Evaluation of in vivo genotoxicity induced by N‐ethyl‐N‐nitrosourea,benzo[a]pyrene,and 4‐nitroquinoline‐1‐oxide in the Pig‐a and gpt assays

The recently developed Pig‐a mutation assay is based on flow cytometric enumeration of glycosylphosphatidylinositol (GPI) anchor‐deficient red blood cells caused by a forward mutation in the Pig‐a gene. Because the assay can be conducted in nontransgenic animals and the mutations accumulate with repeat dosing, we believe that the Pig‐a assay could be integrated into repeat‐dose toxicology studies and provides an alternative to transgenic rodent (TGR) mutation assays. The capacity and characteristics of the Pig‐a assay relative to TGR mutation assays, however, are unclear. Here, using transgenic gpt delta mice, we compared the in vivo genotoxicity of single oral doses of N‐ethyl‐N‐nitrosourea (ENU, 40 mg/kg), benzo[a]pyrene (BP, 100 and 200 mg/kg), and 4‐nitroquinoline‐1‐oxide (4NQO, 50 mg/kg) in the Pig‐a (peripheral blood) and gpt (bone marrow and liver) gene mutation assays. Pig‐a assays were conducted at 2, 4, and 7 weeks after the treatment, while gpt assays were conducted on tissues collected at the 7‐week terminal sacrifice. ENU increased both Pig‐a and gpt mutant frequencies (MFs) at all sampling times, and BP increased MFs in both assays but the Pig‐a MFs peaked at 2 weeks and then decreased. Although 4NQO increased gpt MFs in the liver, only weak, nonsignificant increases (two‐ or threefold above control) were detected in the bone marrow in both the Pig‐a and the gpt assay. These findings suggest that further studies are needed to elucidate the kinetics of the Pig‐a mutation assay in order to use it as an alternative to the TGR mutation assay. Environ. Mol. Mutagen. 54:747–754, 2013. © 2013 Wiley Periodicals, Inc.     

Comparison of the Central Adrenal Vein and the Common Trunk of the Left Adrenal Vein for Adrenal Venous Sampling

PurposeTo compare left adrenal venous sampling (AVS) in two locations: the central adrenal vein and the common trunk.Materials and MethodsA total of 22 patients (12 men and 10 women; mean age, 50 y; range, 26–65 y) who were suspected of having primary aldosteronism (PA) and underwent successful AVS with cortisol concentration measurement and/or venography between November 2010 and August 2011 were retrospectively analyzed. In regard to the left adrenal vein, collections were done at two locations: at the common trunk below the confluence of the inferior phrenic vein and at the central adrenal vein, which was above the confluence. The effects of the inflow from the inferior phrenic vein on plasma aldosterone and cortisol levels were analyzed.ResultsEight patients had bilateral hypersecreting lesions and 13 had a unilateral lesion. One was diagnosed as having secondary hypertension other than PA. The median cortisol levels below and above the confluence were 129 μg/dL (range, 21–400 μg/dL) and 215 μg/dL (range, 21–690 μg/dL), respectively. The median aldosterone levels were 2,120 pg/mL (range, 164–42,700 pg/mL) and 4,275 pg/mL (range, 119–59,000 pg/mL), respectively. The median aldosterone/cortisol (A/C) ratios were 244 (range, 34–2,401) and 278 (range, 25–2,251), respectively. Cortisol and aldosterone levels were significantly higher above the confluence (P = .0050 and P = .0003, respectively), whereas the A/C ratio showed no significant difference (P = .12).ConclusionsAlthough higher levels of cortisol and aldosterone were obtained upstream, A/C ratio was not significantly different between the central adrenal vein and the common trunk.     

Characteristic hydrolyzing of megalosaccharide by human salivary α-amylase and small intestinal enzymes,and its bioavailability in healthy subjects

The digestibility of Megalosaccharide® (newly developed carbohydrate comprising α-1,4-glucosaccharide) was investigated in vitro and in vivo. Isomaltosyl-megalosaccharide® (IMS) and nigerosyl-megalosaccharide® (NMS) contain 20% and 50% of the megalosaccharide fraction (degree of polymerization (DP) 10–35), respectively. IMS was hydrolyzed readily by α-amylase to oligosaccharides (DP?≤?7), and a small amount of glucose was produced from oligosaccharides by small intestinal enzymes (SIEs). NMS was partially hydrolyzed by α-amylase to oligosaccharides, and a small amount of glucose produced by SIEs. When IMS and NMS were treated by SIEs after treatment with human saliva α-amylase for a few minutes, IMS and NMS were hydrolyzed readily to glucose. Plasma levels of glucose and insulin upon ingestion of 50?g of IMS or NMS were elevated the same as those for 50?g of glucose, and breath hydrogen was not excreted. These results suggest that IMS and NMS are digestible carbohydrates.     

