Study of resveratrol against bone loss by using in-silico and in-vitro methods / Estudo do resveratrol contra a perda óssea usando métodos in silico e in vitro

By applying the in-silico method, resveratrol was docked on those proteins which are responsible for bone loss. The Molecular docking data between the resveratrol and Receptor activator of nuclear factor-kappa-Β ligand [RANKL] receptors proved that resveratrol binds tightly to the receptors, showed the highest binding affinities of −6.9, −7.6, −7.1, −6.9, −6.7, and −7.1 kcal/mol. According to in-vitro data, Resveratrol reduced the osteoclasts after treating Marrow-Derived Macrophages [BMM] with Macrophage colony-stimulating factor [MCSF] 20ng / ml and RANKL 50ng / ml, with different concentrations of resveratrol (2.5, 10 μg / ml) For 7 days, the cells were treated with MCSF (20 ng / ml) and RANKL (40 ng / ml) together with concentrated trimethyl ether and resveratrol (2.5, 10 μg / ml) within 12 hours. Which, not affect cell survival. After fixing osteoclast cells with formaldehyde fixative on glass coverslip followed by incubation with 0.1% Triton X-100 in PBS for 5 min and after that stain with rhodamine phalloidin staining for actin and Hoechst for nuclei. Fluorescence microscopy was performed to see the distribution of filaments actin [F.actin]. Finally, resveratrol reduced the actin ring formation. Resveratrol is the best bioactive compound for drug preparation against bone loss.(AU)

Com a aplicação do método in-silico, o resveratrol foi ancorado nas proteínas responsáveis pela perda óssea. Os dados de docking molecular entre o resveratrol e o ligante do receptor ativador do fator nuclear kappa-Β [Receptor Activator of Nuclear Factor kappa-B Ligant (RANKL)] provaram que o resveratrol se liga fortemente aos receptores, mostraram as afinidades de ligação mais altas de −6,9, −7,6, −7,1, −6,9, - 6,7 e -7,1 kcal / mol. De acordo com dados in-vitro, o resveratrol reduziu os osteoclastos após o tratamento de macrófagos derivados da medula óssea [Bone Marrow derived Macrophage (BMM)] com fator estimulador de colônias de macrófagos [Macrophage Colony-Stimulating Factor (MCSF)] 20ng / ml e RANKL 50ng / ml, com diferentes concentrações de resveratrol (2,5, 10 μg / ml). Durante sete dias, as células foram tratadas com MCSF (20 ng / ml) e RANKL (40 ng / ml) juntamente com éter trimetílico concentrado e resveratrol (2,5, 10 μg / ml) em 12 horas, processo que não afeta a sobrevivência celular. Após a fixação de células de osteoclastos com fixador de formaldeído em lamela de vidro seguido de incubação com 0,1% Triton X-100 em PBS por 5 min, foi realizado posteriormente o procedimento para corar com rodamina faloidina a actina e Hoechst os núcleos. A microscopia de fluorescência foi realizada para ver a distribuição dos filamentos de actina [F.actina]. Finalmente, o resveratrol reduziu a formação do anel de actina. O resveratrol é o melhor composto bioativo para o preparo de medicamentos contra a perda óssea.(AU)

Study of resveratrol against bone loss by using in-silico and in-vitro methods / Estudo do resveratrol contra a perda óssea usando métodos in silico e in vitro

Abstract By applying the in-silico method, resveratrol was docked on those proteins which are responsible for bone loss. The Molecular docking data between the resveratrol and Receptor activator of nuclear factor-kappa-Β ligand [RANKL] receptors proved that resveratrol binds tightly to the receptors, showed the highest binding affinities of −6.9, −7.6, −7.1, −6.9, −6.7, and −7.1 kcal/mol. According to in-vitro data, Resveratrol reduced the osteoclasts after treating Marrow-Derived Macrophages [BMM] with Macrophage colony-stimulating factor [MCSF] 20ng / ml and RANKL 50ng / ml, with different concentrations of resveratrol (2.5, 10 μg / ml) For 7 days, the cells were treated with MCSF (20 ng / ml) and RANKL (40 ng / ml) together with concentrated trimethyl ether and resveratrol (2.5, 10 μg / ml) within 12 hours. Which, not affect cell survival. After fixing osteoclast cells with formaldehyde fixative on glass coverslip followed by incubation with 0.1% Triton X-100 in PBS for 5 min and after that stain with rhodamine phalloidin staining for actin and Hoechst for nuclei. Fluorescence microscopy was performed to see the distribution of filaments actin [F.actin]. Finally, resveratrol reduced the actin ring formation. Resveratrol is the best bioactive compound for drug preparation against bone loss.

