Publisher's Note: Search for Majorana Neutrinos Near the Inverted Mass Hierarchy Region with KamLAND-Zen [Phys. Rev. Lett. 117, 082503 (2016)].

This corrects the article DOI: 10.1103/PhysRevLett.117.082503.

Search for Majorana Neutrinos Near the Inverted Mass Hierarchy Region with KamLAND-Zen.

We present an improved search for neutrinoless double-beta (0νßß) decay of ^{136}Xe in the KamLAND-Zen experiment. Owing to purification of the xenon-loaded liquid scintillator, we achieved a significant reduction of the ^{110m}Ag contaminant identified in previous searches. Combining the results from the first and second phase, we obtain a lower limit for the 0νßß decay half-life of T_{1/2}^{0ν}>1.07×10^{26} yr at 90% C.L., an almost sixfold improvement over previous limits. Using commonly adopted nuclear matrix element calculations, the corresponding upper limits on the effective Majorana neutrino mass are in the range 61-165 meV. For the most optimistic nuclear matrix elements, this limit reaches the bottom of the quasidegenerate neutrino mass region.

Virological response and safety of 24-week telaprevir alone in Japanese patients infected with hepatitis C virus subtype 1b.

Hepatitis C virus (HCV) subtype 1b, which infects approximately 70% of Japanese carriers, is likely to be more eradicable by a telaprevir regimen than subtype 1a because of the higher genetic barrier of Val(36) and Arg(155) substitutions. The aims of this exploratory study were to evaluate the virological response and safety of 24-week oral administration of telaprevir alone in chronic HCV subtype 1b infection. Fifteen treatment-naïve patients were treated with telaprevir 750 mg every 8 h for 24 weeks. All patients were Japanese whose median age was 58.0 years (range: 45-68), and six patients (40%) were men. Median baseline HCV RNA level was 6.80 log(10) IU/mL (range: 3.55-7.10). The HCV RNA levels decreased to undetectable in five patients (33%) within 8 weeks. Three patients (20%) with negative HCV RNA by Week 4 achieved end of treatment response. One patient (7%) who achieved sustained virological response had a low baseline viraemia of 3.55 log(10) IU/mL. Most of the adverse events including anaemia and skin disorders were mild to moderate. Developed variants were T54A and A156V/T/F/Y with or without secondary substitutions rather than V36M ± R155K. Telaprevir alone for 24 weeks in Japanese patients with HCV subtype 1b resulted in an sustained viral response rate of 7% (1/15) and was well tolerated for 24 weeks. These results will support the implementation of further studies on oral combination of telaprevir with other direct-acting antiviral agents in patients infected with HCV subtype 1b.

Cutaneous drainage lymphatic map with interstitial multidetector-row computed tomographic lymphography using iopamidol: preliminary results.

We performed preliminary tests of the feasibility of multi-detector computed tomographic lymphography (MDCT-LG) with interstitial injection of iopamidol for mapping cutaneous lymphatic drainage pathways. MDCT-LG images were obtained following cutaneous injection of a total of 1ml iopamidol bilaterally into hind legs of 10 dogs. The locations of the first draining lymph nodes were marked on the skin under MDCT-LG guidance. Five dogs served for postmortem examination of lymphatic anatomy, and the remaining 5 underwent MDCT-LG after ligation of the afferent lymphatic vessels of the first draining popliteal nodes. Clinically, MDCT-LG was attempted in 6 patients with cutaneous malignant melanoma and compared with Tc-99m-human serum albumin lymphoscintigraphy. MDCT-LG clearly visualized the first draining lymph nodes and their afferent lymphatic vessels draining from the contrast injection sites with detailed underlying anatomy in all dogs. At surgery, all these first draining nodes could be found at predicted locations under MDCT-LG guidance. MDCT-LG showed rerouting of lymphatic vessels after ligation of the afferent lymph vessels of the popliteal nodes in the second 5 dogs. Clinically, MDCT-LG also allowed accurate mapping and biopsy of the first draining nodes from primary tumors at predicted locations, with minimal skin incision. Lymphoscintigraphy failed to identify these nodes due to overlapping radioactivity of clustered nodes or transport of the radiotracer to subsequent distant nodes in 4 patients. Although a more extensive study is warranted for further validation, preoperative interstitial MDCT-LG appears to have the potential feasibility for accurate sentinel lymph node mapping and biopsy in patients with cutaneous melanoma.

