Frequency of Common Copy-Number Variations at 15q11.2q13 in Sperm of Healthy Men.

The genomic region at 15q11.2q13 represents a hotspot for copy-number variations (CNVs) due to nonallelic homologous recombination. Previous studies have suggested that the development of 15q11.2q13 deletions in sperm may be affected by seasonal factors because patients with Prader-Willi syndrome resulting from 15q11.2q13 deletions on paternally derived chromosomes showed autumn-dominant birth seasonality. The present study aimed to determine the frequency of 15q11.2q13 CNVs in sperm of healthy men and clarify the effects of various environmental factors, i.e., age, smoking status, alcohol intake, and season, on the frequency. Thirty volunteers were asked to provide semen samples and clinical information once in each season of a year. The rates of 15q11.2q13 CNVs were examined using 2-color FISH. The results were statistically analyzed using a generalized estimating equation with negative binomial distribution and a log link function. Consequently, informative data were obtained from 83 samples of 26 individuals. The rates of deletions and duplications ranged from 0.04 to 0.48% and from 0.08 to 0.30%, respectively. The rates were not correlated with the age, smoking status, or alcohol intake. Sperm produced in winter showed 1.2 to 1.4-fold high rates for both deletions and duplications as compared with sperm produced in the other seasons; however, there was no significant difference. These results demonstrate high and variable CNV rates at 15q11.2q13 in sperm of healthy men. These CNVs appear to occur independent of the age, smoking status, or alcohol intake, while the effect of season remains inconclusive. Our results merit further validation.

[Effectiveness of indocyanine green fluorescence in endoscopic marking in gastric cancer surgery:a feasibility study for replacing India ink].

When injected, indocyanine green (ICG) immediately combines with lipoproteins to fluoresce. Here, we studied whether ICG fluorescence is effective for endoscopic marking in gastric cancer surgery using a photodynamic eye (PDE) camera and fluorescent endoscope. An ICG solution was endoscopically injected into the submucosal layer of the gastric tumor 3 days before surgery. We observed the lesions using both a PDE camera and a fluorescent endoscope during laparotomy and laparoscopy, respectively;we also observed the fluorescent luminance and fluorescent size of the resected lesions. We could intraoperatively detect the size of the resected lesions in eight patients with early gastric cancer and six patients with advanced gastric cancer. We believe that the use of ICG fluorescence in endoscopic marking requires additional information, such as the volume of the ICG solution and the timing of the ICG injection.

Oral aversion to dietary sugar, ethanol and glycerol correlates with alterations in specific hepatic metabolites in a mouse model of human citrin deficiency.

Mice carrying simultaneous homozygous mutations in the genes encoding citrin, the mitochondrial aspartate-glutamate carrier 2 (AGC2) protein, and mitochondrial glycerol-3-phosphate dehydrogenase (mGPD), are a phenotypically representative model of human citrin (a.k.a., AGC2) deficiency. In this study, we investigated the voluntary oral intake and preference for sucrose, glycerol or ethanol solutions by wild-type, citrin (Ctrn)-knockout (KO), mGPD-KO, and Ctrn/mGPD double-KO mice; all substances that are known or suspected precipitating factors in the pathogenesis of human citrin deficiency. The double-KO mice showed clear suppressed intake of sucrose, consuming less with progressively higher concentrations compared to the other mice. Similar observations were made when glycerol or ethanol were given. The preference of Ctrn-KO and mGPD-KO mice varied with the different treatments; essentially no differences were observed for sucrose, while an intermediate intake or similar to that of the double-KO mice was observed for glycerol and ethanol. We next examined the hepatic glycerol 3-phosphate, citrate, citrulline, lysine, glutamate and adenine nucleotide levels following forced enteral administration of these solutions. A strong correlation between the simultaneous increased hepatic glycerol 3-phosphate and decreased ATP or total adenine nucleotide content and observed aversion of the mice during evaluation of their voluntary preferences was found. Overall, our results suggest that the aversion observed in the double-KO mice to these solutions is initiated and/or mediated by hepatic metabolic perturbations, resulting in a behavioral response to increased hepatic cytosolic NADH and a decreased cellular adenine nucleotide pool. These findings may underlie the dietary predilections observed in human citrin deficient patients.

