua capacidade de inibir a formação de biofilme Xoo em uma placa de microtitulação de 96 poços. Os resultados mostraram que os controles negativos tiveram o maior valor de OD do que os outros tratamentos, indicando a intensa formação de biofilme. Isso foi seguido do controle positivo (sulfato de estreptomicina, com concentração de 0,2 mg/mL, e extrato de folha de AMMH, com concentração de 3,13 mg/mL), que não apresentou diferenças significativas nos seus valores OD (1,96 e 1,57, respectivamente). Todos os outros tratamentos com concentrações de 6,25, 9,38, e 12,5 mg/mL não tiveram diferenças significativas nos seus valores OD (0,91, 0,79, e 0,53, respectivamente). Para percentagens de inibição, o tratamento com concentração 12,5 mg/mL apresentou o maior resultado (81,25%), seguido do tratamento em concentrações de 6,25 e 9,38 mg/mL, que não mostraram diferenças significativas na sua percentagem de inibição (67,75 e 72,23%, respectivamente). Concentração 3,13 mg/mL resultou em 44,49% de inibição do biofilme, e o controle positivo resultou em 30,75% de inibição do biofilme. Análise por microscopia confocal de leitura a laser de inibição e separação de biofilme Xoo revelou a presença de células Xoo não viáveis e alterações no tamanho da agregação por causa do aumento na concentração de extrato de folha de AMMH. Slides de controle mostraram a ausência de células Xoo mortas.

HIV-2 Infection in Malaysia: Current situation and the use of in-house real-time reverse transcription PCR for HIV-2

@#HIV-2 surveillance has been carried out in Malaysia for more than 25 years ago. Tests to discriminate HIV-1 and HIV-2 are available but the options of test are limited and the need to develop a new in-house HIV-2 real-time reverse transcription PCR (RT-PCR) is crucial. A study was done on 29 samples from hospitals in Malaysia which were found to be positive screening for HIV-2 antibodies by the commercial Western Blot assay. These samples were further tested by a Western Blot assay that detects specific antibodies to HIV-2. Detection of HIV-2 genome was then performed by using a commercial kit. Fifteen samples were evaluated by using in-house real-time RT-PCR for HIV-2. Ninety-three percent (27/29) of samples have positive results for HIV-2 on HIV-2 Western Blot with only 2 samples showing indeterminate results. All samples showed negative results for HIV-2 genomes by using a PCR commercial kit and the 15 samples that were subjected to our in-house real-time RT-HIV-2 PCR were also tested negative for HIV-2 RNA. Results of HIV-2 Western Blot did not reflect the actual positivity as both HIV-1 and HIV-2 antibodies may cross-react with either viral proteins. None of the samples was confirmed positive for HIV-2 by the commercial and in-house real-time RTPCR. In-house real-time RT-HIV-2 PCR assay can be further used to confirm the presence of HIV-2 genome. Up to the year 2015, Malaysia is still free from HIV-2 infection.

Pesticide Application, Dermal Exposure Risk And Factors Influenced Distribution On Different Body Parts Among Agriculture Workers

@#Agriculture sector accounts significant numbers of injuries and fatalities in the workplace particularly related to pesticide management. Among three main pathways of pesticide exposure, dermal contact is the most common route, which exposure usually occurs during pesticide mixing/loading, application, harvesting and other farming activities. This review aims to present and discuss several vital components of pesticide dermal exposure among agriculture workers, as well as pesticide application in agriculture sector in Malaysia involving different commodity agriculture sub-sectors. Pesticide exposure was discussed from perspective of three pesticide management activities (i.e. preparation, application and cleaning) that contribute to the risk of exposure through three routes (i.e. emission, deposition, transfer). Moreover, this paper also discussed pesticide dermal exposure risk assessment methods which can be defined into exposure assessment and effect assessment. The exposure rate was affected by various factors such as application equipment, application rate and duration, type of pesticide formulation, pesticide management stage, usage of personal protective equipment, training and aptitude of the applicator as well as environmental factors (i.e. temperature, humidity, wind speed and direction). The factors mention earlier have been used to explain the exposure distribution over different parts of the body and support the fact that pesticide type was not a major factor in total exposure.

