Optimization and Validation of HS-SPME-GCMS Method for Determination of Multifumigant Residues in Grain, Oilseeds, Nuts, and Dry Fruit.

Background: Fumigants are approved in many countries and used to treat food, feed, and seed. The amount of residue that remains in fumigated materials is measured and reported. Objective: The optimization and validation of a high-sensitivity headspace solid-phase microextraction gas chromatograph mass spectrometer (HS-SPME-GCMS) method for determination of eight fumigant residues-phosphine, methyl bromide, cyanogen, sulfuryl fluoride, ethylene oxide, propylene oxide, ethyl bromide and ethyl formate-in a range of food matrices were developed. The food matrices included grain, oilseed, dried fruit, and nut. Methods: The new method was used to determine residue levels from a fumigant complex in food matrices by monitoring the change of the absorption of spiked standards and desorption of fumigant from fumigated samples. Results: Based on the observation, the process of physical sorption and chemisorption was defined. The equilibrium time of the sample analysis was chosen at 5 hours. The LODs of the fumigants were in the range of 0.03 to 1.99 ng/g. Response to a range of diluted authentic standards gave significant linear regressions (r² > 0.9983), and the RSDs were ≤8.7% at the 3 ng/g level of aged spiking standard, except for sulfuryl fluoride, for which the LOD was 1.99 ng/g and the RSD value was 39.7% (6.64 ng/g). Conclusions: The performance of the HS-SPME-GCMS method was more sensitive than the use of a gas syringe, except for sulfuryl fluoride. Highlights: An innovative multifumigant residue detection method based on HS-SPME sampling technology with gas chromatograph-mass selective detection (GC-MSD) analysis was established, including phosphine, methyl bromide, cyanogen, sulfuryl fluoride, ethylene oxide, propylene oxide, ethyl bromide, and ethyl formate; The first report of a sorption study of the process of physical sorption and chemisorption of residual fumigant in fruit and food matrices by an HS-SPME-GCMS method; Consider the effect of time on fumigant behavior in dried fruit, grain, and nut; The LODs of the fumigants were in the range of 0.03 to 1.99 ng/g.

Cold Response of the Mediterranean Fruit Fly (Ceratitis capitata) on a Lab Diet.

Cold treatment at 0.0 °C with different exposure durations (0⁻12 days) was applied to the Mediterranean fruit fly Ceratitis capitata (Wiedemann) fed on a lab diet. The examined developmental stages were early eggs (<6 h), late eggs (>42 h), first instar, second instar and third instar larvae. Pupation, adult emergence and sex ratios of survived flies were investigated to study the C. capitata responses to this low temperature treatment. Our results showed that exposure time at low temperature has a clear effect on pupation and adult emergence. Based on pupation ratios, the first and third instar are the most cold tolerant stages, with LT99 = 7.3 for both of them. Cold tolerance at both stages are very close and no significant differences were detected. There were no significant differences on C. capitata sex ratios among different stages after treatment. This study improves our understanding of C. capitata responses to cold treatment, which may assist in the improvement of the current treatment strategies to control this destructive horticulture pest species.

Coffee Intake and Liver Enzyme Association in Korean Immigrants and Japanese: A Comprehensive Cross-Sectional Study.

OBJECTIVES: Significant inverse association between coffee intake and the levels of liver enzymes has been reported. We demonstrated higher prevalence of metabolic syndrome in Korean immigrants (KIs) than in indigenous Japanese (IJs). The aim of this study was to investigate whether the association between coffee intake and liver enzyme levels was different between the 2 ethnic groups. METHODS: This study is a cross-sectional study including a total of 966 subjects comprising KIs and IJs. The association between the quintiles of coffee intake and dichotomous values of liver enzymes was evaluated by logistic regression analysis in KIs, IJs, a high-risk group (current smokers or alcohol drinkers ≥45 g/day), and a low-risk group (non-smokers and alcohol drinkers <45 g/day). RESULTS: In KIs, a significant inverse association between coffee intake and serum aspartate aminotransferase (AST) levels was observed. In the IJs, a significant inverse association between coffee intake and serum alanine aminotransferase levels was observed. In the high-risk group, a significant inverse association between coffee intake and serum AST and gamma-glutamyltransferase levels was observed. CONCLUSION: No difference was observed between KIs and IJs regarding the association between coffee and liver enzymes. Coffee might inhibit hepatic damage by alcohol drinking and smoking.