Development of a new enzyme-linked immunosorbent assay for the detection of anti-M2 in primary biliary cirrhosis.

We developed a new enzyme-linked immunosorbent assay (ELISA) for the detection of antimitochondrial antibody (AMA)-M2 in sera from patients with primary biliary cirrhosis (PBC), using 2-oxo-acid dehydrogenase complex (2-OADC) purified from porcine myocardium as the antigen source. The immunoreactivity was tested in a total of 354 sera, including 63 sera from patients with PBC by our ELISA. In the sera, indirect immunofluorescence for AMA, former ELISA for anti-pyruvate dehydrogenase complex (PDC) and immunoblot assay were performed, respectively. Of the 63 sera from patients with PBC, 51 sera (81.0%) were positive for anti-M2 in the new ELISA. Thirty-eight of the 63 sera (60.3%) were positive for anti-PDC in the former ELISA; the difference was significant between them (P=0.011). None of the 291 control sera from healthy volunteers showed reactivity against 2-OADC in the new ELISA. Moreover, in comparison with the results of immunoblot analysis, sensitivity and specificity in our ELISA to the sera from patients with PBC were 100 and 92.3%, respectively. Our results indicate that the new ELISA for anti-M2 using 2-OADC is simple, rapid and sensitive enough for the detection of AMA specific to PBC.

Detection of hypermethylation of the p16(INK4A) gene promoter in chronic hepatitis and cirrhosis associated with hepatitis B or C virus.

BACKGROUND/AIM: Inactivation of the p16(INK4A) (p16) tumour suppressor gene by promoter region hypermethylation has been demonstrated not only in many types of tumours, including hepatocellular carcinoma (HCC), but also in early preneoplastic lesions in the lung, colon, oesophagus, and pancreas. The aim of this study was to examine the methylation status of the p16 promoter in pre- and/or non-neoplastic liver diseases. PATIENTS/SUBJECTS/METHODS: The methylation status of p16 was evaluated in 22 HCC, 17 cirrhosis, 17 chronic hepatitis, nine primary biliary cirrhosis (PBC), eight autoimmune hepatitis, seven drug induced liver disease, six fatty liver, and three normal liver tissues using methylation specific polymerase chain reaction (MSP). p16 protein expression was also examined by immunohistochemical staining. RESULTS: Methylation of the p16 promoter was detected in HCC (72.7%, 16/22) and also in cirrhosis (29.4%, 5/17) and chronic hepatitis (23.5%, 4/17), all of which were positive for hepatitis B or C virus infections. Methylation was not detected in any of the other samples. All methylation positive HCC, cirrhosis, and chronic hepatitis samples showed loss of p16 expression, and a significant correlation was found between methylation and loss of expression. Analysis of serial samples from individual patients with methylation positive HCC revealed that loss of p16 expression with promoter methylation occurred in 18 of 20 patients at the stage of chronic hepatitis without clinically detectable carcinoma. CONCLUSIONS: Our results suggest that methylation of the p16 promoter and the resulting loss of p16 protein expression are early events in a subset of hepatocarcinogenesis and that their detection is useful in the follow up of patients with a high risk of developing HCC, such as those with hepatitis B or C viral infections.

Frequent hypermethylation of CpG islands and loss of expression of the 14-3-3 sigma gene in human hepatocellular carcinoma.

