Increase of <Emphasis Type="Italic">Drosophila melanogaster</Emphasis> lifespan due to <Emphasis Type="Italic">D</Emphasis>-<Emphasis Type="Italic">GADD45</Emphasis> overexpression in the nervous system

The GADD45 protein family plays an important role in stress signaling and participates in the integration of cellular response to environmental and physiological factors. GADD45 proteins are involved in cell cycle control, DNA repair, apoptosis, cell survival and aging, and inflammatory response by complicated protein–protein interactions. In Drosophila melanogaster a single D-GADD45 ortholog (GG1086) has been described. Our data show that overexpression of the D-GADD45 gene in the nervous system leads to a significantly increase of Drosophila lifespan without a decrease in fecundity and locomotor activity. The lifespan extension effect is more pronounced in males than in females, which agrees with the sex-dependent expression of this gene. The longevity of D. melanogaster with D-GADD45 overexpression is apparently due to more efficient recognition and repair of DNA damage, as the DNA comet assay showed that the spontaneous DNA damage in the larva neuroblasts is reduced with statistical significance.     

Gene Transfer of <Emphasis Type="Italic">c</Emphasis>-<Emphasis Type="Italic">met</Emphasis> Confers Protection Against <Emphasis Type="SmallCaps">d</Emphasis>-Galactosamine/Lipopolysaccharide-Induced Acute Liver Failure

Background  

Acute liver failure (ALF) is characterized by impaired regeneration of hepatocytes, partially resulting from the defective signaling between elevated levels of hepatocyte growth factor (HGF) and the downregulation of its receptor, c-met.     

Positron emission tomography with 2-deoxy-2-[<Superscript>18</Superscript>F] fluoro-<Emphasis Type="SmallCaps">d</Emphasis>-glucose for diagnosis of intraductal papillary mucinous tumor of the pancreas with parenchymal invasion

We used positron emission tomography with 2-deoxy-2-[¹⁸F]fluoro-d-glucose (FDG-PET) in the diagnosis of two cases of malignant intraductal papillary mucinous tumor (IPMT) of the pancreas. A 56-year-old man and a 72-year-old man, both with tumors in the pancreatic head, were referred to Akita University Medical Center. Computed tomography revealed tumors with multiple cystic components in both patients. FDG-PET images showed markedly high FDG uptake in the area corresponding to a solid component found in one patient and diffuse faint uptake, higher than that of the surrounding tissue, in the other patient, who had no solid component. Histological examination of the resected specimens after pancreatectomy showed invasive carcinoma involving the pancreatic parenchyma in both patients. Although our experience is limited and preliminary, FDG-PET seems to be useful for the detection of malignancy in IPMT, especially in patients not showing any solid component on conventional diagnostic images such as computed tomography.     

Colistin resistant <Emphasis Type="Italic">Escherichia coli</Emphasis> carrying <Emphasis Type="Italic">mcr</Emphasis>-<Emphasis Type="Italic">1</Emphasis> in urban sludge samples: Dhaka,Bangladesh

Of 48 bacteria belonging to the family Enterobacteriaceae tested from urban sludge samples, one Escherichia coli isolate was resistant to colistin and possessed the resistance marker gene mcr-1 found for the first time from Bangladesh. The colistin resistant E. coli was multidrug resistant showing resistance to 11 different antibiotics tested.     

<Emphasis Type="Italic">Fas/FasL,Bcl2</Emphasis> and <Emphasis Type="Italic">Caspase</Emphasis>-<Emphasis Type="Italic">8</Emphasis> gene polymorphisms in Chinese patients with rheumatoid arthritis

Apoptosis signals are necessary for maintaining homeostasis and an adequate immune response. Dysregulation of apoptosis-related genes in the immune system has an important impact on autoimmune diseases such as rheumatoid arthritis (RA). Thus, we investigated the association between Fas rs2234767 G/A, FasL rs763110 C/T, Bcl2 rs12454712 T/C, Bcl2 rs17757541 C/G, and Caspase-8 rs1035142 G/T polymorphisms and RA susceptibility in a Chinese population. These five single-nucleotide polymorphisms (SNPs) were studied in a Chinese population consisting of 615 patients with RA and 839 controls. Genotyping was performed using a custom-by-design 48-Plex SNP scan TM kit. Furthermore, we undertook a meta-analysis between FasL rs763110 C/T and RA. This study indicated that Fas rs2234767 and Bcl2 rs17757541 polymorphisms were risk factors for RA. No association was observed between FasL rs763110 C/T, Bcl2 rs12454712 T/C, and Caspase-8 rs1035142 G/T polymorphisms and RA in this study. The results of this meta-analysis suggested no significant association between FasL rs763110 C/T and RA. However, stratification analysis of this meta-analysis indicated that FasL rs763110 C/T increased the risk of Caucasian RA patients. In conclusion, this study demonstrated that Fas rs2234767 G/A and Bcl2 rs17757541 T/C polymorphisms might be associated with an increased risk of RA. This meta-analysis revealed that FasL rs763110 C/T was associated with an increased risk of Caucasian RA patients.     

