Study on chronic pancreatitis and pancreatic cancer using MRS and pancreatic juice samples

AIM:To investigate the markers of pancreatic diseases and provide basic data and experimental methods for the diagnosis of pancreatic diseases.METHODS:There were 15 patients in the present study,among whom 10 had pancreatic cancer and 5,chronic pancreatitis.In all patients,pancreatic cancer or chronic pancreatitis was located on the head of the p-a-ncrea-s.Pa-thology da-ta-of a-ll p-a-tients wa-s confirmed by biopsy and surgery.Among the 10 patients with pancreatic cancer,3 people had a medical history of l...     

In vitro studies of gastric juice in patients with food-cobalamin malabsorption

Food-cobalamin absorption depends on the initial release of cobalamin from its binders in food. Therefore, the characterization of patients' gastric juices and their behavior in this process was undertaken. Pentagastrin-stimulated gastric juice specimens from three patients with severe food-cobalamin malabsorption, six patients with mild malabsorption, and five patients with normal absorption were tested for pH, pepsin, intrinsic factor content, and anin vitro method that quantitates transfer of cobalamin from egg yolk to gastric R binder. Transfer of cobalamin correlated best within vivo egg yolk-cobalamin absorption test results in the 14 patients (r=0.731,P<0.005). Transfer also correlated inversely with gastric juice pH (r=–0.619,P<0.02). Basal gastric juice specimens, with their higher pH, from the same subjects failed to promote cobalamin transfer until their pH was lowered to 1.0–1.3. Pepsin levels did not correlate within vitro transfer or with absorptionin vivo; nevertheless, raising the low pepsin concentration of one stimulated gastric juice improved transfer, while inhibiting pepsin activity with pepstatin A inhibited transfer. Mixing experiments with selected stimulated gastric juices demonstrated that poorin vitro transfer, which in a few cases seemed unrelated to pH or pepsin levels, was not due to any inhibitory activity of such gastric juices. These studies confirm that gastric acid and pepsin play a central role in releasing food-bound cobalamin and transferring it to R binder, but suggest that other, still unidentified gastric defects occasionally contribute to impaired transfer; the latter defects are not inhibitory in nature but seem to involve the absence of a permissive activity. The finding that the ability of a gastric juice to promote the transfer of cobalaminin vitro was the best overall indicator of a patient's ability to absorb food cobalaminin vivo suggests that gastric juice defects are responsible for most cases of food-cobalamin malabsorption. The phenomenon may also provide a practicalin vitro estimate of a patient's ability to absorb food cobalamin.This study was supported by grant DK 32640 from the National Institutes of Health.     

Clinical significance of cathepsin E in pancreatic juice in the diagnosis of pancreatic ductal adenocarcinoma

BACKGROUND: It has been reported that cathepsin E (CTSE) is a non-secretory and intracellular aspartic proteinase found in the superficial epithelial cells of the stomach and that it is also expressed in pancreatic ductal adenocarcinoma. We evaluated the diagnostic value of CTSE in the pancreatic juice in the diagnosis of pancreatic ductal adenocarcinoma compared with that of CA19-9, carcinoembryonic antigen (CEA) and K-ras mutations. METHODS: One hundred and one patients (25 with pancreatic ductal adenocarcinoma and 76 with chronic pancreatitis) were examined for the diagnostic significance of CTSE in the pancreatic juice in the diagnosis of pancreatic ductal adenocarcinoma. Forty of 101 patients (15 with pancreatic ductal adenocarcinoma and 25 with chronic pancreatitis) were examined to compare the diagnostic value of various tumor markers in the pancreatic juice, namely CA19-9, CEA, K-ras mutations and CTSE. RESULTS: The detection frequency of CTSE was significantly higher in patients with pancreatic ductal adenocarcinoma (64.0%) than in patients with chronic pancreatitis (7.9%; chi2 = 34.76; P < 0.0001). The sensitivity, specificity and diagnostic accuracy of CTSE in the pancreatic juice for pancreatic ductal adenocarcinoma was 66.7, 92.0 and 82.5%, respectively. These values were more efficient in comparison with those of CA19-9, CEA and K-ras mutations. The main cause of the detection failure of CTSE in pancreatic ductal adenocarcinoma was obstruction of the main pancreatic duct. Sensitivity was 85.7% in patients without obstruction of the main pancreatic duct. CONCLUSIONS: Cathepsin E in the pancreatic juice is a novel marker for a definitive diagnosis of pancreatic ductal adenocarcinoma.     