Resumo Com a aplicação do método in-silico, o resveratrol foi ancorado nas proteínas responsáveis ​​pela perda óssea. Os dados de docking molecular entre o resveratrol e o ligante do receptor ativador do fator nuclear kappa-Β [Receptor Activator of Nuclear Factor kappa-B Ligant (RANKL)] provaram que o resveratrol se liga fortemente aos receptores, mostraram as afinidades de ligação mais altas de −6,9, −7,6, −7,1, −6,9, - 6,7 e -7,1 kcal / mol. De acordo com dados in-vitro, o resveratrol reduziu os osteoclastos após o tratamento de macrófagos derivados da medula óssea [Bone Marrow-derived Macrophage (BMM)] com fator estimulador de colônias de macrófagos [Macrophage Colony-Stimulating Factor (MCSF)] 20ng / ml e RANKL 50ng / ml, com diferentes concentrações de resveratrol (2,5, 10 μg / ml). Durante sete dias, as células foram tratadas com MCSF (20 ng / ml) e RANKL (40 ng / ml) juntamente com éter trimetílico concentrado e resveratrol (2,5, 10 μg / ml) em 12 horas, processo que não afeta a sobrevivência celular. Após a fixação de células de osteoclastos com fixador de formaldeído em lamela de vidro seguido de incubação com 0,1% Triton X-100 em PBS por 5 min, foi realizado posteriormente o procedimento para corar com rodamina faloidina a actina e Hoechst os núcleos. A microscopia de fluorescência foi realizada para ver a distribuição dos filamentos de actina [F.actina]. Finalmente, o resveratrol reduziu a formação do anel de actina. O resveratrol é o melhor composto bioativo para o preparo de medicamentos contra a perda óssea.

Study of resveratrol against bone loss by using in-silico and in-vitro methods / Estudo do resveratrol contra a perda óssea usando métodos in silico e in vitro

By applying the in-silico method, resveratrol was docked on those proteins which are responsible for bone loss. The Molecular docking data between the resveratrol and Receptor activator of nuclear factor-kappa-Β ligand [RANKL] receptors proved that resveratrol binds tightly to the receptors, showed the highest binding affinities of −6.9, −7.6, −7.1, −6.9, −6.7, and −7.1 kcal/mol. According to in-vitro data, Resveratrol reduced the osteoclasts after treating Marrow-Derived Macrophages [BMM] with Macrophage colony-stimulating factor [MCSF] 20ng / ml and RANKL 50ng / ml, with different concentrations of resveratrol (2.5, 10 μg / ml) For 7 days, the cells were treated with MCSF (20 ng / ml) and RANKL (40 ng / ml) together with concentrated trimethyl ether and resveratrol (2.5, 10 μg / ml) within 12 hours. Which, not affect cell survival. After fixing osteoclast cells with formaldehyde fixative on glass coverslip followed by incubation with 0.1% Triton X-100 in PBS for 5 min and after that stain with rhodamine phalloidin staining for actin and Hoechst for nuclei. Fluorescence microscopy was performed to see the distribution of filaments actin [F.actin]. Finally, resveratrol reduced the actin ring formation. Resveratrol is the best bioactive compound for drug preparation against bone loss.

Com a aplicação do método in-silico, o resveratrol foi ancorado nas proteínas responsáveis pela perda óssea. Os dados de docking molecular entre o resveratrol e o ligante do receptor ativador do fator nuclear kappa-Β [Receptor Activator of Nuclear Factor kappa-B Ligant (RANKL)] provaram que o resveratrol se liga fortemente aos receptores, mostraram as afinidades de ligação mais altas de −6,9, −7,6, −7,1, −6,9, - 6,7 e -7,1 kcal / mol. De acordo com dados in-vitro, o resveratrol reduziu os osteoclastos após o tratamento de macrófagos derivados da medula óssea [Bone Marrow derived Macrophage (BMM)] com fator estimulador de colônias de macrófagos [Macrophage Colony-Stimulating Factor (MCSF)] 20ng / ml e RANKL 50ng / ml, com diferentes concentrações de resveratrol (2,5, 10 μg / ml). Durante sete dias, as células foram tratadas com MCSF (20 ng / ml) e RANKL (40 ng / ml) juntamente com éter trimetílico concentrado e resveratrol (2,5, 10 μg / ml) em 12 horas, processo que não afeta a sobrevivência celular. Após a fixação de células de osteoclastos com fixador de formaldeído em lamela de vidro seguido de incubação com 0,1% Triton X-100 em PBS por 5 min, foi realizado posteriormente o procedimento para corar com rodamina faloidina a actina e Hoechst os núcleos. A microscopia de fluorescência foi realizada para ver a distribuição dos filamentos de actina [F.actina]. Finalmente, o resveratrol reduziu a formação do anel de actina. O resveratrol é o melhor composto bioativo para o preparo de medicamentos contra a perda óssea.