Suppression of Xanthomonas oryzae pv. oryzae biofilm formation by Acacia mangium methanol leaf extract

Abstract Xanthomonas oryzae pv. oryzae (Xoo), a pathogen responsible for rice bacterial leaf blight, produces biofilm to protect viable Xoo cells from antimicrobial agents. A study was conducted to determine the potency of Acacia mangium methanol (AMMH) leaf extract as a Xoo biofilm inhibitor. Four concentrations (3.13, 6.25, 9.38, and 12.5 mg/mL) of AMMH leaf extract were tested for their ability to inhibit Xoo biofilm formation on a 96-well microtiter plate. The results showed that the negative controls had the highest O.D. values from other treatments, indicating the intense formation of biofilm. This was followed by the positive control (Streptomycin sulfate, 0.2 mg/mL) and AMMH leaf extract at concentration 3.13 mg/mL, which showed no significant differences in their O.D. values (1.96 and 1.57, respectively). All other treatments at concentrations of 6.25, 9.38, and 12.5 mg/mL showed no significant differences in their O.D. values (0.91, 0.79, and 0.53, respectively). For inhibition percentages, treatment with concentration 12.5 mg/mL gave the highest result (81.25%) followed by treatment at concentrations 6.25 and 9.38 mg/mL that showed no significant differences in their inhibition percentage (67.75% and 72.23%, respectively). Concentration 3.13 mg/mL resulted in 44.49% of biofilm inhibition and the positive control resulted in 30.75% of biofilm inhibition. Confocal laser scanning microscopy (CLSM) analysis of Xoo biofilm inhibition and breakdown showed the presence of non-viable Xoo cells and changes in aggregation size due to increase in AMMH leaf extract concentration. Control slides showed the absence of Xoo dead cells.

Resumo Xanthomonas oryzae pv. oryzae (Xoo), um patogênico responsável pela influência bacteriana na folha do arroz, produz biofilme para proteger células Xoo viáveis de agentes antimicrobianos. Foi conduzido um estudo para determinar a potência do extrato de folha de Acacia mangium methanol (AMMH) como um inibidor de biofilme Xoo. Quatro concentrações (3,13, 6,25, 9,38 e 12,5 mg/mL) de extrato de folha de AMMH foram testadas quanto à sua capacidade de inibir a formação de biofilme Xoo em uma placa de microtitulação de 96 poços. Os resultados mostraram que os controles negativos tiveram o maior valor de OD do que os outros tratamentos, indicando a intensa formação de biofilme. Isso foi seguido do controle positivo (sulfato de estreptomicina, com concentração de 0,2 mg/mL, e extrato de folha de AMMH, com concentração de 3,13 mg/mL), que não apresentou diferenças significativas nos seus valores OD (1,96 e 1,57, respectivamente). Todos os outros tratamentos com concentrações de 6,25, 9,38, e 12,5 mg/mL não tiveram diferenças significativas nos seus valores OD (0,91, 0,79, e 0,53, respectivamente). Para percentagens de inibição, o tratamento com concentração 12,5 mg/mL apresentou o maior resultado (81,25%), seguido do tratamento em concentrações de 6,25 e 9,38 mg/mL, que não mostraram diferenças significativas na sua percentagem de inibição (67,75 e 72,23%, respectivamente). Concentração 3,13 mg/mL resultou em 44,49% de inibição do biofilme, e o controle positivo resultou em 30,75% de inibição do biofilme. Análise por microscopia confocal de leitura a laser de inibição e separação de biofilme Xoo revelou a presença de células Xoo não viáveis e alterações no tamanho da agregação por causa do aumento na concentração de extrato de folha de AMMH. Slides de controle mostraram a ausência de células Xoo mortas.

Antifungal efficacy of Moringa oleifera leaf and seed extracts against Botrytis cinerea causing gray mold disease of tomato (Solanum lycopersicum L.)

