High resolution melting analysis for the detection of SLC25A13 gene mutations in Taiwan

BackgroundCitrin, encoded by SLC25A13 gene, is a mitochondrial solute transporter with a crucial role in urea, nucleotide and protein synthesis. SLC25A13 mutations cause two phenotypes, adult-onset type II citrullinemia and neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD). This study aimed to develop a high resolution melting (HRM) analysis for SLC25A13 mutation scanning and determine the carrier rate in Taiwan.MethodsDNAs from healthy subjects (n = 479), and patients with hepatocellular carcinoma (HCC, n = 100) and NICCD (n = 5) were scanned in exons 6, 9, 11, 16, and 17 and parts of introns of SLC25A13 using HRM analysis. All mutations detected by HRM analysis were further confirmed by TaqMan method and/or direct sequencing.ResultsIn healthy subjects, seventeen carriers with mutants c.851_854del (n = 10), c.1638_1660dup, c.615 + 5G > A (n = 4), and two novel mutants, c.475C > T and c.1658G > A, were detected. The frequency of carriers was about 1/28. In patients with HCC, there were only 2 carriers with c.851_854del mutant. Patients with NICCD (n = 5) diagnosed during 2007 and 2008, harbored compound heterozygous mutations c.851_854del/c.1177 + 1G > A, c.851_854del/c.1638_1660dup (n = 2), c.851_854del/c.615 + 5G > A, and c.1638_1660dup/c.615 + 5G > A.ConclusionsHRM analysis is a simple, rapid and robust method for detecting SLC25A13 mutations in clinical laboratories. SLC25A13 mutations may not be a major contributor to the pathogenesis of HCC in Taiwan.     

Low serum testosterone and frailty in older men and women

Background

Frail older persons are at high risk of morbidity and mortality, and are characterized by body composition alterations. Serum testosterone, which regulates body composition, declines with age. We investigated the relation between serum testosterone level and physiological frailty in both older men and women.

Methods

This was a cross-sectional study of 108 adults 65 years old or older. Frailty status was determined by hand-grip strength, weight change, walking speed, exhaustion, and activity levels, and was classified as frail (3 or more deficits), pre-frail (1 or 2 deficits), or robust (no deficit) according to the Fried criteria. Serum total testosterone (TT) and sex-hormone-binding globulin were measured while free testosterone (FT) was estimated.

Results

Median (range) TT and FT were lower in frail than in pre-frail and robust men (TT: (frail) 15.7 [2.4–26.9] vs. (pre-frail) 19.4 [7.2–39.9] and (robust) 25.9 [13.2–35.2] nmol/L, P = 0.03; FT: 230.0 [35.9–299.0] vs. 272.0 [86.7–411.0] and 303.0 [267.0–396.0] pmol/L, P = 0.02) and women (TT: 0.31 [0.10–0.51] vs. 0.47 [0.14–1.55] and 0.45 [0.36–1.25] nmol/L, P = 0.02; FT: 4.59 [0.46–6.63] vs. 4.66 [1.57–15.10] and 6.65 [3.91–21.00] pmol/L, P = 0.03). After adjusting for age, comorbidities, body mass index, and serum albumin in ordinal logistic regression model, odds ratios of being frail were significantly higher for those participants whose TT and FT levels were in the lowest tertile compared to the highest tertile in men (TT: odds ratio [OR] 3.29, 95% confidence interval [CI] 1.14–9.50; FT: OR 3.44, 95% CI 1.05–11.22) and in women (TT: OR 6.69, 95% CI 1.84–24.31; FT: OR 4.86, 95% CI 1.31–18.08).

Conclusions

Low serum testosterone levels were independently associated with frailty in the elderly Taiwanese.     

Enhanced dissolution of gallstone by combining ethanol with two commonly used cholelitholytic solvents.

BACKGROUND/AIMS: Contact dissolution therapy is one of the non-surgical treatments for patients with gallstone. Among the various solvents, methyl tert-butyl ether (MTBE) is used for cholesterol gallstone, while tetrasodium ethyl-dimethyl tetraacetate (EDTA-4Na) solution is used to dissolve calcium bilirubinate stones. However, the contents of gallstone cannot be precisely predicted while they are still present in the human body. This study was designed to test if the MTBE and EDTA can be mixed together and to test the solubility of different kinds of gallstone in each original solution and mixture. METHODOLOGY: Each 0.1 gm of mixed cholesterol stone, brown stone and pigment stone from 18 patients was used. Pure ethanol was chosen to enhance the miscibility between the organic phase of MTBE and the aqueous phase of EDTA. The contents of gallstone after dissolution were examined with scanning electromicroscopy. RESULTS: We found the mixture of ethanol, MTBE and EDTA to be the most efficient solvent in gallstone dissolution in comparison with the other two original solvents. The mixture reached a dissolution percentage of 97.96 +/- 1.00, 88.96 +/- 6.51 and 67.75 +/- 14.26 for cholesterol, brown and black pigment gallstone, respectively. CONCLUSIONS: We concluded that ethanol can be used to mix the MTBE and EDTA with good preservation in their litholytic effects on gallstone. The ethanol-MTBE-EDTA solvent is, therefore, a promising universal cholelitholytic agent which deserves further tests for its safety and efficacy in the in vivo study.     

