Overexpression of poplar GSTU51 confers selective tolerance to both mercury and methyl viologen but not to CDNB or cadmium in transgenic poplars

Glutathione S-transferases belong to a large ancient gene family and are thought to be one of the effective detoxification systems. To elucidate the function of the gene in poplar, a tau class gst gene (PatgGSTU51) was cloned from poplar cell suspension cDNA library and its expression was examined. The gene was not expressed in normal conditions, but significantly induced by toxic heavy metals like cadmium and inorganic mercury. However, the highest expression was observed when treated with its synthetic substrate, 1-chloro-2,4-dinitrobenzene (CDNB). Several transgenic poplar lines harboring a chimeric p35S-PatgGSTU51 were developed to understand its function in plant defense. Real-time quantitative PCR, western blot and cellular GST activities consistently showed the transgene was highly expressed in the transgenic lines. The transgenic lines showed increased tolerance to methyl viologen as well as to inorganic mercury but not to cadmium. Furthermore, they did not show any significant tolerance against CDNB, the electrophilic substrate of GST isozymes. Thus, the results suggest that, although PatgGSTU51 is induced by a number of stress agents, it confers a selective tolerance to the toxicity of heavy metal mercury over those of other stress agents and also to oxidation stress caused by MV. Tolerance to CDNB that induces the gene to high level may require such an extra supplemental gene action as vacuolar sequestration.     

Lysophosphatidylcholine stimulates EGF receptor activation and mesangial cell proliferation: regulatory role of Src and PKC

Lysophosphatidylcholine (LPC), a major component of oxidized-low density lipoproteins (ox-LDL), modulates various pathobiological processes involved in vascular and glomerular diseases. Although several studies have shown increased plasma concentrations of ox-LDL as well as LPC in patients with renal disease, the role of LPC in mesangial cell proliferation and associated signaling mechanisms are not clearly understood. In this study, we have shown that LPC induced the phosphorylation of epidermal growth factor receptor (EGFR), as well as the p42/44 MAP kinases. LPC activated Src-kinase and protein kinase C (PKC), and both Src kinase inhibitor PP-2 and PKC inhibitor inhibited the activation of EGFR by LPC. LPC (5-25 microM) stimulated human mesangial cell proliferation by 4-5 fold. Preincubation of mesangial cells with the Src inhibitor (PP-2), or PKC inhibitor (bisindolylmaleimide GF109203-X), or EGF receptor kinase inhibitor (AG1478), or MEK inhibitor (PD98059) significantly inhibited LPC-mediated mesangial cell proliferation. The data suggest that LPC, by activating Src and PKC signaling pathways, stimulates EGF receptor transactivation and down-stream MAP kinase signaling resulting in mesangial hypercellularity, which is a characteristic feature of diverse renal diseases.     

Deletion of phospholipase D1 decreases bone mass and increases fat mass via modulation of Runx2, β-catenin-osteoprotegerin,PPAR-γ and C/EBPα signaling axis

In osteoporosis, mesenchymal stem cells (MSCs) prefer to differentiate into adipocytes at the expense of osteoblasts. Although the balance between adipogenesis and osteogenesis has been closely examined, the mechanism of commitment determination switch is unknown. Here we demonstrate that phospholipase D1 (PLD1) plays a key switch in determining the balance between bone and fat mass. Ablation of Pld1 reduced bone mass but increased fat in mice. Mechanistically, Pld1^/? MSCs inhibited osteoblast differentiaion with diminished Runx2 expression, while osteoclast differentiation was accelerated in Pld1^?/? bone marrow-derived macrophages. Pld1^?/? osteoblasts showed decreased expression of osteogenic makers. Increased number and resorption activity of osteoclasts in Pld1^?/? mice were corroborated with upregulation of osteoclastogenic markers. Moreover, Pld1^?/? osteoblasts reduced β-catenin mediated-osteoprotegerin (OPG) with increased RANKL/OPG ratio which resulted in accelerated osteoclast differentiation. Thus, low bone mass with upregulated osteoclasts could be due to the contribution of both osteoblasts and osteoclasts during bone remodeling. Moreover, ablation of Pld1 further increased bone loss in ovariectomized mice, suggesting that PLD1 is a negative regulator of osteoclastogenesis. Furthermore, loss of PLD1 increased adipogenesis, body fat mass, and hepatic steatosis along with upregulation of PPAR-γ and C/EBPα. Interestingly, adipocyte-specific Pld1 transgenic mice rescued the compromised phenotypes of fat mass and adipogenesis in Pld1 knockout mice. Collectively, PLD1 regulated the bifurcating pathways of mesenchymal cell lineage into increased osteogenesis and decreased adipogenesis, which uncovered a previously unrecognized role of PLD1 in homeostasis between bone and fat mass.     