Abstract Drawbacks associated with the use of chemical fungicides to control plant pathogenic fungi such as Botrytis cinerea stimulate the need for alternatives. Therefore, the present study was carried out to determine the antifungal potentials of Moringa oleifera extracts against B. cinerea. Phytochemical analysis using qualitative chemical tests revealed the presence of huge amount of crucial phytochemicals compounds like phenolic compounds, alkaloids and saponins in the M. oleifera leaf extract. Antifungal bioassay of the crude extracts indicated better mycelial growth inhibition by methanol leaf extract (99%). The minimum inhibitory concentration (MIC) was 5 mg/ml with 100% spore germination inhibition and minimum fungicidal concentration (MFC) was 10 mg/ml with 98.10% mycelial growth inhibition using broth micro dilution and poisoned food techniques. Gas chromatographymass spectrometry (GC-MS) analysis led to the identification of 67 volatile chemical compounds in the leaf extract with 6-decenoic acid (Z)- (19.87%) was the predominant compound. Further chemical elucidation of the crude extracts performed by liquid chromatography with tandem mass spectrometry (LC-MS/MS) showed the presence of non-volatile chemical compounds, mostly flavones, flavonoids and phenolic acids (i.e. quercetin and kaempferol). Scanning electron microscopy and transmission electron microscopy analysis showed positive effect of M. oleifera leaf extract on the treated conidia and mycelium of B. cinerea. Findings revealed that irreversible surface and ultra-structural changes with severe detrimental effects on conidia and mycelium morphology compared to control treatment. Overall findings suggested that M. oleifera leaf extract is a promising candidate for biological control of fungal pathogens, thus limiting overdependence on chemical fungicides.

Resumo As desvantagens associadas ao uso de fungicidas químicos para controlar fungos fitopatogênicos, como Botrytis cinerea, estimulam a necessidade de alternativas. Portanto, o presente estudo foi realizado para determinar o potencial antifúngico de extratos de Moringa oleifera contra B. cinerea. A análise fitoquímica usando testes químicos qualitativos revelou a presença de uma grande quantidade de compostos fitoquímicos cruciais, como compostos fenólicos, alcaloides e saponinas no extrato da folha de M. oleifera. O bioensaio antifúngico dos extratos brutos indicou melhor inibição do crescimento micelial pelo extrato de folhas de metanol (99%). A concentração inibitória mínima (MIC) foi de 5 mg/mL com 100% de inibição da germinação de esporos e a concentração fungicida mínima (MFC) foi de 10 mg/mL com 98,10% de inibição do crescimento micelial usando microdiluição em caldo e técnicas de alimentos envenenados. A análise por cromatografia gasosa-espectrometria de massa (GC-MS) levou à identificação de 67 compostos químicos voláteis no extrato da folha, sendo o ácido 6-decenoico (Z) (19,87%) o composto predominante. Elucidação química adicional dos extratos brutos realizada por cromatografia líquida com espectrometria de massa em tandem (LC-MS/MS) mostrou a presença de compostos químicos não voláteis, principalmente flavonas, flavonoides e ácidos fenólicos (ou seja, quercetina e kaempferol). As análises de microscopia eletrônica de varredura e microscopia eletrônica de transmissão mostraram efeito positivo do extrato de folhas de M. oleifera sobre os conídios e micélios tratados de B. cinerea. Os resultados revelaram a superfície irreversível e alterações ultraestruturais com graves efeitos prejudiciais sobre os conídios e a morfologia micelial, em comparação com o tratamento de controle. Os resultados gerais sugeriram que o extrato da folha de M. oleifera é um candidato promissor para o controle biológico de patógenos fúngicos, limitando assim a dependência excessiva de fungicidas químicos.