Study of resveratrol against bone loss by using in-silico and in-vitro methods

Abstract By applying the in-silico method, resveratrol was docked on those proteins which are responsible for bone loss. The Molecular docking data between the resveratrol and Receptor activator of nuclear factor-kappa- ligand [RANKL] receptors proved that resveratrol binds tightly to the receptors, showed the highest binding affinities of 6.9, 7.6, 7.1, 6.9, 6.7, and 7.1 kcal/mol. According to in-vitro data, Resveratrol reduced the osteoclasts after treating Marrow-Derived Macrophages [BMM] with Macrophage colony-stimulating factor [MCSF] 20ng / ml and RANKL 50ng / ml, with different concentrations of resveratrol (2.5, 10 g / ml) For 7 days, the cells were treated with MCSF (20 ng / ml) and RANKL (40 ng / ml) together with concentrated trimethyl ether and resveratrol (2.5, 10 g / ml) within 12 hours. Which, not affect cell survival. After fixing osteoclast cells with formaldehyde fixative on glass coverslip followed by incubation with 0.1% Triton X-100 in PBS for 5 min and after that stain with rhodamine phalloidin staining for actin and Hoechst for nuclei. Fluorescence microscopy was performed to see the distribution of filaments actin [F.actin]. Finally, resveratrol reduced the actin ring formation. Resveratrol is the best bioactive compound for drug preparation against bone loss.

Resumo Com a aplicação do método in-silico, o resveratrol foi ancorado nas proteínas responsáveis pela perda óssea. Os dados de docking molecular entre o resveratrol e o ligante do receptor ativador do fator nuclear kappa- [Receptor Activator of Nuclear Factor kappa-B Ligant (RANKL)] provaram que o resveratrol se liga fortemente aos receptores, mostraram as afinidades de ligação mais altas de 6,9, 7,6, 7,1, 6,9, - 6,7 e -7,1 kcal / mol. De acordo com dados in-vitro, o resveratrol reduziu os osteoclastos após o tratamento de macrófagos derivados da medula óssea [Bone Marrow-derived Macrophage (BMM)] com fator estimulador de colônias de macrófagos [Macrophage Colony-Stimulating Factor (MCSF)] 20ng / ml e RANKL 50ng / ml, com diferentes concentrações de resveratrol (2,5, 10 g / ml). Durante sete dias, as células foram tratadas com MCSF (20 ng / ml) e RANKL (40 ng / ml) juntamente com éter trimetílico concentrado e resveratrol (2,5, 10 g / ml) em 12 horas, processo que não afeta a sobrevivência celular. Após a fixação de células de osteoclastos com fixador de formaldeído em lamela de vidro seguido de incubação com 0,1% Triton X-100 em PBS por 5 min, foi realizado posteriormente o procedimento para corar com rodamina faloidina a actina e Hoechst os núcleos. A microscopia de fluorescência foi realizada para ver a distribuição dos filamentos de actina [F.actina]. Finalmente, o resveratrol reduziu a formação do anel de actina. O resveratrol é o melhor composto bioativo para o preparo de medicamentos contra a perda óssea.

