Study of GPCR levels in Iraqi diabetic and diabetic nephropathy

Abstract Aim of the study is to determined  G-protein coupled receptor levels in diabetic and diabetic nephropathy patients and compare the results with control group. Study also, aimed   to find relationship  of GPCR with HbA1c , cholesterol and Triglyceride  in these patients ,in order to  that GPCR could be used as a marker combat diabetes and its complication . One hundred fifty subjects were involved  in this study that divided into three groups as follows:- Control group (G1) consists of 50 healthy individuals. Diabetic group (G2) consists of 50 patients and diabetic Nephropathy group (G3) consists of 50 patients . levels of HbA1c % , urea , createnine , albumin , TC, TG, HDL-c, LDL-c , VLDL-c and GPCR were determined in all subjects. Conclusion could be drawn from study that the difference between GPCR levels among  groups indicate GPCR may be used as a marker in development of diabetic nephropathy as well as their  are a significant relation for GPCR with TC and TG . Keywords: GPCR , diabetic and diabetic nephropath

GLP-1, DPP-4 and GPCR Levels in Diabetic and Diabetic neuropathy Patient`s

Background: Neuropathy is a common disorder in DM patients which results from poor glycaemic control and long duration of diabetes. Higher body mass index, smoking, hypertension, hypercholesterolemia and hypertriglyceridemia are associated with the incidence of neuropathy. Objective: The aim of this study is to estimate glucagon like Peptide-1, DPP-4 and G-Protein coupled Receptor levels in diabetic and diabetic neuropathy and to compares the results with control group. Also, to find the correlation of GPCR with GLP-1 and DPP-4 in all studied groups.Subjects and Methods: ninety subjects were enrolled in this study with aged ranged (40-65) years and BMI with (30-35) Kg/m2 that divided into three groups as follows: group one (G1) consists of 30 healthy individuals as a control group, group two (G2) consists of 30 patients with diabetic and group three (G3) consists of 30 patients with diabetic and neuropathy as complication. ESC(Feet Mean), ESC(Hand Mean), ESC(Risk of neuropathies), HbA1C, GLP-1, DPP-4 and GPCR  were determined.Results: The result showed highly significant elevation in HbA1C levels in patients groups comparing to control group. Results, also a highly significant elevation in GLP-1 and DPP-4 levels in G2 comparing to G1, and in G3 comparing to G2 and G1 were noticed. A highly significant elevation in GPCR levels in G2 comparing to G1and G3, while no significant increased noticed in G3 comparing to G2. A high significant positive correlation was found between GLP-1 and DPP-4 in G1, while highly significant negative correlation was found in G2 and G3. A high significant positive correlation was observed between GLP-1 and GPCR in G1. Moreover, No significant positive correlation was observed in G2, but a significant positive correlation was observed in G3. High significant negative correlation was found between GPCR and DPP-4 in G1 and G2, while a high significant positive correlation was found in G3. Conclusion: the conclusion is obtained from this study that GLP-1and GPCR have important role in development of neuro injury in neurodiabetic patients. Also, correlation was found between GPCR with GLP-1 and DPP-4. Keywords: Diabetic neuropathy, GLP-1, DPP-4 and GPCR

