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.     

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.     

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.     

两种方法检测胃液潜血的比较

目的:探讨胶体金法和邻甲苯胺法检测空腹患者胃液潜血的差异.方法:收集58例大于24 h未进饮食(为肠外营养)、3 d内未吃肉类食品、未服用Vc、铁剂等影响邻甲苯胺法检测药物的胃插管患者的胃液标本,用胶体金法和邻甲苯胺法同时检测潜血.结果:胶体金法有31例阳性,阳性率为53.4%,邻甲苯胺法有44例阳性,阳性率为75.9%,两者有显著性差异(P<0.005).结论:邻甲苯胺法检测空腹患者胃液潜血优于胶体金法.     

Gastric juice for the diagnosis of H pylori infection in patients on proton pump inhibitors

AIM： To determine the efficacy of gastric juice polymerase chain reaction （PCR） for the detection of H pylori infection in comparison with histology and gastric antral biopsy PCR in patients on a proton pump inhibitor （PPI）. METHODS： Eighty-five consecutive patients with dyspeptic symptoms were enrolled. Gastric biopsies for histology, PCR and gastric juice were collected at endoscopy for PCR of the H pylori urease C gene （ure C）. Sensitivity, specificity, positive predictive value （PPV）, negative predictive value （NPV）, accuracy, positive and negative likelihood ratio for PCR of gastric juice for the H pylori ure C gene was compared to histology and gastric antral biopsy H pylori ure C PCR in patients with and without PPI. RESULTS： Gastric juice PCR was positive in 66 （78%） patients. Histology showed H pylori associated gastritis in 57 （67%）. Gastric biopsy PCR was positive in 72 （85%）. In patients not taking PPI, the sensitivity, specificity, PPV, NPV, accuracy and positive and negative likelihood ratio for gastric juice PCR were 89%, 72%, 91%, 67%, 90%, 85%, 3.1 and 0.1 respectively. In patients on PPI these values were 86%, 100%%, 100%, 29%, 86%, 9.5 and 1.4, respectively. CONCLUSION： Gastric juice PCR for the diagnosis of H pylori infection has increased sensitivity compared to histology with PPI. The use of gastric juice PCR is recommended to confirm H pylori status in patients taking PPIs.     

Förster resonance energy transfer (FRET)-Labeled nanoprobe enables real-time diagnosis of pancreatic juice activation due to postoperative pancreatic fistula

ObjectivesPostoperative pancreatic fistula (POPF) subsequent to pancreatectomy often causes activation of pancreatic juice, resulting in serious complications. In POPF, the types of pancreatic juices found are active and inactive, and the identification of these two types of pancreatic juice greatly contributes to the development of postoperative management after pancreatectomy. This study reports favorable results of the clinical application of the Förster resonance energy transfer (FRET) nanoprobe that was independently developed to distinguish between the active and inactive types of pancreatic juice.MethodsThe FRET nanoprobe developed was a nanoprotein capsule. It exuded a red color when the capsule structure was maintained. When activated protease in the pancreatic juice acts on it, the capsules are reduced quantitatively and FRET is abolished, resulting in a change in color from red to green. Pancreatic juice activation can be measured by the FRET signal. A total of 117 drainage fluid samples from 16 postpancreatoduodenectomy cases were obtained and evaluated.ResultsThe diagnosis of pancreatic juice activation was possible using the FRET signal with a cut-off value of 1.6. Pancreatic juice activation was not associated with drainage fluid amylase (AMY) levels. The results demonstrated that pancreatic juice was activated when drainage fluid was infected.ConclusionThe use of a FRET nanoprobe enabled real-time detection of the presence or absence of pancreatic juice activation in pancreatic fistula after pancreatic surgery. There was an adequate correlation between infection and pancreatic juice activation regardless of drain AMY levels.     

Pre-Exercise Nutrition: The Role of Macronutrients,Modified Starches and Supplements on Metabolism and Endurance Performance

Endurance athletes rarely compete in the fasted state, as this may compromise fuel stores. Thus, the timing and composition of the pre-exercise meal is a significant consideration for optimizing metabolism and subsequent endurance performance. Carbohydrate feedings prior to endurance exercise are common and have generally been shown to enhance performance, despite increasing insulin levels and reducing fat oxidation. These metabolic effects may be attenuated by consuming low glycemic index carbohydrates and/or modified starches before exercise. High fat meals seem to have beneficial metabolic effects (e.g., increasing fat oxidation and possibly sparing muscle glycogen). However, these effects do not necessarily translate into enhanced performance. Relatively little research has examined the effects of a pre-exercise high protein meal on subsequent performance, but there is some evidence to suggest enhanced pre-exercise glycogen synthesis and benefits to metabolism during exercise. Finally, various supplements (i.e., caffeine and beetroot juice) also warrant possible inclusion into pre-race nutrition for endurance athletes. Ultimately, further research is needed to optimize pre-exercise nutritional strategies for endurance performance.     