Drug resistance mutations among virological failure HIV-1 infected patients in Malaysia

The determination of HIV drug resistance mutations (DRMs) towards antiretroviral (ARV) drugs among HIV-1 treated patients with virological failure is crucial for further management of the patient. This study aimed to assess the most common genomic mutation and to analyse subtypes among the HIV-1 patients with viral load level > 1,000 copies/mL. A total of 101 virological failure HIV-1 patients from four different regions of Peninsular Malaysia with a viral load measurement facility were included in the study. Majority of patients (89.1%) have at least 1 mutation associated with clinical resistance to either protease inhibitors (PIs), nucleoside reverse transcriptase inhibitors (NRTIs) or nonnucleoside reverse transcriptase inhibitors (NNRTIs). Major resistance mutations among the patients towards NRTIs and NNRTIs were 70.3% and 18.8%, respectively. The most common mutation for NRTIs was M184V while K103N mutation was detected in the majority of patients who were treated with NNRTIs. The most commonly observed mutations for major PI and minor PI seen among the study population were V82A/T and L10V, respectively. In HIV-1 subtype analysis, CRF33_01B was the most predominant HIV-1 subtype in this study group. The vast detection of DRMs in this study emphasized the importance of genotypic resistance test in the management of HIV patients as DRMs can alter patient’s susceptibility towards ARV drugs. Further study on larger number of samples is essential for the development of a database on HIV-1 DRMs among patients that experience virological failure in Malaysia.

Evaluation of peripapillary choroidal and retinal nerve fiber layer thickness in eyes with tilted optic disc / Avaliação espessura da camada de fibras nervosas peripapilares da retina e coroide em olhos com disco óptico inclinado

Purpose: This study was performed to evaluate the retinal nerve fiber layer (RNFL) and peripapillary choroidal thickness in eyes with tilted optic disc in order to identify characteristic RNFL and peripapillary choroid patterns verified by optical coherence tomography (OCT). Methods: Twenty-nine eyes of 29 patients with tilted optic discs were studied with spectral-domain (SD)-OCT and compared with age and sex-matched control subjects in a prospective design. The imaging of RNFL was performed using circular scans of a diameter of 3.4 mm around the optic disc using OCT. For measurements of peripapillary choroidal thickness, the standar d protocol for RNFL assessment was performed. Results: SD-OCT indicated significantly lower superotemporal (p<0.001), superonasal (p=0.001), and global (p=0.005) RNFL thicknesses in the tilted disc group than those of the control group. Peripapillary choroid was significantly thicker at the site of the elevated rim of eyes with tilted disc (p<0.001). Conclusion: This study demonstrated a clinical characterization of the main tilted disc morphologies that may be helpful in differentiating a tilted disc from other altered disc morphologies. Further studies are recommended to study the comparison between glaucoma and tilted disc groups. .

Objetivo: Avaliar camada de fibras nervosas da retina (RNFL) e a espessura da coroide peripapilar em olhos com disco óptico inclinado a fim de identificar as características da RNFL e os padrões de coroide peripapilar verificados pela tomografia de coerência óptica (OCT). Métodos: Vinte e nove olhos de 29 pacientes com discos ópticos inclinadas foram estudados prospectivamente com OCT de domínio espectral (SD) e comparados com controles pareados por sexo e idade. As imagens da RNFL foi obtidas por meio de varreduras circulares com um diâmetro de 3,4 mm em torno do disco óptico usando OCT. Para as medições de espessura da coroide peripapilar, o protocolo padrão para avaliação da RNFL foi realizado. Resultados: O OCT SD indicou diminuição das espessuras significativas da RNFL superotemporal (p<0,001), superonasal (p=0,001), e global (p=0,005) no grupo disco inclinado em relação aos do grupo controle. A coroide peripapilar foi significativamente mais espessa no local da borda elevada dos olhos com disco inclinado (p<0,001). Conclusões: Este estudo demonstrou que a caracterização clínica das principais morfologias disco inclinado pode ser útil na diferenciação entre um disco inclinado de outras alterações morfológicas de disco. Seria importante a comparação entre grupos com glaucoma e com discos inclinados, estudos futuros. .

