Polydopamine-PEG-Folic Acid Conjugate Film Engineered TiO2 Nanotube Arrays for Photoelectrochemical Sensing of Folate Binding Protein.

Serum-soluble folate binding protein (FBP) is an important tumor marker, and the development of a simple biosensing method is highly needed. In this work, a photoelectrochemical (PEC) biosensor for the detection of FBP was proposed based on the construction of an antifouling interface and the unique ligand-protein recognition. The PEC sensing platform was prepared by the biomimetic polydopamine (PDA) coating on TiO2 nanotubes arrays (NTAs). A significant PEC enhancement effect was obtained due to the macroporous structures. Excellent antifouling performance was achieved by conjugation of amino-group-terminated 8-arm poly(ethylene glycol) (PEG). The incorporation of folic acid (FA) retains the antifouling property and shows recognition abilities toward FBP. The fabricated PEC biosensor shows good analytical performance. The combination of ligand-protein recognition and a PEC antifouling interface provides a good consideration for the development of FBP biosensors.

FBP2 and Talin-1 are potential protein markers for Mongolian medicine symptom evaluation in viral infectious diseases.

BACKGROUND: Influenza, measles, and mumps are common viral infectious diseases in Mongolia. The traditional Mongolian medicine (TMM) classified them as warm disease, and still plays a major role in the diagnoses and treatments. METHODS: To interpret the connotation of the complex theoretical system in TMM with scientific technique, in this study, a high throughput mass spectrometry was used to identify potential protein markers of TMM symptom types. Fifty venous blood samples were drawn from influenza, measles and mumps patients. Differential proteins between samples of patients diagnosed as immature and mature heat in TMM were detected by mass spectrometry. RESULTS: After proteomics analysis, 1500 proteins and 7619 polypeptides were identified and 1323 in total showed differential expression between those 2 symptom types; then enrichment analysis of the differentially expressed proteins revealed the significant biological functions related to the differentially expressed proteins, including cardiomyopathy, several bacterial and parasitic infections, bacterial invasion of epithelial cells, insulin signaling pathway, and regulation of actin cytoskeleton. The network analysis showed that FBP2 and Talin-1 were critical points and might determine the evolution directions of TMM warm disease symptom. CONCLUSIONS: This study suggests that the identified core differential proteins may be regarded as potential biomarkers, and benefit to evaluate the evolutionary tendency of TMM warm disease symptoms.

[Severe lactic acidosis requiring continuos haemodiafiltration in a young patient with unrecognized metabolic abnormality. Case report].

BACKGROUND: Lactic acidosis (LA) is the most common form of metabolic acidosis, defined by lactate values greater than 5 mmol/L and pH<7.34. The pathogenesis of LA involves hypoxic causes (type A) and non-hypoxic (type B), often coexisting. Identification and removal of the trigger are mandatory in the therapeutic management of LA. The case: A 38 years-old male patient entered the Emergency Ward for dyspnea, fever, vomiting and hyporexia. An important respiratory distress with hyperventilation due to severe LA was found, together with severe hypoglicemia, without renal impairment. Past medical history unremarkable, except for reported episodic hypoglicemia in the childhood, with fructose "intolerance", without any other data. No evidence of intoxications, septic shock or significant cytolysis. No drugs causing LA. The patient underwent orotracheal intubation, glucose infusion, and continuous haemodiafiltration for 36-hrs. A rapid general improvement was obtained with stabilization of acid-base balance. A diagnosis of fructose-1,6-diphosphatase deficiency was made. It is an autosomical recessive gluconeogenesis abnormality, with recurrent episodes of hypoglicemia and lactic acidosis after fasting, potentially lethal. The therapy is based on avoiding prolonged fasts, glucose infusion, and a specific diet, rich in glucose without fructose intake. CONCLUSIONS: The presence of not-otherwise-explained lactic acidosis in young patients has to place the suspect of an underlying and unknown metabolic derangement; in these cases, the involvement of the nephrologist appears to be pivotal for the differential diagnosis of the abnormalities of the acid-base balance, and for setting the best treatment.

Improved glycemic control, pancreas protective and hepatoprotective effect by traditional poly-herbal formulation "Qurs Tabasheer" in streptozotocin induced diabetic rats.