Associations of <Emphasis Type="Italic">TIM</Emphasis>-<Emphasis Type="Italic">1</Emphasis> Genetic Polymorphisms with Asthma: A Meta-analysis

Purpose

Recently, the roles of TIM-1 genetic polymorphisms in asthma have been extensively studied, with conflicting results. Therefore, we performed the present meta-analysis to better assess potential associations of TIM-1 genetic polymorphisms with asthma.

Methods

Eligible articles were searched in PubMed, Medline, EMBASE, Google Scholar, and CNKI up to December 2016. Odds ratios (ORs) and 95% confidence intervals (CIs) were used to detect any potential associations between TIM-1 genetic polymorphisms and asthma.

Results

A total of 12 articles including 3120 asthma patients and 2825 control subjects were analyzed. The overall and subgroup analyses revealed that TIM-1-416G>C single nucleotide polymorphism was significantly associated with asthma for the Asian population in the codominant (G/G vs. G/C, p = 0.0003, OR 1.86, 95% CI 1.33–2.60) and dominant (G/G vs. G/C + C/C, p < 0.0001, OR 1.94, 95% CI 1.40–2.69) genetic models. Nevertheless, we failed to detect any significant associations between TIM-1-416G>C single nucleotide polymorphism and asthma in Caucasians. Additionally, according to our analyses, TIM-1 5383_5397 insertion/deletion polymorphism was not correlated with asthma in both Asians and Caucasians.

Conclusions

In conclusion, our findings suggest that TIM-1-416G>C single nucleotide polymorphism is associated with asthma susceptibility for the Asian ethnicity in certain genetic models. However, TIM-1 5383_5397 insertion/deletion polymorphism may not be correlated with the risk of asthma.

     

Alterations of tumor suppressor gene <Emphasis Type="Italic">p16</Emphasis><Superscript><Emphasis Type="Italic">INK4a</Emphasis></Superscript>in pancreatic ductal carcinoma

Background  

Cell cycle inhibitor and tumor suppressor gene p16 / MTS-1 has been reported to be altered in a variety of human tumors. The purpose of the study was to evaluate primary pancreatic ductal adenocarcinomas for potentially inactivating p16 alterations.     

<Emphasis Type="SmallCaps">L</Emphasis>-Carnitine inhibits protein glycation <Emphasis Type="Italic">in vitro</Emphasis> and <Emphasis Type="Italic">in vivo</Emphasis>: evidence for a role in diabetic management

Glycation-initiated changes in tissue proteins are suggested to play an important role in the development of diabetes-related pathological changes. The purpose of this study was to examine the anti-glycating effect of L-carnitine (CA) in vivo in the high-fructose diet-fed rat and to determine the potential of CA to inhibit in vitro glycation. Additionally the glucose-disposal efficiency of CA in the rat diaphragm was investigated. High-fructose diet (60 g/100 g diet)-fed rats were treated with CA (300 mg/kg/day i.p.) for 60 days. The effect of CA on glucose, fructose and fructosamine in plasma, methyl glyoxal and glycated haemoglobin in whole blood and skin and tail tendon collagen glycation were determined. The inhibitory effect of CA on the glycation of bovine serum albumin in vitro was compared with that of aminoguanidine (AG), a known antiglycation agent. Glucose utilisation induced by insulin in the control rat diaphragm was monitored in the presence and absence of CA. High-fructose feeding induced hyperglycaemia and glycation of haemoglobin and skin and tail tendon collagen. In CA-administered fructose-fed rats glycation was significantly reduced. In vitro glycation and accumulation of advanced glycation end products were mitigated by CA. CA was more effective than AG in inhibiting glycation in vitro. CA also enhanced the utilisation of glucose in the rat diaphragm. The findings of the study reveal that CA not only has antiglycation effect but also enhances glucose disposal in the rat diaphragm. These findings provide evidence for the therapeutic utility of CA in diabetes and associated complications.     

Math1/Atoh1 Contributes to Intestinalization of Esophageal Keratinocytes by Inducing the Expression of <Emphasis Type="Italic">Muc2</Emphasis> and <Emphasis Type="Italic">Keratin</Emphasis>-<Emphasis Type="Italic">20</Emphasis>

Background  

Esophageal intestinal metaplasia, also known as Barrett’s esophagus, is the replacement of the normal epithelium with one that resembles the intestine morphologically. Generally, this includes intestinal mucin-secreting goblet cells. Barrett’s esophagus is an important risk factor for adenocarcinoma development. In-vitro models for Barrett’s esophagus have not, to date, focused on the induction of goblet cells in Barrett’s epithelium.     