Seroprevalence of toxoplasma gondii in women in Najran City, Saudi Arabia

To evaluate the seroprevalence of Toxoplasma gondii among Saudi pregnant women in Najran City, as well as to measure the performance of the diagnostic tests used. A total of 96 women attending prenatal special clinics [Oteafyn special clinic] in Najran Province, Saudi Arabia, over a one year period, from September 2012 to September 2013 were screened for the presence of Toxoplasma antibody in their blood serum using an indirect hemagglutination assay [IHA]. Specific immunoglobulin [Ig] G and IgM antibodies were evaluated using an enzyme-linked immunosorbent assay [ELISA]. Out of the 96 samples of sera tested using IHA, 20 [20.8%] were found to be positive with a titer ranging from 1:80 to 1:320, while 29 [29.2%] and 3 [3.1%] revealed Toxoplasma IgG and Toxoplasma IgM. A positive relationship was found between the seroprevalence of toxoplasmosis and age of tested women, especially in the age group of 21-30 years old [54.7%] by using ELISA-IgG, and 31-40 years old [4.5%] by using ELISA-IgM. The seroprevalence of Toxoplasmosis among pregnant women was found to be comparatively high, compared with previous reports from Saudi Arabia. Serologic checkup before and during pregnancy for seronegative women is recommended

Extracorporeal shock-wave lithotripsy success rate and complications: initial experience at Sultan Qaboos university hospital

To assess the efficacy and safety of extracorporeal shock wave lithotripsy with Modularis Vario Siemens in the management of patients with renal and ureteral stones. Between 2007 and 2009, 225 outpatients were treated with Siemens Modularis Vario lithotripter at Sultan Qaboos University Hospital. Stone size, location, total number of shockwaves, stone-free rate, complications and adjunctive interventions were investigated. Chi-Square and Logistic Regression analyses were used, with p<0.05 set as the level of significance. Of the 225 initial consecutive patients who underwent extracorporeal shock wave lithotripsy, 192 [85%] had renal stones and 33 [15%] had ureteric stones. The mean +/- SD stone size was 11.3 +/- 4.5 mm, while the mean age of the patients was 39.9 +/- 12.8 years with 68.5% males. The mean renal stone size was 11.6 +/- 4.7 mm; a mean of 1.3 sessions was required. The mean ureteric stone size was 9.9 +/- 3 mm; and a mean of 1.3 sessions was required. Treatment success [defined as complete clearance of ureteric stones, stone-free or clinically insignificant residual fragments of <4 mm for renal stones] was 74% for renal stones and 88% for ureteric stones. Additional extracorporeal shock wave lithotripsy and ureteroscopy were the most adjunctive procedures used for stone clearance. Complications occurred in 74 patients [38.5%] with renal stones and 13 patients [39.4%] with uretetric stones. The most common complication was loin pain [experienced by 16.7% with renal stones and 21% with ureteric stones]. Severe renal colic mandating admission occurred in 2% of patients with renal stones and 6% of patients with ureteric stones. In patients with renal stone, steinstrasse occurred in 3.6% and infection post extracorporeal shock wave lithotripsy in 0.5%. Using Multivariate Logistic Regression analysis, factors found to have significant effect on complete stone clearance were serum creatinine [p=0.004] and the number of shockwaves [p=0.021]. Siemens Modularis Vario lithotripter is a safe and effective tool for treating renal and ureteric stones