Impact of alien fishes on the distribution pattern of indigenous freshwater fishes of Punjab, Pakistan / Impacto de peixes invasores no padrão de distribuição de peixes de água doce em Punjab, Paquistão

Pakistan is the country with mega freshwater fish diversity of native and alien species. In the presence of enormous native fishes varies species of exotic fishes have been introduced into the freshwater bodies of Pakistan which are competing with native freshwater fish fauna and making them deprive of food and habitat as well. Intentional or accidental introductions of animals, plants and pathogens away from their native niche is considered as major leading problem for biodiversity of invaded habitat. Three years study (from January 2017 to December 2019) was conducted in freshwater bodies of Province Punjab, Pakistan. Study was designed to know exotic species impacts on the distribution of native species of fishes. During current survey a total of 68 (indigenous and exotic) fish species belonging to 14 families were recorded from head Qadirabad, head Baloki, Islam headworks and Rasul barrage. Statistical analysis showed that Shannon-Wiener diversity index was the highest (1.41) at both Rasul barrage and Islam headworks but, invasive species were present in very less number in these study sites i.e. Oreochromis mozambicus, O. niloticus, Hypophthalmichthys nobilis, Carassius auratus, Ctenophryngdon idella, Cyprinus carpio and H. molitrix. The diversity index showed that comparatively low diversity was present in both area head Qadirabad (1.30) and head Baloki (1.4) due to high pressure of invasive species. Direct observation of species and statistical analysis showed that aliens species produced negative impact on the local fish fauna diversity, evenness, and numbers. It is concluded that many factors are impacts on the diversity of native fishes, but alien (invasive) species also play a major role to reduction of native species, because aliens species produce competition for native species. So it is urgent need to analysis the aliens and native food web and roosting sites in Pakistan, in future.(AU)

O Paquistão é um país com ampla diversidade de espécies nativas e exóticas de peixes de água doce. Contudo, diversas espécies de peixes exóticos estão sendo introduzidas nos corpos de água doce paquistanês, as quais estão competindo com a fauna nativa de peixes, tornando-os privados de alimento e habitat. A introdução intencional ou acidental de animais, plantas e patógenos fora de seu nicho nativo é considerada o principal problema da biodiversidade do habitat recém-invadido. Assim, foi realizado um estudo de três anos, de janeiro de 2017 a dezembro de 2019, em corpos de água doce na província de Punjab, Paquistão. A pesquisa foi desenvolvida para conhecer os impactos de espécies exóticas na distribuição de espécies nativas de peixes. Durante a pesquisa, foi registrado um total de 68 espécies de peixes (nativas e exóticas), pertencentes a 14 famílias, nas nascentes dos rios Qadirabad, Baloki e Islam e na barragem de Rasul. A análise estatística mostrou que o índice de diversidade de Shannon-Wiener foi mais alto (1,41) na barragem Rasul (RB) e na nascente do rio Islam (IH); e que a maioria das espécies introduzidas estava presente em um número muito menor, isto é, Oreochromis mossambicus, Oreochromis niloticus, Hypophthalmichthys nobilis, Carassius auratus, Ctenopharyngodon idella, Cyprinus carpio e Hypophthalmichthy smolitrix nessa área de estudo. Por outro lado, o índice Shannon-Wiener mostrou que a diversidade estava baixa nas nascentes dos rios Qadirabad (1,30) e Baloki (1,4) em razão da alta pressão de espécies invasoras. A observação direta das espécies e a análise estatística indicaram que as espécies invasoras produziram impacto negativo na diversidade, uniformidade e números de peixes da fauna local. Conclui-se que muitos fatores impactam na diversidade de peixes nativos, mas as espécies exóticas (invasoras) também desempenham um papel importante na redução das espécies nativas, em razão da competição existente entre elas. Portanto, é [...].(AU)

Impact of alien fishes on the distribution pattern of indigenous freshwater fishes of Punjab, Pakistan / Impacto de peixes invasores no padrão de distribuição de peixes de água doce em Punjab, Paquistão

Pakistan is the country with mega freshwater fish diversity of native and alien species. In the presence of enormous native fishes varies species of exotic fishes have been introduced into the freshwater bodies of Pakistan which are competing with native freshwater fish fauna and making them deprive of food and habitat as well. Intentional or accidental introductions of animals, plants and pathogens away from their native niche is considered as major leading problem for biodiversity of invaded habitat. Three years study (from January 2017 to December 2019) was conducted in freshwater bodies of Province Punjab, Pakistan. Study was designed to know exotic species impacts on the distribution of native species of fishes. During current survey a total of 68 (indigenous and exotic) fish species belonging to 14 families were recorded from head Qadirabad, head Baloki, Islam headworks and Rasul barrage. Statistical analysis showed that Shannon-Wiener diversity index was the highest (1.41) at both Rasul barrage and Islam headworks but, invasive species were present in very less number in these study sites i.e. Oreochromis mozambicus, O. niloticus, Hypophthalmichthys nobilis, Carassius auratus, Ctenophryngdon idella, Cyprinus carpio and H. molitrix. The diversity index showed that comparatively low diversity was present in both area head Qadirabad (1.30) and head Baloki (1.4) due to high pressure of invasive species. Direct observation of species and statistical analysis showed that aliens’ species produced negative impact on the local fish fauna diversity, evenness, and numbers. It is concluded that many factors are impacts on the diversity of native fishes, but alien (invasive) species also play a major role to reduction of native species, because aliens’ species produce competition for native species. So it is urgent need to analysis the aliens and native food web and roosting sites in Pakistan, in future.