Seroepidemiology of hepatitis a infection in northeastern china, Korea, and Japan.

OBJECTIVES: The epidemiological patterns of endemic hepatitis A virus (HAV) are unclear in northeastern Asia depending on the ethnicity of the country in question. The purpose of this study was to investigate the seroprevalence of HAV in northeastern China, South Korea, and Japan. METHODS: A total of 1,500 serum samples were collected from five groups of inhabitants (300 each) who were over 40 years of age (Korean Chinese, indigenous Chinese, South Korean, Korean living in Japan, and indigenous Japanese). The samples were screened for antibodies to HAV using an enzyme-linked immunosorbent assay. RESULTS: Positivity for HAV antibodies was 93.7% (95% confidence interval [CI]: 90.9-96.4) in Koreans living in northeastern China, 99.7% (95% CI: 99.0-100.3) in indigenous Chinese, 98.0% (95% CI: 96.4-99.6) in indigenous Koreans, 33.3% (95% CI: 28.0-38.7) in Koreans living in Japan, and 20.4% (95% CI: 15.8-25.0) in indigenous Japanese persons. The overall anti-HAV prevalence was not significantly different between northeastern China and South Korea, but it was different in Japan. CONCLUSIONS: These results indicate that differences in seroprevalence can be attributed to geological, environmental, and socioeconomic conditions rather than ethnicity.

Comparison of prevalence of metabolic syndrome between korean emigrants and host country residents in Japan and china-the korean emigrant study.

OBJECTIVES: This study aims to compare the prevalence of metabolic syndrome between Korean emigrants (KEs) and their host country residents in Japan and China. METHODS: The Korean Emigrant Study (KES) is a cohort study initiated in 2005 to elucidate the effect of genetic susceptibility and environmental change on hypertension, diabetes, and metabolic syndrome. Equal numbers of KEs and host country residents, aged 30 or over, were recruited from three regions; Kobe-Osaka in Japan (total number=965), Yanbian in China (n=1,019), and Changchun in China (n=949). RESULTS: The age-adjusted prevalences of metabolic syndrome among KEs in Kobe-Osaka were significantly higher than those among Japanese (in men 24.0% vs. 15.6%, p=0.04, in women 8.4% vs. 2.7%, p=0.01), while the age-adjusted prevalences among KEs in Changchun were similar to those among Chinese (in men 11.7% vs. 16.1%, p=0.37, in women 28.3% vs. 30.1%, p=0.91). The age-adjusted prevalences were generally higher in Yanbian than other regions, and KEs had higher prevalence than Chinese in men but not in women (in men 37.9% vs. 28.3%, p=0.03, women 46.0% vs. 50.6%, p=0.44). The components with significant ethnic differences in prevalence were high blood pressure and abdominal obesity in Japan, and triglyceride in China. The most influential component in diagnosing metabolic syndrome was abdominal obesity in men and triglyceride in women. CONCLUSION: The prevalence of metabolic syndrome was higher in KEs than in host country residents in Japan but not in China. Abdominal obesity and triglyceride are both discriminating and influential components in metabolic syndrome.

Double-filtration plasmapheresis plus IFN for HCV-1b patients with non-sustained virological response to previous combination therapy: early viral dynamics.

Double-filtration plasmapheresis (DFPP) was approved in Japan in April 2008 for the retreatment of chronic hepatitis C patients with genotype 1b and high viral loads, whose hepatitis C virus was not eradicated by earlier IFN therapy or by pegylated IFN plus ribavirin (PEG-IFN/RBV) combination therapy. In this study, we assessed the early viral dynamics of 9 patients with non-sustained virological response to the combination therapy. The overall viral dynamics of DFPP plus IFN treatment with or without RBV for 4 weeks showed a reduction of > or =1 log in the viral load in 22% (2 of 9 patients), 55.6% (5/9), 77.8% (7/9) and 77.8% (7/9) at 24 h, 1, 2 and 4 weeks after the start of treatment. By contrast, DFPP plus consecutive intravenous IFN-beta for 4 weeks reduced the viral load by > or = 1 log in 33% (2/6), 50% (3/6), 83.3% (5/6) and 83.3% (5/6) at 24 h, 1, 2 and 4 weeks. The viral load declined by > or = 2 log in 50% (3/6) at 4 weeks after the start of treatment. DFPP plus consecutive intravenous IFN-beta for 4 weeks is a promising treatment for non-sustained virological response patients.