Clinical Decision Making Ability of Nursing Students in a Tertiary Hospital

Decision making in nursing is one of the most important skills nurses must apply and utilize in their nursing practice. The aim of this study was to determine the perception of clinical decision making ability among nursing students. A descriptive crosssectional study was conducted in a tertiary hospital. A total of 54 nursing students were recruited using a modified version of Clinical Decision Making in Nursing Scale (CDMNS) adapted from Jenkins (1985). The findings showed good CDMNS score with mean and standard deviation of (124.24±12.713). The four sub-scales of CDMNS were: searching for alternative (33.24±4.821), canvassing (28.74±3.514), evaluation and re-evaluation (31.43±3.922), searching for information(30.83±4.765). Nineteen (35%) of the participants chose nursing as their first choice, whereas 35 participants (65%) did not. Thirthy seven (69%) participants were satisfied with their nursing competency, 17 (31%) were unsatisfied. There were significant differences between searching for alternatives, evaluation and re-evaluation, and nursing as their first choice (p=<0.05). There were also significant differences between searching for alternatives and satisfaction with nursing competency (p=<0.05). There was significant difference between education level and searching for alternatives and information (p=<0.05). The nursing students possessed adequate clinical decision making ability. Although most of the nursing students did not choose nursing as their first choice, they sought for alternatives and evaluated-reevaluated during their decision making process. Nursing students’ satisfaction also contributed to appropriate clinical decision making in the critical care setting.

Comparision of cluster formation by mahalanobis’ D2 and metroglyph analyses in groundnut (Arachis Hypogaea L.).

Fifty genotypes of groundnut were classified into five and seven clusters in D2 and Metroglyph methods of analyses respectively. Harvest Index and 100-seed weight were the main contributors to total divergence as per metroglyph analysis. The characters on X and Y axes in scattered metroglyph diagram determine the grouping of genotypes, hence it poses a problem for judicious selection of these characters. The comparative study of these methods indicated that metroglyph analysis would be suitable for preliminary grouping when large number of genotypes are considered at a time. The differences between the two methods, the number of clusters formed, inter cluster distance, characters considered for grouping and superimposition of genotypes within the cluster indicated that D2 analysis was more effective compared to metroglyph analysis.

Character association and path analysis in groundnut (arachis hypogaea l.).

Correlation and path co-efficient analysis were carried out for pod yield and its component characters in 50 genotypes of groundnut. The genotypic correlation co-efficients were found to be of relatively higher magnitude than the corresponding phenotypic correlation co-efficients, indicating strong inherent association between the characters. Pod yield displayed significant positive association with kernel yield per plant, mature pods per plant, total pods per plant, harvest index, 100-seed weight, root weight, plant height and shoot weight. Path co-efficient analysis revealed high direct effects of kernel yield per plant and harvest index on pod yield. Hence, it would be rewarding to give due importance on the selection of these characters for rapid improvement in pod yield of groundnut.

The mosquito dunk (Bacillus thuringensis israelienses) as a control agent against mosquito larvae in Khartoum State, Sudan Abdalmagi

Background: During the mid-1970s, larvicides have become available that are highly effective, yet selective in action, and therefore environmentally safe to non-target organisms, as well as for human exposure(1). Objectives: The small field trial was carried out from 12th of January to 16th of February 2008 in Khartoum State to evaluate the efficacy and persistence mosquito dunk® (Bti) against mosquito larvae and to measure the effect of physic-chemical properties on mosquito dunk. Material & Methods: The efficacy and persistence of Bacillus thuringiensis subspecies israelensis (mosquito dunk®) as a biological control agent against mosquito larvae was conducted in Khartoum State. Twelve ponds were used as natural breeding habitats of mosquitoes; six of them were treated with dunk at a rate of 1 dunk per 100 square feet and six ponds left untreated (control). Results: The study revealed that more than 80% reduction in immature stages density was observed up to 5, 3 and 2 weeks for the 3rd, 2nd and 4th instars of Anopheline spp., respectively. However, the study showed that the mosquito dunk was noteffective (under 80% mortality) against 1st, instar larvae and pupae of Anopheline species as well as Culex developmental stages. Conclusion & Interpretations: The dunk was very effective in controlling 3rd and 4th instars of Anopheles spp; for 2 weeks interval. Therefore we propose a surface application regime of once every 2 weeks for mosquito dunk

