Blood profile of rusa deer (Cervus timorensis).

The objective of the study was to establish the reference range for blood and coagulation parameters in normal, healthy male and female rusa deer (Cervus timorensis) of different ages. The study was conducted at Taman Pertanian Universiti, Universiti Putra Malaysia. The sample population comprised of 40 rusa deer, which was divided into 4 groups of 13 young (≤1-year-old) males, 5 young females, 13 adult males and 9 adult females. Jugular venous blood samples were collected to determine concentrations of blood and coagulation parameters. The data obtained were normally distributed. However, the analytical results revealed that significant (p0.05) in blood or coagulation parameters was observed between sex in the young deer. Between age group, adult deer had significantly (p<0.05) higher mean cell volume, plasma protein and globulin concentrations than young rusa deer. Thus, it is necessary to take into account the age and sex when using blood reference values for the diagnosis of diseases in the rusa deer

Application of explanatory sequential design of mixed methods research in conciliating qualitative and quantitative findings on social stressors and to examine the social problem solving orientation in substance abuse population

Substance abuse has been a problem with every society and across every generation. The increasing number of substance abusers in Hong Kong causes a huge impact to our healthcare system. With the consideration in the complexity of psychosocial nature of this population, the utility of both qualitative and quantitative research methods have been becoming increasingly accepted in health sciences. This is a pioneer project in Hong Kong that employed mixed methods research for substance abuse populations which incorporates evidence of pre-treatment individual characteristics and other specific-tailored treatment factors in promoting changes over time. This study aimed at developing a comprehensive understanding of how individual factors, different social stressors and social problem solving orientation in shaping their behavior. This study covered two phases. The first one was to collect quantitative data in subjects’ ability in problem recognition, treatment readiness, social values and their orientation in social problem solving. A regression model of relapse prediction was constructed, in which, social problem solving was the most prominent factor, followed by treatment readiness, problem recognition and emotional problem. In the second phase of the study, individual semi-structure interview, and qualitative focus group activities of free-listing and pile-sorting were employed to collect qualitative data on the impact of various social stressors like stress from peers, from family, and from work or study. In the final step, we interpreted findings from these two subsets of quantitative and qualitative data. Results indicated stress from peers was the most prominent single stressor of substance abusers, which yielded similar impact as the combination of cross-product of stress from family and from work or study. Moreover, most subjects adopted negative orientation in their social problem solving, in which, stress from work or study noted with the highest response rate in negative orientation in their social problem solving

An upregulation in the expression of vanilloid transient potential channels 2 enhances hypotonicity-induced cytosolic Ca²⁺ rise in human induced pluripotent stem cell model of Hutchinson-Gillford Progeria.

Hutchinson-Gillford Progeria Syndrome (HGPS) is a fatal genetic disorder characterized by premature aging in multiple organs including the skin, musculoskeletal and cardiovascular systems. It is believed that an increased mechanosensitivity of HGPS cells is a causative factor for vascular cell death and vascular diseases in HGPS patients. However, the exact mechanism is unknown. Transient receptor potential (TRP) channels are cationic channels that can act as cellular sensors for mechanical stimuli. The aim of this present study was to examine the expression and functional role of TRP channels in human induced pluripotent stem cell-derived endothelial cells (iPSC-ECs) from the patients with HGPS. The mRNA and protein expression of TRP channels in HGPS and control (IMR90) iPSC-ECs were examined by semi-quantitative RT-PCRs and immunoblots, respectively. Hypotonicity-induced cytosolic Ca²⁺ ([Ca²⁺](i)) rise in iPSC-ECs was measured by confocal microscopy. RT-PCRs and immunoblots showed higher expressional levels of TRPV2 in iPSC-ECs from HGPS patients than those from normal individuals. In functional studies, hypotonicity induced a transient [Ca²⁺](i) rise in iPSC-ECs from normal individuals but a sustained [Ca²⁺](i) elevation in iPSC-ECs from HGPS patients. A nonselective TRPV inhibitor, ruthenium red (RuR, 20 µM), and a specific TRPV2 channel inhibitor, tranilast (100 µM), abolished the sustained phase of hypotonicity-induced [Ca²⁺](i) rise in iPSC-ECs from HGPS patients, and also markedly attenuated the transient phase of the [Ca²⁺](i) rise in these cells. Importantly, a short 10 min hypotonicity treatment caused a substantial increase in caspase 8 activity in iPSC-ECs from HGPS patients but not in cells from normal individuals. Tranilast could also inhibit the hypotonicity-induced increase in caspase 8 activity. Taken together, our data suggest that an up-regulation in TRPV2 expression causes a sustained [Ca²⁺](i) elevation in HGPS-iPSC-ECs under hypotonicity, consequently resulting in apoptotic cell death. This mechanism may contribute to the pathogenesis of vascular diseases in HGPS patients

