Phytoremediation of toxic heavy metals in polluted soils and water of Dargai District Malakand Khyber Pakhtunkhwa, Pakistan.

The contamination of natural resources with heavy metals released from steel mills is the primary cause of soil and water pollution in the Dargai Malakand, located on the northern side of Pakistan. Therefore, the present study was aimed to determine the level of heavy metals in soil and water samples of this area. The wild plant growing (nine native plants: Pteris vittata, Populus nigra, Eucalyptus camaldulensis, Persicaria maculosa, Arundo donax, Xanthium strumarium, Verbascum thapsus, Ricinus communis and Parthenium hysterophorus) there were then tested for their phytoremediation capabilities which is an environmentally friendly, generally utilized, and low-cost approach to eliminate heavy metals from polluted soils and water. Soil, water, and effluent samples were taken from the contaminated sites of seven steel mills in Dargai District Malakand and subjected to heavy metals analysis. Based on bioconcentration factor (BCF) and translocation factor (TF) calculated, The highest BCF for zinc was recorded for Pteris vittata roots (3.93), while the lowest value was observed for Verbascum thapsus leaves (0.306). Pteris vittata root showed the highest BCF for iron (1.618), while Ricinus communis leaves showed the lowest (0.023). The highest BCF value for chromium was highest for Populus nigra roots (0.717), while the lowest value was recorded for Persicaria maculosa leaves (0.031). For the selected metals; Fe, Zn and Cr the highest TF were recorded for Pteris vittata (0.988), Verbascum thapsus (0.944) and Xanthium strumairum (0.968) respectively. Therefore, it is recommended that these plants should be grown near to steel mills to reclaim heavy metals from industrial effluent, polluted soil as well as from polluted water.

Response of Rhizobacterial strains and organic amendments on chickpea growth.

Plant Growth Promoting Rhizobacteria (PGPR) are beneficial bacteria that colonize plant roots and promote plant growth through a variety of mechanisms such as phosphate solubilization, phytohormones production, antifungal activity and also improve plant growth and yield. Field experiment was carried out to investigate the residual effect of organic amendments plus soil microbes along with integrated nutrient management. (PGPR) are important soil organism that promotes plant growth and yield root colonization is an example of a direct and indirect mechanism. The treatments included control, (inorganic fertilizer and no organic fertilization).Five bacterial strains were identified morphologically and biochemically screened from the rhizospheres of chickpea, lentil, barseem, mungbean, and sesame. The experiment was conducted at the Arid Zone Research Center in D.I.Khan (Pakistan). The majority of isolates resulted in significant increase in shoot length, root length, and dry matter production of Cicer arietinum seedlings' shoot and root. The experiment represented that isolates treated plots with rhizobium strain inoculation resulted in greater plant height (35.000 cm) and nodule count (38.00) No of pods per plant-1 (44.66) when compared to the control treatment, While (Mesorhizobium cicero) along with organic amendments showed significant response the greater root length (50 cm) was observed in T4 treatment. The Performance of rhizobial strains on chickpea germination in an arid environment was found to significantly increase crop germination percentage. This combination thus increases nitrogen and phosphorus uptake in inoculation treated plots. The study found that plots with inoculation treatments yielded significantly higher than non-treated plots Treatment with Mesorhizobium Cicero and compost resulted in a higher grain yield (8%) as compared to the control. The greater grain yield was observed in Treatment T4 (183.67).The result showed that use of PGPR have the potential to increase nutrient absorption from soil while improved growth of chickpea.

Statistical modeling for analyzing grain yield of durum wheat under rainfed conditions in Azad Jammu Kashmir, Pakistan / Modelagem estatística para analisar o rendimento de grãos de trigo durum sob condições de chuva em Azad Jammu Kashmir, Paquistão

