OBJECTIVES: The aim of this study was to investigate the clinical and biochemical profiles according to homeostasis model assessment of insulin resistance (HOMA-IR) in Korean polycystic ovary syndrome (PCOS) patients. METHODS: In 458 PCOS patients diagnosed by the Rotterdam European Society for Human Reproduction and Embryology (ESHRE) criteria, measurements of somatometry, blood test of hormones, glucose metabolic and lipid profiles, and transvaginal or transrectal ultrasonogram were carried out. HOMA-IR was then calculated and compared with the clinical and biochemical profiles related to PCOS. The patients were divided into 4 groups by quartiles of HOMA-IR. RESULTS: The mean level of HOMA-IR was 2.18 +/- 1.73. Among the four groups separated according to HOMA-IR, body weight, body mass index (BMI), waist-to-hip ratio (WHR), triglyceride (TG), total cholesterol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, lipid accumulation product (LAP) index, high-sensitivity C-reactive protein (hs-CRP), Apoprotein B, free testosterone, and sex hormone binding globulin (SHBG) were found to be significantly different. TG, LAP index, glucose metabolic profiles, and hs-CRP were positively correlated with HOMA-IR after adjustment for BMI. CONCLUSION: Our results suggest that the clinical and biochemical profiles which are applicable as cardiovascular risk factors are highly correlated with HOMA-IR in Korean women with PCOS.
Apoproteins ; Body Mass Index ; Body Weight ; C-Reactive Protein ; Cardiovascular Diseases ; Cholesterol ; Embryology ; Female ; Glucose ; Hematologic Tests ; Homeostasis* ; Humans ; Insulin Resistance ; Lipid Accumulation Product ; Lipoproteins ; Metabolome ; Polycystic Ovary Syndrome* ; Reproduction ; Risk Factors ; Sex Hormone-Binding Globulin ; Testosterone ; Triglycerides ; Ultrasonography ; Waist-Hip Ratio

This study is to optimize the preparation process of fusion protein Fv-LDP which was expressed in the form of inclusion body and consisted of lidamycin apoprotein LDP and single-chain Fv antibody (scFv) directed against type IV collagenase. The preparation and the dissolution of inclusion body, the immobilized metal affinity chromatography of the target protein and the renaturization by stepwise dialysis were optimized by single-factor analysis or orthogonal design. In addition, the refolded fusion protein Fv-LDP was refined by Sephadex G-75 chromatography followed by fluorescence-activated cell sorter (FACS)-based saturation binding assay to measure its antigen-binding activity. After optimization of the process, the purity of fusion protein Fv-LDP existed in the inclusion body was 63.9% and the corresponding solubility was 95.7%; Under denaturing conditions, the purity of fusion protein Fv-LDP was more than 95% after the purification process. The percentage of monomeric fusion protein Fv-LDP was 60% after the refolding process, while it was further refined to 85% which was 5.6-fold higher than that of the initial refolding condition. The refined fusion protein Fv-LDP could bind to human lung adenocarcinoma PAa cells and human hepatoma BEL-7402 cells with the dissociation constants (Kd) of 0.176 micromol x L(-1) and 0.904 micromol x L(-1), respectively. The preparation process of fusion protein Fv-LDP has been successfully optimized, which provides the experimental basis for the production and future development of fusion protein Fv-LDP, and might serve as a relatively practical system for the preparation of other scFv-based proteins expressed in the form of inclusion body.
Adenocarcinoma ; metabolism ; pathology ; Aminoglycosides ; chemistry ; metabolism ; Antibiotics, Antineoplastic ; chemistry ; metabolism ; Apoproteins ; chemistry ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Collagenases ; immunology ; Enediynes ; chemistry ; metabolism ; Escherichia coli ; chemistry ; metabolism ; Humans ; Inclusion Bodies ; chemistry ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Lung Neoplasms ; metabolism ; pathology ; Protein Binding ; Recombinant Fusion Proteins ; chemistry ; metabolism ; Single-Chain Antibodies ; chemistry ; metabolism