O Paquistão é um país com ampla diversidade de espécies nativas e exóticas de peixes de água doce. Contudo, diversas espécies de peixes exóticos estão sendo introduzidas nos corpos de água doce paquistanês, as quais estão competindo com a fauna nativa de peixes, tornando-os privados de alimento e habitat. A introdução intencional ou acidental de animais, plantas e patógenos fora de seu nicho nativo é considerada o principal problema da biodiversidade do habitat recém-invadido. Assim, foi realizado um estudo de três anos, de janeiro de 2017 a dezembro de 2019, em corpos de água doce na província de Punjab, Paquistão. A pesquisa foi desenvolvida para conhecer os impactos de espécies exóticas na distribuição de espécies nativas de peixes. Durante a pesquisa, foi registrado um total de 68 espécies de peixes (nativas e exóticas), pertencentes a 14 famílias, nas nascentes dos rios Qadirabad, Baloki e Islam e na barragem de Rasul. A análise estatística mostrou que o índice de diversidade de Shannon-Wiener foi mais alto (1,41) na barragem Rasul (RB) e na nascente do rio Islam (IH); e que a maioria das espécies introduzidas estava presente em um número muito menor, isto é, Oreochromis mossambicus, Oreochromis niloticus, Hypophthalmichthys nobilis, Carassius auratus, Ctenopharyngodon idella, Cyprinus carpio e Hypophthalmichthy smolitrix nessa área de estudo. Por outro lado, o índice Shannon-Wiener mostrou que a diversidade estava baixa nas nascentes dos rios Qadirabad (1,30) e Baloki (1,4) em razão da alta pressão de espécies invasoras. A observação direta das espécies e a análise estatística indicaram que as espécies invasoras produziram impacto negativo na diversidade, uniformidade e números de peixes da fauna local. Conclui-se que muitos fatores impactam na diversidade de peixes nativos, mas as espécies exóticas (invasoras) também desempenham um papel importante na redução das espécies nativas, em razão da competição existente entre elas. Portanto, é [...].

Impact of alien fishes on the distribution pattern of indigenous freshwater fishes of Punjab, Pakistan

Abstract Pakistan is the country with mega freshwater fish diversity of native and alien species. In the presence of enormous native fishes varies species of exotic fishes have been introduced into the freshwater bodies of Pakistan which are competing with native freshwater fish fauna and making them deprive of food and habitat as well. Intentional or accidental introductions of animals, plants and pathogens away from their native niche is considered as major leading problem for biodiversity of invaded habitat. Three years study (from January 2017 to December 2019) was conducted in freshwater bodies of Province Punjab, Pakistan. Study was designed to know exotic species impacts on the distribution of native species of fishes. During current survey a total of 68 (indigenous and exotic) fish species belonging to 14 families were recorded from head Qadirabad, head Baloki, Islam headworks and Rasul barrage. Statistical analysis showed that Shannon-Wiener diversity index was the highest (1.41) at both Rasul barrage and Islam headworks but, invasive species were present in very less number in these study sites i.e. Oreochromis mozambicus, O. niloticus, Hypophthalmichthys nobilis, Carassius auratus, Ctenophryngdon idella, Cyprinus carpio and H. molitrix. The diversity index showed that comparatively low diversity was present in both area head Qadirabad (1.30) and head Baloki (1.4) due to high pressure of invasive species. Direct observation of species and statistical analysis showed that aliens species produced negative impact on the local fish fauna diversity, evenness, and numbers. It is concluded that many factors are impacts on the diversity of native fishes, but alien (invasive) species also play a major role to reduction of native species, because aliens species produce competition for native species. So it is urgent need to analysis the aliens and native food web and roosting sites in Pakistan, in future.