Monocyte Chemoattractant Protein-1 (MCP-1) as a Potential Therapeutic Target and a Noninvasive Biomarker of Liver Fibrosis Associated With Transient Myeloproliferative Disorder in Down Syndrome.

Liver fibrosis is one of the common complications of transient myeloproliferative disorder (TMD) in Down syndrome (DS), but the exact molecular pathogenesis is largely unknown. We herein report a neonate of DS with liver fibrosis associated with TMD, in which we performed the serial profibrogenic cytokines analyses. We found the active monocyte chemoattractant protein-1 expression in the affected liver tissue and also found that both serum and urinary monocyte chemoattractant protein-1 concentrations are noninvasive biomarkers of liver fibrosis. We also showed a prospective of the future anticytokine therapy with herbal medicine for the liver fibrosis associated with TMD in DS.

Highly Enantioselective Bromocyclization of Allylic Amides with a P/P=O Double-Site Lewis Base Catalyst.

The enantioselective bromocyclization of allylic amides catalyzed by phosphorus-containing Lewis bases was examined in detail. A series of control experiments and NMR studies showed that a partially oxidized bis-phosphine generated in situ serves as the actual enantioselective catalyst. The reaction mechanism involves distinct roles of two Lewis basic sites, P and P=O, with P+ Br serving as a fine-tuning element for substrate fixation in the chiral environment, and P+ OBr as the Br+ transfer agent to the olefin. Catalyst loading could be reduced to as little as 1 mol %, and the reaction affords enantioenriched oxazolines with up to >99.5 % ee.

Minimally Required Iodine Dose for the Detection of Hypervascular Hepatocellular Carcinoma on 80-kVp CT.

OBJECTIVE: The objective of our study was to determine the iodine dose per unit of body weight (BW) or body surface area (BSA) that is minimally required to detect hypervascular hepatocellular carcinoma (HCC) on 80-kVp CT. SUBJECTS AND METHODS: One hundred eleven patients (78 men and 33 women; mean age, 68 years; age range, 43-85 years) with chronic hepatitis were randomized into three groups with different iodine loads (0.5, 0.4, and 0.3 g I/kg BW) and underwent contrast-enhanced CT at 80 kVp. Enhancement of the liver and of hypervascular HCCs was quantitatively and qualitatively assessed on hepatic arterial, portal venous, and equilibrium phase images and compared between the groups. Values for iodine dose per unit of BSA (g I/m(2)) were also computed and analyzed. RESULTS: No significant differences in the contrast-to-noise ratio (CNR) of hypervascular HCCs in any phase were found between the groups (p = 0.34-0.99). In the portal venous phase, the mean increase in hepatic contrast enhancement (ΔHU) of the 0.5 g I/kg group (80.3 HU) was higher than those of the 0.4 g I/kg (63.4 HU) and 0.3 g I/kg (53.3 HU) groups (p < 0.001). Linear correlation equations for the increase in hepatic contrast enhancement were as follows: ΔHU = 5.9 + 150.0 × IL(BW) (r = 0.69, p < 0.001), where IL(BW) is the iodine load per unit of BW (g I/kg), and ΔHU = 13.0 + 3.68 × IL(BSA) (r = 0.66, p < 0.001), where IL(BSA) is the iodine load pre unit of BSA (g I/m(2)). CONCLUSION: The minimal iodine dose required to achieve a tumor-to-liver CNR that is acceptable for the detection of hypervascular HCCs on 80-kVp CT was 0.3 g I/kg BW or 11.0 g I/m(2) BSA.

MR imaging findings of uterine pyomyoma: radiologic-pathologic correlation.

A 69-year-old postmenopausal female with a spontaneously occurring uterine pyomyoma was described with emphasis on the MR imaging findings. On unenhanced T1- and T2-weighted MR images, a huge mottled mass suspected to contain blood products, necrotic tissue, or purulent or viscous fluid was demonstrated within anterior myometrial wall of uterine body. The mass was surrounded by a peripheral rim that was hyperintense on T1-weighted images and hypointense on T2-weighted images. On gadolinium-enhanced MR images, most of the mass was unenhanced, but the peripheral rim was equally enhanced with the surrounding myometrium. Pathological examination revealed an intramural uterine pyomyoma surrounded by fibrous capsules with abundant lymphocytes and neutrophils. Our findings indicate that pyomyoma should be considered when MR images demonstrate a myometrial cystic lesion accompanied by a peripheral rim.