Eleven novel microsatellite loci for Japanese whiting (Sillago japonica) and cross amplification in the endangered small-scale sillago (Sillago parvisquamis)

Japanese whiting (Sillago japonica) are a relatively common species that inhabit coastal shallow waters in Japan and are the target species in an important recreational fishery. We isolated eleven candidate microsatellite loci from a small insert genomic DNA library of S. japonica. We screened for polymorphisms in the eleven loci using wild individuals (n = 48) collected from Suounada Sound, in the Seto Inland Sea, Japan. The number of alleles per locus ranged from 6 to 26 with no evidence of linkage disequilibrium. Observed heterozygosity ranged from 0.58 to 0.98 with one locus exhibiting a significant departure from Hardy–Weinberg equilibrium. A test for cross-amplification using the closely related species, Sillago parvisquamis yielded scoreable peaks and a high level of polymorphism in four loci. These polymorphic microsatellites can be used to identify population structure in S. japonica and provide potential markers for the endangered S. parvisquamis.     

Changes of NK Cells in Preeclampsia

The regulation of uterine and circulating peripheral blood natural killer (NK) cells has been associated with reproductive immunology such as recurrent pregnancy losses, implantation failures, or preeclampsia. Preeclampsia is a hypertensive disorder of pregnancy characterized by increased blood pressure accompanied by proteinuria and is a major cause of maternal and fetal mortality. Natural cytotoxicity receptors (NCRs) are unique markers, which regulate NK cell cytotoxicity and cytokine production. The relation of NCRs to reproduction is not fully characterized yet. The different profile of NCRs expression may suggest presence of abnormal regulation of NK cell in women with reproductive failures. Pregnant women with preeclampsia carry immunological abnormalities of NCRs on peripheral blood NK cells during pregnancy. The lower expression of NKp46⁺ NK cells in women with preeclampsia may account for the higher production of NK1 cytokine that is known as NK1 shift in pregnant women with preeclampsia. Evaluation of NKp46 on peripheral blood NK cells may be applicable to find the onset of preeclampsia. In this review, various expressions of NK cell surface markers including NCRs on NK cells, NK cell cytotoxicity, and production of cytokines and angiogenic factors by NK cells were reviewed in relation to preeclampsia.     

Long‐term analysis of phase II studies of single‐agent lenalidomide in relapsed/refractory mantle cell lymphoma

Mantle cell lymphoma (MCL) is a type of non‐Hodgkin lymphoma (NHL) with aggressive disease characteristics resulting in multiple relapses after initial treatment. Lenalidomide is an immunomodulatory agent approved in the US for patients with relapsed/refractory MCL following bortezomib based on results from 3 multicenter phase II studies (2 including relapsed/refractory aggressive NHL and 1 focusing on MCL post‐bortezomib). The purpose of this report is to provide longer follow‐up on the MCL‐001 study (follow‐ups were 6.8 [NHL‐002], 7.6 [NHL‐003], and 52.2 [MCL‐001] months). The 206 relapsed MCL patients treated with single‐agent lenalidomide (25 mg/day PO, days 1 to 21 every 28‐days) had a median age of 67 years (63% ≥65 years), 91% with stage III/IV disease, and 50% with ≥4 previous treatment regimens. With a median follow‐up of X, the combined best overall response rate (ORR) was 33% (including 11% with complete remission [CR]/CR unconfirmed CRu). Lenalidomide produced rapid and durable responses with a median time to response of 2.2 months and median duration of response (DOR) of 16.6 months (95% CI: 11.1%‐29.8%). The safety profile was consistent and manageable; myelosuppression was the most common adverse event (AE). Overall, single‐agent lenalidomide showed consistent efficacy and safety in multiple phase II studies of heavily pretreated patients with relapsed/refractory MCL, including those previously treated with bortezomib.