One of the most important traits that plant breeders aim to improve is grain yield which is a highly quantitative trait controlled by various agro-morphological traits. Twelve morphological traits such as Germination Percentage, Days to Spike Emergence, Plant Height, Spike Length, Awn Length, Tillers/Plant, Leaf Angle, Seeds/Spike, Plant Thickness, 1000-Grain Weight, Harvest Index and Days to Maturity have been considered as independent factors. Correlation ,regression, and principal component analysis (PCA) are used to identify the different durum wheat traits, which significantly contribute to the yield. The necessary assumptions required for applying regression modeling have been tested and all the assumptions are satisfied by the observed data. The outliers are detected in the observations of fixed traits and Grain Yield. Some observations are detected as outliers but the outlying observations did not show any influence on the regression fit. For selecting a parsimonious regression model for durum wheat, best subset regression, and stepwise regression techniques have been applied. The best subset regression analysis revealed that Germination Percentage, Tillers/Plant, and Seeds/Spike have a marked increasing effect whereas Plant thickness has a negative effect on durum wheat yield. While stepwise regression analysis identified that the traits, Germination Percentage, Tillers/Plant, and Seeds/Spike significantly contribute to increasing the durum wheat yield. The simple correlation coefficient specified the significant positive correlation of Grain Yield with Germination Percentage, Number of Tillers/Plant, Seeds/Spike, and Harvest Index. These results of correlation analysis directed the importance of morphological characters and their significant positive impact on Grain Yield. [...].

Uma das características mais importantes que os produtores de plantas visam melhorar é o rendimento de grãos, que é uma particularidade altamente quantitativa e controlada por várias características agromorfológicas. Foram considerados 12 traços morfológicos como fatores independentes, como Porcentagem de Germinação, Dias para Emergência da Espiga, Altura da Planta, Comprimento da Espiga, Comprimento da Aresta, Perfilhos /Planta, Ângulo da Folha, Sementes /Espiga, Espessura da Planta, Peso de 1000 Grãos, Índice de Colheita e Dias até a Maturidade. A correlação, regressão e análise de componentes principais (em inglês Principal Component Analysis (PCA)) são usadas para identificar as diferentes características do trigo duro, que contribuem significativamente para o rendimento. As suposições necessárias exigidas para a aplicação da modelagem de regressão foram testadas e todas as suposições são adequadas de acordo com os dados observados. Os outliers são detectados nas observações de características fixas e rendimento de grãos. Algumas observações são detectadas como outliers, mas as observações outliers não mostraram qualquer influência no ajuste da regressão. Para selecionar um modelo de regressão parcimonioso para o trigo duro, foram aplicadas tanto a melhor regressão de subconjunto quanto as técnicas de regressão stepwise. A melhor análise de regressão de subconjunto revelou que a porcentagem de germinação, perfilhos /planta e sementes /espiga tem um efeito de aumento acentuado, enquanto a espessura da planta tem um efeito negativo sobre o rendimento do trigo duro. Enquanto a análise de regressão passo a passo identificou que as características, porcentagem de germinação, perfilhos/planta e sementes /espiga contribuem significativamente para aumentar a produtividade do trigo duro. O coeficiente de correlação simples especificou a correlação positiva significativa do [...].

Statistical modeling for analyzing grain yield of durum wheat under rainfed conditions in Azad Jammu Kashmir, Pakistan

Abstract One of the most important traits that plant breeders aim to improve is grain yield which is a highly quantitative trait controlled by various agro-morphological traits. Twelve morphological traits such as Germination Percentage, Days to Spike Emergence, Plant Height, Spike Length, Awn Length, Tillers/Plant, Leaf Angle, Seeds/Spike, Plant Thickness, 1000-Grain Weight, Harvest Index and Days to Maturity have been considered as independent factors. Correlation, regression, and principal component analysis (PCA) are used to identify the different durum wheat traits, which significantly contribute to the yield. The necessary assumptions required for applying regression modeling have been tested and all the assumptions are satisfied by the observed data. The outliers are detected in the observations of fixed traits and Grain Yield. Some observations are detected as outliers but the outlying observations did not show any influence on the regression fit. For selecting a parsimonious regression model for durum wheat, best subset regression, and stepwise regression techniques have been applied. The best subset regression analysis revealed that Germination Percentage, Tillers/Plant, and Seeds/Spike have a marked increasing effect whereas Plant thickness has a negative effect on durum wheat yield. While stepwise regression analysis identified that the traits, Germination Percentage, Tillers/Plant, and Seeds/Spike significantly contribute to increasing the durum wheat yield. The simple correlation coefficient specified the significant positive correlation of Grain Yield with Germination Percentage, Number of Tillers/Plant, Seeds/Spike, and Harvest Index. These results of correlation analysis directed the importance of morphological characters and their significant positive impact on Grain Yield. The results of PCA showed that most variation (70%) among data set can be explained by the first five components. It also identified that Seeds/Spike; 1000-Grain Weight and Harvest Index have a higher influence in contributing to the durum wheat yield. Based on the results it is recommended that these important parameters might be considered and focused in future durum wheat breeding programs to develop high yield varieties.