OBJECTIVE: Several studies showed that increased arterial stiffness is an independent risk factor for cardiovascular disease. Pulse wave velocity (PWV) is known as a marker for large vessel stiffness. Recent studies show that serum cystatin C is associated with PWV and may predict future cardiovascular events, even in subjects with normal renal function. However, there have been few studies for the relationship between cystatin C and arterial stiffness in patients with type 2 diabetes (T2DM). In this study, we investigated the relationship between serum cystatin C and branchial-ankle PWV in T2DM patients with normal renal function. METHODS: Patients with urinary albumin/creatinine ratio (ACR) higher than 300 microg albumin/mg creatinine, estimated glomerular filtration rate (eGFR) less than 60 mL/min were excluded. A total of 88 patients (47 male/41 female; age, 59+/-2 years; ACR, 33+/-5 microg/mg) were included. Doppler-derived aortic PWV and serum cystatin C were measured. RESULTS: Cystatin C is significantly related to age (r=0.51, P<0.001), hemoglobin A1c (r=-0.23, P<0.05), high density lipoprotein-cholesterol (r=-0.22, P<0.05), apoprotein A (r=-0.22, P<0.05), and eGFR (r=-0.56, P<0.001). Aortic PWV is significantly associated with age (r=0.29, P<0.01), cystatin C (r=0.33, P<0.005), and eGFR (r=-0.24, P<0.05). In multiple regression analysis, there is significant association between aortic PWV and serum cystatin C levels. CONCLUSION: Serum cystatin C is significantly associated with arterial stiffness in T2DM patients with normal renal function. Our results suggest that cystatin C could be a marker for early atherosclerosis in T2DM patients.
Apoproteins ; Atherosclerosis ; Cardiovascular Diseases ; Creatinine ; Cystatin C ; Diabetes Mellitus, Type 2 ; Glomerular Filtration Rate ; Glycosaminoglycans ; Hemoglobins ; Humans ; Pulse Wave Analysis ; Risk Factors ; Vascular Stiffness

OBJECTIVELidamycin (LDM) can be dissociated to an apoprotein (LDP) and an active enediyne chromophore (AE). The detached AE can reassemble with its LDP-containing fusion protein to endow the latter with potent antitumor activity. However, the reassembly of AE with LDP is affected by several factors. Our aim was to optimize the assembly efficiency of the AE with a LDP-containing fusion protein and investigate the influence of several factors on the assembly efficacy.

METHODSA method based on RP-HPLC was developed to analyze the assembly rate, and an orthogonal experimental design L(9) (3(4)) was used to investigate the effects of temperature, assembly time, pH and molecular ratio of LDP-containing fusion protein to AE on the assembly rate. Furthermore, the determined optimum conditions for the assembly rate of the LDP-containing fusion protein with AE were applied and evaluated.

RESULTSA calibration curve based on the LDM micromolar concentration against the peak-area of AE by HPLC was obtained. The order in which individual factors in the orthogonal experiment affected the assembly rate were temperature>time>pH>molar ratio of AE to protein and all were statistically significant (P<0.01). The optimal assembly conditions were temperature at 10°C, time of 12 h, pH 7.0, and the molar ratio of AE: protein of 5:1. The assembly rate of AE with a LDP-containing fusion protein was improved by 23% after condition optimization.

CONCLUSIONThe assembly rate of chromophore of lidamycin with its LDP-containing fusion protein was improved after condition optimization by orthogonal design, and the optimal conditions described herein should prove useful for the development of this type of LDP-containing fusion protein.