Career urgency and turnover intention among young adult workers: a comparison by gender and employment status.

BACKGROUND: This study aimed to analyze, by grouping young adult workers by gender and employment status, the model that states that the tendency for impatience in situations such as "stagnation in career exploration," "low evaluation from affiliation," "upward comparison of careers between friends and acquaintances," and "lack of work-life balance" leads to turnover intention through career urgency such as the "feeling of being pressurized," having the "urge to develop one's career," and having "concern for one's career." METHODS: An online survey was conducted targeting 400 young adult workers. A simultaneous multi-population analysis was performed. RESULTS: For both male and female regular employees, the tendency for impatience when their career exploration stagnated led to their turnover intention by the "feeling of being pressurized." However, for both male and female non-regular employees, although the tendency for impatience promotes the "feeling of being pressurized" upon stagnation in career exploration, it does not lead to turnover intention. Further, the results showed that in the case of female non-regular employees, the tendency for impatience when comparing their own career to those of friends and acquaintances, who are in a more desirable state than their own, leads to turnover intention through "concern for one's career." CONCLUSIONS: Future research should consider marital status and the presence or absence of children in addition to gender and employment. Future studies should consider whether non-regular employees are of the involuntary type and whether they wish to change their status as regular employees.

Effects of supplementation on food intake, body weight and hepatic metabolites in the citrin/mitochondrial glycerol-3-phosphate dehydrogenase double-knockout mouse model of human citrin deficiency.

The C57BL/6:Slc23a13(-/-);Gpd2(-/-) double-knockout (a.k.a., citrin/mitochondrial glycerol 3-phosphate dehydrogenase double knockout or Ctrn/mGPD-KO) mouse displays phenotypic attributes of both neonatal intrahepatic cholestasis (NICCD) and adult-onset type II citrullinemia (CTLN2), making it a suitable model of human citrin deficiency. In the present study, we show that when mature Ctrn/mGPD-KO mice are switched from a standard chow diet (CE-2) to a purified maintenance diet (AIN-93M), this resulted in a significant loss of body weight as a result of reduced food intake compared to littermate mGPD-KO mice. However, supplementation of the purified maintenance diet with additional protein (from 14% to 22%; and concomitant reduction or corn starch), or with specific supplementation with alanine, sodium glutamate, sodium pyruvate or medium-chain triglycerides (MCT), led to increased food intake and body weight gain near or back to that on chow diet. No such effect was observed when supplementing the diet with other sources of fat that contain long-chain fatty acids. Furthermore, when these supplements were added to a sucrose solution administered enterally to the mice, which has been shown previously to lead to elevated blood ammonia as well as altered hepatic metabolite levels in Ctrn/mGPP-KO mice, this led to metabolic correction. The elevated hepatic glycerol 3-phosphate and citrulline levels after sucrose administration were suppressed by the administration of sodium pyruvate, alanine, sodium glutamate and MCT, although the effect of MCT was relatively small. Low hepatic citrate and increased lysine levels were only found to be corrected by sodium pyruvate, while alanine and sodium glutamate both corrected hepatic glutamate and aspartate levels. Overall, these results suggest that dietary factors including increased protein content, supplementation of specific amino acids like alanine and sodium glutamate, as well as sodium pyruvate and MCT all show beneficial effects on citrin deficiency by increasing the carbohydrate tolerance of Ctrn/mGPD-KO mice, as observed through increased food intake and maintenance of body weight.

Uneven haustra on CT colonography: a clue for the detection of transperitoneal invasion from gastric cancer.