Resumo Uma das características mais importantes que os produtores de plantas visam melhorar é o rendimento de grãos, que é uma particularidade altamente quantitativa e controlada por várias características agromorfológicas. Foram considerados 12 traços morfológicos como fatores independentes, como Porcentagem de Germinação, Dias para Emergência da Espiga, Altura da Planta, Comprimento da Espiga, Comprimento da Aresta, Perfilhos /Planta, Ângulo da Folha, Sementes /Espiga, Espessura da Planta, Peso de 1000 Grãos, Índice de Colheita e Dias até a Maturidade,. A correlação, regressão e análise de componentes principais (em inglês Principal Component Analysis (PCA)) são usadas para identificar as diferentes características do trigo duro, que contribuem significativamente para o rendimento. As suposições necessárias exigidas para a aplicação da modelagem de regressão foram testadas e todas as suposições são adequadas de acordo com os dados observados. Os outliers são detectados nas observações de características fixas e rendimento de grãos. Algumas observações são detectadas como outliers, mas as observações outliers não mostraram qualquer influência no ajuste da regressão. Para selecionar um modelo de regressão parcimonioso para o trigo duro, foram aplicadas tanto a melhor regressão de subconjunto quanto as técnicas de regressão stepwise. A melhor análise de regressão de subconjunto revelou que a porcentagem de germinação, perfilhos /planta e sementes /espiga tem um efeito de aumento acentuado, enquanto a espessura da planta tem um efeito negativo sobre o rendimento do trigo duro. Enquanto a análise de regressão passo a passo identificou que as características, porcentagem de germinação, perfilhos/planta e sementes /espiga contribuem significativamente para aumentar a produtividade do trigo duro. O coeficiente de correlação simples especificou a correlação positiva significativa do rendimento de grãos com a porcentagem de germinação, número de perfilhos/planta, sementes / espiga e índice de colheita. Esses resultados da análise de correlação direcionaram a importância dos caracteres morfológicos e seu impacto positivo e significativo no rendimento de grãos. Os resultados da PCA mostraram que a maior parte da variação (70%) entre o conjunto de dados pôde ser explicada pelos cinco primeiros componentes. Também identificou que Sementes / Espiga, Peso de 1000 Grãos e Índice de Colheita têm uma maior influência na contribuição para o rendimento do trigo duro. Com base nos resultados, recomenda-se que esses importantes parâmetros possam ser considerados e focados em futuros programas de melhoramento de trigo duro para desenvolver variedades de alto rendimento.

Statistical modeling for analyzing grain yield of durum wheat under rainfed conditions in Azad Jammu Kashmir, Pakistan / Modelagem estatística para analisar o rendimento de grãos de trigo durum sob condições de chuva em Azad Jammu Kashmir, Paquistão