Aminoglycosides ; administration & dosage ; chemical synthesis ; chemistry ; pharmacology ; Antibiotics, Antineoplastic ; administration & dosage ; chemical synthesis ; chemistry ; pharmacology ; Apoproteins ; chemistry ; Cell Line, Tumor ; Cell Survival ; Chromatography, High Pressure Liquid ; Drug Design ; Enediynes ; administration & dosage ; chemical synthesis ; chemistry ; pharmacology ; Humans ; Recombinant Fusion Proteins ; chemistry ; Single-Chain Antibodies ; chemistry

This study is to investigate the binding capability of lidamycin apoprotein (LDP), an enediyne-associated apoprotein of the chromoprotein antitumor antibiotic family, to human breast cancer and normal tissues, the correlation of LDP binding capability to human breast cancer tissues and the expression of tumor therapeutic targets such as VEGF and HER2. In this study, the binding capability of LDP to human breast cancer tissues was detected with tissue microarray. The correlation study of LDP binding capability to human breast tumor tissues and relevant therapeutic targets was performed on breast cancer tissue microarrays. Immunocytochemical examination was used to detect the binding capability of LDP to human breast carcinoma MCF-7 cells. As a result, tissue microarray showed that LDP staining of 73.2% (30/41) of breast cancer tissues was positive, whereas that of 48.3% (15/31) of the adjacent normal breast specimens was positive. The difference between the tumor and normal samples was significant (Chi2 = 4.63, P < 0.05). LDP immunoreactivity in breast cancer correlated significantly with the overexpression of VEGF and HER2 (P < 0.001 and < 0.01, r = 0.389 and 0.287, respectively). Determined with confocal immunofluorescent analysis, LDP showed the binding capability to mammary carcinoma MCF-7 cells. It is demonstrated that LDP can bind to human breast cancer tissues and there is significant difference between the breast cancer tissues and the corresponding normal tissues. Notably, the binding reactivity shows positive correlation with the expression of VEGF and HER2 in breast carcinoma tissues. The results imply that LDP may have a potential use as targeting drug carrier in the research and development of new anticancer therapeutics. This study may provide reference for drug combination of LDM and other therapeutic agents.
Aminoglycosides ; metabolism ; Antibiotics, Antineoplastic ; metabolism ; Apoproteins ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Enediynes ; metabolism ; Female ; Humans ; Protein Binding ; Receptor, ErbB-2 ; metabolism ; Tissue Array Analysis ; methods ; Vascular Endothelial Growth Factor A ; metabolism

Apoproteins ; analysis ; Epithelium ; chemistry ; ultrastructure ; Humans ; Lung ; chemistry ; pathology ; Nuclear Proteins ; analysis ; Pulmonary Sclerosing Hemangioma ; chemistry ; pathology ; Pulmonary Surfactant-Associated Proteins ; analysis ; Secretory Component ; analysis ; Thyroid Nuclear Factor 1 ; Transcription Factors ; analysis

The prevalence and cerebral hemorrhage of cerebral amyloid angiopathy(CAA) are age-related. It is rare in young adults. The authors report on CAA coexisting with an arteriovenous malformation(AVM) in a 30-year-old male, who present with the sudden onset of headache and vomiting. Magnetic resonance imaging revealed a cerebral hemorrhage with an AVM. The AVM was completely removed through the hematoma and the histological section obtained from the periphery of the hematoma showed the typical findings of CAA. The epsilon4 allele of apoprotein E(apoE) was identified in genotype determination.
Adult ; Alleles ; Amyloid ; Apolipoproteins E ; Apoproteins ; Arteriovenous Malformations* ; Cerebral Amyloid Angiopathy* ; Cerebral Hemorrhage ; Genotype ; Headache ; Hematoma ; Humans ; Magnetic Resonance Imaging ; Male ; Pathology ; Prevalence ; Vomiting ; Young Adult*

Phycoerythrocyanin(PEC) lyase-isomerase PecE/PecF from Mastigocladus laminosus is the specific enzyme for biosynthesis of PEC alpha-subunit(alpha-PEC). In this work, the specificity of PecE/PecF on substrate apoproteins was reported. PecE/PecF could catalyse the reconstitution of phycocyanobilin(PCB) with apoproteins of alpha-PEC from two different subspecies of Mastigocladus laminosus, as well the site-directed mutated apoprotein of alpha-PEC with Trp at 128 to Phe in vitro, but could not catalyse the reconstitution of PCB with apoprotein of phycocyanin alpha-subunit(alpha-CPC) from Mastigocladus laminosus. The surfactant Triton X-100 had no effect for the reconstitution of alpha-PEC, while it could improve the reconstitution of PCB with apoprotein of alpha-CPC.
Apoproteins ; metabolism ; Bacterial Proteins ; Catalysis ; Cyanobacteria ; enzymology ; Light-Harvesting Protein Complexes ; Lyases ; metabolism ; Octoxynol ; pharmacology ; Proteins ; metabolism ; Substrate Specificity