Insulin and Glucagon Levels in Diabetic, Diabetic Neuropathy Patient`s

Background: Diabetic neuropathy affects 30–50% of patients with diabetes mellitus. It encompasses several neuropathic syndromes, the commonest being distal symmetrical polyneuropathy or ‘diabetic peripheral neuropathy’ (DPN). Risk factors for DPN include poor glycaemic control and drivers of macrovascular disease including hypertension.Objective: The study aimed to determination of insulin, glucagon levels in diabetic and diabetic neuropathy patient`s and compare the results with control group. Also, Insulin/Glu ratio calculated that may be used in early diagnosis of diabetic neuropathy.Subjects and Methods: ninety subjects were enrolled in this study with aged ranged (40-65) years and BMI with (30-35) Kg/m2 that divided into three groups as follows: group one (G1) consists of 30 healthy individuals as a control group, group two (G2) consists of 30 patients with diabetic and group three (G3) consists of 30 patients with diabetic and neuropathy as complication. ESC(Feet Mean), ESC(Hand Mean), ESC(Risk of neuropathies), FBS, insulin, Glucagon were determined. Also, HOMA-IR and insulin/Glu ratio were calculated.Results: The results of this study revealed highly significant differences in ESC (Feet Mean), ESC (Hand Mean) and ESC (Risk of neuropathies) between G2 and G3 comparing with G1 and in G3 comparing to G2. Results revealed a highly significant increased in FBS, insulin and HOMA-IR levels in G2 comparing to G1 and in G3 comparing to G2 and G1. Results, also, showed a high significant increased in glucagon levels in G2 and G3 comparing to G1 while no significant increased was found in G3 comparing to G2. Results, also, showed no significant reduction was found in In/Glu ratio in G2 comparing to G1, while a high significant elevation was observed in G3 comparing to G2 and G1.Conclusion: The conclusion could be drawn from this study that insulin and glucagon increased in patients with neurodiabetic when comparing to diabetic patients. Also, In/Glu ratio increased significantly in neurodiabetic patients when comparing to diabetic and control group that may be considered important marker in diagnosis of development of neuropathy risk. Keywords: Diabetic Neuropathy, Insulin, Glucagon, Insulin/ Glu Ratio

A Study of Visfatin Levels and Other Related Parameters in Sera of Iraqi Non Diabetic Male with Hyperlipidemia

The aim of the present study is to evaluate the levels of visfatin in sera of non-diabetic male with hyperlipidemia classified according to Frederickson classification  .At the same time to find the correlation of visfatin with related parameters in the studied groups. The study included (22) healthy Iraqi male as control group (G1) and (62) non-diabetic male diagnosed with hyperlipidemia which divided into three groups as follows: group(2)consist of (20)male with hypercholesterolemia[Type I] ,group (3)consist of (20) male with hypertriglyceridemia [Type IIa] and group (4):consist of (22)male with hypercholesterolemia and hypertriglyceridemia [Type IIb] .The age of all studied groups ranged between (21-50) years and BMI with (19.8-24.3) Kg/m2 .Serum was used for determination of FBG, lipid profile(HDL,LDL,VLDL,TG) ,insulin ,CRP and visfatin .Whole  blood was used for determination of  HbA1C. The results revealed  no significant elevation in FBS and HbA1C levels were seen in patients groups when comparing to healthy control . The results indicates a significant elevation in TC and LDL in G2and G4 comparing to G1. Also, there are significant elevation in G3 comparing to G 2 and G4 comparing to G3,while no significant elevation was found in TC and VLDL in G3 with G1and G4 with G2.The results ,also, illustrated significant elevation in TG and VLDL in G3 and G4 when comparing to G1, and in G3,G4 comparing to G2 . No significant elevation was noticed in G2 comparing to G1and G4 comparing to G3 . HDL levels showed significant decrease in G2,G3 and G4 comparing to G1. No significant decrease seen in G3 and G4 comparing to G2 and in G4 comparing to G3.A high significant elevation in insulin and HOMA-IR levels in G2,G3 and G4 was found when comparing with G1 , while a significant elevation  was noticed in G3 and G4 when comparing with G2 and in G4 when comparing with G3. The  results, also ,showed significant increase  in CRP  and visfatin levels for  G2,G3 ,G4 when comparing to G1  and in G4 when comparing to G2 and  G3 . But no significant elevation was found in G3 when comparing to G2 .A significant positive correlation was found between visfatin levels and TC in G1,G2 and G4 while significant negative correlation was observed in G3 between visfatin and TC. A high significant positive correlation coefficients was found between visfatin and TG in four studied groups .The results , also, showed a significant negative correlation between visfatin levels and HDL for all studied groups .The results  illustrated significant positive correlation in visfatin levels with LDL in G1 and G4 while there are significant negative correlation in visfatin levels with LDL in G2 and G3. A significant negative correlation between visfatin and insulin ,HOMA-IR levels was noticed in all studied groups. The conclusion could be drown from this study that visfatin increased in  G4 more than other groups comparing to G1 . Also, there are significant correlation between visfatin levels with insulin , HOMA-IR and CRP levels. Keywords : visfatin , Insulin , CRP , Hyperlipoproteinemi

