Introducing differential expression of human heat shock protein 27 in hepatocellular carcinoma: moving toward identification of cancer biomarker.

Previously, it has to be acknowledged that overexpressed heat shock protein B27 (HSPB27) have been implicated in the etiology of wide range of human cancers. However, the molecular mechanism leading to the disease initiation to progression in liver cancer is still unknown. Present work was undertaken to investigate the differentially expressed HSPB27 in association with those damages that lead to liver cancer development. For the identification of liver cancer biomarker, samples were subjected to comparative proteomic analysis using two-dimensional gel electrophoresis (2-DE) and were further validated by Western blot and immunohistochemical analysis. After validation, in silico studies were applied to demonstrate the significantly induced phosphorylated and S-nitrosylated signals. The later included the interacting partner of HSPB27, i.e., mitogen-activated protein kinase-3 and 5 (MAPK3 and 5), ubiquitin C (UBC), v-akt murine thymoma viral oncogene homolog 1 (AKT1), mitogen-activated protein kinase 14 (MAPK14), and tumor protein p53 (TP53), which bestowed with critical capabilities, namely, apoptosis, cell cycling, stress activation, tumor suppression, cell survival, angiogenesis, proliferation, and stress resistance. Taking together, these results shed new light on the potential biomarker HSPB27 that overexpression of HSPB27 did lead to upregulation of their interacting partner that together demonstrate their possible role as a novel tumor progressive agent for the treatment of metastasis in liver cancer. HSPB27 is a promising diagnostic marker for liver cancer although further large-scale studies are required. Also, molecular profiling may help pave the road to the discovery of new therapies.

Protein expression profiling of nuclear membrane protein reveals potential biomarker of human hepatocellular carcinoma.

BACKGROUND: Complex molecular events lead to development and progression of liver cirrhosis to HCC. Differentially expressed nuclear membrane associated proteins are responsible for the functional and structural alteration during the progression from cirrhosis to carcinoma. Although alterations/ post translational modifications in protein expression have been extensively quantified, complementary analysis of nuclear membrane proteome changes have been limited. Deciphering the molecular mechanism that differentiate between normal and disease state may lead to identification of biomarkers for carcinoma. RESULTS: Many proteins displayed differential expression when nuclear membrane proteome of hepatocellular carcinoma (HCC), fibrotic liver, and HepG2 cell line were assessed using 2-DE and ESI-Q-TOF MS/MS. From the down regulated set in HCC, we have identified for the first time a 15 KDa cytochrome b5A (CYB5A), ATP synthase subunit delta (ATPD) and Hemoglobin subunit beta (HBB) with 11, 5 and 22 peptide matches respectively. Furthermore, nitrosylation studies with S-nitrosocysteine followed by immunoblotting with anti SNO-cysteine demonstrated a novel and biologically relevant post translational modification of thiols of CYB5A in HCC specimens only. Immunofluorescence images demonstrated increased protein S-nitrosylation signals in the tumor cells and fibrotic region of HCC tissues. The two other nuclear membrane proteins which were only found to be nitrosylated in case of HCC were up regulated ATP synthase subunit beta (ATPB) and down regulated HBB. The decrease in expression of CYB5A in HCC suggests their possible role in disease progression. Further insight of the functional association of the identified proteins was obtained through KEGG/ REACTOME pathway analysis databases. String 8.3 interaction network shows strong interactions with proteins at high confidence score, which is helpful in characterization of functional abnormalities that may be a causative factor of liver pathology. CONCLUSION: These findings may have broader implications for understanding the mechanism of development of carcinoma. However, large scale studies will be required for further verification of their critical role in development and progression of HCC.

Role of fructose concentration on cataractogenesis in senile diabetic and non-diabetic patients.