Outcome and early viral dynamics with viral mutation in PEG-IFN/RBV therapy for chronic hepatitis in patients with high viral loads of serum HCV RNA genotype 1b.

We investigated whether sustained virological response (SVR) and non-SVR by chronic hepatitis C patients to pegylated interferon plus ribavirin (PEG-IFN/RBV) combination therapy are distinguishable by viral factors such as the IFN/RBV resistance-determining region (IRRDR) and by on-treatment factors through new indices such as the rebound index (RI). The first RI (RI-1st; the viral load at week 1 divided by the viral load at 24 h) and the second RI (RI-2nd; the viral load at week 2 divided by the viral load at 24 h) were calculated. The subject patients were divided into 3 groups based on RI-1st and RI-2nd: an RI-A group (RI-1st < or = 1.0), an RI-B group (RI-1st >1.0 and RI-2nd <0.7) and an RI-C group (RI-1st >1.0 and RI-2nd > or = 0.7). The SVR rate was 71.4% (10/14) in the RI-A group, 46.2% (6/13) in the RI-B group and 20.0% (3/15) in the RI-C group (p = 0.005 between the RI-A group and the RI-C group). In IRRDR > or = 6 and IRRDR < or = 5 the SVR rate was 81.3% (13/16) and 23.1% (6/26) (p = 0.0002), respectively. By combining RI and IRRDR as a predicting factor, the SVR rate was 87.5% (7/8) in the RI-A group (> or = 6 mutations in the IRRDR) and 7.7% (1/13) in the RI-C group (< or = 5 IRRDR mutations) (p = 0.0003).

Well-differentiated hepatocellular carcinoma smaller than 15 mm in diameter totally eradicated with percutaneous ethanol injection instead of radiofrequency ablation.

We describe three cases of well-differentiated hepatocellular carcinoma (HCC) smaller than 15 mm in diameter completely eradicated with percutaneous ethanol injection (PEI) instead of using radiofrequency ablation (RFA). Ultrasound (US) examination revealed one nodule each in segment 2 (hypoechoic, near bile ducts, 10 mm), in segment 5 (hyperechoic, near the gall bladder, 15 mm), and in segment 7 (hypoechoic, near the diaphragm, 15 mm). Although imaging studies revealed isovascular (case 1) and hypervascular (cases 2 and 3) nodules, histological analysis of US-guided biopsy tissue revealed well-differentiated HCC. In consideration of the location of the nodules, PEI, instead of RFA, was administered and the nodules were rendered necrotic. Although RFA is superior to PEI in the treatment of small HCCs from the viewpoint of treatment response and long survival, PEI is strongly recommended for HCCs located near bile ducts, the gall bladder, and the diaphragm, especially when the nodules are smaller than 15 mm in diameter.

Epidemiology of hepatitis E in Northeastern China, South Korea and Japan.

OBJECTIVES: The seroprevalence of hepatitis E virus (HEV) infection in Northeastern Asia is unknown. This study was conducted to gain insight into the epidemiology of HEV that has been obscure in Northeastern China, South Korea and Japan. METHODS: A total of 1500 samples of serum were collected (300 each) from 5 groups of inhabitants over 40 years of age (Korean Chinese, indigenous Chinese, South Koreans, Koreans living in Japan, and indigenous Japanese) and screened for antibodies to HEV by the antigen-antibody-antigen sandwich Enzyme Linked Immunosorbent Assay system. RESULTS: The positivity for HEV antibodies was 50.7% (95%CI: 45.0-56.3) in Korean Chinese, 47.7% (95%CI: 42.1-53.3) in indigenous Chinese, 34% (95%CI: 28.9-39.5) in South Koreans, 14.3% (95%CI: 10.8-18.8) in Koreans living in Japan, and 6.0% (95%CI: 3.8-9.3) in indigenous Japanese. CONCLUSIONS: This result emphasizes that HEV is endemic in Northeastern Asia and tends to accumulate in developing countries. Further studies are needed to elucidate the genotype of HEV circulating in these areas and its transmission route-water-borne outbreaks, smaller outbreaks or sporadic forms attributed to zoonosis-with reference to past epidemics, food culture, and sanitary conditions.