Detection of c-Ki- ras point mutation from pancreatic juice

Summary Cytological diagnosis of pancreatic carcinoma sometimes poses difficulties in distinguishing malignant from benign cells. Recent molecular study of pancreatic carcinoma has revealed a very high incidence of a point mutation of the c-Ki-ras oncogene at codon 12 in this neoplasm. To take advantage of this technique for the diagnosis of pancreatic carcinoma, we attempted to amplify the c-Ki-ras gene from endoscopically obtained pancreatic juice by isolation of DNA and polymerase chain reaction (PCR) coupled with restriction fragment length polymorphism (RFLP). PCR was possible in approx 70% of the cases. A point mutation was nonradioisotopically detected in 4 of 6 pancreatic carcinomas and in one intraductal papillary neoplasm, whereas no mutation was detected in other cases. Thus, this method was thought to be useful for the diagnosis of pancreatic carcinoma.     

Grapefruit Juice and its Constituents Augment Colchicine Intestinal Absorption: Potential Hazardous Interaction and the Role of P-Glycoprotein

Purpose  To investigate the potential interaction between grapefruit juice (GFJ) and the oral microtubule polymerization inhibitor colchicine, a P-gp and CYP3A4 substrate. Methods  Colchicine intestinal epithelial transport was investigated across Caco-2 cell monolayers in both AP–BL and BL–AP directions, in the absence/presence of known P-gp inhibitors (verapamil and quinidine). The concentration-dependent effects of GFJ and its major constituents (6′-7′-dihydroxybergamottin, naringin and naringenin) on colchicine Caco-2 mucosal secretion were examined. The effect of GFJ on colchicine intestinal-permeability was then investigated in-situ in the rat perfusion model, in both jejunum and ileum. Results  Colchicine exhibited 20-fold higher BL–AP than AP–BL Caco-2 permeability, indicative of net mucosal secretion, which was reduced by verapamil/quinidine. Colchicine AP–BL permeability was increased and BL–AP was decreased by GFJ in a concentration-dependent manner (IC₅₀ values of 0.75% and 0.46% respectively), suggesting inhibition of efflux transport, rather than metabolizing enzyme. Similar effects obtained following pre-experiment incubation with GFJ, even though the juice was not present throughout the transepithelial study. 6′-7′-Dihydroxybergamottin, naringin and naringenin displayed concentration-dependent inhibition on colchicine BL–AP secretion (IC₅₀ values of 90, 592 and 11.6 μM respectively). Ten percent GFJ doubled colchicine rat in-situ ileal permeability, and increased 1.5-fold jejunal permeability. Conclusion  The data suggest that GFJ may augment colchicine oral bioavailability. Due to colchicine narrow therapeutic-index and severely toxic side-effects, awareness of this interaction is prudent.     

Inhibitory effect of grapefruit juice on the genotoxicity induced by hydrogen peroxide in human lymphocytes

By means of the comet assay we demonstrated a strong effect by hydrogen peroxide (HP) and no damage by grapefruit juice (GJ) in human lymphocytes. Cells exposed to HP and treated with three concentrations of GJ (10-90 min) showed an increase of DNA damage by HP over the control level, and a decrease of such damage by GJ. With the comet assay plus formamidopyrimidine-DNA-glycosylase we found the strongest increase of DNA damage by HP over the control level, and the strongest reduction of such damage by GJ. By applying the comet/FISH method we determined 98% of the p53 gene signals in the comet head of control cells along the experiment (10-90 min), in contrast with about 90% signals in the comet tail of cells exposed to HP. Cells treated with both agents showed a significant, concentration/time dependent return of p53 signals to the head, suggesting enhancement of the gene repair. Finally, with the annexin V assay we found an increase in apoptosis and necrosis by HP, and no effect by GJ; when GJ was added to HP treated cells no modification was observed in regard to apoptosis, although a decrease of necrosis was observed.     

Genotypic identification of Penicillium expansum and the role of processing on patulin presence in juice

This work aimed at isolation and identification of patulin producing fungi and to follow the presence of patulin during apple juice processing. Among 34 Penicillium isolates, eight isolates (five from healthy appeared apples and 12 from rot spotted apples) were considered as patulin producers using thin-layer chromatography. These isolates were classically identified as a Penicillium expansum. PCR utilizing primers based on the polygalacturonase gene of P. expansum was applied for detecting this mold. The PCR amplified a 404-bp DNA product from all tested P. expansum isolates, but not in other common food spoilage Penicillium species. RAPD technique using P1 or M13 primers was applied to determine the similarity of the P. expansum isolates. RAPD results revealed that the tested strains showed high percentage of similarity and no correlation was observed between cluster analysis and the sources of isolation. Patulin could not be detected in healthy appeared apples and their extracted juice during different stages of juice process. In apple juice made from the healthy parts of apples decayed by P. expansum contained patulin which may present health hazard. The obtained results assured that patulin is known to be stable in apple juice even after pasteurization. In conclusion, the removal of the rotten part from the fruit is not sufficient to eliminate the mycotoxin patulin from apple juice. Although, the enzyme treatment (pectinase and amylase) and pasteurization (95 °C for 7 min) significantly (p < 0.05) reduced patulin level, its level is still higher than the level of <50 μg/kg considered by Codex alimentarius when the apple juice processed from the healthy parts of rot spotted fruits.     