One of the most important traits that plant breeders aim to improve is grain yield which is a highly quantitative trait controlled by various agro-morphological traits. Twelve morphological traits such as Germination Percentage, Days to Spike Emergence, Plant Height, Spike Length, Awn Length, Tillers/Plant, Leaf Angle, Seeds/Spike, Plant Thickness, 1000-Grain Weight, Harvest Index and Days to Maturity have been considered as independent factors. Correlation, regression, and principal component analysis (PCA) are used to identify the different durum wheat traits, which significantly contribute to the yield. The necessary assumptions required for applying regression modeling have been tested and all the assumptions are satisfied by the observed data. The outliers are detected in the observations of fixed traits and Grain Yield. Some observations are detected as outliers but the outlying observations did not show any influence on the regression fit. For selecting a parsimonious regression model for durum wheat, best subset regression, and stepwise regression techniques have been applied. The best subset regression analysis revealed that Germination Percentage, Tillers/Plant, and Seeds/Spike have a marked increasing effect whereas Plant thickness has a negative effect on durum wheat yield. While stepwise regression analysis identified that the traits, Germination Percentage, Tillers/Plant, and Seeds/Spike significantly contribute to increasing the durum wheat yield. The simple correlation coefficient specified the significant positive correlation of Grain Yield with Germination Percentage, Number of Tillers/Plant, Seeds/Spike, and Harvest Index. These results of correlation analysis directed the importance of morphological characters and their significant positive impact on Grain Yield. The results of PCA showed that most variation (70%) among data set can be explained by the first five components. It also identified that Seeds/Spike; 1000-Grain Weight and Harvest Index have a higher influence in contributing to the durum wheat yield. Based on the results it is recommended that these important parameters might be considered and focused in future durum wheat breeding programs to develop high yield varieties.

Uma das características mais importantes que os produtores de plantas visam melhorar é o rendimento de grãos, que é uma particularidade altamente quantitativa e controlada por várias características agromorfológicas. Foram considerados 12 traços morfológicos como fatores independentes, como Porcentagem de Germinação, Dias para Emergência da Espiga, Altura da Planta, Comprimento da Espiga, Comprimento da Aresta, Perfilhos /Planta, Ângulo da Folha, Sementes /Espiga, Espessura da Planta, Peso de 1000 Grãos, Índice de Colheita e Dias até a Maturidade,. A correlação, regressão e análise de componentes principais (em inglês Principal Component Analysis (PCA)) são usadas para identificar as diferentes características do trigo duro, que contribuem significativamente para o rendimento. As suposições necessárias exigidas para a aplicação da modelagem de regressão foram testadas e todas as suposições são adequadas de acordo com os dados observados. Os outliers são detectados nas observações de características fixas e rendimento de grãos. Algumas observações são detectadas como outliers, mas as observações outliers não mostraram qualquer influência no ajuste da regressão. Para selecionar um modelo de regressão parcimonioso para o trigo duro, foram aplicadas tanto a melhor regressão de subconjunto quanto as técnicas de regressão stepwise. A melhor análise de regressão de subconjunto revelou que a porcentagem de germinação, perfilhos /planta e sementes /espiga tem um efeito de aumento acentuado, enquanto a espessura da planta tem um efeito negativo sobre o rendimento do trigo duro. Enquanto a análise de regressão passo a passo identificou que as características, porcentagem de germinação, perfilhos/planta e sementes /espiga contribuem significativamente para aumentar a produtividade do trigo duro. O coeficiente de correlação simples especificou a correlação positiva significativa do rendimento de grãos com a porcentagem de germinação, número de perfilhos/planta, sementes / espiga e índice de colheita. Esses resultados da análise de correlação direcionaram a importância dos caracteres morfológicos e seu impacto positivo e significativo no rendimento de grãos. Os resultados da PCA mostraram que a maior parte da variação (70%) entre o conjunto de dados pôde ser explicada pelos cinco primeiros componentes. Também identificou que Sementes / Espiga, Peso de 1000 Grãos e Índice de Colheita têm uma maior influência na contribuição para o rendimento do trigo duro. Com base nos resultados, recomenda-se que esses importantes parâmetros possam ser considerados e focados em futuros programas de melhoramento de trigo duro para desenvolver variedades de alto rendimento.

Statistical modeling for analyzing grain yield of durum wheat under rainfed conditions in Azad Jammu Kashmir, Pakistan.