Genome editing a mouse locus encoding a variant histone,H3.3B,to report on its expression in live animals

Chromatin remodeling via incorporation of histone variants plays a key role in the regulation of embryonic development. The histone variant H3.3 has been associated with a number of early events including formation of the paternal pronucleus upon fertilization. The small number of amino acid differences between H3.3 and its canonical counterparts (H3.1 and H3.2) has limited studies of the developmental significance of H3.3 deposition into chromatin due to difficulties in distinguishing the H3 isoforms. To this end, we used zinc‐finger nuclease (ZFN) mediated gene editing to introduce a small C‐terminal hemagglutinin (HA) tag to the endogenous H3.3B locus in mouse embryonic stem cells (ESCs), along with an internal ribosome entry site (IRES) and a separately translated fluorescent reporter of expression. This system will allow detection of expression driven by the reporter in cells, animals, and embryos, and will facilitate investigation of differential roles of paternal and maternal H3.3 protein during embryogenesis that would not be possible using variant‐specific antibodies. Further, the ability to monitor endogenous H3.3 protein in various cell lineages will enhance our understanding of the dynamics of this histone variant over the course of development. genesis 52:959–966, 2014. © 2014 Wiley Periodicals, Inc.     

Restriction of Francisella novicida Genetic Diversity during Infection of the Vector Midgut

The genetic diversity of pathogens, and interactions between genotypes, can strongly influence pathogen phenotypes such as transmissibility and virulence. For vector-borne pathogens, both mammalian hosts and arthropod vectors may limit pathogen genotypic diversity (number of unique genotypes circulating in an area) by preventing infection or transmission of particular genotypes. Mammalian hosts often act as “ecological filters” for pathogen diversity, where novel variants are frequently eliminated because of stochastic events or fitness costs. However, whether vectors can serve a similar role in limiting pathogen diversity is less clear. Here we show using Francisella novicida and a natural tick vector of Francisella spp. (Dermacentor andersoni), that the tick vector acted as a stronger ecological filter for pathogen diversity compared to the mammalian host. When both mice and ticks were exposed to mixtures of F. novicida genotypes, significantly fewer genotypes co-colonized ticks compared to mice. In both ticks and mice, increased genotypic diversity negatively affected the recovery of available genotypes. Competition among genotypes contributed to the reduction of diversity during infection of the tick midgut, as genotypes not recovered from tick midguts during mixed genotype infections were recovered from tick midguts during individual genotype infection. Mediated by stochastic and selective forces, pathogen genotype diversity was markedly reduced in the tick. We incorporated our experimental results into a model to demonstrate how vector population dynamics, especially vector-to-host ratio, strongly affected pathogen genotypic diversity in a population over time. Understanding pathogen genotypic population dynamics will aid in identification of the variables that most strongly affect pathogen transmission and disease ecology.     

Psychological Factors as Predictors of Suicidal Ideation among Adolescents in Malaysia

Background

There has been a drastic increase in the rate of suicides over the past 45 years in Malaysia. The statistics show that adolescents aged between 16 and 19 years old are at high risk of committing suicide. This could be attributed to issues relating to the developmental stage of adolescents. During this stage, adolescents face challenges and are exposed to various stressful experiences and risk factors relating to suicide.