Study of IL-35 levels in Multiple Myeloma patients and its Relation with Immunoglobulins

Multiple myeloma (MM) is a malignant disorder characterized by the proliferation of plasma cellsand the second most common hematological standing next to lymphoma. Multiple myeloma patients commonly present with defects in numbers and function of various   immune cells including dendritic cells ,B cells,Tcells and natural kill cells[23].Interleukin- 35 (IL-35) is a novel anti-inflammatory cytokine suppressing the immune response through the expansion of regulatory T cells and suppression of Th17 cell development.The present study aimedto investigate the possible role of IL-35 in pathogenesis of MM and its relation withimmunoglobulins such as IgA,IgG and IgM.Forty Iraqis patients with multiple myeloma(G1,G2) and twenty healthy individuals as a control group (G1)were enrolled in this study.Whole blood used for determination of  hemoglobin. Serum samples were used for determination of albumin,total protein using standard procedures of the biochemistry laboratory of hospital. Also immunoglobulin's (IgA,IgG,IgM) and IL-35 were determined in serum.The results revealed a significant increasing in urine total protein ,ESR,creatinine ,uric acid ,BUN, and TP in  patients groups(G2,G3) comparing to control group(G1) while a   significant decrease in albumin and Hb  were found in  patients groups comparing to control .Also a significant differences  was observed in calcium concentration in G3  compared to G2 while there are no differences  was observed between G2 and G1 . The results shown highly significant increase in IgA,IgG, IgM and  IL-35,in  patients groups comparing to control group. Also a significant  differences were noticed in G3 comparing to G2. A highly significant positive correlation between IL-35 and IgA was noted in G2  and G3. Also a highly significant positive correlation between IL-35 and IgG levels was noticed in G2 and G3. Moreover a significant negative correlation founded between IL-35  andIgM in G2  and  G3 .A conclusion could be drown from this study for the first time that MM patients showed increasing in IL-35  levels so, it may be considered as a novel cytokine involved in the pathogenesis of MM. Keyword:IL-35,Multiple Myeloma,Immunoglobulines

Purification of G-Protein Coupled Receptor from Whole Cell of Local Strain of Saccharomyces cerevisiae