Oxidative markers in children with severe obesity following low‐calorie diets supplemented with mandarin juice

Aim: To evaluate the effect of supplementing a hypocaloric diet with mandarin juice, a food with a high content of antioxidants (vitamin C, flavonoids and carotenoids), on biomarkers of oxidant/antioxidant status of severe obese children. Methods: Forty obese children were randomized into two groups pair‐wise in a 4‐week controlled intervention study. Both groups followed a hypocaloric diet. One group received additionally a supplementation of 500 mL of 100% mandarin juice daily. Clinical data, anthropometry, dietary intake and fasting blood samples were collected at baseline and after the intervention. Lipid peroxidation was assessed by circulating levels of malondialdehyde, and protein oxidation was determined by the concentration of plasma carbonyl groups. The antioxidant defence was evaluated by red cell‐reduced glutathione and plasma levels of α‐tocopherol and vitamin C. Results: The supplemented group experienced a decrease in the levels of malondialdehyde (?9.6%, p = 0.014) and carbonyl groups (?36.1%, p = 0.006) and an increase in antioxidants (α‐tocopherol +16.1%, p = 0.006, glutathione +36.1%, p < 0.0001, and vitamin C + 94.6%, p < 0.0001). Conclusion: The mandarin juice consumption with a reduced calorie diet positively affects the antioxidant defence and produces a decrease in biomarkers of oxidative stress in obese children.     

Red fruit juice quality and authenticity control by HPLC

The present work used International Federation of Fruit-Juice Producers (IFU) Method No. 71 with minor modifications for the analysis of anthocyanins, betacyanins, synthetic red pigments, hydroxycinnamic acids, hydroxybenzoic acids and catechins present in red fruit and vegetable juices and red-purple soft-drinks. The proposed HPLC method has been implemented with simultaneous UV-Visible and fluorescence detection and offers unambiguous composition results for 9 red fruit and vegetable juices: strawberry, red raspberry, blueberry, European cranberry, blackcurrant, sour cherry, red grape, purple carrot and purple prickly pear. Twenty-eight anthocyanins, 4 betacyanins, 1 natural and 6 synthetic pigments, 11 hydroxycinnamic acids, 6 hydroxybenzoic acids and 2 catechins were determined in a 30-min chromatogram. This method is useful for quality and authentication analyses of red fruit and vegetable juices, and red-purple soft-drinks. The use of a unique analysis method for polyphenol analysis is encouraged as a helpful tool to build up an unambiguous polyphenol composition database of foods.     

The effects of Xanthigen™ in the weight management of obese premenopausal women with non-alcoholic fatty liver disease and normal liver fat

Aim: To investigate the effects of Xanthigen (brown marine algae fucoxanthin + pomegranate seed oil (PSO)) on body weight, body fat, liver lipids, and blood biochemistry; and Xanthigen and its individual components on resting energy expenditure (REE) in obese, non-diabetic female volunteers with non-alcoholic fatty liver disease (NAFLD) and normal liver fat (NLF) content.
Methods: Sixteen-week, double-blind, randomized, placebo-controlled study. Food record data, body composition, REE (only 41 volunteers with NAFLD) and blood sample analysis were assessed weekly for 16 weeks in 151 non-diabetic, obese premenopausal women with liver fat content above 11% (NAFLD) n = 113, and below 6.5% (NLF) n = 38.
Results: Xanthigen-600/2.4 mg (300 mg PSO + 300 mg brown seaweed extract containing 2.4 mg fucoxanthin) resulted in statistically significant reduction of body weight (5.5 ± 1.4 kg NAFLD group and 4.9 ± 1.2 kg NLF group, p < 0.05), waist circumference (NAFLD group only), body (3.5 ± 1.9 kg NAFLD group, p < 0.001; 3.6 ± 0.7 kg NLF group, p < 0.05) and liver fat content, liver enzymes (NAFLD group only), serum triglycerides and C-reactive protein. Weight loss and reduction in body and liver fat content occurred earlier in patients with NLF than in patients with NAFLD. Fucoxanthin ( > 2.4 mg) and Xanthigen-400/1.6 mg (200 mg PSO + 200 mg brown seaweed extract containing 1.6 mg fucoxanthin) significantly increased REE in NAFLD subjects compared to placebo.
Conclusions: Xanthigen promoted weight loss, reduced body and liver fat content, and improved liver function tests in obese non-diabetic women. Xanthigen and Fucoxanthin also increased REE. This product may be considered a promising food supplement in the management of obesity.