BACKGROUND: Fructose intake has increased steadily during the past 2 decades. Fructose, like other reducing sugar, can react with proteins, which may account for aging and cataract formation. Fructose participates in glycation (fructation) and advanced glycation endproducts (AGE) formation some ten times faster than glucose. This study aims to determine the fructose concentration and correlate with antioxidant status in senile diabetic and non-diabetic cataract patients. METHODS: The study included 124 subjects. Of them, 31 were normal senile subjects, 33 were senile diabetic patients without cataract, 30 were senile diabetic patients with cataract, and 30 were senile non-diabetic patients with cataract. The patients were selected on clinical grounds from Eye Ward, Jinnah Postgraduate Medical Centre, Karachi, Pakistan. RESULTS: Serum fructose was significantly increased (P < 0.001) in senile diabetic patients with and without cataract and senile non-diabetic patients with cataract as compared with senile control subjects. Negative significant correlation was observed between serum fructose and serum total antioxidant status in diabetic and non-diabetic patients with cataract. Positive significant correlation was observed between serum fructose and s-AGEs in diabetic and non-diabetic patients with cataract. Serum total antioxidant status was found to be significantly decreased (P < 0.001) in senile diabetic patients with and without cataract and senile non-diabetic patients with cataract as compared with senile control subjects. Fasting blood glucose, HbA(1C) and serum fructosamine were significantly increased (P < 0.001) in senile diabetic patients with or without cataract as compared with senile non-diabetic patients with cataract and senile control subjects. CONCLUSIONS: The results indicate that the increased fructose concentration which induces oxidative stress in diabetic and non-diabetic patients with cataract may be a predictor for cataractogenesis.

Could oxidative stress associate with age products in cataractogenesis?

BACKGROUND: Oxidative stress has been reported to contribute to aging and cataract formation in the lens. The aim was to determine the association of oxidative stress with advanced glycation end products (AGEs) in elderly diabetic and non-diabetic patients with cataract. METHODS: In the present study, malondialdehyde, vitamin E, serum AGEs, and glycemic control were investigated. The study included 156 subjects. Out of them, 30 were normal elderly subjects, 31 were elderly diabetic patients without cataract, 33 were elderly diabetic patients with cataract, 32 were elderly non-diabetic with cataract, and 30 were normal young subjects. The patients were selected on clinical grounds from Eye Ward, Jinnah Postgraduate Medical Centre, Karachi, Pakistan. RESULTS: Positive significant correlation was observed between s-AGEs and malondialdehyde in elderly diabetic and non-diabetic patients with cataract. Negative significant correlation was observed between s-AGEs and vitamin E in elderly diabetic and non-diabetic patients with cataract. However, the malondialdehyde and serum AGEs were found to be significantly increased (p < 0.001) in elderly diabetic and non-diabetic patients with and without cataract compared with elderly control subjects. In contrast to all four senile groups, the serum AGEs was significantly lower (p < 0.001) in young control subjects. Serum vitamin E was found to be significantly decreased (p < 0.001) in elderly diabetic patients with and without cataract compared with elderly control subjects. Fasting blood glucose, HbA(1C) and serum fructosamine levels were significantly increased (p < 0.001) in elderly diabetic patients with and without cataract compared with non-diabetic elderly patients with cataract and elderly control subjects. CONCLUSIONS: This study revealed that increased AGEs were associated with oxidative stress in the elderly groups. AGE, as a result of oxidative stress, might have a role in cataract formation, which, in diabetic patients, occurs vigorously as compared with non-diabetic cataract patients.

Advanced glycation end-products in senile diabetic and non-diabetic patients with cardiovascular complications.

Advanced glycation end products (AGEs) have been reported to contribute to aging and cardiovascular complications. In the present study, the immunoreactivity of AGEs in human serum samples of healthy older subjects (n = 31), senile diabetic patients without cardiovascular complications (n = 33), senile diabetic patients with cardiovascular complications (n = 32), senile non-diabetic patients with cardiovascular complications (n = 30) ,and healthy young subjects (n = 31) were investigated. The patients were selected on clinical grounds from the National Institute of Cardiovascular Disease, Karachi and the Jinnah Postgraduate Medical Centre, Karachi, Pakistan. Fasting blood glucose, HbA(1C) and serum fructosamine levels were significantly (P < 0.001) increased in senile diabetic patients with and without cardiovascular complications as compared to non-diabetic senile patients with cardiovascular complications and healthy older subjects. Additionally, serum AGEs were found to be significantly (P < 0.001) increased in senile diabetic patients with cardiovascular complications and senile non-diabetic patients with cardiovascular complications, followed by diabetic patients without cardiovascular complications as compared to healthy older subjects and young control subjects. However, no significant difference was found in the senile diabetic patients without cardiovascular complications and senile non-diabetic patients with cardiovascular complications. In contrast to all four senile groups, serum AGEs were significantly (P < 0.001) lower in young control subjects. The AGEs distribution in the senile groups corroborates the hypothesis that the advanced glycation process might play a role in the development of cardiovascular complications, which are more severe in diabetic patients compared with non-diabetic patients with cardiovascular complications.