Pegylated interferon plus ribavirin combination therapy for chronic hepatitis C with high viral load of serum hepatitis C virus RNA, genotype 1b, discontinued on attaining sustained virological response at week 16 after onset of acute pancreatitis.

Recent clinical trials have shown that pegylated interferon-alpha (PEG-IFN-alpha) in combination with ribavirin (RBV) improves the rate of sustained virological response (SVR), with over 50% of patients demonstrating a positive response to treatment. However, no SVR has been reported when PEG-IFN/RBV combination therapy is discontinued by week 16, especially in cases of chronic hepatitis with a high viral load of serum hepatitis C virus (HCV) RNA, genotype 1b. Here, we describe SVR in a 67-year-old woman whose PEG-IFN/RBV combination therapy for chronic hepatitis C with a high viral load of serum HCV RNA, genotype 1b, was discontinued after 16 weeks because of the onset of PEG-IFN plus RBV-induced acute pancreatitis. Among viral factors, substitution of amino acid 70 (Arg) and 91 (Leu) in the core region and HCV RNA negativity were observed after 8 weeks. Host factors including low body weight, no alcohol consumption, no coinfection with hepatitis B virus, slight fibrosis, and viral factors including early viral clearance, double wild type in the core region, may have contributed to the SVR irrespective of the discontinuation of the combination therapy at week 16. Moreover, PEG-IFN plus RBV-induced acute pancreatitis might have been related to the SVR.

Acute Pancreatitis Associated with Pegylated Interferon and Ribavirin Treatment of Chronic Hepatitis C, Genotype 1b with High Viral Load.

Acute pancreatitis, an uncommon side effect of pegylated interferon α (PEG-IFN α) and ribavirin (RBV) combination therapy, has rarely been reported in the English language literature. Here, acute pancreatitis associated with PEG-IFN plus RBV treatment is described in three patients with chronic hepatitis C, genotype 1b with high serum hepatitis C virus RNA levels. The patients had been started on weekly subcutaneous injections of PEG-IFN α (60, 80, and 90 µg) plus a daily oral dose of RBV (600 mg). The therapy was discontinued, however, because of the onset of acute pancreatitis (after 15 weeks, 48 weeks, and 3 weeks respectively). The drug-induced pancreatitis was diagnosed on the basis of elevated levels of amylase and lipase and the absence of other identifiable causes. High tumor necrosis factor-α was found in one patient and high interleukin-6 in the other two. The immune system stimulated by PEG-IFN and RBV combination therapy might have caused the acute pancreatitis. Further study is needed to clarify the mechanism of the onset of drug-induced pancreatitis by PEG-IFN and RBV combination therapy.

Focal nodular hyperplasia-like lesion with venous washout in alcoholic liver cirrhosis.

A case of 22 mm hypervascular nodule in segment two of the liver but without hepatitis B or C virus infection in a 32-year-old Japanese woman with a history of alcohol abuse is presented. Imaging studies such as contrast-enhanced ultrasound, computed tomography and magnetic resonance imaging showed hypervascularity in the early phase and venous washout in the late phase. Histologically, stellate scar-like fibrous septa, pericellular fibrosis, fatty change, neutrophilic infiltration, slight increase of cell density, and diffuse capillarization of the sinusoids together with small unpaired arteries were observed. The nodule was diagnosed as focal nodular hyperplasia-like lesion in alcoholic liver cirrhosis.

Usefulness of a new immunoradiometric assay of HCV core antigen to predict virological response during PEG-IFN/RBV combination therapy for chronic hepatitis with high viral load of serum HCV RNA genotype 1b.