Resumo O Paquistão é um país com ampla diversidade de espécies nativas e exóticas de peixes de água doce. Contudo, diversas espécies de peixes exóticos estão sendo introduzidas nos corpos de água doce paquistanês, as quais estão competindo com a fauna nativa de peixes, tornando-os privados de alimento e habitat. A introdução intencional ou acidental de animais, plantas e patógenos fora de seu nicho nativo é considerada o principal problema da biodiversidade do habitat recém-invadido. Assim, foi realizado um estudo de três anos, de janeiro de 2017 a dezembro de 2019, em corpos de água doce na província de Punjab, Paquistão. A pesquisa foi desenvolvida para conhecer os impactos de espécies exóticas na distribuição de espécies nativas de peixes. Durante a pesquisa, foi registrado um total de 68 espécies de peixes (nativas e exóticas), pertencentes a 14 famílias, nas nascentes dos rios Qadirabad, Baloki e Islam e na barragem de Rasul. A análise estatística mostrou que o índice de diversidade de Shannon-Wiener foi mais alto (1,41) na barragem Rasul (RB) e na nascente do rio Islam (IH); e que a maioria das espécies introduzidas estava presente em um número muito menor, isto é, Oreochromis mossambicus, Oreochromis niloticus, Hypophthalmichthys nobilis, Carassius auratus, Ctenopharyngodon idella, Cyprinus carpio e Hypophthalmichthy smolitrix nessa área de estudo. Por outro lado, o índice Shannon-Wiener mostrou que a diversidade estava baixa nas nascentes dos rios Qadirabad (1,30) e Baloki (1,4) em razão da alta pressão de espécies invasoras. A observação direta das espécies e a análise estatística indicaram que as espécies invasoras produziram impacto negativo na diversidade, uniformidade e números de peixes da fauna local. Conclui-se que muitos fatores impactam na diversidade de peixes nativos, mas as espécies exóticas (invasoras) também desempenham um papel importante na redução das espécies nativas, em razão da competição existente entre elas. Portanto, é urgente uma análise futura das espécies invasoras e da teia alimentar nativa no Paquistão..

Impact of alien fishes on the distribution pattern of indigenous freshwater fishes of Punjab, Pakistan / Impacto de peixes invasores no padrão de distribuição de peixes de água doce em Punjab, Paquistão

Pakistan is the country with mega freshwater fish diversity of native and alien species. In the presence of enormous native fishes varies species of exotic fishes have been introduced into the freshwater bodies of Pakistan which are competing with native freshwater fish fauna and making them deprive of food and habitat as well. Intentional or accidental introductions of animals, plants and pathogens away from their native niche is considered as major leading problem for biodiversity of invaded habitat. Three years study (from January 2017 to December 2019) was conducted in freshwater bodies of Province Punjab, Pakistan. Study was designed to know exotic species impacts on the distribution of native species of fishes. During current survey a total of 68 (indigenous and exotic) fish species belonging to 14 families were recorded from head Qadirabad, head Baloki, Islam headworks and Rasul barrage. Statistical analysis showed that Shannon-Wiener diversity index was the highest (1.41) at both Rasul barrage and Islam headworks but, invasive species were present in very less number in these study sites i.e. Oreochromis mozambicus, O. niloticus, Hypophthalmichthys nobilis, Carassius auratus, Ctenophryngdon idella, Cyprinus carpio and H. molitrix. The diversity index showed that comparatively low diversity was present in both area head Qadirabad (1.30) and head Baloki (1.4) due to high pressure of invasive species. Direct observation of species and statistical analysis showed that aliens' species produced negative impact on the local fish fauna diversity, evenness, and numbers. It is concluded that many factors are impacts on the diversity of native fishes, but alien (invasive) species also play a major role to reduction of native species, because aliens' species produce competition for native species. So it is urgent need to analysis the aliens and native food web and roosting sites in Pakistan, in future.