Two patients with gastric carcinoma underwent CT colonography (CTC) for preoperative work-up. Although no obvious peritoneal nodules were seen on axial CT images, colonic wall deformities were noted on three-dimensional (3D) air images. Multiplanar-reformatted images revealed corresponding colonic wall thickening at the deformities, and in addition, dense cordlike structures connecting the primary gastric cancer and colonic wall thickening were also observed. In one patient, cordlike indurations consistent with peritoneal invasion were found to connect the primary gastric cancer, gastrocolic ligament, and transverse mesocolon during exploratory surgery, and in the other, colonic scars consistent with peritoneal invasion after chemotherapy were observed. These observations suggest that CTC could be of potential use for the differentiation of transperitoneal colonic invasion and gastric cancer.

Simultaneous determination of eight catechins and four theaflavins in bottled tea by liquid chromatography-tandem mass spectrometry for forensic analysis.

Tea, and particularly bottled tea, is widely consumed worldwide and is often encountered at crime scenes in poisoning cases or used in place of urine in drug abuse monitoring. Tea is a rich source of polyphenols, such as catechins and theaflavins, and these compounds are useful for identification of trace quantities of tea samples. However, information on the contents of catechins and theaflavins in bottled tea is limited. In this study, a method was developed for simultaneous analysis of eight catechins and four theaflavins in tea using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The concentrations of these polyphenols were determined in bottled black, oolong, and green teas after a simple pretreatment process by the standard addition method. The developed LC-MS/MS method was rapid and all tested polyphenol compounds were separated within ~14 min. All tea types contained all the catechins, at varying concentrations, but not all the theaflavins were present in all the tea types. This indicates that the theaflavin composition reflects the degree of the fermentation and could be used for discrimination among different types of tea. All the green tea samples contained all eight catechins; however, the concentrations of these compounds varied among the tea samples. Principal component analysis and hierarchical cluster analysis were useful for discrimination of samples. It has been unclear whether the variations of chemical components are useful for forensic discrimination. Our results demonstrate that, in addition to identification of tea varieties, catechins and theaflavins can be used for the discrimination of bottled tea samples.

Metabolomic analysis reveals hepatic metabolite perturbations in citrin/mitochondrial glycerol-3-phosphate dehydrogenase double-knockout mice, a model of human citrin deficiency.

The citrin/mitochondrial glycerol-3-phosphate dehydrogenase (mGPD) double-knockout mouse displays phenotypic attributes of both neonatal intrahepatic cholestasis and adult-onset type II citrullinemia, making it a suitable model of human citrin deficiency. In the present study, we investigated metabolic disturbances in the livers of wild-type, citrin (Ctrn) knockout, mGPD knockout, and Ctrn/mGPD double-knockout mice following oral sucrose versus saline administration using metabolomic approaches. By using gas chromatography/mass spectrometry and capillary electrophoresis/mass spectrometry, we found three general groupings of metabolite changes in the livers of the double-knockout mice following sucrose administration that were subsequently confirmed using liquid chromatography/mass spectrometry or enzymatic methods: a marked increase of hepatic glycerol 3-phosphate, a generalized decrease of hepatic tricarboxylic acid cycle intermediates, and alterations of hepatic amino acid levels related to the urea cycle or lysine catabolism including marked increases in citrulline and lysine. Furthermore, concurrent oral administration of sodium pyruvate with sucrose ameliorated the hyperammonemia induced by sucrose, as had been shown previously, as well as almost completely normalizing the hepatic metabolite perturbations found. Overall, we have identified additional metabolic disturbances in double-KO mice following oral sucrose administration, and provided further evidence for the therapeutic use of sodium pyruvate in our mouse model of citrin deficiency.

Phase I study of tamibarotene monotherapy in pediatric and young adult patients with recurrent/refractory solid tumors.