One of the most important traits that plant breeders aim to improve is grain yield which is a highly quantitative trait controlled by various agro-morphological traits. Twelve morphological traits such as Germination Percentage, Days to Spike Emergence, Plant Height, Spike Length, Awn Length, Tillers/Plant, Leaf Angle, Seeds/Spike, Plant Thickness, 1000-Grain Weight, Harvest Index and Days to Maturity have been considered as independent factors. Correlation, regression, and principal component analysis (PCA) are used to identify the different durum wheat traits, which significantly contribute to the yield. The necessary assumptions required for applying regression modeling have been tested and all the assumptions are satisfied by the observed data. The outliers are detected in the observations of fixed traits and Grain Yield. Some observations are detected as outliers but the outlying observations did not show any influence on the regression fit. For selecting a parsimonious regression model for durum wheat, best subset regression, and stepwise regression techniques have been applied. The best subset regression analysis revealed that Germination Percentage, Tillers/Plant, and Seeds/Spike have a marked increasing effect whereas Plant thickness has a negative effect on durum wheat yield. While stepwise regression analysis identified that the traits, Germination Percentage, Tillers/Plant, and Seeds/Spike significantly contribute to increasing the durum wheat yield. The simple correlation coefficient specified the significant positive correlation of Grain Yield with Germination Percentage, Number of Tillers/Plant, Seeds/Spike, and Harvest Index. These results of correlation analysis directed the importance of morphological characters and their significant positive impact on Grain Yield. The results of PCA showed that most variation (70%) among data set can be explained by the first five components. It also identified that Seeds/Spike; 1000-Grain Weight and Harvest Index have a higher influence in contributing to the durum wheat yield. Based on the results it is recommended that these important parameters might be considered and focused in future durum wheat breeding programs to develop high yield varieties.

A randomized adaptive phase II/III study of buparlisib, a pan-class I PI3K inhibitor, combined with paclitaxel for the treatment of HER2- advanced breast cancer (BELLE-4).

Background: Phosphatidylinositol 3-kinase (PI3K) pathway activation in preclinical models of breast cancer is associated with tumor growth and resistance to anticancer therapies, including paclitaxel. Effects of the pan-Class I PI3K inhibitor buparlisib (BKM120) appear synergistic with paclitaxel in preclinical and clinical models. Patients and methods: BELLE-4 was a 1:1 randomized, double-blind, placebo-controlled, adaptive phase II/III study investigating the combination of buparlisib or placebo with paclitaxel in women with human epidermal growth factor receptor 2-negative locally advanced or metastatic breast cancer with no prior chemotherapy for advanced disease. Patients were stratified by PI3K pathway activation and hormone receptor status. The primary endpoint was progression-free survival (PFS) in the full and PI3K pathway-activated populations. An adaptive interim analysis was planned following the phase II part of the study, after ≥125 PFS events had occurred in the full population, to decide whether the study would enter phase III (in the full or PI3K pathway-activated population) or be stopped for futility. Results: As of August 2014, 416 patients were randomized to receive buparlisib (207) or placebo (209) with paclitaxel. At adaptive interim analysis, there was no improvement in PFS with buparlisib versus placebo in the full (median PFS 8.0 versus 9.2 months, hazard ratio [HR] 1.18), or PI3K pathway-activated population (median PFS 9.1 versus 9.2 months, HR 1.17). The study met protocol-specified criteria for futility in both populations, and phase III was not initiated. Median duration of study treatment exposure was 3.5 months in the buparlisib arm versus 4.6 months in the placebo arm. The most frequent adverse events with buparlisib plus paclitaxel (≥40% of patients) were diarrhea, alopecia, rash, nausea, and hyperglycemia. Conclusions: Addition of buparlisib to paclitaxel did not improve PFS in the full or PI3K pathway-activated study population. Consequently, the trial was stopped for futility at the end of phase II.

Plant species affect colonization patterns and metabolic activity of associated endophytes during phytoremediation of crude oil-contaminated soil.