The aim of study To purify GPCR from a local strain of S. cerevisiae using Ion exchange and gel filtrationchromatography techniques , by packing materials for columns which will be chosen of low cost comparing to the alreadyused in published researches, which depend on the costly affinity chromatography and other expensive methods ofpurification. Local strain of S. cerevisiae chosen for extraction and purification of G-protein coupled receptor (GPCR).The strains were obtained from biology department in Al- Mosul University, Iraq. The isolated colony was activated onYeast Extract Pepton Dextrose Broth (YEPDB) and incubated at 30 C° for 24 h .Loop fully of the yeast culture wastransferred to (10ml) of yeast extract peptone glucose agar (YEPGA) slant , then incubated at 30C°for 24h , after that itwas stored at 4C° ,the yeast cultures were reactivated and persevered after each two weeks period. S.cerevisiae wasidentified by morphological, microscopic characterization and biochemical test . The GPCR that extract from whole cellof S.cerevisiae was purified by ion exchange chromatography using DEAE-Sepharose ,the bound proteins (negativelycharged) were then eluted using gradient concentration of NaCl ranged between( 0.1 -0.5M). Gel filtrationchromatography using Sepharose 6B was applied as a second step of purification. The optical density for each fractionwas measured at 280 nm by UV-VS spectrophotometer then the GPCR concentration was determined by using ELISAKit . The fractions which gave the highest absorbance and concentration of GPCR were collected .The molecular weightof GPCR was determined by gel filtration chromatography using blue dextrin solution. Standard curve was plottedbetween log of molecular weight for standard protein and the ratio of Ve/Vo of GPCR . The purity of the GPCR thatextracted and purified from whole cell of S, cerevisiae were carried out by using SDS-PAGE electrophoresis . In ionexchange chromatography the fraction were collected with 5 ml tube at a flow rate 0.5 ml/ min and eluted with gradient(0.1-0.5M) of sodium chloride solution. Two proteins peaks appeared after eluted by the gradient concentration ofsodium chloride, while no protein peaks appeared in the washing fractions. The GPCR concentration was measured inthe fractions of these two protein peaks, data indicated that GPCR located in the first protein peak (eluted at 0.1M ofNaCl) at fraction numbers between 3 and 9, the maximum concentration of GPCR was 9.281 with specific activity71.58(ng/mg)protein , 3.125 purification folds and72.9(%) yield while the second peaks (eluted at 0.4 M of NaCl) don'tgive any concentration for GPCR, thus its neglected. Gel filtration chromatography was used as second step ofpurification which applied by using sepharose 6B. Results show single active protein peaks appeared that identical withthe peak of GPCR at fractions numbers(29-35). The maximum concentration of GPCR was 9.082 (ng/ml)was observed inthese fractions. The specific activity for these fractions was 151.37 (ng/mg) protein with 6.608 purification folds and39.64 (%) yield. The present study a chive a relatively high purification of GPCR from whole cell of a local strain S.cerevisiae with fold purification 6.608 and a yield of 39.64 % and molecular weight about~33KD.Key words: GPCR purification, S.Cerevisea, whole cel

Synthesis of Novel Acetylinc Derivative of Metformin as a DPP-4 Inhibitors and Study its Effects on Sera of Rabbits with induced Diabetes

The study aimed to prepare novel derivative of metformin similar to the work of the enzyme inhibitors DPP-4 and so the need for new inhibitors may be a side effect with less addition to linking types of drugs have a stronger effect on patients with diabetes and to study the impact of this derivative inside the living cell has been prepared derived through interaction metformin with propargyl chloride. It was to make sure the chemical structure by using analytical and spectral methods (FT-IR, 1H-NMR, and 13C-NMR), and the results confirming the obtained structures,  then purified by column chromatography by using silica gel as stationary phase and methanol as a mobile phase. The study is derived on the impact of rabbits where they were taking the 40 rabbits with similar weights and were divided into four groups (10 rabbits per group) weredivided as follows, the first group G1 obtained as a control group, which did not gave any things. The second group G2 has injected by aloxane a concentration of 120 mg / kg using syringes medical capacity of 3 ml to inject rabbits in the vein ear and after two hours of injection they were given glucose solution of 10%, the confirmed they injured rabbits diabetes by measuring blood sugar to 10 rabbits have been selected randomly and then it was taken two sets of this group, the third group G3 were given a drug sitagliptin with concentration of 10 mg / kg, and the fourth group G4, were given the prepared derivative with concentration of 8 mg / kg for 3 days and pulled blood samples after the last dose on the third day. Serum used in determination of lipid profile,DPP-4 activity,alanine transaminase(ALT),Aspartate transaminase(AST) and insulin. Results of statistical analysis showed a significant decrease in the level of FBG and  DPP-4 activity  of  the prepared derivative, and also showed a decrease in the level of cholesterol, triglyceride, LDL and VLDL, while  results showed increase in HDL level in G4 comparing to G1. Keywords : Diabetes Mellitus , Dipeptidylpeptidase -4  , metformin .