O Paquistão é um país com ampla diversidade de espécies nativas e exóticas de peixes de água doce. Contudo, diversas espécies de peixes exóticos estão sendo introduzidas nos corpos de água doce paquistanês, as quais estão competindo com a fauna nativa de peixes, tornando-os privados de alimento e habitat. A introdução intencional ou acidental de animais, plantas e patógenos fora de seu nicho nativo é considerada o principal problema da biodiversidade do habitat recém-invadido. Assim, foi realizado um estudo de três anos, de janeiro de 2017 a dezembro de 2019, em corpos de água doce na província de Punjab, Paquistão. A pesquisa foi desenvolvida para conhecer os impactos de espécies exóticas na distribuição de espécies nativas de peixes. Durante a pesquisa, foi registrado um total de 68 espécies de peixes (nativas e exóticas), pertencentes a 14 famílias, nas nascentes dos rios Qadirabad, Baloki e Islam e na barragem de Rasul. A análise estatística mostrou que o índice de diversidade de Shannon-Wiener foi mais alto (1,41) na barragem Rasul (RB) e na nascente do rio Islam (IH); e que a maioria das espécies introduzidas estava presente em um número muito menor, isto é, Oreochromis mossambicus, Oreochromis niloticus, Hypophthalmichthys nobilis, Carassius auratus, Ctenopharyngodon idella, Cyprinus carpio e Hypophthalmichthy smolitrix nessa área de estudo. Por outro lado, o índice Shannon-Wiener mostrou que a diversidade estava baixa nas nascentes dos rios Qadirabad (1,30) e Baloki (1,4) em razão da alta pressão de espécies invasoras. A observação direta das espécies e a análise estatística indicaram que as espécies invasoras produziram impacto negativo na diversidade, uniformidade e números de peixes da fauna local. Conclui-se que muitos fatores impactam na diversidade de peixes nativos, mas as espécies exóticas (invasoras) também desempenham um papel importante na redução das espécies nativas, em razão da competição existente entre elas. Portanto, é urgente uma análise futura das espécies invasoras e da teia alimentar nativa no Paquistão.

Study of resveratrol against bone loss by using in-silico and in-vitro methods.

By applying the in-silico method, resveratrol was docked on those proteins which are responsible for bone loss. The Molecular docking data between the resveratrol and Receptor activator of nuclear factor-kappa-Β ligand [RANKL] receptors proved that resveratrol binds tightly to the receptors, showed the highest binding affinities of -6.9, -7.6, -7.1, -6.9, -6.7, and -7.1 kcal/mol. According to in-vitro data, Resveratrol reduced the osteoclasts after treating Marrow-Derived Macrophages [BMM] with Macrophage colony-stimulating factor [MCSF] 20ng / ml and RANKL 50ng / ml, with different concentrations of resveratrol (2.5, 10 µg / ml) For 7 days, the cells were treated with MCSF (20 ng / ml) and RANKL (40 ng / ml) together with concentrated trimethyl ether and resveratrol (2.5, 10 µg / ml) within 12 hours. Which, not affect cell survival. After fixing osteoclast cells with formaldehyde fixative on glass coverslip followed by incubation with 0.1% Triton X-100 in PBS for 5 min and after that stain with rhodamine phalloidin staining for actin and Hoechst for nuclei. Fluorescence microscopy was performed to see the distribution of filaments actin [F.actin]. Finally, resveratrol reduced the actin ring formation. Resveratrol is the best bioactive compound for drug preparation against bone loss.

Impact of alien fishes on the distribution pattern of indigenous freshwater fishes of Punjab, Pakistan.

Pakistan is the country with mega freshwater fish diversity of native and alien species. In the presence of enormous native fishes varies species of exotic fishes have been introduced into the freshwater bodies of Pakistan which are competing with native freshwater fish fauna and making them deprive of food and habitat as well. Intentional or accidental introductions of animals, plants and pathogens away from their native niche is considered as major leading problem for biodiversity of invaded habitat. Three years study (from January 2017 to December 2019) was conducted in freshwater bodies of Province Punjab, Pakistan. Study was designed to know exotic species impacts on the distribution of native species of fishes. During current survey a total of 68 (indigenous and exotic) fish species belonging to 14 families were recorded from head Qadirabad, head Baloki, Islam headworks and Rasul barrage. Statistical analysis showed that Shannon-Wiener diversity index was the highest (1.41) at both Rasul barrage and Islam headworks but, invasive species were present in very less number in these study sites i.e. Oreochromis mozambicus, O. niloticus, Hypophthalmichthys nobilis, Carassius auratus, Ctenophryngdon idella, Cyprinus carpio and H. molitrix. The diversity index showed that comparatively low diversity was present in both area head Qadirabad (1.30) and head Baloki (1.4) due to high pressure of invasive species. Direct observation of species and statistical analysis showed that aliens' species produced negative impact on the local fish fauna diversity, evenness, and numbers. It is concluded that many factors are impacts on the diversity of native fishes, but alien (invasive) species also play a major role to reduction of native species, because aliens' species produce competition for native species. So it is urgent need to analysis the aliens and native food web and roosting sites in Pakistan, in future.