Plants coupled with endophytic bacteria hold great potential for the remediation of polluted environment. The colonization patterns and activity of inoculated endophytes in rhizosphere and endosphere of host plant are among the primary factors that may influence the phytoremediation process. However, these colonization patterns and metabolic activity of the inoculated endophytes are in turn controlled by none other than the host plant itself. The present study aims to determine such an interaction specifically for plant-endophyte systems remediating crude oil-contaminated soil. A consortium (AP) of two oil-degrading endophytic bacteria (Acinetobacter sp. strain BRSI56 and Pseudomonas aeruginosa strain BRRI54) was inoculated to two grasses, Brachiaria mutica and Leptochloa fusca, vegetated in crude oil-contaminated soil. Colonization patterns and metabolic activity of the endophytes were monitored in the rhizosphere and endosphere of the plants. Bacterial augmentation enhanced plant growth and crude oil degradation. Maximum crude oil degradation (78%) was achieved with B. mutica plants inoculated with AP consortium. This degradation was significantly higher than those treatments, where plants and bacteria were used individually or L. fusca and endophytes were used in combination. Moreover, colonization and metabolic activity of the endophytes were higher in the rhizosphere and endosphere of B. mutica than L. fusca. The plant species affected not only colonization pattern and biofilm formation of the inoculated bacteria in the rhizosphere and endosphere of the host plant but also affected the expression of alkane hydroxylase gene, alkB. Hence, the investigation revealed that plant species can affect colonization patterns and metabolic activity of inoculated endophytic bacteria and ultimately the phytoremediation process.

Chaos control via feeding switching in an omnivory system.

Tanabe and Namba (Ecology, 86, 3411-3414) studied a three species Lotka-Volterra model with omnivory and explored that omnivory can create chaos. It is well documented that predator switching is a similar biological phenomenon to omnivory and likely to occur simultaneously. In the present paper, the tri-trophic Lotka-Volterra food web model with omnivory and predator switching is re-investigated. We observe that if we incorporate predator switching in the system and the intensity of predator switching increases above a threshold value, then the system will be stable from chaotic dynamics. To study the global dynamics of the system extensive numerical simulations are performed. Our analytical and numerical results suggest that predator switching mechanism enhances the stability and the persistence of a food chain system.

Suicide and attempted suicide trends in Mianwali, Pakistan: social perspective.

This study aimed to examine the epidemiology and potential causes of suicides and attempted suicide cases that were reported to Rescue 1122 Mianwali in Pakistan. The data were collected prospectively for 2011. One hundred and eight suicides were reported during the study period. Four (4%) victims were found dead at the scene and 104 (96%) were taken to hospital. There were 84(78%) men and 24(22%) women and 98 (90%) were aged 11-30 years. Toxic substances (36%), pesticides (31%) and drug overdose (11%) were the most common materials/methods used. Depression/mental illness (33%), socioeconomic conditions (24%) and unemployment (21%) were the major reasons for attempting suicide. Suicide is a sensitive and multifaceted problem that needs to be addressed in Pakistan.

Investigation of impact of environmental changes on precipitation pattern of Pakistan.

In this paper, variability in precipitation pattern of Pakistan due to environmental and climatic changes has been studied. Maps have been generated to depict global precipitation variation. Precipitation data of 25 stations of Pakistan have been used. These data were taken from Meteorological Department, Islamabad, Pakistan. The results of two global climate models, namely Australia's Commonwealth Scientific and Industrial Research's third generation general circulation model and National Center for Atmospheric Research's first generation precipitation circulation model for A2 scenario have been applied to investigate the changes. It is observed that precipitation pattern will change significantly in the future. The occurrence of precipitation in all seasons for Pakistan is expected to increase with almost uniform distribution across a season. Average annual precipitation of the country will undergo an increase in the range of +57 to +71 % as compared to average of the base period.

Detection and phylogenetic analysis of peste des petits ruminants virus isolated from outbreaks in Punjab, Pakistan.

Peste des Petits Ruminants (PPR) is an important viral disease of small ruminants and is endemic in Pakistan. In the following study, samples from two outbreaks of PPR in goats have been subjected to laboratory investigations. The Peste des Petits Ruminants virus (PPRV) genome was detected using both conventional and real-time PCR. Genetic characterization of the local PPRV field isolates was conducted by sequencing 322 bp of the fusion (F) gene and 255 bp of the nucleoprotein (N) gene. The phylogenetic tree based on the F gene clustered samples from both outbreaks into lineage 4 along with other Asian isolates, specifically into subcluster 1 along with isolates from Middle East. Analysis of N gene revealed a different pattern. In this case, the Pakistani samples clustered with Chinese, Tajikistani and Iranian isolates, which probably represents the true geographical pattern of virus circulation. This is the first report presenting the phylogenetic tree based on N gene as well as performing a parallel comparison of the trees of F and N gene together from Pakistani isolates. The results of this study shed light on the PPRV population in Pakistan and emphasize the importance of using molecular methods to understand the epidemiology. Such understanding is essential in any efforts to control the number and impact of outbreaks that are occurring in endemic countries such as Pakistan, especially in the current scenario where OIE and FAO are eager to control and subsequently eradicate PPR from the globe, as has been achieved for Rinderpest.