Comparison of adiponectin, leptin and blood lipid levels in normal and obese postmenopausal women.

OBJECTIVE: To determine adiponectin, leptin and blood lipid levels in normal and obese postmenopausal women and to compare them. METHODS: Eighty postmenopausal women were selected which included 40 normal controls and 40 obese women matched for age and height. Adiponectin and leptin levels were determined by ELISA. Triglycerides, cholesterol, LDL cholesterol and HDL cholesterol were determined by standard kit methods using Clinicon 4010. RESULTS: Serum adiponectin level decreased and leptin level increased significantly (P < 0.001) in obese women compared to controls. Similarly triglycerides, cholesterol and LDL cholesterol were increased significantly (P < 0.001) in obese subjects compared with controls. No significant change in HDL cholesterol was observed in both groups. Significant correlation of leptin with lipid profile was observed when both groups were combined. CONCLUSION: Serum adiponectin level decreased and leptin level increased significantly in obese women compared to controls. Significant correlation exists between leptin and lipid profile in combined group.

Serum thiocyanate levels in smokers, passive smokers and never smokers.

OBJECTIVE: To compare the serum thiocyanate levels in smokers, passive smokers and never smokers. METHODS: Serum thiocyanate was analyzed on 135 self reported smokers, never smokers and passive smokers by UV-Spectrophotometry (Manual method of Bowler). RESULTS: The mean serum thiocyanate concentration (112 +/- 36.6 micromol/L) of smokers was significantly higher (p value < 0.001) as compared to passive (55 +/- 16.1 micromol/L) and never smokers (41 +/- 6.6 micromol/L). The serum thiocyanate of female smokers (133 + 28.7 micromol/L) was higher (p value < 0.05) than male smokers (103 +/- 36.3 miccromol/L). Serum thiocyanate showed 91% sensitivity, 80% specificity and 84% accuracy at > or = 60 micromol/L for smokers. CONCLUSION: Serum thiocyanate is significantly higher in smokers than passive and never smokers. Serum thiocyanate at > or = 60 micromol/L may be considered as cutoff value for distinguishing smokers from non-smokers for future studies.

Leptin and blood lipid levels in postmenopausal diabetic women with and without complication of ischemic heart disease.

BACKGROUND: Recent studies have provided evidence that leptin has significant effects on vascular development and repair. The aim was to determine the levels of leptin and lipid profile in diabetic postmenopausal women with and without the complication of ischemic heart disease and to develop correlation between them. Moreover, the relationship between leptin levels and extent of ischemic changes were determined. MATERIAL/METHODS: One hundred twenty postmenopausal subjects between the ages of 45 and 60 years were included in the study. They were divided into three groups of forty subjects each. The first group comprised normal healthy controls, the second diabetic type 2 patients with no history of ischemic heart disease (I1ID), and the third diabetic patients with IHD. Serum leptin levels were determined by a Kit obtained from DRG and samples were analyzed on ELISA. Fasting and random blood glucose was determined by the glucose oxidase method, cholesterol, triglycerides, LDL cholesterol, and HDL cholesterol were also determined by kits obtained from Merck. RESULTS: The results show that leptin and serum lipid levels increased significantly in diabetic patients with IHD compared with diabetic patients without IHD as well as normal subjects. Moreover; the sertum leptin level increased significantly in the diabetic patients with IHD who had positive findings in myocardial perfusion scan compared with those having negative findings. CONCLUSIONS: Hyperleptinemia in diabetic patients shows that leptin contributes to the development of cardiovascular disease in diabetic patients.