Pancreatic enzymes in uremic patients with or without dialysis

One hundred thirty blood samples from 87 patients with renal failure, but without abdominal pain, were analyzed for blood urea nitrogen (BUN), creatinine, amylase, p-isoamylase, and lipase simultaneously. We found that 74, 78, and 80% of the patients had hyperamylasemia, hyperisoamylasemia, and hyperlipasemia. None had amylase higher than five times the upper limit. A few patients (2.3%) had lipase elevated to more than 10 times the upper limit. No significant change of pancreatic enzyme level was noted as a result of hemodialysis, but a significant amount of amylase was removed from the circulation in patients receiving intermittent peritoneal dialysis. Significantly lower pancreatic enzyme levels were observed in patients with less impairment of renal function. We conclude that elevation of pancreatic enzymes in uremic patients is more frequent and more extensive than most articles indicate, and that the extent of increase is related more to renal function than to the modalities of dialysis the patients received.     

The assessment of biliary CA 125, CA 19-9 and CEA in diagnosing cholangiocarcinoma--the influence of sampling time and hepatolithiasis

BACKGROUND/AIMS: This study was conducted to assess the clinical value of biliary CA 19-9, CA 125 and CEA sampled in different situations for the diagnosis of cholangiocarcinoma. METHODOLOGY: Bile was obtained from patients with bile duct obstruction on the day of biliary drainage and 3 days later separately. The etiology of biliary obstruction included choledocholithiasis (N = 51), hepatolithiasis (N = 19) and cholangiocarcinoma (N = 28). Patients of the former two conditions were all complicated with cholangitis. RESULTS: The sensitivity of biliary CEA, CA 19-9 and CA 125, whenever checked were all less than 70%. The biliary CEA and CA 19-9 were elevated in the presence of cholangitis. In addition, the biliary CEA was also increased in the patients with hepatolithiasis. The specificity of CA 125 was better than those of CEA and CA 19-9 (75.7% vs. 33.3% and 60%, respectively) on the day of biliary drainage. The diagnostic efficiency was slightly improved when combining biliary CA 125 and CEA. CONCLUSIONS: As the biliary CA 125 was less affected by inflammation and hepatolithiasis, it is more useful than CEA and CA 19-9 in the diagnosis of cholangiocarcinoma. The single tumor marker test of CA 125 or combined tumor marker test of CEA and CA 125 may be used as a useful complement to other investigative methods to differentiate benign from malignant causes of the bile duct obstruction.     

Student nurses in Taiwan at high risk for needlestick injuries

PURPOSE: To describe the prevalence and characteristics of needlestick injuries (NSI) in student nurses in Taiwan. METHODS: A self-administered questionnaire was completed by 931 student nurses from 16 hospitals randomly selected from the 132 accredited hospitals. RESULTS: The questionnaire was completed by 708 of 931 students who were contacted for participation in this study. NSI during internship was reported by 61.9% (438/708) of students, of whom 14.2% (62/438) made a formal report. The majority (70.1%) of NSI occurred in the patient's room. Hollow-bored needles contributed to half (219/438) of the NSIs of which 86.8% were syringe needles. Just over half (53.2%) of those items involved in NSIs had been used on patients. Of the hollow-bored needles involved in NSIs, 21.5% had been used on a patient with an infectious disease. Vaccination against hepatitis B virus (HBV) was lacking in 47.6% of students. CONCLUSIONS: NSIs and non-reporting of NSIs were highly prevalent in nursing students. More intensive education programs should be directed at students to increase their awareness of and compliance with Universal Precautions (UP) before commencing their practical work experience. Students need to practice prompt post-exposure evaluation so that the need for early intervention can be assessed. In addition, any public health and infection control strategy should include a universal catch-up HBV vaccination program among students before commencement of internship.     