BACKGROUND: The present study was undertaken to evaluate the antihyperglycemic, antihyperlipidemic and hepatoprotective effect of a traditional unani formulation "Qurs Tabasheer" in streptozotocin (STZ) induced diabetic wistar rats. Up till now no study was undertaken to appraise the efficacy of "Qurs Tabasheer" in the diabetic rats. Qurs Tabasheer is a unani formulation restraining preparations from five various herbs namely Tukhme Khurfa (Portulaca oleracea seed), Gule Surkh (Rosa damascena flower), Gulnar (Punica granatum flower), Tabasheer (Bambusa arundinasia dried exudate on node), Tukhme Kahu (Lactuca sativa Linn seed). METHODS: Effect of Qurs Tabasheer was assessed in STZ (60 mg/kg, i.p single shot) induced diabetic wistar rats. STZ produced a marked increase in the serum glucose, Total Cholesterol, LDL cholesterol, VLDL Cholesterol, Triglycerides and trim down the HDL level. We have weighed up the effect of Qurs Tabasheer on hepatic activity through estimating levels of various liver enzymes viz. Hexokinase, Glucose-6-Phosphatase and Fructose-1-6-biphosphatase in STZ diabetic wistar rats. RESULTS: In STZ-induced diabetic wistar rats level of Hexokinase, and Glucose-6-Phosphatase was decreased to a significant level while the level of fructose-1-6-biphophatase was augmented. Therapy with Qurs Tabasheer for 28 days to STZ-induced diabetic rats significantly reduces the level of serum glucose, total cholesterol, triglycerides, glucose-6-phosphatase and fructose-1-6-biphosphatase, while magnitude of HDL cholesterol and hexokinase was amplified. CONCLUSION: Antihyperglycemic, antihyperlipidemic activity of Qurs Tabasheer extract in STZ- induced wistar rats was found to be more effective than standard oral hypoglycemic drug Glimepiride.

Effect of pterostilbene on hepatic key enzymes of glucose metabolism in streptozotocin- and nicotinamide-induced diabetic rats.

The purpose of this study was to investigate the effect of pterostilbene and its effect on key enzymes of glucose metabolism. Diabetic rats were orally administered with pterostilbene (10, 20, 40 mg/kg) for 2, 4 and 6 weeks on glucose was determined. Administration of pterostilbene at 40 mg/kg significantly decreases plasma glucose. Based on these data, the higher dose, 40 mg/kg pterostilbene, was selected for further evaluation. Oral administration of pterostilbene for 6 weeks on glucose, insulin levels and hepatic enzymes in normal and streptozotocin (STZ)-nicotinamide-induced diabetic rats. A significant decrease in glucose and significant increase in plasma insulin levels were observed in normal and diabetic rats treated with pterostilbene. Treatment with pterostilbene resulted in a significant reduction of glycosylated hemoglobin and an increase in total hemoglobin level. The activities of the hepatic enzymes such as hexokinase was significantly increased whereas glucose-6-phosphatase, fructose-1,6-bisphosphatase were significantly decreased by the administration of pterostilbene in diabetic rats. A comparison was made between the action of pterostilbene and the antidiabetic drug--metformin.

Antihyperglycaemic effect of Casearia esculenta root extracts in streptozotocin-induced diabetic rats.

The present study was carried out to evaluate the antihyperglycaemic effect of Casearia esculenta root extract and to study the activities of liver hexokinase and gluconeogenic enzymes such as glucose-6-phosphatase and fructose-1,6-bisphosphatase in liver and kidney of normal and streptozotocin-induced diabetic rats. Oral administration of aqueous extract of root (300 mg/kg body weight) for 45 days resulted in a significant reduction in blood glucose from 250.79 +/- 12.65 to 135.70 +/- 8.90 and in a decrease in the activities of glucose-6-phosphatase and fructose-1,6-bishosphatase and an increase in the activity of liver hexokinase. However, in the case of 200 mg/kg body weight of extract, less activity was observed. The study clearly shows that the root extract of C. esculenta possesses potent antihyperglycaemic activity but weaker than that of glibenclamide.

Discrimination between periportal and pericentral necrosis of rat liver by determination of glutamine synthetase and other enzyme activities in serum.

Periportal or pericentral necrosis of rat liver was produced by injection of allyl-alcohol or bromobenzene, respectively. Activities of predominantly periportal and perivenous enzymes were determined in serum during maximal necrosis. Aspartate aminotransferase, which is more or less homogeneously distributed in the liver acinus, exhibited similar activities in serum after periportal and pericentral injury. Serum activities of the mainly periportal enzymes alanine aminotransferase and fructose 1,6-bisphosphatase were 1.5- to 2-fold higher after periportal as compared to pericentral necrosis. Serum activity of the mainly pericentral glutamate dehydrogenase was 3-fold higher after pericentral than after periportal damage. However, due to individual variations necrosis could not be definitively localized in any case by measurement of these enzyme activities. Better discrimination between periportal and pericentral necrosis was achieved by the serum activity of the exclusively pericentral enzyme glutamine synthetase, which was 8-fold higher after pericentral as compared to periportal necrosis. Conclusive discrimination was obtained by the activity ratio fructose 1,6-bisphosphatase/glutamine synthetase in serum.