Effect of hypotonicity and ATP on [Ca²⁺]_i in IMR90-iPSC-ECs and HGPS-iPSC-ECs.

<p>(A and B), Representative traces (A) and data summary (B) showing the effect of hypotonicity (210 mOsm) on [Ca²⁺]_i (fluorescence ratio F1/F0) in IMR90-iPSC-ECs and HGPS-iPSC-ECs bathed in isotonic solution. n  =  6-7 experiments. (C), Representative traces showing the effect of hypotonic solution (210 mOsm) on [Ca²⁺]_i in cells bathed in Ca²⁺-free isotonic saline. n  =  8 experiments. D. Basal [Ca²⁺]_i level in IMR90-iPSC-ECs and HGPS-iPSC-ECs as determined by Fura-2 dye. n  =  8. ^**<i>p</i><0.01 unpaired <i>t</i>-test compared with the sustained [Ca²⁺]_i level in IMR90-iPSC-EC group in B or compared with basal [Ca²⁺]_i level in D. (E and F), Representative traces showing the effect of ATP (1 µM) on [Ca²⁺]_i in cells bathed in normal physiological saline. Representative from 3 experiments.</p

Effect of ruthenium red (RuR) on hypotonicity-induced [Ca²⁺]_i rise.

<p>(A-B), Representative traces showing the effect of RuR (20 µM) on hypotonicity-induced [Ca²⁺]_i rise in IMR90-iPSC-ECs (A) and HGPS-iPSC-ECs (B). (C and D), Summarized data showing the effect of RuR on transient phase (C) and sustained phase (D) of hypotonicity-induced [Ca²⁺]_i rise. n  =  6–7 independent experiments, 5–10 cells per experiment. ^**<i>p</i><0.01 or ^##<i>p</i><0.01 unpaired <i>t</i>-test compared with corresponding vehicle control.</p

Effect of tranilast on hypotonicity-stimulated caspase-8 activity.

<p>Data summary showing the inhibitory effect of tranilast on hypotonicity-induced activation of caspase-8 in HGPS-iPSC-ECs. n  =  6 independent experiments for each group. ^**<i>p</i><0.01 unpaired <i>t</i>-test compared with isotonicity. ^##<i>p</i><0.01 unpaired t-test compared with hypotonicity without tranilast.</p

Expression of TRP channel transcripts in IMR90-iPSC-ECs and HGPS-iPSC-ECs.

<p>Shown were the expressional levels of transcripts for TRPV (A), TRPC (B), TRPM (C) and TRPP (D) in IMR90-iPSC-ECs (I) and HGPS-iPSC-ECs (H). n  =  4 independent experiments. ^*<i>p</i><0.05 unpaired <i>t</i>-test compared with IMR90-iPSC-ECs.</p

Effect of tranilast on hypotonicity-induced [Ca²⁺]_i rise.

<p>(A-B), Representative traces showing the effect of tranilast (100 µM) on hypotonicity-induced [Ca²⁺]_i rise in IMR90-iPSC-ECs (A) and HGPS-iPSC-ECs (B). (C and D), Summarized data showing the effect of tranilast on transient phase (C) and sustained phase (D) of hypotonicity-induced [Ca²⁺]_i rise. n  =  6–7 independent experiments, 5–10 cells per experiment. ^**<i>p</i><0.01 unpaired <i>t</i>-test compared with vehicle control.</p