Oxidative Stress in Vivax Malaria

Malaria is still a leading cause of morbidity and mortality. The increase in lipid peroxidation reported in malaria infection and antioxidant status may be a useful marker of oxidative stress during malaria infection. The aim of this study was to investigate the role of antioxidant enzymes against toxic reactive oxygen species in patients infected with Plasmodium vivax and healthy controls. Malondialdehyde levels, superoxide dismutase, and glutathione peroxidase activities were determined in 91 P. vivax patients and compared with 52 controls. Malondialdehyde levels, superoxide dismutase, and glutathione peroxidase activities were 8.07+/-2.29 nM/ml, 2.69+/-0.33 U/ml, and 49.6+/-3.2 U/g Hb in the patient group and 2.72+/-0.50 nM/ml, 3.71+/-0.47 U/ml, and 62.3+/-4.3 U/g Hb in the control group, respectively. Malondialdehyde levels were found statistically significant in patients with vivax malaria higher than in healthy controls (P<0.001). On the other hand, superoxide dismutase and glutathione peroxidase activities were found to be significantly lower in vivax malaria patients than in controls (P<0.05). There was an increase in oxidative stress in vivax malaria. The results suggested that antioxidant defense mechanisms may play an important role in the pathogenesis of P. vivax.

Prevalence of sen virus infection in hemodialysis and blood diseased patients in Assiut university hospitals

SENV is a blood- borne, circular ss DNA virus and possessing nine genotypes [A to I]. Among nine genotypes, SENV-D and SENV-H genotypes have the strong link with patients with non [A-E] hepatitis infections .Recently, the identification of SEN virus [SENV] as a possible etiologic agent of parenteral transmission hepatitis let to the study of the prevalence of such agent. This study compared SENV prevalence and its two important genotypes [D and H] which might be pathogenic in high risk subjects including blood diseased patients and hemodialysis patients and low risk subjects as healthy blood donors. This study included 75 multitransfused patients, 60 of them were hemodialysis and the remaining were blood diseased including hemophilics, anemics and leukemics. The study included also 25 healthy blood donors as a control They were received consecutively at Department of Internal Medicine, Assiut University Hospital. The sera were separated and SENV DNA was detected by polymerase chain reaction. The results showed a higher prevalence of SENV infection in patients group than blood donors [46.7% versus 20%].No significant relation was found between SENV infection and age, duration of hemodialysis or liver enzymes. However, there was significant difference between SENV positive and negative patients as regards gender and number of blood transfusions. SENV is commonly present in blood diseased and hemodialysis patients attended to Assiut University Hospitals as well as in blood donors at variable rates. SENV infection has been found in only 20% of blood donors but in 46.7% of patients. The results also indicated that other possible routes of SENV infection other than blood transfusion may be included. Its pathogenic role in causing hepatitis is not documented, so far it can be considered as simple guest till further studies have been done

Prevalence of transfusion transmitted virus [TTV] infection in patients with liver diseases attending to Assiut University Hospital

In 1997 Transfusion Transmitted Virus [TTV] was isolated from the serum of a patient with post transfusion hepatitis of unknown etiology, in Japan. It's considered as a causative agent of non A to G hepatitis. To assess the prevalence of TTV infection among patients with liver diseases compared with healthy controls and the significance of TTV infection in patients with liver disease. This investigation was conducted on 76 patients with liver diseases, classified into four groups: Acute hepatitis group [20 patients], chronic liver diseases [30 patients], Liver cirrhosis [18 patients] and hepatocellular carcinoma [8 patients]. In addition to the patient groups, the fifth group of 24 healthy blood donors as control group was included within the study. All patients and control groups were examined for the detection of TTV DNA by PCR. Thirty seven had history of blood transfusion and 23 patients were subjected to surgical manipulation. TTV DNA was detected in 57.9% [44/76] of patients with liver diseases and in 45.7% [11/24] of healthy blood donors. The prevalence of TTV in the studied groups were 60%, 46.7%, 66,7% and 75% in acute hepatitis, chronic liver diseases, liver cirrhosis and hepatocellular carcinoma respectively. TTV is commonly present in patients with liver disease attended to Assiut University Hospitals as well as in blood donors. High prevalence of TTV in blood donors may indicate other routes of transfusion of this virus such as fecal-oral and sexual routes beside transfusion of blood and blood products. The blood transfusion and operative intervention are a major risk factor for transmission of TTV