Cellular localization and function of the antiviral protein, ovine Mx1 (oMx1): II. The oMx1 protein is a regulator of secretion in an ovine glandular epithelial cell line

PROBLEM: Embryonic loss is a major contributor to infertility. Understanding factors affecting embryonic loss will help increase fertility. METHOD OF STUDY: We investigated if ovine Mx1 (oMx1) mediated secretion by ovine glandular epithelial (oGE) cells using small interfering RNA (siRNA). Effects on secretion were examined through the conventional endoplasmic reticulum-Golgi pathway using beta2- microglobulin (beta2MG) as a marker, and interferon-stimulated gene 15 (ISG15) as a marker for unconventional secretion. RESULTS: Mx1 siRNA reduced oMx1 mRNA levels at 12 and 24 hr after IFN-tau treatment (P < 0.05), without affecting levels of oMx2, ISG15, 2',5'-oligoadenylate synthetas or beta2MG. Mx1 siRNA reduced Mx1 protein levels at 48 and 120 hr after treatment (P < 0.05) and protein levels remained low at 120 hr. Transient oMx1 knock-down reduced secretion of oMx1 (P < 0.01). ISG15 protein in secretions was reduced without affecting intracellular levels (P < 0.05). Levels of beta2MG in secretions were not affected by Mx1 siRNA. CONCLUSION: We showed that oMx1 protein is secreted by oGE cells and that reduction in oMx1 protein levels by siRNA reduced secretion of ISG15, but not beta2MG. Results support the hypothesis that oMx1 is a regulator of secretion through unconventional secretory pathway(s).     

The EUROHIS-QOL 8-item index: psychometric results of a cross-cultural field study

Background: Survey research including multiple health indicatorsrequires brief indices for use in cross-cultural studies, whichhave, however, rarely been tested in terms of their psychometricquality. Recently, the EUROHIS-QOL 8-item index was developedas an adaptation of the WHOQOL-100 and the WHOQOL-BREF. Theaim of the current study was to test the psychometric propertiesof the EUROHIS-QOL 8-item index. Methods: In a survey on 4849European adults, the EUROHIS-QOL 8-item index was assessed across10 countries, with equal samples adjusted for selected sociodemographicdata. Participants were also investigated with a chronic conditionchecklist, measures on general health perception, mental health,health-care utilization and social support. Results: Findingsindicated good internal consistencies across a range of countries,showing acceptable convergent validity with physical and mentalhealth measures, and the measure discriminates well betweenindividuals that report having a longstanding condition andhealthy individuals across all countries. Differential itemfunctioning was less frequently observed in those countriesthat were geographically and culturally closer to the UK, butacceptable across all countries. A universal one-factor structurewith a good fit in structural equation modelling analyses (SEM)was identified with, however, limitations in model fit for specificcountires. Conclusions: The short EUROHIS-QOL 8-item index showedgood cross-cultural field study performance and a satisfactoryconvergent and discriminant validity, and can therefore be recommendedfor use in public health research. In future studies the measureshould also be tested in multinational clinical studies, particularlyin order to test its sensitivity.     

Age and sex-related differences in dose-dependent hemodynamic response to landiolol hydrochloride during general anesthesia

Objective Landiolol hydrochloride is a new ultra-short-acting β₁-adrenergic receptor blocking agent that is used for patients with tachycardia during general anesthesia. The hemodynamic response to a β-adrenergic receptor blocking agent is generally dependent on the subject. In the present investigation we investigated the effects of age and sex on the hemodynamic response to different doses of landiolol. Methods Following a persistence of tachycardia for more than 1 min, landiolol was infused at 0.03125, 0.0625 and 0.125 mg kg⁻¹ min⁻¹ for 1 min followed by 0.01, 0.02 and 0.04 mg kg⁻¹min⁻¹ for 10 min in groups L, M and H, respectively. Heart rate (HR), systolic (sBP) and diastolic blood pressure (dBP) were recorded each minute during the procedure. The respective changes were evaluated using logistic analysis with the equation , where p, q, m and k indicate the upper asymptote, lower asymptote, maximum slope and time at the maximum slope. Results Parameters q and k for HR in group H were smaller than those in group L, whereas the parameters for sBP and dBP were unchanged among the three groups. Parameter q for HR in group H decreased with age of the patient. There was no significant difference in the landiolol-induced change in HR between males and females. Conclusion The hemodynamic response to landiolol is reliably modeled by the logistic function, especially in terms of HR. Landiolol causes a rapid and dose-dependent decrease in HR, whereas landiolol-induced changes in sBP and dBP are independent of dose. The landiolol-induced decrease in HR becomes larger with aging, but shows no sex difference. The logistic model may be useful for studying hemodynamic responses to landiolol based on age and sex differences, and may allow development of an improved monitoring system. Implication statement The logistic function reliably represents the hemodynamic responses to landiolol. Landiolol reduces HR rapidly and in a dose-dependentl manner whereas landiolol-induced changes in sBP and dBP are independent of dose. A larger decrease in HR is caused by landiolol in elderly patients, but there is no sex difference in the landiolol-induced change in HR.     