Diagnosis of fructose-1,6-bisphosphatase deficiency using leukocytes: normal leukocyte enzyme activity in three female patients.

In the past 5 years we have discovered 8 boys and 3 girls who suffer from fructose-1,6-bisphosphatase deficiency. Although they all showed the typical symptoms of the deficiency such as frequent vomiting, hypoglycemia, lactic acidosis, and hepatomegaly, the age at diagnosis varied from 2 months to 4 years. All the boys revealed the deficient enzyme activity in leukocytes but none of the girls. The liver biopsy was investigated in six patients to confirm the diagnosis. These results suggest the existence of tissue-specific isoenzymes for fructose-1,6-bisphosphatase possibly with a different gene origin.

Rat serum fructose 1,6 bisphosphatase: modifications in different experimental conditions.

Fructose 1,6 bisphosphatase activity was measured in rat serum. The activity of the enzyme presented the following characteristics: pH optimum alkaline, between 8-8.5 and inhibited by AMP. The activity was measured in different experimental situations, such as streptozotocin diabetes, hepatocellular injury produced by carbon tetrachloride (CCl4), and bile-duct ligation. In diabetic rat, serum activity increased 2-fold with respect to the control values. In animals treated with CCl4 the activity increased 10-14 fold. On the contrary bile duct ligation decreased activity according to the cholestasis time. The results obtained in this study show that fructose 1,6 bisphosphatase is an enzyme measurable in serum, which changes according to different situations of liver cell injury.

Evaluation of key gluconeogenic enzymes in experimental biliary obstruction.

In order to evaluate the usefulness of key gluconeogenic enzymes, in relation to the markers commonly used (alkaline phosphatase and gamma-glutamyl transpeptidase) for the diagnose of cholestasis the serum activity of phosphoenolpyruvate carboxykinase, fructose 1,6-bisphosphatase and glucose-6-phosphatase has been measured in rats with bile-duct ligation. Among the gluconeogenic enzymes studied only phosphoenolpyruvate carboxykinase activity increased significantly in the first 48 hours after cholestasis, decreasing thereafter to normal values. Both alkaline phosphatase and gamma-glutamyl transpeptidase activities showed a very significant increase which persisted throughout the experiment. These results seem to indicate that in spite of the high organ specificity of these enzymes they do not appear to be useful for the diagnosis of cholestasis.

Serum activity of the key gluconeogenic enzymes in carbon-tetrachloride-induced experimental hepatotoxicity.

Serum activity has been measured in three of the key enzymes in the gluconeogenic pathway in rats subjected to experimental hepatotoxicity after intraperitoneal administration of carbon tetrachloride. The levels of phosphoenolpyruvate carboxykinase (PEPCK) and fructose-1,6-biphosphatase (FBPase) showed a similar behavior to the transaminase (AST and ALT), increasing markedly with respect to the controls at 12 h after administration of the poison, reaching their maximum peak of activity at between 24 and 36 h, and returning to normal values at 96 h. The activity of glucose-6-phosphatase was not significantly modified throughout the treatment. These results seem to demonstrate that the determination of the serum activity of PEPCK and FBPase could be a sensitive and specific marker of hepatic cytolysis.

Fructose bisphosphatase activity in the serum of rats treated with carbon tetrachloride.

In this work, fructose bisphosphatase activity in the serum of rats treated by different carbon tetrachloride doses was measured. Fructose bisphosphatase activity increased very significantly with respect to the control animals in all groups assayed. The severe reduction of the activity measured in the presence of adenosine-5'-monophosphate and its stability when measured in the presence of 1-p-bromotetramisole oxalate support its specific origin. These data suggest that serum FBPase activity measurement could be used as a biochemical marker in the diagnosis of hepatocellular injury.

[Hypoglycemia and hyperlactacidemia in relation with a new case of fructose-1,6-diphosphatase deficiency. Determination of enzymatic activity in leukocytes]. / Hypoglycémie et hyperlactacidémie en rapport avec un nouveau cas de déficit en fructose-1,6-diphosphatase. Détermination de l'activité enzymatique dans les leucocytes.

A 11.5 month-old girl with recurrent episodes of hypoglycemia and lactic acidosis was identified as having fructose-1,6-diphosphatase deficiency. The diagnosis may be realised with a simple way by an oral fructose tolerance test and the activity dosage of this enzyme in white blood cells. The fructose and sucrose-free diet and avoidance of prolonged fasting resulted in a decrease of hepatomegaly and normal values of lactate between the episodes.

Detection of heterozygotes for fructose 1,6-diphosphatase deficiency by measuring fructose 1,6-diphosphatase activity in their cultured peripheral lymphocytes.

