Claudin-4 differentiates biliary tract cancers from hepatocellular carcinomas.

The recently identified claudins are dominant components of tight junctions, responsible for cell adhesion, polarity and paracellular permeability. Certain claudins have been shown to have relevance in tumor development, with some of them, especially claudin-4, even suggested as future therapeutic target. The aim of the present study was to analyze the expression of claudin-4 in the biliary tree, biliary tract cancers and hepatocellular carcinomas. A total of 107 cases were studied: 53 biliary tract cancers, 50 hepatocellular carcinomas, 10 normal liver and 10 normal extrahepatic biliary duct samples. Immunohistochemical analysis was performed on conventional specimens and on tissue microarrays as well. Claudin-4 was further investigated by Western blot analysis and real-time RT-PCR. Intense membranous immunolabeling was found for claudin-4 in all biliary tract cancers unrelated to the primary site of origin, namely intrahepatic, extrahepatic or gallbladder cancers. Normal biliary epithelium showed weak positivity for claudin-4. In contrast, normal hepatocytes and tumor cells of hepatocellular carcinomas did not express claudin-4. The results of Western immunoblot analysis and real-time RT-PCR were in correlation with the immunohistochemical findings. Cytokeratins, as CK7 (92%) and CK19 (83%) were mostly positive in biliary tract cancers, however, one-third of hepatocellular carcinomas also expressed CK7 (34%). HSA antibody (HepPar1) reacted with the majority of hepatocellular carcinomas (86%), while being positive in a low percentage of the biliary tract cancers (8%). In conclusion, this is the first report of a significantly increased claudin-4 expression in biliary tract cancers, which represents a novel feature of tumors of biliary tract origin. Claudin-4 expression seems to be a useful marker in differentiating biliary tract cancers from hepatocellular carcinomas and could well become a potential diagnostic tool.     

Urinary excretion of beta-aminoisobutyric acid in hematological diseases

The level of beta-aminoisobutyric acid (beta-AIB), a thymine catabolite, has been measured in urine samples of 160 healthy individuals, 28 patients with renal, 27 patients with cardiovascular and 27 patients with hematological diseases and of 36 tumor patients. No significant difference in the prevalence of high excretors of beta-AIB between patients with cancer, renal and cardiovascular diseases and the healthy group was found, whereas all but two patients with hematological diseases were high excretors. Urinary beta-AIB shows a reverse correlation with the hemoglobin level and erythrocyte count in the cases of anemia, and appears to be directly correlated with the leukocyte count and blast cell content in the cases of leukemia, with its amount decreasing two to five-fold with the return of the hematological markers to normal levels after medicinal treatment. Therefore the beta-AIB concentration in urine may be used in combination with hematological indicators in assessing the disease status and in monitoring of the treatment response.     

Effect of PJ-34 PARP-inhibitor on rat liver microcirculation and antioxidant status

BACKGROUND: Ischemia-reperfusion (I-R) injury during liver resection leads to the production of toxic free radicals and oxidants that influence the microcirculation. DNA single-strand breaks can be induced by these reactive species. In response to excessive DNA damage, PARP [poly(ADP-ribose) polymerase] becomes overactivated, which can lead to cellular ATP depletion and cell death. The aim of our study was to evaluate whether PARP is expressed in post-ischemic liver, and to examine the effect of the administration of PJ-34 PARP inhibitor on liver function, histopathology, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) reaction, and the oxidative state of the liver after injury. METHODS: Male Wistar rats (weighing 250 g) underwent 60 min of normothermic, segmental liver ischemia followed by 30 min of reperfusion. The animals (n = 45) were divided into three groups: sham operated; I-R (control) treated with saline; and PJ-34 pre-treated (10 mg/kg i.v.). Hepatic microcirculation was monitored by a laser Doppler flowmeter. The reperfusion was characterized as the integral of the reperfusion area (RA) and the maximal plateau (PM). Histological alterations, TUNEL-reaction, serum, and liver tissue antioxidant levels, as well as serum ALT and AST levels were measured. RESULTS: Upon reperfusion, the PJ-34 group had significantly (P < 0.05) higher flow rates than control groups (PM(PJ-34): 58%, PM(control): 37%; RA(PJ-34.): 48%, RA(control): 25%). At the end of the 30 min reperfusion, PJ-34 resulted in significantly (P < 0.05) lower serum ALT and AST levels and chemiluminescent intensity (free radicals) of the liver. The liver's free SH-group concentration and H-donor ability of the plasma was elevated in the PARP-inhibitor treated group. Positive staining for TUNEL, after PJ-34 pre-treatment was significantly increased (P < 0.05); in contrast, the control tissues were less positively stained for TUNEL but necrotic tissue was abundant. CONCLUSION: PARP plays a pathogenetic role in the deterioration of the hepatic microcirculation and promotes hepatocellular necrosis in liver reperfusion injury.     