Molecular characterisation of foot-and-mouth disease viruses from Pakistan, 2005-2008.

Foot-and-mouth disease (FMD), an economically important disease of cloven-hoofed animals, is endemic in Pakistan where three virus serotypes are present (O, A and Asia 1). Fifty-eight clinical samples collected between 2005 and 2008 from animals with suspected FMD in various locations in Pakistan were subjected to virus isolation on primary cell culture, antigen ELISA and real-time RT-PCR (rRT-PCR). Viruses were isolated from 32 of these samples and identified as FMDV type O (n = 31) or type A (n = 1). Foot-and-mouth disease virus (FMDV) genome was detected in a further 11 samples by real-time RT-PCR. Phylogenetic analyses of the VP1 nucleotide sequences showed that all of the type O viruses belonged to the MIDDLE EAST-SOUTH ASIA topotype with the majority belonging to the PanAsia-2 lineage; a single example of the older PanAsia lineage was identified. The single FMDV type A virus belonged to the ASIA topotype, but did not cluster with known strains that are currently circulating (such as Iran-05) and was not closely related to other type A viruses from the region. These findings demonstrate the widespread distribution of O-PanAsia-2 in Pakistan and the presence of undisclosed novel type A lineages in the region.

Evaluation of irradiation in foods using DNA Comet assay.

Comet assay is a rapid, inexpensive and sensitive biological technique to detect DNA damage in food stuffs by irradiation. In this study the Comet assay is applied on foods of plant and animal origins. Samples were irradiated by using Co-60 gamma-radiation source. The applied doses were 2, 6 and 10 kGy for food of plant origin and 0.5, 1 and 2 kGy for meat items. The un-irradiated and irradiated samples were clearly differentiated on the basis of DNA fragmentation. During the electrophoresis study, it was found that in un-irradiated cells DNA remained intact and appeared as Comets without tail whereas in irradiated cells Comets with tails were visible due to stretching of fragmented DNA. Moreover, it was also revealed that the DNA tail length was dose dependent. Dry food stuffs (seeds) showed good results as compared to moist foods (meat, fruits and vegetables) due to the absence of background damage.

DNA damage in Pakistani pesticide-manufacturing workers assayed using the Comet assay.

The production and use of chemical pesticides has increased in recent years. Although the increased use of pesticides may benefit agriculture, they are also the potential source of environmental pollution, and exposure to pesticides can have negative consequences for human health. In the present study, we have assessed DNA damage in blood leukocytes from 29 Pakistani pesticide-factory workers and 35 controls of similar age and smoking history. The workers were exposed to various mixtures of organophosphates, carbamates, and pyrethroids. DNA damage was measured with the single cell gel electrophoresis (SCGE) assay or Comet assay, using the mean comet tail length (microm) as the DNA damage metric. Exposed workers had significantly longer comet tail lengths than the controls (mean +/- SD 19.98 +/- 2.87 vs. 7.38 +/- 1.48, P < 0.001). Of the possible confounding factors, smokers had significantly longer mean comet tail lengths than nonsmokers and exsmokers for both the workers (21.48 +/- 2.58 vs.18.37 +/- 2.28, P < 0.001) and the controls (8.86 +/- 0.56 vs. 6.79 +/- 1.31, P < 0.001), while age had a minimal effect on DNA damage (P > 0.05 and P < 0.05 for workers and controls, respectively). The results of this study indicate that occupational exposure to pesticides causes DNA damage.

Cytogenetic analysis of Pakistani individuals occupationally exposed to pesticides in a pesticide production industry.