OBJECTIVE: The purpose was to test the hypothesis that the common missense mutation of 5,10-methylenetetrahydrofolate reductase(MTHFR) is more prevalent among preeclamptic women compared with control and also was to determine whether homocysteine and other lipid profile is changed in pregnant women with preeclampsia. METHODS: We measured plasma homocysteine, cholesterol, HDL, LDL, triglyceride, apoprotein B, vitamin B12, and folate in 48 pregnant women without preeclampsia and 22 women with preeclampsia. And the MTHFR genotype was determined with a polymerase chain reaction/restriction fragment length polymorphism method. Results were analyzed with a X2 contingency table and T-test. RESULTS: The prevalence of the MTHFR C677T mutation was not significantly different between the population studied. There was no significant difference in the level of plasma homocysteine, cholesterol, HDL, LDL, triglyceride, apoprotein B, and folate between controls and preeclamptic women. Furthermore, there was a significant difference in the level of plasma vitamin B12 between the population studied. CONCLUSION: These data suggest that the MTHFR C677T mutation is not a risk factor for preeclampsia in this population. Plasma homocysteine, cholesterol, HDL, LDL, triglyceride, apoprotein B, and folate level are not elevated in preeclamptic women. However, the plasma vitamin B12 level is elevated in preeclamptic women. Further studies are necessary to determine why the plasma vitamin B12 level is elevated in preelamptic women although they did not have vitamin drug.
Apoproteins ; Cholesterol, HDL ; Female ; Folic Acid ; Genotype ; Homocysteine* ; Humans ; Methylenetetrahydrofolate Reductase (NADPH2)* ; Mutation, Missense ; Plasma ; Polymorphism, Genetic ; Pre-Eclampsia* ; Pregnant Women ; Prevalence ; Risk Factors ; Triglycerides ; Vitamin B 12 ; Vitamins

OBJECTIVE: The purpose was to test the hypothesis that the common missense mutation of 5,10-methylenetetrahydrofolate reductase(MTHFR) is more prevalent among preeclamptic women compared with control and also was to determine whether homocysteine and other lipid profile is changed in pregnant women with preeclampsia. METHODS: We measured plasma homocysteine, cholesterol, HDL, LDL, triglyceride, apoprotein B, vitamin B12, and folate in 48 pregnant women without preeclampsia and 22 women with preeclampsia. And the MTHFR genotype was determined with a polymerase chain reaction/restriction fragment length polymorphism method. Results were analyzed with a X2 contingency table and T-test. RESULTS: The prevalence of the MTHFR C677T mutation was not significantly different between the population studied. There was no significant difference in the level of plasma homocysteine, cholesterol, HDL, LDL, triglyceride, apoprotein B, and folate between controls and preeclamptic women. Furthermore, there was a significant difference in the level of plasma vitamin B12 between the population studied. CONCLUSION: These data suggest that the MTHFR C677T mutation is not a risk factor for preeclampsia in this population. Plasma homocysteine, cholesterol, HDL, LDL, triglyceride, apoprotein B, and folate level are not elevated in preeclamptic women. However, the plasma vitamin B12 level is elevated in preeclamptic women. Further studies are necessary to determine why the plasma vitamin B12 level is elevated in preelamptic women although they did not have vitamin drug.
Apoproteins ; Cholesterol, HDL ; Female ; Folic Acid ; Genotype ; Homocysteine* ; Humans ; Methylenetetrahydrofolate Reductase (NADPH2)* ; Mutation, Missense ; Plasma ; Polymorphism, Genetic ; Pre-Eclampsia* ; Pregnant Women ; Prevalence ; Risk Factors ; Triglycerides ; Vitamin B 12 ; Vitamins