Apoptotic Effects of Melandryum firmum Root Extracts in Human SH-SY5Y Neuroblastoma Cells

Melandryum firmum is a biennial plant that has been used in traditional medicine for treatment of bacterial and fungal infection. Here, we investigated molecular mechanisms underlying apoptotic effects of Melandryum firmum root extract (MFRE) in neuroblastoma cells, since the effect of this natural compound on cancer cells has not been fully clarified. The root extract of M. firmum reduced cell proliferation, as revealed by cell viability assay. However, MFRE-treated cells exhibited morphological changes including cell rounding, neurite retraction and membrane blebbing. These alterations of cellular shape suggest this morphological change might be due to the apoptosis which shows fragmented DNA. In addition, MFRE up-regulated the pro-apoptotic protein Bax and down-regulated the anti-apoptotic protein Bcl-2 and Mcl-1, which also finally activated cleaved caspase-3 in a dose-dependent manner, as determined by western blot analyses. Together, these findings demonstrate that apoptotic and cytotoxic effects of MFRE on SH-SY5Y cells are mediated by intrinsic mitochondria-mediated caspase pathway and that this natural extract might be effective as an anticancer agent for neuroblastoma malignancies.

Antiproliferative and Cytotoxic Effects of Resveratrol in Mitochondria-Mediated Apoptosis in Rat B103 Neuroblastoma Cells

Resveratrol, a natural compound, has been shown to possess anti-cancer, anti-aging, anti-inflammatory, anti-microbial, and neuroprotective activities. In this study, we examined the antiproliferative and cytotoxicity properties of resveratrol in Rat B103 neuroblastoma cells; although it's molecular mechanisms for the biological effects are not fully defined. Here, we examined the cellular cytotoxicity of resveratrol by cell viability assay, antiproliferation by BrdU assay, DNA fragmentation by DNA ladder assay, activation of caspases and Bcl-2 family proteins were detected by western blot analyses. The results of our investigation suggest that resveratrol increased cellular cytotoxicity of Rat B103 neuroblastoma cells in a dose-and time-dependent manner with IC50 of 17.86 microM at 48 h. On the other hand, incubation of neuroblastoma cells with resveratrol resulted in S-phase cell cycle arrests which dose-dependently and significantly reduced BrdU positive cells through the downregulation of cyclin D1 protein. In addition, resveratrol dose-dependently and significantly downregulated the expression of anti-apoptotic protein includes Bcl-2, Bcl-xL and Mcl-1 and also activates cleavage caspase-9 and-3 via the downregulation of procaspase-9 and -3 in a dose-dependent manner which indicates that involvement of intrinsic mitochondria-mediated apoptotic pathway. In conclusion, resveratrol increases cellular cytotoxicity and inhibits the proliferation of B103 neuroblastoma cells by inducing mitochondria-mediated intrinsic caspase dependent pathway which suggests this natural compound could be used as therapeutic purposes for neuroblastoma malignancies.

Prevalence and determinants of diabetic retinopathy in Al Hasa region of Saudi Arabia: primary health care centre based cross-sectional survey, 2007-2009

To evaluate the prevalence of diabetic retinopathy [DR] in the urban and rural areas of Al Hasa region of Saudi Arabia and to determine risk factors related to DR. This study was conducted on patient attending primary health care centers between July 2007 and June 2009. A retrospective chart review was conducted on subjects with diabetes mellitus greater than 18 years old. Ophthalmologists examined DR status through dilated pupils by using direct, indirect, and slit lamp bio-microscopy. Frequencies, percentage, and their 95% confidence intervals [CIs] were calculated. Odd's ratio was used to associate DR with possible risk factors. A P value less than 0.05 was considered statistically significant. The prevalence of DR among 473 diabetic subjects was 30% [95% CI:25.80-34.20]. The odd ratios [ORs] of DR among diabetic residing in an urban area was significantly higher than diabetics residing in rural areas [OR = 1.94 [95% CI of OR 0.82-2.89]]. DR was associated to the duration of diabetes [adjusted OR= 1.70], uncontrolled blood sugar level [adjusted OR=1.96], hyperlipidemia [adjusted OR= 2.04], and hypercholesterolemia [adjusted OR = 2.80]. DR appears to be a public health problem in the Al Hasa district of Saudi Arabia, and a planned approach is required to avoid severe visual impairment in patients with diabetes mellitus. Primary prevention and early detection could be implemented through primary health centers and non-ophthalmologists

Comparison of blood protein levels between diabetic and non diabetic patients with retinopathy