The 14-3-3 sigma gene has been implicated in G2/M cell cycle arrest by p53. Frequent inactivation of the 14-3-3 sigma gene by hypermethylation of CpG islands has recently been reported in human breast carcinoma. The aim of this study was to examine the methylation status of CpG islands of the 14-3-3 sigma gene in hepatocellular carcinoma (HCC). The methylation status of the 14-3-3 sigma gene was evaluated in four normal liver tissues and 19 paired specimens of carcinoma and adjacent non-tumorous liver tissues using bisulfite-single strand conformation polymorphism (bisulfite-SSCP), a combination of sodium bisulfite modification and fluorescence-based polymerase chain reaction (PCR)-SSCP. The 14-3-3 sigma protein expression was examined by immunohistochemical staining. Hypermethylation of CpG islands of the 14-3-3 sigma gene was detected in 89% (17/19) of the HCC tissues but not in any of the four normal liver tissues. All of the 14 methylation-positive HCC samples analysed by immunohistochemistry showed loss of 14-3-3 sigma expression, while both of the methylation-negative HCC samples retained the expression, and a significant correlation was found between methylation and loss of expression. Lower levels of methylation were detected in adjacent non-tumorous liver tissues (6/16 in cirrhotic tissues and 1/3 in chronic hepatitis tissues), but the 14-3-3 sigma expression was retained in all of these tissues. In a methylation-positive HCC cell line, HLE, 5-aza-2'-deoxycytidine (5-aza-dC)-induced demethylation of CpG islands led to reactivation of gene expression, indicating that hypermethylation plays a causal role in inactivation of the 14-3-3 sigma gene in HCC. Hypermethylation and the resulting loss of expression of the 14-3-3 sigma gene corresponds to one of the most common abnormalities reported to date in HCC, suggesting their crucial role in the development and/or progression of HCC.

Gastric varices with splenic vein occlusion treated by splenic arterial embolization.

A 53-year-old man was admitted to our hospital in August 1997 with enlarged gastric varices. Computed tomography (CT) showed splenic vein occlusion, gastric varices, and extra-gastric wall collateral veins. Color flow images of gastric varices were clearly visualized, and the velocity in the gastric varices was 19.6 cm/s via endoscopic color Doppler ultrasonography (ECDUS). The patient was diagnosed with gastric varices according to angiographic findings of splenic vein occlusion, and splenic arterial embolization was performed. Two weeks after the splenic arterial embolization, CT showed peripheral areas of low attenuation in the spleen, due to splenic infarction, with 70% of the spleen volume showing low attenuation. Eight months after the splenic arterial embolization, ECDUS revealed a decrease in gastric variceal color flow images, with the velocity in the gastric varices being 10.3 cm/s.

Infrequent widespread microsatellite instability in hepatocellular carcinomas.

Widespread or high-frequency microsatellite instability (MSI) due to the defective DNA mismatch repair (MMR) occurs in the majority of hereditary non-polyposis colorectal cancer and a subset of sporadic malignant tumors. The incidence of MSI and underlying DNA MMR defects have been well characterized in gastrointestinal carcinogenesis, but not in hepatocarcinogenesis. To address the issue, we analyzed 55 Japanese hepatocellular carcinomas using several indicators of DNA MMR defects, such as microsatellite analysis, loss of heterozygosity (LOH) and mutation analysis of MMR genes, methylation of hMLH1 promoter, and frameshift mutations of mononucleotide repeat sequences within possible target genes. Mutation of beta2-microglobulin gene, which is presumably involved in MSI-positive tumor cell escape from immune surveillance was also examined. Some of these analyses were also carried out in 9 human liver cancer cell lines. None of the 3 quasi-monomorphic mononucleotide markers sensitive for MSI, BAT26, BAT25, and BAT34C4 presented shortened unstable alleles in any of the carcinoma, cirrhosis, chronic hepatitis tissues, or cell lines. LOH at MMR genes was infrequent (4.4 approximately 7.1%), and no mutations were detected. Neither hMLH1 hypermethylation nor frameshift mutation in the target genes was detected. No mutations were found in beta2-microglobulin. Widespread MSI due to the defective DNA MMR appears to play little if any part in Japanese hepatocarcinogenesis.

Early mutation of precore (A1896) region prior to core promoter region mutation leads to decrease of HBV replication and remission of hepatic inflammation.