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

OBJECTIVE: To compare serum protein levels between diabetic and non-diabetic patients with retinopathy. STUDY DESIGN: Comparative study. PLACE AND DURATION: Ziauddin Medical University and Jinnah Postgraduate Medical Centre, Karachi, Pakistan, from 2000 to 2002. PATIENTS AND METHODS: 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. RESULTS: 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 non-diabetic patients with retinopathy, diabetic patients without complication and control subjects. Gamma globulin was significantly decreased in diabetic and non-diabetic patients with retinopathy. CONCLUSION: 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.

Relationship between adiponectin, glycemic control and blood lipids in diabetic type 2 postmenopausal women with and without complication of ischemic heart disease.

BACKGROUND: Recently major adipocytokine, adiponectin, are thought to play important roles in the regulation of cardiovascular and metabolic homeostasis. We compared serum adiponectin concentrations and lipid profile in diabetic patients with and without complication of cardiovascular disease in postmenopausal women. METHODS: We included 120 female subjects between the age of 45 and 60 years. The subjects were divided into 3 groups and each group comprised of 40 subjects. First group comprised of normal healthy controls, second group diabetic type 2 patients with no history of ischemic heart disease (IHD) and the third group diabetic patients with IHD. Serum adiponectin concentrations, serum triglycerides, cholesterol, LDL cholesterol and HDL cholesterol concentrations were determined. RESULTS: Values of fasting and random blood glucose, serum triglycerides, cholesterol, LDL cholesterol were significantly increased (P<0.001) in diabetic patients with IHD as compared with normal control subjects except for serum adiponectin and HDL cholesterol concentrations, which were significantly decreased (P<0.001) in diabetic patients and diabetic patients with IHD group. When values of cholesterol, triglycerides, LDL cholesterol, HDL cholesterol, fasting and random blood glucose and adiponectin were compared among diabetic and diabetic patients with IHD, it was observed that they were significantly increased (P<0.001) except for adiponectin, which was decreased significantly (P<0.001) and random blood glucose, LDL and HDL cholesterol in which no change was observed between the 2 groups. CONCLUSIONS: Serum adiponectin concentrations may be a predictor for development of cardiovascular disease in postmenopausal diabetic patients.

Changes in glycosylated proteins in type-2 diabetic patients with and without complications.

BACKGROUND: Diabetes mellitus constitutes one of the most important problems in developing and developed countries. Increased glycosylation of various proteins in diabetic patients has been reported by many authors. The present study describes the changes in protein glycosylation in diabetic patients with and without diabetic complication. METHODS: The study included one hundred and three subjects. Among them 21 were type 2 diabetic patients without any clinical evidence of chronic diabetic complications, 21 were type 2 diabetic patients with cardiovascular complications, 20 were type 2 diabetic patients with cataract, 20 were type 2 diabetic patients with retinopathy and 21 apparently normal, age, sex and weight matched controls. The patients were selected from Ziauddin Medical University Hospital, Karachi and Jinnah Postgraduate Medical Centre, Karachi. RESULTS: Fasting plasma glucose was increased in all diabetic patients and correlated significantly with glycosylated hemoglobin, glycosylated plasma proteins and serum fructosamine concentrations. There was no significant difference in the levels of fasting plasma glucose, glycosylated plasma proteins, glycosylated hemoglobin, serum fructosamine, hexosamine or sialic acid between diabetic patients with or without chronic complications. Alpha-1 and alpha-2 globulin fraction were significantly increased in diabetic patients without complications, diabetic patients with cardiovascular complications and diabetic patients with cataract. Albumin was found to be decreased in diabetic patients with cataract while gamma globulin was increased in diabetic patients with cardiovascular complications and diabetic patients with cataract. CONCLUSIONS: In uncomplicated diabetic patients alpha-1 and alpha-2 glycoproteins were increased. In diabetic patients with cardiovascular complications alpha-1, alpha-2 and gamma globulin were increased while in diabetic patients with cataract alpha-1, alpha-2 and gamma globulin were increased but serum albumin was significantly decreased.

Efficacy of daily vs. weekly supplementation of iron in schoolchildren with low iron status.