Short-term alanyl-glutamine dipeptide pretreatment in liver ischemia-reperfusion model: effects on microcirculation and antioxidant status in rats

BACKGROUND & AIMS: Ischemia-reperfusion (I-R) injury is responsible for the morbidity associated with liver surgery. Production of toxic free radicals influences the microcirculation. The aim of our study was to examine the effect of glutamine (Gln) supplementation--adminstered in alanyl-glutamine dipeptide form--on liver function, immuno/histopathology and the oxidative state of the liver after injury. METHODS: Two-hundred and fifty grams male Wistar rats underwent normothermic, 60 min, segmental liver ischemia followed by 6 h of reperfusion. The animals (n = 45) were divided into three groups: sham operated, I-R and parenteral Gln pretreatment. Hepatic microcirculation was monitored by laser Doppler flowmetry. At the 6 h of reperfusion, histological alterations, TUNEL reaction, active caspase-3 reaction, serum and liver tissue antioxidant levels, serum ALAT, ASAT and TNF-alpha levels were measured. RESULTS: Upon reperfusion, the Gln group had significantly (p<0.05) higher flow rates than the I-R group and, at the end of the 6h of reperfusion, significantly (p<0.05) lower serum ALAT and ASAT levels. The liver chemiluminescent intensity was lower, free SH-groups were elevated, while the reducing power was decreased in the Gln-pretreated group. Positive staining for caspase-3 after Gln pretreatment was significantly increased in contrast to the control tissues. CONCLUSION: Glutamine pretreatment is beneficial in supporting hepatic microcirculation and can prevent hepatocellular necrosis in liver reperfusion injury.     

Nutrient compositions of culinary-medicinal mushroom fruiting bodies and mycelia

Mushrooms (including fruiting bodies and mycelia) are a food with high nutritional value. This article summarizes the results of proximate composition studies of 38 fruiting bodies and 19 mycelia of 32 species of culinary-medicinal mushrooms from genera Agaricus, Agrocybe, Antrodia, Auricularia, Boletus, Clitocybe, Coprinus, Cordyceps, Trametes, Dictyophora, Flammulina, Ganoderma, Grifola, Hericium, Hypsizygus, Inonotus, Lentinus, Morchella, Pleurotus, Sparassis, Termitomyces, Tremella, and Tricholoma. Based on the proximate composition, most fruiting bodies and mycelia are low in fat and rich in protein and dietary fiber (DF); however, some are rich in soluble polysaccharides and others are rich in crude fiber. Due to the high amount of DF present, the energy provided by 100 g of dry fruiting bodies and mycelia is 46.96-292.37 kcal and 195.84-373.22 kcal, respectively. The energy (100 g) is classified into four levels: first level of >300 kcal, second level of 200-300 kcal, third level of 100-200 kcal, and fourth level of <100 kcal. Most fruiting bodies are listed in the third level; nine mycelia are listed in the first level and ten in the second level. Overall, the information about the proximate composition and energy are of great interest for fruiting bodies and mycelia to be used as foods or food-flavoring materials or in the formulation of health foods.     