We investigated the clinical usefulness of a new immunoradiometric (IRM) assay of hepatitis C virus (HCV) core antigen in predicting virological response during pegylated interferon plus ribavirin (PEG-IFN/RBV) combination therapy for chronic hepatitis with high viral loads of serum HCV RNA genotype 1b. Thirty-nine patients received a regimen of PEG-IFNalpha-2b (1.5 microg/kg/week s.c.) in combination with RBV (600-1,000 mg/day). Of the 39 patients, 18 (46.2%) achieved sustained virological response (SVR), 11 (28.2%) attained partial response (PR) and 10 (25.6%) showed no response (NR). Four weeks after the start of therapy, 1- and 2-log reductions in the amount of HCV core antigen were observed in 20 (2/10) and 0% (0/10) showing NR, 91 (10/11) and 63.6% (7/11) with PRs, and 88.9 (16/18) and 55.6% (10/18) of patients with SVR, respectively. The 1- and 2-log reductions 4 weeks after the start of therapy were not a defining condition for PR and SVR. The amount of HCV core antigen was significantly different between SVR and PR patients on days 1 and 7, and between patients with NR and SVR at all points of time. In conclusion, this new IRM assay is useful in predicting virological response during PEG-IFN/RBV therapy.

Long-term outcome of percutaneous ethanol injection therapy for minimum-sized hepatocellular carcinoma.

AIM: To evaluate long-term follow-up of minimum-sized hepatocellular carcinoma (HCC) treated with percutaneous ethanol injection (PEI). METHODS: PEI was applied to 42 lesions in 31 patients (23 male and eight female) with HCC < 15 mm in diameter, over the past 15 years. RESULTS: Overall survival rate was 74.1% at 3 years, 49.9% at 5 years, 27.2% at 7 years and 14.5% at 10 years. These results are superior to, or at least the same as those for hepatic resection and radiofrequency ablation. Survival was affected only by liver function, but not by sex, age, etiology of Hepatitis B virus or Hepatitis C virus, alpha-fetoprotein levels, arterial and portal blood flow, histological characteristics, and tumor multiplicity or size. Patients in Child-Pugh class A and B had 5-, 7- and 10-years survival rates of 76.0%, 42.2% and 15.8%, and 17.1%, 8.6% and 0%, respectively (P = 0.025). CONCLUSION: Treatment with PEI is best indicated for patients with HCC < 15 mm in Child-Pugh class A.

Case of adult genotype C HBV carrier after acute hepatitis B, losing HBsAg and acquiring HBsAb after IFN and lamivudine treatment.

Here we report the case of a 48-year-old man, carrier of genotype C HBV for longer than 6 months after contracting sexually transmitted acute hepatitis B, who eventually lost HBsAg and acquired HBsAb by IFN/lamivudine therapy. The patient had been negative for HBsAg in 2001, but, during his stay in China from January to July in 2003, he developed acute hepatitis B after having an extra-marital sexual contact there. HBsAg was still positive and a liver biopsy indicated chronic hepatitis when he was admitted to our hospital in December 2003 for detailed examination of liver dysfunction. HBV DNA in his serum, revealed to segregate to genotype C by sequencing on admission, decreased to undetectable levels at the end of a 3-month IFN therapy, and remained undetectable during and after the successive 6-month lamivudine therapy. HBeAg seroconverted to HBeAb during the therapy, and HBsAb appeared after the therapy. To our knowledge, this is the first case of genotype C chronic hepatitis B occurring after acute hepatitis.

Well-differentiated HCC manifesting hyperattenuation on CT during arterial portography.

A rare case of well-differentiated minute hepatocellular carcinoma (HCC) with hepatitis C virus-related cirrhosis, with unusual radiologic features, is presented. A 10-mm hypoechoic nodule disclosed by ultrasound in segment six showed hypoattenuation on computed tomography hepatic arteriography and hyperattenuation on computed tomography during arterial portography, indicating that the portal vein may have been the dominant vascularity of the nodule. Contrast-enhanced ultrasound revealed hypovascularity in the early arterial phase, isovascularity in the late vascular phase, and the same perfusion as that surrounding the liver parenchyma in the post-vascular phase, with the same pattern observed on the two imaging techniques. These findings were considered not compatible with those of well-differentiated HCC. Ultrasound-guided biopsy showed histological features of well-differentiated HCC with over two-fold the cellularity of the non-tumorous area with a high nuclear/cytoplasmic ratio, increased cytoplasmic eosinophilia, slight atypia and fatty change with an irregular thin trabecular pattern. Further studies may provide insights into the correlation between tumor neovascularity in multistep hepatocarcinogenesis and dual hemodynamics, including the artery and the portal vein.

Primary sclerosing cholangitis and hepatitis C virus infection.