PURPOSE: Tamibarotene is a synthetic retinoid that inhibits proliferation and induces differentiation of malignant cells by binding to the retinoic acid receptor α/ß. Previous in vitro studies have shown that some pediatric solid tumors with retinoic acid receptors differentiate in response to retinoic acid. We conducted a phase I dose-escalation study to determine the recommended dose of tamibarotene for further study in pediatric and young adult patients with recurrent/refractory solid tumors. METHODS: Pediatric and young adult patients with recurrent/refractory solid tumors were administered tamibarotene at 4, 6, 8, 10, and 12 mg/m2/day for 14 or 21 days of a 28 day cycle. Safety, efficacy, and pharmacokinetics of tamibarotene were evaluated. RESULTS: Twenty-two patients (median age 8 years) were enrolled in this study. No dose-limiting toxicity (DLT) was encountered, and tamibarotene was generally well tolerated. Two patients experienced severe adverse events (AEs), leading to discontinuation of the treatment. One grade 4 venous thrombosis and one grade 2 erythema multiforme were observed, which promptly resolved after tamibarotene discontinuance. The grade 4 venous thrombosis was a severe AE but not DLT because it occurred after the evaluation period. Pharmacokinetic analyses showed a dose-dependent increase in the maximum drug concentration (Cmax) and area under the concentration-time curve (AUC). None of the patients achieved a complete response or partial response. Seven patients had stable disease lasting longer than 18 weeks. CONCLUSIONS: The recommended dose for phase II study of tamibarotene in pediatric and young adult patients with refractory solid tumors is 12 mg/m2/day for 21 days in a 28 day cycle.

Rationale and design of the BRAVERY study (EPOC1701): a multicentre phase II study of eribulin in patients with BRAF V600E mutant metastatic colorectal cancer.

Background: BRAF V600E mutations are associated with aggressive biology and limited response to standard chemotherapy, especially during second-line and beyond therapies. BRAF V600E mutant and wild-type colorectal cancers (CRCs) differ in their expression profiles, and preclinical evidence suggests that microtubule inhibitors have an antitumour effect on xenograft models of BRAF V600E mutant CRCs. Eribulin has the best growth inhibitory activity in vitro of the microtubule inhibitors. Also, we have evidenced a hint of activity for patients with BRAF V600E mutant metastatic CRC (mCRC) with tumour shrinkage following eribulin treatment. Trial design: The BRAVERY study is a multicentre phase II study to evaluate the efficacy and safety of eribulin in patients with BRAF V600E mutant mCRC detected in either tumour tissues (primary analysis part) or circulating tumour DNA assays (liquid biopsy part). Key eligibility criteria are refractoriness and intolerance to at least one regimen (including irinotecan or oxaliplatin) containing fluoropyrimidine and Eastern Cooperative Oncology Group performance status of 0-1. Eribulin is to be administered intravenously at a dose of 1.4 mg/m2 on days 1 and 8 and repeated every 21 days. The primary endpoint is the confirmed objective response rate (ORR) by investigator's assessment. We calculated the sample size of the primary analysis part at 27 patients using a two-stage design with 25% ORR deemed promising and 5% unacceptable (one-sided α, 0.05; ß, 0.1). Secondary endpoints include disease control rate, progression-free survival, overall survival and adverse events. Moreover, we will collect pretreated tissue and serial blood samples for biomarker analyses, focusing on gene expression associated with BRAF mutant-like CRC to find predictive markers and acquired gene alterations to detect resistance mechanisms to eribulin. We initiated patient enrolment in March 2018, completed the primary analysis on May 2019, and are currently continuing with the liquid biopsy part. Trial registration number: UMIN000031221 and 000031552.

[In vitro chromosome aberration test and in vivo micronucleus test of Ca-type Garcinia extract].

The induction of chromosome aberration of Ca-type Garcinia cambogia extract containing about 65% (-)-hydroxycitric acid was investigated by use of the chromosome aberration test in cultured Chinese hamster lung cells (CHL/IU) and the micronucleus test in mice. In the chromosome aberration test, Ca-type Garcinia cambogia extract did not increase the number of cells with structural aberration and/or numerical aberrations. The micronucleus test was carried out with bone marrow cells of Slc : ddY male mice after single oral administration of up to 2,000 microg/kg. There was no significant increase in the frequency of micronucleated polychromatic erythrocytes. These results indicate that Ca-type Garcinia cambogia extract does not induce chromosome aberration.

Intraoperative localization of arteriovenous malformation of a jejunum with combined use of angiographic methods and indocyanine green injection: Report of a new technique.