Expression of Matrilin-2 in Liver Cirrhosis and Hepatocellular Carcinoma

The recently described matrilin protein family is part of the extracellular matrix, their pathophysiological role as well as distribution in liver diseases, however, have not yet been studied. Considering that matrilins have been found to play role in cell growth and tissue remodeling, their possible involvement in carcinogenesis has been raised. The main objective of this study was to investigate the changes in matrilin-2 expression which is one of the main components of basement membranes. Thirty-five cases of surgically resected hepatocellular carcinomas, 35 corresponding surrounding liver tissues and 10 normal liver samples were used for the study. In 15 of 35 cases the tumor developed on the basis of cirrhosis. Matrilin-2 protein expression was detected in normal liver around bile ducts, portal blood vessels, while sinusoids were negative by immunohistochemistry. Cirrhotic surrounding tissue showed intensive matrilin-2 staining along the sinusoids. Tumorous neovasculature was found strongly positive by immunohistochemistry. No differences, however, were detected by morphometry regarding the amount of protein expression based on the grade of hepatocellular carcinomas. Real-time RT-PCR did not show significant differences in matrilin-2 mRNA expression between normal, cirrhotic and tumor samples. This suggests posttranslational modification of matrilin-2 manifesting in altered distribution in liver fibrosis. Our data indicate that matrilin-2 is a novel basement membrane component in the liver, which is synthetised during sinusoidal "capillarization" in cirrhosis and in hepatocellular carcinoma. This is the first report to describe the expression and distribution of matrilin-2 in human normal and cirrhotic liver as well as in hepatocellular carcinoma.     

Prevalence and Determinants of Vitamin D Deficiency in 9595 Mongolian Schoolchildren: A Cross-Sectional Study

Population-based data relating to vitamin D status of children in Northeast Asia are lacking. We conducted a cross-sectional study to determine the prevalence and determinants of vitamin D deficiency in 9595 schoolchildren aged 6–13 years in Ulaanbaatar (UB), the capital city of Mongolia. Risk factors for vitamin D deficiency were collected by questionnaire, and serum 25-hydroxyvitamin D (25[OH]D) concentrations were measured using an enzyme-linked fluorescent assay, standardized and categorized as deficient (25[OH]D <10 ng/mL) or not. Odds ratios for associations between independent variables and risk of vitamin D deficiency were calculated using multivariate analysis with adjustment for potential confounders. The prevalence of vitamins D deficiency was 40.6% (95% CI 39.7% to 41.6%). It was independently associated with female gender (adjusted odds ratio [aOR] for girls vs. boys 1.23, 95% CI 1.11–1.35), month of sampling (aORs for December–February vs. June–November 5.28 [4.53–6.15], March–May vs. June–November 14.85 [12.46–17.74]), lower levels of parental education (P for trend <0.001), lower frequency of egg consumption (P for trend <0.001), active tuberculosis (aOR 1.40 [1.03–1.94]), household smoking (aOR 1.13 [1.02 to1.25]), and shorter time outdoors (P for trend <0.001). We report a very high prevalence of vitamin D deficiency among Mongolian schoolchildren, which requires addressing as a public health priority.     

Squaric acid dibutyl ester for the treatment of alopecia areata: A retrospective evaluation

Squaric acid dibutyl ester (SADBE) as topical immunotherapy is a good alternative in patients with refractory alopecia areata. In this study, we aimed to evaluate the effectiveness of SADBE treatment in alopecia areata (AA) and alopecia totalis/alopecia universalis (AT/AU) patients and determine the prognostic factors affecting treatment response. Data obtained from 34 (AA/AT/AU) patients treated with SADBE were analyzed retrospectively. Of the 34 patients, 16 (47.1%) were female and 18 (52.9%) were male. Sufficient responses were obtained in 19 (55.9%) patients. About 9 of the 19 patients (47.4%) with sufficient response reached a cosmetically acceptable level. As the severity of disease subsided, response to treatment increased. A better response was obtained when the disease onset in the spring and winter. Patients with a disease duration between 1 and 5 years responded better to the SADBE treatment compared to those with a disease shorter than 1 year and longer than 5 years. Severity of the disease, onset season of the disease, number of flares, duration of disease, and low levels of vitamin D in adult patients were observed to affect the SADBE response negatively.