The fructose 1,6-diphosphatase activities in peripheral lymphocytes from the parents of a patient with fructose 1,6-diphosphatase deficiency were lower than the mean value of normal controls, but the value of the mother overlapped lower values for normal controls. The fructose 1,6-diphosphatase activities in lymphocytes of normal adults and the parents increased progressively during in vitro culture, but no enzyme activity could be detected in the lymphocytes of the patient even after culture. None of the values for the parents overlapped those of normal controls on either day 5 or 10 of culture. Thus, it seems probable that heterozygotes for fructose 1,6-diphosphatase deficiency can be distinguished from normal individuals by measuring the fructose 1,6-diphosphatase activity in their cultured lymphocytes.

Fructose 1,6-bisphosphatase in the diagnosis of chronic hepatitis. I. Activity measurements of fructose 1,6-bisphosphatase in human serum.

In this communication, we propose a method for the determination in human serum of fructose 1,6-bisphosphatase based on parallel measurements of enzyme activities in presence of 1-p-bromotetramisole oxalate and adenosine 5'-monophosphate. The employment of these specific inhibitors renders the discrimination between specific and non-specific activities feasible. A regression analysis identifies fructose 1,6-bisphosphatase (EC 3.1.3.11) as the origin of the specific and alkaline phosphatase (EC 3.1.3.1) as the source of the non-specific fructose 1,6-bisphosphate dephosphorylating activities. This procedure lends itself to the diagnosis using serum samples of 'piecemeal' necrosis in liver disease.

Fructose 1,6-bisphosphatase in the diagnosis of chronic hepatitis. II. Classification of chronic hepatitis based on fructose 1,6-bisphosphatase and other laboratory data.

The histomorphology of typical liver cell necroses are here correlated with heterotope distributions of enzymes in liver parenchyma. A variety of findings indicate a congruence between gluconeogenetic areas of the liver and the typical pattern of 'piecemeal' necrosis. We therefore propose a diagnostic index based on fructose 1,6-bisphosphatase activity and the data from the clinical laboratory. This index makes it possible to distinguish between chronic persistent and chronic aggressive hepatitis.

Deficiency of glucose-6-phosphate dehydrogenase found in a case of hepatic fructose-1,6-diphosphatase deficiency.

The first case of fructose-1,6-diphosphatase (FDPase) deficiency in Japan showed a decreased activity of glucose-6-phosphate dehydrogenase (G6PD) in the liver, white, and red blood cells. In the enzymatic study of G6PD which was partially purified from red cells, the following characteristics were observed in the enzyme of the patient. 1) The G6PD activity of the patient was reduced to 17% of normal, but no evidence of a hemolytic episode was found in his past and family history. 2) In the investigation of G6PD of the patient, no abnormalities were observed in its enzymatic parameters such as electrophoretic mobility, Km for G6P and NADP, Ki for NADPH, the utilization of 2-deoxy G6P and deamino NADP, heat-stability, and pH curves. 3) The dissociation constants of red blood cell G6PD for NADP and NADPH, which were obtained from the investigations on the reactivation of cold-inactivated G6PD at 37 degrees C, were about 3 times higher in the patient as compared to the values of the normal controls. Based on these findings, it might be concluded that the G6PD deficiency found in the red blood cells of this case of a FDPase deficiency is a unique variant, which could not be characterized by using only the method recommended by a World Health Organization (WHO) scientific group. Considering that the abnormality observed in the G6PD of this patient was a decrease in the affinity of the enzyme for its coenzymes, the dissociation constants for the coenzymes in reactivation process might be another important kinetic parameter in characterizing the G6PD deficiency.

Effect of starvation on tissue and serum gluconeogenic enzymes, alkaline phosphatase and tissue glycogen in the freshwater catfish, Heteropneustes fossilis (Bloch).

The influence of starvation has been studied on tissue and serum G-6Pase F-D-Pase and alkaline phosphatase activities and on the muscle and liver glycogen content of the freshwater catfish H. fossilis (Bloch). A marked increase in G-6Pase and F-D-Pase activities and a fall in the muscle and liver glycogen content recorded during 40 day starvation. The rise in gluconeogenic enzymes during starvation may be due to glucocorticoid stimulation. Alkaline phosphatase activity was found to decline markedly during starvation. The decline in enzyme activity is attributed to some factors like a fall in the rate of synthesis caused by lowered metabolic demands and to electrolyte imbalance caused by tissue overhydration. The fall in glycogen content may be related to the starved condition of the fish. Elevation in glycogen content and alkaline phosphatase activity and a fall in gluconeogenic enzymes were noted when feeding had been resumed.