To evaluate the relationship between mutations and clinical courses, we investigated precore (preC) and core promoter (CP) mutations and serum HBV DNA levels in HBe-antibody-positive HBV carriers. Fifty-six asymptomatic carriers (ASC), 29 patients with chronic hepatitis who showed normal ALT levels for more than two years (CH-ASC), 31 patients with chronic hepatitis (CH), and 32 patients with hepatocellular carcinoma (HCC) were studied. Almost all patients (99.2%) had mutations in either CP or preC. Mutation only in preC (A1896) was present in 52.2% with ASC, 25.0% with CH-ASC, 16.1% with CH, and 8.0% with HCC, and was significantly higher in ASC (P < 0.01). The patients with only preC mutation showed low HBV DNA levels in each clinical stage. The mutation of preC (A1896) prior to the mutation of CP might control the replication of HBV, which leads to the remission of hepatitis.

Evaluation of magnetic resonance angiography in detection of gastric varices.

We evaluated the detection of gastric varices, inflowing blood vessels to gastric varices, and outflowing blood vessels from gastric varices via magnetic resonance (MR) angiography in 31 patients with gastric varices. Twenty-four patients had F2 type varices and 7 had F3 type, classified according to the Japanese Research Society for Portal Hypertension. Seventeen patients had cardiofornical varices, and 14 had fundal varices. All patients were examined with an MR system operating at 1.5T. MR angiography was performed using the two-dimensional time-of-flight method. With MR angiography, the imaging of gastric varices was clearly delineated in 28 of the 31 patients (90.3%). From the images of MR angiography, flow direction itself cannot be determined. The outflowing blood vessels of gastric varices were reported to be the gastro-renal shunt and the subphrenic vein, and angiographic findings have shown the inflowing blood vessels to be the left gastric vein (LGV), the short gastric vein (SGV), and the posterior gastric vein (PGV). In 25 of the 31 patients (80.7%), the outflowing blood vessels from gastric varices were detected (gastro-renal shunt in 24; subphrenic vein in 1). MR angiography provided clear images of the inflowing blood vessels to gastric varices in 18 of the 31 patients (58.1%). These inflowing vessels were categorized as SGV in 7 patients, LGV in 5, LGV and SGV in 4, and LGV and PGV in 2. We suggest that MR angiography be used as a routine method for detecting and diagnosing collateral veins in patients with gastric varices.

Early loss of serum hepatitis C virus RNA can predict a sustained response to interferon therapy in patients with chronic hepatitis C.

OBJECTIVES: We evaluated whether the loss of serum hepatitis C virus (HCV) RNA early in interferon (IFN) therapy would indicate a subsequent response to IFN therapy. METHODS: One hundred fourteen patients with chronic hepatitis C were treated with IFN-alpha for 24 weeks. All patients were positive for anti-HCV antibodies and serum HCV RNA. Serum HCV RNA was measured by highly sensitive and specific RT-PCR (modified Amplicor HCV). RESULTS: Of 114 patients who were treated with IFN-alpha for 24 weeks, 22 of 29 patients (75.9%) who lost HCV RNA at the first week of treatment, 5 of 14 patients (35.7%) who lost HCV RNA at the second week, and 2 of 16 patients (12.5%) who lost HCV RNA at fourth week were judged as sustained responder (SR). The SR rate was significantly higher in patients who lost HCV RNA at the first week of therapy (p < 0.05). On the contrary, none of 55 patients who retained HCV RNA during the first 4 weeks of IFN therapy were judged as SR. Concerning the patients who lost HCV RNA at the first week of therapy, there were no significant differences in the SR rate in either HCV genotype (1b, 2a, and 2b). CONCLUSIONS: Our study confirms that the early response to IFN (loss of HCV RNA at the end of the first week of IFN therapy) can be a predictor of the subsequent sustained response to IFN therapy. Additionally, positivity of HCV RNA at the fourth week of IFN therapy can be a predictor of the subsequent nonsustained response to IFN therapy.

Nucleotide sequences of the hepatitis C virus core region in patients without anti-core antibody.