Superoxide dismutase activities in long-lived <Emphasis Type="Italic">Drosophila melanogaster</Emphasis> females:<Emphasis Type="Italic"> chico</Emphasis><Superscript>1</Superscript> genotypes and dietary dilution

Superoxide dismutase (SOD) activities were determined for dietary dilution conditions that extend the life span of Drosophila melanogaster. The hypothesis motivating this research was that elevated SOD activity is associated with increased life span resulting from flies being held on a restricted diet. SOD activities were also measured for chico ¹ which is a mutation in the insulin receptor substrate protein gene associated with life span extension. This allowed us to confirm the results of (Clancy et al. 2001) and extend the results by measuring CuZn SOD and Mn SOD activities in addition to the previously determined overall SOD activity. If the same form of SOD activity (CuZn SOD or Mn SOD) was elevated on the dilute diet that extends life span and in the long lived chico ¹ homozygotes, then it would suggest that life span extension by dietary restriction and by insulin signaling mutations has a similar underlying mechanism. However, overall SOD activity, and CuZn SOD or Mn SOD activities did not differ among the diets tested. As observed previously (Clancy et al. 2001), overall SOD activity was elevated in chico ¹ homozygotes compared to the heterozygote or wild type. Results from the present study indicate that elevated CuZn SOD activity, not Mn SOD, is the basis for the relatively high level of SOD activity in the chico ¹ homozygotes.     

<Emphasis Type="Italic">T-</Emphasis>Scores and <Emphasis Type="Italic">Z</Emphasis>-Scores

Bone mineral density (BMD) can be measured by a variety of techniques at several skeletal sites. Once measured, the manufacturers’ software uses the BMD to calculate a T-score and/or Z-score. Both T-scores and Z-scores are derived by comparison to a reference population on a standard deviation scale. The recommended reference group for the T-score is a young gender-matched population at peak bone mass, while the Z-score should be derived from an age-matched reference population. T-scores and Z-scores are widely quoted in scientific publications on osteoporosis and BMD studies, and are the values used for DXA diagnostic criteria and current clinical guidelines for the management of osteoporosis. Errors in BMD measurement, differences in reference populations, and variations in calculation methods used, can all affect the actual T-score and Z-score value. Attempts to standardize these values have made considerable progress, but inconsistencies remain within and across BMD technologies. This can be a source of confusion for clinicians interpreting BMD results. A clear understanding of T-scores and Z-scores is essential for correct interpretation of BMD studies in clinical practice.     

Disorganization of cytoplasmic Ca<Superscript>2+</Superscript> oscillations and pulsatile insulin secretion in islets from <Emphasis Type="Italic">ob</Emphasis>/<Emphasis Type="Italic">ob </Emphasis>mice

AIMS/HYPOTHESIS: In normal mouse islets, glucose induces synchronous cytoplasmic [Ca(2+)](i) oscillations in beta cells and pulses of insulin secretion. We investigated whether this fine regulation of islet function is preserved in hyperglycaemic and hyperinsulinaemic ob/ obmice. METHODS: Intact islets from ob/ ob mice and their lean littermates were used after overnight culture for measurement of [Ca(2+)](i) and insulin secretion. RESULTS: We observed three types of [Ca(2+)](i) responses during stimulation by 9 to 12 mmol/l of glucose: sustained increase, rapid oscillations and slow (or mixed) oscillations. They occurred in 8, 18 and 74% of lean islets and 9, 0 and 91% of ob/ ob islets, respectively. Subtle desynchronisation of [Ca(2+)](i) oscillations between regions occurred in 11% of lean islets. In ob/ ob islets, desynchronisation was frequent (66-82% depending on conditions) and prominent: oscillations were out of phase in different regions because of distinct periods and shapes. Only small ob/ ob islets were well synchronised, but sizes of synchronised lean and desynchronised ob/ ob islets were markedly overlapped. The occurrence of desynchronisation in clusters of 5 to 50 islet cells from ob/ obmice and not from lean mice further indicates that islet hypertrophy is not the only causal factor. In both types of islets, synchronous [Ca(2+)](i) oscillations were accompanied by oscillations of insulin secretion. In poorly synchronised ob/ ob islets, secretion was irregular but followed the pattern of the global [Ca(2+)](i) changes. CONCLUSIONS/INTERPRETATION: The regularity of glucose-induced [Ca(2+)](i) oscillations is disrupted in islets from ob/ ob mice and this desynchronisation perturbs the pulsatility of insulin secretion. A similar mechanism could contribute to the irregularity of insulin oscillations in Type II (non-insulin-dependent) diabetes mellitus.     