INTRODUCTION: The localization of small intestine sources of obscure gastrointestinal bleeding has been a challenge. The use of indocyanine green (ICG) is effective in aiding intraoperative localization if a bleeding lesion is identified on angiography. CASE PRESENTATION: A 95-year-old Japanese man presented with hematochezia. Selective angiography of the superior mesenteric artery (SMA) established an arteriovenous malformation (AVM). ICG injection into the feeding vessel was administered intraoperatively, and the demarcated segment of the jejunum was resected. DISCUSSION: Diluted ICG was injected in the SMA by intraoperative angiography, and the region could be easily and clearly visualized by the ICG fluorescence imaging; small patchy poolings of ICG were recognized. Ultimately, the region was diagnosed as an AVM of the jejunum. To the best of our knowledge, this is the first reported description of this technique. CONCLUSION: Our new technique of combining selective angiography with intraoperative ICG injection and focused enterectomy is a safe, accurate, and cost-effective treatment.

F-18 FDG uptake on positron emission tomography as a predictor for lymphovascular invasion in patients with lung adenocarcinoma.

PURPOSE: To evaluate the contributory value of Fluorine-18 fluorodeoxyglucose (F-18 FDG) positron emission tomography/computed tomography (PET/CT) in the prediction of lymphovascular tumor invasion in patients with lung adenocarcinoma. MATERIALS AND METHODS: We evaluated F-18 FDG-PET/CT images in 84 patients with histopathologically proven lung adenocarcinoma (37 men and 47 women, age range 39-83 years, mean age 67.0 ± 8.9 years). The maximum standardized uptake values (SUVmax) of the carcinomas were measured from the PET images. The Mann-Whitney U test was conducted to compare the median SUVmax between the tumor groups with and without lymphovascular invasion. In the subgroup patients with no lymph-node metastasis, we also compared the median SUVmax between the tumor groups with and without lymphatic invasion. RESULTS: The tumors with lymphovascular invasion had a significantly (P < 0.0001) greater median SUVmax than those without invasion. In the subgroup patients with no lymph-node metastasis, the median SUVmax was higher in tumors with lymphatic invasion than those without (P = 0.0004). The sensitivity, specificity, and area under the receiver operating characteristic curve for the detection of tumors with lymphovascular invasion were 89, 75 %, and 0.82, respectively, with a cutoff SUVmax value of 2.32. CONCLUSION: The SUVmax of lung adenocarcinoma is a potential imaging biomarker for predicting tumor lymphovascular invasion.

BRAF V600 mutations in Langerhans cell histiocytosis with a simple and unique assay.

BACKGROUND: BRAF (V-raf murine sarcoma viral oncogene homolog B1) is a serine-threonine protein kinase involved in cell survival, proliferation, and differentiation. The most common missense mutation of BRAF (mainly V600E) contributes to the incidence of various cancers, including Langerhans cell histiocytosis (LCH). BRAF inhibitors molecularly targeting the V600E mutation have been developed to counteract the effect of the mutation. To ensure the administration of effective pharmacotherapy, it is therefore imperative to develop an effective assay to screen LCH patients for the V600E mutation. However, tumor tissues of LCH typically contain many inflammatory cells which make a correct judgement of the mutation status difficult in the DNA sequence analysis. RESULTS: In this study, we present a new, highly sensitive analyzing method combining PCR, restriction enzyme digestion, and a sequencing assay using DNA extracted from formalin-fixed paraffin-embedded (FFPE) tissue specimens. TspRI is a restriction enzyme that cleaves the sequence encompassing the wild-type BRAF codon 600 into two fragments, which cannot be used as a template for subsequent BRAF PCR amplification. We therefore evaluated the sensitivity of BRAF V600 mutation detection by amplifying the primary PCR product digested with TspRI and sequencing the secondary PCR products. The V600E mutation was detected in FFPE tissue samples from 32 LCH patients; our assay was able to identify mutations in four samples that gave inconclusive results, and ten that were negative, according to standard PCR and sequencing. CONCLUSIONS: We presented a new and highly sensitive method to detect BRAF V600 mutations. This screening method is expected to play an important role to select the most effective therapies.

Enantioselective bromocyclization of allylic amides catalyzed by BINAP derivatives.

A highly enantioselective bromocyclization of allylic amides with N-bromosuccinimide (NBS) was developed with DTBM-BINAP as a catalyst, affording chiral oxazolines with a tetrasubstituted carbon center in high yield with up to 99% ee. By utilizing the bromo substituent as a handle, the obtained compounds were converted to synthetically useful chiral building blocks.