To compare serum protein levels between diabetic and non-diabetic patients with retinopathy. Comparative study. Ziauddin Medical University and Jinnah Postgraduate Medical Centre, Karachi, Pakistan, from 2000 to 2002. Sixty patients were selected. Among them, 21 were diabetic patients without any clinical evidence of chronic diabetic complications; 20 were diabetic patients with retinopathy and 19 were non-diabetic patients with retinopathy. Twenty-one apparently normal, age, gender and weight-matched control subjects were also inducted. All these patients were selected on clinical grounds. Blood values, fasting plasma glucose, glycosylated hemoglobin, serum fructosamine, glycosylated plasma protein, hexosamine, sialic acid and total serum proteins were determined and compared. Fasting plasma glucose was high in all diabetic patients and correlated significantly with glycosylated hemoglobin, glycosylated plasma proteins and serum fructosamine concentrations. Fasting plasma glucose, glycosylated hemoglobin, glycosylated plasma proteins, serum fructosamine, sialic acid, hexosamine and total serum protein were increased in diabetic patients with retinopathy and diabetic patients without any complications. These values were not different in diabetic patients with retinopathy and diabetic patients without chronic complications as compared with control subjects. Alpha-1 and alpha-2 globulins were significantly increased in diabetic patients with retinopathy, diabetic patients without complications and non-diabetic patients with retinopathy as compared with control subjects. Beta globulin was significantly increased in diabetic patients with retinopathy as compared with nondiabetic patients with retinopathy, diabetic patients without complication and control subjects. Gamma globulin was significantly decreased in diabetic and non-diabetic patients with retinopathy. Fasting plasma glucose, glycosylated hemoglobin, glycosylated plasma proteins, serum fructosamine, sialic acid, hexosamine and total serum protein were increased in diabetic patients with and without complications but these parameters remained within normal limits in non-diabetic patients with retinopathy. The decrease in gamma globulins may be associated with a retinopathy

Adiponectin! obesity and coronary artery disease

Plasma adiponectin rapidly accumulates in the subendothelial space of the injured human artery. Plasma concentrations of adiponectin are reduced in the setting of obesity in patient with non-insulin dependent diabetes mellitus, and in patients with coronary artery diseas [CAD] Several studies have been carried out on animals and recently in humans also to it vestigate the role of adiponectin in obesity and insulin resistance leading to coronary artery disease

Role of nicotine, cotinine and thiocyanate in cigarette smoking [active and passive]

The studies have been continuously carried out on serum nicotine, cotinine and thiocyanate but Pakistan is still deficient of such studies. Tobacco use is very common in Pakistan and about 34% of men and 12.5% of women use some form of tobacco on a regular basis. In addition to cigarettes, tobacco is smoked in unique local ways, which include "Bidi" [tobacco rolled in dryes] and huqqa [Hubble-Bubble]. Tobacco smoke contains more than 4000 chemicals including 43 that are known to cause cancer. Active smoking can cause respiratory disorders, cardiovascular hazards, cerebral and peripheral vascular diseases and cancers. Passive smoking also increases risk for higher and lower respiratory tract illness, ischemic heart diseases and lung cancer. The hazards of passive smoking have yet to be clearly understood so biochemical measurements of appropriate markers are particularly valuable in assessing exposure to environmental tobacco smoke because individuals may differ in their awareness of the extent and duration of such exposures. Markers of exposure to cigarette smoke include thiocyanate ions, nicotine and cotinine, which is a primary metabolite of nicotine. Nicotine, the most tobacco specific component of cigarette smoke responsible for tobacco addiction and can be measured in both active and passive smokers. At present, cotinine is generally regarded as the best marker for monitoring tobacco exposure in either actively or passively exposed individuals. Thiocyanate is a major metabolic product of hydrocyanic acid. Three fold higher concentration of thiocyanate is found in the sera, urine and saliva of smokers as compared to non-smokers; because of cyanide provided by cigarette smoke

Role of diet on the enterohepatic recycling of estrogen in women taking contraceptive pills

The effect of diet on the entrohepatic recycling of estrogen after oral administration of 1 mg non-radioactive estriol [E[3]] was studied in six women using contraceptive pills. The women were followed for two consecutive menstrual cycles, firstly on a high fibre diet [HFD] and then changing on to low fibre diet [LFD] during the next cycle, hence each subject acted as its own control. The extent of enterohepatic recycling of estriol [E[3]] during the early follicular phase of menstrual cycle was assessed by monitoring during 48 hours, the urinary excertion of its two major metabolites, i.e., estriol-3-glucuronide [E[3]-3-G] and estriol 16 alpha-glucuronide [E[3]-16alpha-G]. An increase in values of the variables including E[3]-3-G/E[3]-16alpha-G output ratio, E[3]-3-G output as percent of total [E[3]-3-G+E[3]-16alpha-G] excretion and total E[3] [E[3]-3-G+E[3]-16alpha-G] output as percent dose was noted as a result of change from high to low fibre diet. The urinary excretion of E[3] in the form of its metabolites was also delayed as a result of dietary change. These findings reveal that extent of enterohepatic recycling of estrogen containing contraceptives could be higher in women on LFD as compared to those who are on HFD