Method

The present study examined psychological factors (i.e., depression, anxiety and stress) as predictors for suicidal ideation among adolescents. A cross-sectional study was conducted on 190 students (103 males and 87 females), aged 15 to 19 years old from two different schools in Kuala Lumpur. The Depression Anxiety Stress Scale 21-item version (DASS-21) was used to measure depression, anxiety and stress among the students, and the Beck Scale for Suicide Ideation (BSS) to measure suicidal ideation. The data were analysed using Pearson''s correlation and multiple regression analysis.

Results

The results show that 11.10%, 10.00%, and 9.50% of the students reported that they were experiencing severe depression, anxiety and stress, respectively. There were significant correlations between depression, anxiety, and stress with suicidal ideation. However, only depression was identified as a predictor for suicidal ideation.

Conclusion

Hence, this study extends the role of depression in predicting suicidal ideation among adolescents in the Malaysian context. The findings imply that teenagers should be assisted in strengthening their positive coping strategies in managing distress to reduce depression and suicidal ideation.     

Gender Differences and Socioeconomic Status in Relation to Overweight among Older Korean People

Background

The ever-increasing older population and its association with serious overweight problems have garnered much attention. The correlation between being overweight and socioeconomic status factors could be helpful for understanding the inequalities among the overweight population. We examined the correlation between being overweight and some key variables, such as demographics, socioeconomic status, general health status, and health behavior in a large sample of older individuals, by each gender.

Methods

We used data from the 2008 Korean Longitudinal Study of Aging and it included 8,157 participants who were 45 years or older. To understand the relationship between the overweight participants in accordance to demographic and socioeconomic characteristics, health status, and health behaviors, a weighted chi-square test and logistic regression analysis were conducted by separating variables related to overweight, according to the genders.

Results

The number of people in the normal group was 6,347 (77.8%), while the people who were considered overweight were 1,810 (22.2%). Women (n = 4,583) constituted 52.7% of the subject, 24.9% of whom were classified as overweight. Meanwhile, 20.6% of the 47.3% (n = 3,574) of the sample who were men were classified as overweight. Participants between the ages of 45 and 64 with chronic diseases were more likely to be overweight. Men in the 4th quartile of household income were more likely to be overweight than those who were in the 1st quartile, in contrast, while unemployed women with lower education levels and urban residents were at greater risk for being overweight.

Conclusions

Among the men, health status and health behavior appeared to show a correlation with being overweight; however, among women, socioeconomic status factors were strongly related to being overweight. These findings appear to support the association of gender-specifics with the prevalence of being overweight.     

Prevalence of and Factors Associated with Lens Opacities in a Korean Adult Population with and without Diabetes: The 2008–2009 Korea National Health and Nutrition Examination Survey

Objective

We examined the prevalence of and factors associated with lens opacities in a Korean adult population with and without diabetes.

Research Design and Methods

Among the 11,163 adults (≥19 years old) from the fourth Korea National Health and Nutrition Examination Survey in 2008–2009, the data from laboratory tests, nutritional surveys, and slit-lamp examinations of 10,248 persons (4,397 men, 5,851 women) were examined. Cataract was defined as the presence of any nuclear, cortical, subcapsular, or mixed cataract in at least one eye, using the Lens Opacities Classification System III.

Results

The weighted prevalence of cataracts were 23.5% [95% confidence interval (CI), 21.7–25.4] in a Korean adult population (19–39 years old, 1.8% [1.3–2.5], 40–64 years old, 25.2% [22.5–28.1],≥65 years old, 87.8% [85.4–89.9])and 54.7% [50.1–59.2] in a diabetic population(19–39 years old, 11.6% [4.5–26.5], 40–64 years old, 41.1% [35.4–47.0], ≥65 years old, 88.3% [83.5–91.8]). In a logistic regression analysis, age, myopia, and the presence of diabetes were independent risk factors. For young (age 19–39 years) and middle aged (age 40–65 years) adults with diabetes, the OR of having a lens opacity is 5.04 [1.41–17.98] and 1.47 [1.11–1.94], respectively, as those without diabetes, whereas for adults aged 65 and older, there was no difference in the prevalence of cataract.

Conclusions

According to these national survey data, ∼ 24% of Korean adults and ∼ 55% of people with diabetes have cataracts. The presence of diabetes was independently associated with cataracts in young and middle aged adults.