Although several cytogenetic biomonitoring studies on workers exposed to pesticides have been reported, there is only limited information on this topic from developing countries where pesticides have been widely used over the years. People in developing countries are at higher risk from exposure, due to poor working conditions and a lack of awareness of the potential hazards during manufacturing and application of the pesticides. The present study has assessed the genotoxic effects of pesticides on workers involved in the pesticide manufacturing industry. Subjects in the exposed group (29) were drawn from workers at a pesticide production plant in district Multan (Pakistan). The control group (unexposed) composed of 35 individuals from the same area but was not involved in pesticide production. Liver enzymes, serum cholinesterase (SChE), micronucleus assay and some haematological parameters were used as biomarkers in this study. A statistically significant (P < 0.001) increase in levels of alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase was detected in exposed workers with respect to the control group. There was a significant (P < 0.001) decrease in the level of SChE in the exposed group. Exposed individuals exhibited cytogenetic damage with increased frequencies (P < 0.001) of binucleated cells with micronuclei and total number of micronuclei in binucleated lymphocytes in comparison with subjects of the control group. A decrease (P < 0.001) in cytokinesis block proliferation index similarly demonstrates a genotoxic effect due to pesticide exposure. The results indicate that the pesticide industry workers have experienced significant genotoxic exposure. This study highlights the risk to workers in the pesticide manufacturing industries of developing countries such as Pakistan and the need for implementation of suitable safety measures to prevent/limit exposure to harmful toxins.

Effect of prolonged exercise and carbohydrate ingestion on type 1 and type 2 T lymphocyte distribution and intracellular cytokine production in humans.

The present study was undertaken to examine the role of the exercise-induced stress hormone response on the regulation of type 1 and type 2 T lymphocyte intracellular cytokine production. Subjects performed 2.5 h of cycling exercise at 65% maximal O2 uptake while ingesting a 6.4% carbohydrate (CHO) solution, 12.8% CHO solution, or a placebo. Peripheral whole blood samples were stimulated and stained for T lymphocyte surface antigens (CD4 and CD8). Cells were then permeabilized, stained for intracellular cytokines, and analyzed using flow cytometry. Exercise resulted in a decrease (P < 0.05) in the number and percentage of IFN-gamma positive CD4+ and CD8+ T lymphocytes. These stimulated cells produced less IFN-gamma immediately postexercise (P < 0.05) and 2-h postexercise (P < 0.05) compared with preexercise. However, CHO ingestion, which attenuated the exercise-induced stress hormone response compared with placebo (P < 0.05), prevented both the decrease in the number and percentage of IFN-gamma-positive CD4+ and CD8+ T lymphocytes and the suppression of IFN-gamma production from stimulated CD4+ and CD8+ T lymphocytes. There was no effect of exercise on the number of, or cytokine production from, IL-4-positive CD4+ or CD8+ T lymphocytes. These data provide support for the role of exercise-induced elevations in stress hormones in the regulation of type 1 T lymphocyte cytokine production and distribution.

The phosphatidylinositol-3 kinase pathway regulates bladder cancer cell invasion.

OBJECTIVES: To investigate the role of the phosphatidylinositol (PI)-3 kinase pathway in the invasion of bladder cancer cell lines, and to assess the activation of this pathway in primary human bladder tumours. MATERIALS AND METHODS: Human bladder cancer cells were treated with pathway specific inhibitors or were transfected with PI-3 kinase pathway components. The invasion of cultured bladder cancer cells was analysed by an invasion assay. Bladder cancer cells lines and primary human bladder tumours were analysed for pathway activation by western blotting. RESULTS: A specific inhibitor of PI-3 kinase enzyme activity, Ly294002, potently suppressed the invasive properties of three highly invasive bladder tumour cell lines. Restoration of the PTEN gene to invasive UM-UC-3 bladder tumour cells or expression of a dominant-negative version of the PI-3 kinase target, Akt, also potently inhibited invasion, indicating a central role for the PI-3 kinase/Akt pathway in this process. In addition, 55% of primary tumours from patients with bladder cancer had markedly high levels of phosphorylated Akt. CONCLUSION: Pharmacological or biochemical inhibition of the PI-3 kinase pathway drastically reduced the invasive capacity of bladder cancer cell lines; over half of primary human bladder tumours had high Akt phosphorylation, suggesting that the aberrant activation of this pathway may contribute to the invasion of a significant subset of bladder cancers.