Influence of Moringa oleifera leaf meal used as phytogenic feed additive on the serum metabolites and egg bioactive compounds in commercial layers

Phytogenic feed additives have been increasingly used in the last decade, and several plants and their metabolites have been investigated for the said purpose. In this context, present study aimed at evaluating the effects of Moringa oleifera as feed additive on layer performance, and egg bioactive compound levels and nutrient profile. HyLine W36 layers (n=200), 50 weeks of age, were randomly distributed in four treatments with five replicates of ten birds each. Four isocaloric (2725 kcal/kg) and isonitrogenous (CP 16%) diets were formulated and supplemented with 0, 0.5, 1.0, or 1.5% (w/w) of dried Moringa oleifera leaf powder (MLM). The results showed positive effects of MLM on egg production, egg mass, and feed conversion ratio, but negative effects on egg quality (p≤0.05). The contents of bioactive compounds, like β-carotene, quercetin, and selenium, in the diet and in the egg yolk were significantly (p≤0.05) higher in the group fed1.5% MLM, with values of 8.90, 48.88, and 0.54 mg/kg feed and 4906, 241 and 56.82 µg/100g yolk, respectively. Creatinine and glucose serum levels and cholesterol levels (serum and eggs) linearly increased as a function of increasing MLM dietary levels (p≤0.05). Antibody titers against Newcastle Disease significantly improved (p≤0.05) in the group fed the diet supplemented with 1.5% MLM. It was concluded that Moringa oleifera used as phytogenic feed additive enriches eggs with bioactive and functional compounds, and improves the production performance and the health status of layers.(AU)

Influence of Moringa oleifera leaf meal used as phytogenic feed additive on the serum metabolites and egg bioactive compounds in commercial layers

Phytogenic feed additives have been increasingly used in the last decade, and several plants and their metabolites have been investigated for the said purpose. In this context, present study aimed at evaluating the effects of Moringa oleifera as feed additive on layer performance, and egg bioactive compound levels and nutrient profile. HyLine W36 layers (n=200), 50 weeks of age, were randomly distributed in four treatments with five replicates of ten birds each. Four isocaloric (2725 kcal/kg) and isonitrogenous (CP 16%) diets were formulated and supplemented with 0, 0.5, 1.0, or 1.5% (w/w) of dried Moringa oleifera leaf powder (MLM). The results showed positive effects of MLM on egg production, egg mass, and feed conversion ratio, but negative effects on egg quality (p≤0.05). The contents of bioactive compounds, like β-carotene, quercetin, and selenium, in the diet and in the egg yolk were significantly (p≤0.05) higher in the group fed1.5% MLM, with values of 8.90, 48.88, and 0.54 mg/kg feed and 4906, 241 and 56.82 µg/100g yolk, respectively. Creatinine and glucose serum levels and cholesterol levels (serum and eggs) linearly increased as a function of increasing MLM dietary levels (p≤0.05). Antibody titers against Newcastle Disease significantly improved (p≤0.05) in the group fed the diet supplemented with 1.5% MLM. It was concluded that Moringa oleifera used as phytogenic feed additive enriches eggs with bioactive and functional compounds, and improves the production performance and the health status of layers.

Reduced osmolarity oral rehydration solution for persistent diarrhea in infants: a randomized controlled clinical trial.

OBJECTIVE: We evaluated and compared the efficacy of the World Health Organization (WHO) oral rehydration solution (ORS) and 2 different formulations of reduced osmolarity ORSs in infants with persistent diarrhea. STUDY DESIGN: Infants with persistent diarrhea (n = 95) were randomized to 1 of the 3 ORSs: WHO-ORS (control, n = 32), a glucose-based reduced osmolarity ORS (RORS-G, n = 30), or a rice-based reduced osmolarity ORS (RORS-R, n = 31) for replacement of ongoing stool losses for up to 7 days. Major outcome measures were stool volume and frequency, ORS intake, and resolution of diarrhea. RESULTS: Although there were variations from one study day to another, the stool volume was approximately 40% less in the reduced osmolarity ORS groups; consequently, these children required less ORS (22% for RORS-G and 27% for RORS-R groups). A higher proportion of children in the RORS-R groups also had resolution of diarrhea during the study period. No children in any of the treatment groups had hyponatremia. CONCLUSION: Reduced osmolarity ORS is clinically more effective than WHO-ORS and may thus be advantageous for use in the treatment of children with persistent diarrhea.