Study the Levels of GPCR , GLP-1 and Related Hormones Controlled and Uncontrolled in Diabetic Patient's

The aim of the present study is to evaluate the change in the levels of glucagon, GLP-1 and GPCR in diabetic patient's and diabetic with dyslipidemia as metabolic syndrome. The study included 75 male aged ranged (30-50) years and with BMI (25-29) kg/m2 which divided into three groups as follows: group one (G1): consist of 25 subjects as healthy control group. Group two (G2): consist of 25 patient's   with diabetes mellitus and group three (G3): consist of 25 patient's with diabetic and dyslipidemia as metabolic syndrome. Serum was used in determination of FBG, lipid profile, insulin, glucagon, GLP-1 and GPCR. Whole blood was determination of HbA1c.  The results revealed significant elevation in FBG and HbA1c in G2 and G3 comparing to G1. While non -significant elevation was found in FBG and HbA1c in G3 comparing to G2. The results also, showed no significant elevation in each of TC, TG, LDL and VLDL in G2 comparing to G1. Whereas, significant elevation was noticed in these parameters when G3comparing  to G2 and G1. Also, the levels of HDL showed no significant reduction in G2 comparing to G1, while significant reduction was found in G3 comparing to G2 and G1. The results also, revealed no significant elevation in insulin levels in G2 comparing to G1. While significant elevation was found in G3 comparing to G2 and G1. Also, the results illustrated significant elevation in glucagon levels in G2 comparing to G1. While significant reduction was seen in G3 comparing to G2. Significant reduction in GLP-1 and GPCRlevels was found in G2 comparing to G1. While significant elevation in these parameters noticed in G3 comparing to G2 and G1. The conclusion could be drawn from this study that dyslipidemia affecting GLP-1 and GPCR levels that may be these patient's at high risk for cardiovascular disease. Key Words: GPCR, GLP-1, Diabetic patient's with Metabolic Syndrome

Atherogenic Index of Plasma Levels in Patients with Diabetic and Neurodiabetic

ABSTRACT: Objective: To determined the levels of lipid profile (TC, TG, HDL-c, LDL-C, VLDL) in diabetic and diabetic neuropathy patients and compare the results with control group. Also, to compare Atherogenic Index of Plasma (AIP) levels in these groups that may be predict prone of patients to cardiovascular disease. Methodology: Ninety subjects were enrolled in this study with aged ranged (40-65) years and BMI with (30-35) Kg/m2 that divided into three groups as follows: group one (G1) consists of 30 healthy individuals as a control group, group two (G2) consists of 30 patients with diabetes and group three (G3) consists of 30 patients with diabetes and neuropathy as complication. Electrochemical Skin Conductance (Feet Mean), Electrochemical Skin Conductance (Hand Mean), Electrochemical Skin Conductance (Risk of neuropathies), FBS, TC, TG and HDL-c were determined. Also, LDL-C, VLDL and AIP were calculated. Results: This study illustrated no significant increased were found in TC and LDL in G2 comparing to G1, while there was highly significant increase in TC and LDL in G2 comparing to G3 and in G3 comparing to G1. Results, also, revealed highly significant increase in TG and VLDL in G2 comparing to G1, G2 comparing to G3, and G3 comparing to G1. Data demonstrated highly significant decrease in HDL-c in G2 and G3 comparing to G1, while no significant decrease observed in G3 comparing to G2. Data, also, demonstrated highly significant increase in AIP levels in G2 and G3 comparing to G1, while no significant increased observed in G3 comparing to G2. Conclusion: The study concluded that AIP is elevated in diabetic and neurodiabetic patients who could be used as a marker for predict CVD in patients groups. Recommendation: Study the genetic analysis for wide range patients with diabetic, neurodiabetic and control as a comprehensive surrey.Keywords: Diabetic Neuropathy, Lipid profile and AIP