<Emphasis Type="Italic">Gcg</Emphasis><Superscript><Emphasis Type="Italic">CreERT2</Emphasis></Superscript> knockin mice as a tool for genetic manipulation in pancreatic alpha cells

Aims/hypothesis

The Cre/loxP system, which enables tissue-specific manipulation of genes, is widely used in mice for diabetes research. Our aim was to develop a new Cre-driver mouse line for the specific and efficient manipulation of genes in pancreatic alpha cells.

Methods

A Gcg ^CreERT2 knockin mouse, which expresses a tamoxifen-inducible form of Cre from the endogenous preproglucagon (Gcg) gene locus, was generated by homologous recombination. The new Gcg ^CreERT2 mouse line was crossed to the Rosa26 ^tdTomato (R26 ^tdTomato ) Cre reporter mouse line in order to evaluate the tissue specificity, efficiency and tamoxifen dependency of Gcg ^CreERT2 -mediated recombination. Cell types of pancreatic islets were identified using immunohistochemistry. Biochemical and physiological data, including blood glucose levels, plasma glucagon and glucagon-like peptide (GLP)-1 levels, and pancreatic glucagon content, were collected and used to assess the overall effect of Gcg gene targeting on Gcg ^CreERT2/w heterozygous mice.

Results

Tamoxifen-treated Gcg ^CreERT2/w ;R26 ^tdTomato/w mice displayed Cre reporter activity, i.e. expression of tdTomato red fluorescent protein (RFP) in all known cells that produce proglucagon-derived peptides. In the adult pancreas, RFP was detected in 94–97% of alpha cells, whereas it was detected in a negligible (~ 0.2%) proportion of beta cells. While more than 98% of cells labelled with tamoxifen-induced RFP were glucagon-positive cells, 14–25% of pancreatic polypeptide (PP)-positive cells were also positive for RFP, indicating the presence of glucagon/PP bihormonal cell population. Tamoxifen-independent expression of RFP occurred in approximately 6% of alpha cells. In contrast to alpha cells and GLP-1-producing neurons, in which RFP expression persisted for at least 5 months after tamoxifen administration (presumably due to rare neogenesis in these cell types in adulthood), nearly half of RFP-positive intestinal L cells were replaced with RFP-negative L cells over the first 2 weeks after tamoxifen administration. Heterozygous Gcg ^CreERT2/w mice showed reduced Gcg mRNA levels in islets, but maintained normal levels of pancreatic and plasma glucagon. The mice did not exhibit any detectable baseline physiological abnormalities, at least in young adulthood.

Conclusions/interpretation

The newly developed Gcg ^CreERT2 knockin mouse shows faithful expression of CreER^T2 in pancreatic alpha cells, intestinal L cells and GLP-1-producing neurons. This mouse line will be particularly useful for manipulating genes in alpha cells, due to highly specific and efficient CreER^T2-mediated recombination in this cell type in the pancreas.

     

<Emphasis Type="Italic">p16</Emphasis><Superscript><Emphasis Type="Italic">INK4A</Emphasis></Superscript> Alterations are accompanied by aberrant protein immunostaining in endometrial carcinomas

Purpose To date, the significance of p16^INK4A tumor suppressor gene inactivation in sporadic endometrial cancer (EC) has only rarely been described. In this study, we examined the alteration type and frequency of gene alterations [point mutations, aberrant promoter methylation and loss of heterozygosity (LOH)] in 50 sporadic ECs, and correlated the genetic findings with the immunohistochemical expression of the p16^INK4A protein and the classical clinicopathological features.Methods Gene mutations were detected by PCR-SSCP-sequencing analysis, promoter hypermethylation by methylation-specific PCR (MSP), and LOH by PCR of the STS-marker c5.1.Results In total, p16^INK4A alterations were found in 14 of 50 (28%) sporadic ECs. In six (12%) cases, two alterations occurred simultaneously. Partial p16^INK4A deletions were found in four of 50 (8%) samples. There was one missense mutation (codon 70; CCCGCC) and one frameshift mutation (1-bp deletion in exon 2). Only 2 of 47 (4.2%) tumors exhibited aberrant promoter methylation. An allelic loss was detected in 12 of 50 (24%) carcinomas with a higher incidence in advanced endometrial carcinomas than in early-stage uterine tumors. p16^INK4A alterations were generally accompanied by gene silencing, confirmed by aberrant protein immunostaining (r=-0.442; P=0.001). There was a significant difference in the frequency of p16^INK4A alterations between early (stage I; 18%) and advanced (stages II–IV; 58%) ECs (P=0.022). One case showed complete protein loss, but absence of genetic alterations.Conclusions Our data indicate that p16^INK4A inactivation plays a role in the tumorigenesis of the subset of sporadic ECs, particularly in cases exhibiting an aggressive clinical behavior. We demonstrate that p16^INK4A methylation can act efficiently and similarly to other genetic alterations as one of the two necessary hits according to the Knudson two-hit hypothesis of tumor suppressor gene inactivation.