Iron deficiency anemia (IDA) is still a major nutritional and public health problem in developing countries. The prevalence among young children and pregnant women is particularly high. Daily oral supplementation with medicinal iron is considered an effective strategy for reducing the incidence of IDA but non-compliance is a major problem with this strategy. We undertook this study to compare the results of once-weekly vs. daily oral iron supplementation in schoolchildren. Sixty children ranging between 5 and 10 years with iron deficiency anemia were selected from a school in Karachi, Pakistan and were divided into two equal groups, i.e., daily and weekly supplementation groups. Hemoglobin (Hb), hematocrit (Hct), serum iron, total iron binding capacity (TIBC), and serum ferritin were determined before the start of the study. Ferrous sulfate (200 mg) was given daily to the daily supplementation group and once-weekly to the weekly supplementation group for 2 months. When post-supplementation values of the above-mentioned parameters were determined, a significant improvement was observed in all parameters in both groups. It is concluded that once-weekly iron supplementation is as effective as daily supplementation for the treatment of iron deficiency anemia. Moreover, weekly iron supplementation is cost effective and has no or fewer side-effects.

Changes in glycosylated proteins in diabetic and non-diabetic patients with and without cardiovascular complications.

The objective of this study was to find the changes in glycoprotein composition in both diabetic and non-diabetic patients with and without cardiovascular complications. The study was carried out in Ziauddin Medical University Karachi, Pakistan. Eighty-three patients and control subjects were selected. Among them twenty-one were diabetic patients without any clinical evidence of chronic diabetic complications, twenty-one were diabetic patients with cardiovascular complications, twenty were non-diabetic patients with cardiovascular complications and twenty-one apparently normal, age, sex and weight matched control subjects were investigated. All these patients were selected on clinical grounds from National Institute of Cardiovascular Disease, Karachi. Fasting plasma glucose was increased in all diabetic patients and correlated significantly with and without cardiovascular complications. Fasting plasma glucose, glycosylated haemoglobin, glycosylated plasma proteins, serum fructosamine, sialic acid, hexosamine and total serum protein and its fractions were increased in diabetic patients with and without cardiovascular complications. Fasting plasma glucose, glycosylated haemoglobin, glycosylated plasma proteins, serum fructosamine, sialic acid and hexosamine were not different in diabetic patients with cardiovascular complications and diabetic patients without chronic complications as compared with control subjects. In conclusion, fasting plasma glucose, glycosylated haemoglobin, glycosylated plasma proteins, serum fructosamine, sialic acid, hexosamine and total serum proteins and its fractions were increased in diabetic patients with and without complications, but these parameters remained within normal limits in non-diabetic patients with cardiovascular complications.

Serum leptin levels in female patients with NIDDM.

OBJECTIVE: To compare serum leptin levels of diabetic and non-diabetic female subjects and also assess the relationship of hyperglycemia with serum insulin, C-peptide and leptin levels. DESIGN: It is a case control study. PLACE AND DURATION OF STUDY: The study was conducted at Medicare Hospital, Family Care Clinic and Baqai Institute of Diabetes and Endocrinology between December 1997 to September 1999. SUBJECT AND METHODS: One hundred and forty female subjects with different body mass indices and fasting blood sugar levels were selected from three different diabetic centers. A venous sample was drawn after an overnight fast (12 hours) for determination of blood parameters in all groups. Glycosylated hemoglobin, hexosamine, fructosamine, insulin and C-peptide were determined only in diabetic patients. Blood glucose, triacylglycerol (TAG), total cholesterol, HDL cholesterol, HbA1C, hexosamine and fructosamine were determined enzymatically. Serum leptin, C-peptide and insulin were measured using enzyme-linked immunoassay. RESULTS: Serum leptin levels of obese diabetic and non-diabetic subjects were significantly higher as compared with lean diabetic patients and non-diabetic subjects (P< 0.05). Leptin levels were positively correlated with serum insulin and C-peptide levels. Serum leptin increased with increase in body mass index and waist hip ratio was strongly related with insulin resistance in NIDDM. CONCLUSION: Leptin levels are increased in obesity and may play a role in development of insulin resistance and NIDDM.