Intraoperative amylase level of pancreatic juice as a simple predictor of pancreatic fistula after pancreaticoduodenectomy

IntroductionA soft remnant texture of the pancreas is commonly accepted as a risk factor for postoperative pancreatic fistula (POPF) after pancreaticoduodenectomy (PD). However, its assessment is subjective. The aim of this study was to evaluate the significance of intraoperative amylase level of the pancreatic juice as a risk factor of POPF after PD.MethodThis study included 75 patients who underwent PD between November 2014 and April 2020 at Jikei University Hospital. We investigated the relationship between pancreatic texture, intraoperative amylase level of pancreatic juice, results of the pathological evaluations, and the incidence of POPF.ResultsTwenty-three patients (31%) developed POPF. The significant predictors of POPF were non-ductal adenocarcinoma (p < 0.01), soft pancreatic remnant (p < 0.01), high intraoperative blood loss (p < 0.01), high intraoperative amylase level of pancreatic juice (p < 0.01), and low pancreatic fibrosis (p < 0.01). Multivariate analysis revealed that the significant independent predictors of POPF were high intraoperative blood loss (p < 0.01) and high intraoperative amylase level of pancreatic juice (p = 0.02). Receiver operating characteristic (ROC) analysis showed that the cut-off value for the intraoperative amylase level of pancreatic juice was 2.17 × 10⁵ IU/L (area under the curve = 0.726, sensitivity = 95.7%, and specificity = 50.0%)ConclusionsThe intraoperative amylase level of pancreatic juice is a reliable objective predictor for POPF after PD.     

Consumption of cranberry beverage improved endogenous antioxidant status and protected against bacteria adhesion in healthy humans: a randomized controlled trial

Consumption of polyphenol-rich foods is associated with lower risk from many chronic diseases. We hypothesized that a single dose of cranberry beverage would improve indices of oxidative stress, inflammation, and urinary antibacterial adhesion activity in healthy humans. Six males and 6 females (18-35 years; body mass index, 19-25 kg/m²) consumed placebo, cranberry leaf extract beverage, or low-calorie cranberry juice cocktail (LCJC) once in a randomized, double-blind, placebo-controlled cross-over experimental design trial. The washout period between beverages was 1 week. Blood was collected 0, 2, 4, 8, and 24 hours after beverage consumption for measuring oxidative and inflammatory biomarkers. Urine was collected at 0, 0 to 3, 3 to 6, 6 to 9, 9 to 12, and 24 hours postintervention to assess antibacterial adhesion activity. Consumption of cranberry leaf extract beverage elevated (P < .05) blood glutathione peroxidase activity, whereas LCJC consumption increased (P < .05) glutathione concentrations and superoxide dismutase activity compared with placebo. Cranberry leaf extract beverage and LCJC consumption had no effect on the inflammatory biomarkers measured as compared with placebo. At 0 to 3 hours postconsumption, urine from participants who consumed cranberry beverages had higher (P < .05) ex vivo antiadhesion activity against P-fimbriated Escherichia coli compared with placebo. An acute dose of cranberry beverages improved biomarkers of antioxidant status and inhibition of bacterial adhesion in urine.     

Study on combined effects of blanching and sonication on different quality parameters of carrot juice

This study was conducted to evaluate the combined effects of blanching and sonication on carrot juice quality. Carrots were blanched at 100?°C for 4?min in normal and acidified water. Juice was extracted and sonicated at 15?°C for 2?min keeping pulse duration 5?s on and 5?s off (70% amplitude level and 20?kHz frequency). No significant effect of blanching and sonication was observed on Brix, pH and titratable acidity except acidified blanching that decreased pH and increased acidity significantly. Peroxidase was inactivated after blanching that also significantly decreased total phenol, flavonoids, tannins, free radical scavenging activity, antioxidant capacity and ascorbic acid and increased cloud and color values. Sonication could improve all these parameters significantly. The present results suggest that combination of blanching and sonication may be employed in food industry to produce high-quality carrot juice with reduced enzyme activity and improved nutrition.