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.

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.

Daily contact with a patient and poor housing affordability as determinants of pulmonary tuberculosis in urban Pakistan

This study aimed to evaluate the factors associated with pulmonary tuberculosis [TB] among individuals aged 15 years or more in urban Karachi, Pakistan. A case-control design was implemented in three major tertiary-care hospitals to select cases [n = 342] with active pulmonary TB [i.e. two sputum smears positive for Mycobacterium tuberculosis with clinical and radiographic evidence of current pulmonary TB and diagnosed between August 2002 and October 2003. Selected controls [n = 342] were surgery patients from the same hospitals at time of recruitment of the cases, without clinical and radiographic evidence of pulmonary TB. Multivariable logistic regression model showed that daily contact with a pulmonary TB patient [adjusted odds ratio [OR[adj]] = 5.07; 95% CI: 3.31, 7.78], and poor housing affordability [i.e. rented vs. owned] [OR[adj] = 1.59; 95% CI: 1.13, 2.26] were significantly associated with pulmonary TB status. The overall adjusted summary population attributable risk [%] for both the risk factors together was 38.7. Reaching out to underprivileged TB patients for delivery of DOTS and focused education of patients and their contacts about M. tuberculosis transmission mode may substantially minimize pulmonary TB risk in this and similar settings

Association between Interleukin-5 and Body Mass Index among Iraqi asthmatic patients

Asthma is an allergic hypersensitivity disease in which many mediators play a role in its pathogenecity. One of these etiological agents are the mediators such as lnterleukin-5 [IL-5]. The aim of this study is to demonstrate the association between IL-5 and obesity in asthma development. One hundred and four sera samples for asthma cases have been studied in comparison with 41 non-asthmatic bronchitis as patient controls beside 30 apparently healthy controls. Cytokine has been estimated using ELISA method in correlation with Body Mass Index [BMI]. This study revealed a significant correlation between IL-5 concentration and BMI [P<0.05], particularly among females. There is certain correlation between obesity and asthma accompanied by IL-5

Possible Role of lnterleukin-6 in Rheumatoid Arthritis

Rheumatoid Arthritis [RA] is an autoimniune disease. Many etiological agents are proposed to play a role in its pathogenecisy. One of these factors is cytokines such as Interleukin6. ELISA method has been used for IL-6 estimation in 75 RA patients in comparison with 67 SLE as patient controls and 39 apparently healthy controls. This study showed that there was an elevation of IL -6 in the sera of RA patients with high significant differences between RA patients and controls [P < 0.001]. Moreover a good correlation between IL-6 level and RE titer were observed However, for most patients with high IL-6 were shown to be HLA-DR4. Interleukin-6 play a crucial role in the disease which may be participate in the severity of RA and subsequently its treatment

Viral load among the sera of Iraqi hepatitis C virus patients

Hepatitis C Virus is the main causative agent of hepatitis among blood transfused patients, in which most chronic cases result in liver carcinoma. Enzyme-Linked Immunosorbent Assay for HCV, FNF-[] and IL-12p40 estimation with Recombinant Immunoblotting Assay [RIBA] as a confirmatory test for HCV, have been applied for HCV detection in 80 HCV patients' samples in comparison with 30 samples for apparently healthy control, while viral load has been estimated using Branched-DNA [b-DNA] technique for 32 randomly selected positive cases for HCV. Liver function test has been applied for patients' sera in comparison with control. This study reveals highly specificity and sensitivity of ELISA technique for HCV detection which results in 100% positivity by RIBA methods. Moreover, Viral load estimation shows that [71.9%] of HCV sera samples with viral load >615 IU /ml and only [28.1%] with viral load <615 IU/ml with highly significant difference between them [P = 0.013]. Furthermore, there is a highly significant variations between liver function test in comparison with control group [P< 0.01], while SGPT is the only parameter which significantly affected by viral load [P= 0.011]. Beside that, neither INF-[] nor IL-12p40 level has been affected by viral loads more or less 615 IU /ml. It was concluded that ELISA technique is still the best accurate reliable method for viral detection and SGPT is a good marker for highly viral loaded samples

Possible role for lnterleukin-5 in asthma

Asthma is an allergic disease characterized by airway obstruction as a result of cellular accumulation due to the liberation of certain mediators. Among those mediators are the cytokines such as IL-5. lnterleukin-5 concentration has been estimated in 94 sera samples of Asthmatic patients in comparison with 41 non-asthmatic bronchitis as patient controls in addition to 30 apparent healthy control group using ELISA method. There is highly significant elevation of IL-5 in the asthmatic cases in comparison with healthy controls [P< 0.001]. We conclude that this cytokine may play the major role in asthmatic attack and it may be a good marker for the disease

Neonatal infection in the neonatal unit at Baghdad Teaching Hospital, Iraq

In Iraq, the true magnitude of the neonatal among newborns receiving care is not known as health system was badly eroded in the last two decades. This work was carried out to find factors associated with neonatal infection among newborns receiving care in intensive unit. The records of a total of 723 neonates admitted to the neonatal intensive unit at Baghdad teaching hospital during the period Nov. 2003 to Oct. 2004 were reviewed. Information regarded birth weight, gestational age, medical history of the mothers, bacterial infections and management procedures were collected. Out of the total neonates, 9.3% had bacterial infection. Low birth weight, prematurity and prolonged rupture of membrane were factors significantly associated with bacterial infection [p = 0.001, 0.002 and 0.03, respectively]. Oxygen therapy, invasive procedures, and the nasogastric tube [p = 0.01] were significantly associated with infection. S. aureus, enterbacter and E. coli were the commonest pathogens. The study provides useful information for future surveillance in association with preventive program as there were several factors associated with neonatal infection which could be prevented

Evaluation of knowledge and practices for kidney transplantation recipients

Knowledge and practices concerning medcation signs and symptoms of rejection, follow-up, Diet, personal hygiene, Environmental health, Measurement of body temperature were evaluated in a purposive sample of [150] kidney transplantation recipients who were followed for the period between th beginning of Feb, 2000 to the end of Dec, 2000 in two teaching hospitals which were Baghdad Teaching Hospital and Al-Karama Teaching Hospiotal. The data were analyzed through the application of descriptive and inferential statistics in order to meet the study objectives. Their knowledge about medication side effect, signs and symptoms of rejection has been found unacceptable and inadequate; further more it has been found that 83% of kidney transplantation recipients do not know signs and symptoms of rejection. Concerning their practices, it has been found that personal hygiene was the best practice among them While the follow-up, diet environmental health was unsatisfactory, in addition the measurement of body temperature was the worst practices which showed 96% of them not know how to check their temperature