Carbon monoxide may enhance bile secretion by increasing glutathione excretion and Mrp2 expression in rats

BackgroundNitric oxide (NO) donors have been reported to induce choleresis via an increased excretion of glutathione. The effects of another gas molecule, carbon monoxide (CO), on bile formation are, however, inconsistent among previous reports. We investigated the sequential changes of bile output and the biliary contents in rats with or without CO supplementation to elucidate the mechanism of CO on bile excretion.MethodsDichloromethane (DCM) was gastrically fed to male Sprague–Dawley rats to yield CO by liver biotransformation. The rats were divided into DCM-treated (n = 7), DCM plus L-NAME-treated (n = 6), and corn oil-treated-(n = 8) groups. Bile samples were collected hourly to examine the flow rate and bile content. Serum levels of nitrite and nitrate 4 hours after DCM supplementation with or without NO synthase (NOS) inhibition were measured by capillary electrophoresis. The expression of hepatic inducible NOS was evaluated by Western blotting 6 hours after DCM administration.ResultsLevels of carboxyhemoglobin rose to around 10% at 4 hours after DCM supplementation and were maintained until the end of the experiments. Bile flow increased after DCM supplementation and was associated with a concomitant increase of biliary glutathione and higher hepatic multidrug resistance-associated protein 2 (Mrp2) expression. Hepatic inducible NOS expression and serum nitrate/nitrite levels were also increased. Treatment with an NOS inhibitor (L-NAME) abolished the CO-induced glutathione excretion and choleresis, but not Mrp2 expression.ConclusionThe present study demonstrated that CO enhanced biliary output in conjunction with NO by increasing the biliary excretion of glutathione. The increment in biliary glutathione was associated with an increased expression of hepatic Mrp2.     

High Oxidative Stress Is Correlated with Frailty in Elderly Chinese

OBJECTIVES: To evaluate the relationship between oxidative stress and frailty in elderly people. DESIGN: Cross‐sectional study. SETTING: Community and hospital‐based outpatient clinic. PARTICIPANTS: Ninety participants aged 65 and older. MEASUREMENTS: Frailty status was determined according to the presence of weak handgrip strength, weight loss, slow walking speed, exhaustion, and low activity level and was classified as frail (≥3 criteria), prefrail (1 or 2 criteria), or robust (0 criteria). An oxidative stress marker (serum 8‐hydroxy‐2′‐deoxyguanosine, 8‐OHdG), metabolic markers (body mass index, waist–hip ratio, serum lipids, glucose, and albumin), an inflammatory marker (serum high‐sensitivity C‐reactive protein, hs‐CRP), demographic information, and comorbidities (diabetes mellitus, hypertension, congestive heart failure, osteoarthritis, overweight or obesity, impaired fasting plasma glucose, renal insufficiency, and depression) were assessed. RESULTS: Of the 90 participants, 21 (23.3%) were frail, 56 (62.2%) were prefrail, and 13 (14.4%) were robust. Frail subjects had higher median (range) serum 8‐OHdG (2.5 ng/mL (1.5–6.2 ng/mL) vs 2.3 ng/mL (0.5–8.1 ng/mL) and 1.0 ng/mL (0.5–5.3 ng/mL)) and serum hs‐CRP (2.5 mg/L (0.3–32.1 mg/L) vs 1.8 mg/L (0.3–50.5 mg/L) and 1.7 mg/L (0.3–4.0 mg/L)) levels, lower mean±standard deviation serum albumin levels (4.1±0.4 g/dL vs 4.4±0.4 g/dL and 4.6±0.2 g/dL) and higher mean waist–hip ratios (0.96±0.11 vs 0.91±0.07 and 0.89±0.05)) than prefrail and robust subjects, respectively (P<.05 for all). In multivariable regression analysis, high serum 8‐OHdG level was still significantly associated with frailty after adjusting for age, smoking status, comorbidities, waist–hip ratio, serum albumin level, and hs‐CRP level. CONCLUSION: High oxidative stress, characterized by high serum 8‐OHdG level, was independently associated with frailty in the selected sample of elderly Chinese.     

Depressive,anxiety and post-traumatic stress disorders at six years after occupational injuries

European Archives of Psychiatry and Clinical Neuroscience - The aim of this study is to determine the prevalence rates of depressive, anxiety and PTSDs, and the risk factors for psychological...     

Expression of oncogene products HER2/Neu and Ras and fibrosis-related growth factors bFGF, TGF-beta, and PDGF in bile from biliary malignancies and inflammatory disorders

The expression of several growth factors and K-ras gene mutation in bile were studied to better understand the pathogenesis and improve early diagnosis of bile duct cancers. Bile samples were collected from 12 cholangiocarcinomas (CLC), 10 ampullary cancers (APC), 3 gallbladder cancers (GBC), 7 pancreatic cancers (PNC), 9 biliary tract infection (BTI), 8 biliary stone disease (ST), and 5 normal controls (NC). The highest mean value of TGF- in bile was in patients with BTI; the mean levels of bFGF and PDGF were highest in CLC, and patients with APC and CLC had higher expression of HER2/Neu than other groups. In bile, a K-ras gene codon 12 mutation was found in 5 of 6 (83%) cases of CLC by the PCR-RFLP method. The results suggest overexpression of bFGF, PDGF, and HER2/Neu and the presence of K-ras mutation are important for carcinogenesis of bile duct cancers, and detection of the above abnormalities in bile is helpful for early diagnosis.