Second-generation assays for detection of hepatitis C virus (HCV) infection that include reactivity of antibodies to core, NS3, NS4 are used because of their high sensitivity. Among these antibodies, anti-core antibody seems to be the most sensitive. However, there are some patients without anti-core antibodies, although HCV RNA is detectable by reverse transcription-polymerase chain reaction and branched DNA assay. The mechanism for the absence of anti-core antibody on its own is unclear. We therefore determined the nucleotide and deduced amino acid sequences of the core region obtained from two anti-core antibody-negative patients with HCV RNA (genotype 1b) and compared them with those of four anti-core antibody-positive patients and a previously reported sequence. Amino acids spanning 1-47, which seemed to exist in major B cell epitopes, were found to be completely conserved among these patients. Furthermore, the predictive binding motif to HLA DR4 (a.a 81-90) was completely conserved in both of the anti-core antibody-negative patients. There were various mutations in the residual amino acids spanning 49-108, but specific mutations could not be found in anti-core antibody-negative patients. These data indicate that the absence of anti-core antibody in two patients is not due to the presence of some formerly unknown viral variants, but due to a possible defect in the host's immune system.

Significance of 2',5'-oligoadenylate synthetase activity and HCV genotype in IFN therapy for type C chronic hepatitis.

Although the mechanisms of elimination of HCV by IFN have not been fully elucidated, the 2-5A system was reported to be one of the mechanisms of the anti-viral effect of IFN. Therefore, the relationship between HCV genotype, induction of 2-5AS and clinical effect was investigated. As for the anti-viral effect during IFN therapy, in type III or IV, most patients lost HCV-RNA regardless of serum 2-5AS induction even in high HCV-RNA concentration cases. In contrast, in type II, the negativity rate of HCV-RNA became high along with an increase of serum 2-5AS activity, but in patients with high HCV-RNA concentration, HCV-RNA was persistently positive. As for the long term clinical effect of IFN therapy judged 6 months after completion of IFN therapy, HCV genotypes were closely related to the effect; that is, the patients with type III or type IV HCV genotype showed a higher complete response rate compared with the patients with type II HCV genotype. However, the relationship between the long term clinical effects and induction of serum 2-5AS during IFN therapy was obscure. These results suggest that induction of 2-5AS is closely related to the anti-viral effect during IFN administration, but the viral factors appeared to be related to long term clinical effects after cessation of IFN therapy.

[Classification of hepatitis C virus into two major groups in serologic activity: correlation with response to interferon].

Genotypes of HCV have been reported to be one of the predictors of response to IFN therapy in patients with chronic hepatitis C. HCV genotypes were determined enzyme-linked immunoblot assay based on group I and II specific recombinant peptides of the NS-4 region (amino acid No. 1676-1670). We examined the correlation between HCV groups and response to IFN in patients with chronic hepatitis C. Among the 84 patients with chronic hepatitis C who underwent IFN treatment, 16 of 51 (31.4%) patients with group I and 12 of 23 (52.2%) patients with group II showed complete sustained response. These results suggest that HCV group assay may improve the ability to predict IFN treatment outcomes.

[Interstitial pneumonia induced by interferon therapy in type C hepatitis].

Interstitial pneumonia is known as one of the serious side effect of drugs. Recently, many investigators have reported that interstitial pneumonia (IP) which occurred during interferon (IFN) therapy for type C chronic hepatitis, and its appearance rate is considered to be more than 0.2% of patients who receive IFN. Though the mechanisms of IP during IFN therapy remains to be elucidated, one of the possible explanations is that excessive focal immune response in the lung derived from IFN might leads to the inflammation. For safe treatment, we must recognize that IFN induces IP during IFN therapy and give attention to its occurrence.

[Evaluation of the blood flow in tumors before and after irradiation with color Doppler flow imaging].

The blood flow in tumors of three patients with lung cancer, cancer in oral cavity and metastatic lymphnode was respectively evaluated before and after irradiation with color Doppler flow imaging. The arterial pulsating flow in lung cancer disappeared after 30 Gy irradiation and that in cancer of oral cavity disappeared after 15 Gy irradiation with intraarterial infusion chemotherapy. But the arterial pulsating flow in metastatic lymphnode was detected during irradiation and disappeared after 40 days post- irradiation. Therefore color Doppler flow imaging seemed to be useful for evaluation of the blood flow in tumors before and after irradiation.