Two cases of primary sclerosing cholangitis with hepatic C virus infection in a 62-year-old man and a 60-year-old woman are presented. The infection in the man was eradicated with interferon therapy in 1992. Seven years thereafter, endoscopic retrograde cholangiography revealed a diffuse 2.5-cm-long stenotic lesion in the common bile duct which was consequently resected. Histological examination of the resected specimen revealed proliferation of epithelial cells, plasma cell infiltration, and fibrosis in the submucosal layer of the common bile duct. The human leukocyte antigen DR loci were 2 and 9. In the woman, a 6-month course of interferon therapy in 1992 failed to eradicate the infection. Cholangiography in 1999 revealed multiple narrowings and dilatations of intra- and extrahepatic bile ducts. Ultrasound guided biopsy of the liver in 1992 had revealed onionskin lesions around the bile duct epithelium in the portal tract. The human leukocyte antigen DR locus was 2. From these findings, the 2 cases were diagnosed as primary sclerosing cholangitis. Further studies may provide insights into the relation between the pathogenesis of the disease and the infection.

Effects of physical training on heat loss responses of young women to passive heating in relation to menstrual cycle.

To examine the effects of physical training on cutaneous vasodilation and sweating responses of young women in the follicular and luteal phase, 11 physically trained (T group) and 13 untrained (U group) women were passively heated by lower-leg immersion into hot water of 42 degrees C (ambient temperature of 30 degrees C and 45%RH) for 60 min in their mid-follicular and mid-luteal phases of the menstrual cycle. Female hormones increased significantly from the mid-follicular to the mid-luteal phase in T and U groups, but the degree of increase was significantly lower in T group. Mean body temperature (T(B)) thresholds for cutaneous vasodilation and sweating responses were significantly lower in T group than in U group, in both the menstrual phases, and the differences between the groups were greatest during the mid-luteal phase. The slope of the relationship between frequency of sweat expulsion (F(sw)) and (T(B)), and between local sweating rate and F(sw) was significantly greater in T group, although the slope of the relationship between cutaneous blood flow and (T(B)) did not differ between the groups, regardless of body site or menstrual phase. These results suggest that regular physical activity enhanced sweating and cutaneous vasodilation in young women. The enhancement of sweating was due to both central and peripheral mechanisms, and the enhancement of cutaneous vasodilation was possibly due to a central mechanism. Enhancement of heat loss responses via central mechanisms was greater during the mid-luteal phase than in the mid-follicular phase because the elevation of female reproductive hormone levels during the mid-luteal phase was relatively low in T group.

Prediction of efficacy of interferon treatment of chronic hepatitis C and occurrence of HCC after interferon treatment by a new classification.

One hundred and twenty-one chronic hepatitis C patients were administered interferon (IFN) and divided into two groups: 31 complete responders (CR) with prolonged HCV-RNA negative 1 year after treatment and 90 non-complete responders (non-CR), including partial responders (PR) with transiently negative HCV-RNA and non-responders (NR) with continuously positive HCV-RNA. Liver biopsy specimens were classified into four grades and stages according to the degree of severity and the extent of fibrosis, respectively. No correlation was observed between the rate of IFN efficacy and grading. By staging, however, a difference in the efficacy of IFN was observed between F1 or F2 and F3 (0.05 < p < 0.1 and 0.01 < p < 0.025, respectively). Of the CR, 0% (0/5) were at F0 and 27.9% (24/86) at F1, 42.9% (6/14) at F2, and 6.3% (1/16) at F3. Another group of 118 chronic hepatitis patients (31 CR, 41 PR and 46 NR) followed up for over 2 years after IFN treatment were analyzed. By staging, 7 cases were at F0, 76 at F1, 18 at F2, and 17 at F3. HCC occurred in 1 of the 31 CR cases (representing an annual incidence rate of 1.21%), in 4 of the 41 PR cases (4.08%), and in 4 of the 46 NR cases (3.55%). HCC did not occur in any of the 7 cases at F0 (representing an annual incidence rate of 0%); it occurred in 2 of the 76 cases at F1 (1.01%), in 1 of the 18 cases at F2 (2.28%), and in 6 of the 17 cases at F3 (16.57%). These results suggest that the new classification would be conducive to roughly predicting the efficacy of IFN treatment and the occurrence of HCC after IFN treatment.