Endogenous and exogenous nitric oxide enhance the DNA strand scission induced by tert-butylhydroperoxide in PC12 cells via peroxynitrite-dependent and independent mechanisms, respectively

A short-term exposure to tert-butylhydroperoxide (tB-OOH) promoted a concentration-dependent formation of DNA single-strand breaks in PC12 cells. These events were paralleled by an increase in the cytosolic concentration of Ca2+ that was in part cleared by the mitochondria. Unlike the extent of Ca2+ mobilization and/or mitochondrial Ca2+ clearance, the DNA strand scission evoked by the hydroperoxide was markedly reduced by the nitric oxide (NO) scavenger 2-phenyl-4,4,5,5-tetramethylimidazolin-1-oxyl-3-oxide (PTIO) or by the NO synthase inhibitor N-nitro-L-arginine methylester (L-NAME). Inhibitors of electron transport (rotenone and myxothiazol), ruthenium red (RR, a polycation which inhibits the calcium uniporter of mitochondria), or peroxynitrite scavengers (Trolox and L-methionine) were as effective as PTIO or L-NAME in inhibiting the DNA-damaging response mediated by tB-OOH. Rotenone, RR or peroxynitrite scavengers did not further reduce the residual DNA cleavage observed following treatment with tB-OOH in L-NAME-supplemented cells. Exogenous NO also increased the DNA damage caused by tB-OOH in L-NAME-supplemented cells and this response was blunted by RR or by inhibitors of electron transport but was insensitive to peroxynitrite scavengers. We conclude that both endogenous and exogenous NO enhance the DNA cleavage generated by tB-OOH in PC12 cells. However, only endogenous NO set the bases for an involvement of peroxynitrite in this DNA-damaging response.     

Short Stature and Turner Skeletal Features in an 11-Year-Old Boy with a Ring Y Chromosome Missing the Short Stature Homeobox Containing Gene

We report on an 11-yr-old boy with short stature and Turner skeletal features. Chromosome analysis revealed a 46,X,r(Y)(p11.3q11.2) karyotype, and FISH analysis showed loss of the Short stature homeobox containing gene (SHOX) from the ring Y chromosome. The results are consistent with the association of SHOX haploinsufficiency with short stature and Turner skeletal features, and suggest the importance of SHOX analysis in boys with Turner-like skeletal phenotype.     

Longitudinal Observation of a Patient with Leri-Weill Dyschondrosteosis and SHOX Haploinsufficiency

Haploinsufficiency of the short stature homeobox-containing (SHOX) gene causes Turner skeletal features, a certain proportion of idiopathic short stature and Leri-Weill dyschondrosteosis (LWD). Here we report a Japanese female with LWD. Her physical growth, skeletal deformity, and endocrine status were recorded longitudinally. She exhibited a constant growth rate (average + 6.2 cm/yr) from 6 to 9 yr old, followed by a downward shift at 10 yr old. Her final height was 135 cm (–4.4 SD for an adult female) and weight was 50.5 kg (–0.3 SD) at 12 yr and 10 mo old. Mesomelia and cubitus valgus were noticed from 2 yr old, and metaphyseal lucency and epiphyseal hypoplasia of the medial side of the distal radius were detected at 6 yr old. Madelung deformity was obvious at 10 yr old, when menarche occurred. Fluorescence in situ hybridization (FISH) analysis demonstrated a single copy of the SHOX gene. The short stature of the patient was thought to be exaggerated by the combination of SHOX haploinsufficiency and relatively early puberty.     

Prenatal diagnosis of Down syndrome using cell-free fetal DNA in amniotic fluid by quantitative fluorescent polymersase chain reaction

Backgroud Amniotic fluid （AF） supernatant contains cell-free fetal DNA （cffDNA） fragments.This study attempted to take advantage of cffDNA as a new material for prenatal diagnosis,which could be combined with simple quantitative fluorescent polymerase chain reaction （QF-PCR） to provide an ancillary method for the prenatal diagnosis of trisomy 21 syndrome.Methods AF supernatant samples were obtained from 27 women carrying euploid fetuses and 28 women carrying aneuploid fetuses with known cytogenetic karyotypes.Peripheral blood samples of the parents were collected at the same time.Short tandem repeat （STR） fragments on chromosome 21 were amplified by QF-PCR.Fetal condition and the parental source of the extra chromosome could be determined by the STR peaks.Results The sensitivity of the assay for the aneuploid was 93％ （26/28; confidence interval,CI：77％-98％） and the specificity was 100％ （26/26; CI：88％-100％）.The determination rate of the origin of the extra chromosome was 69％.The sensitivity and the specificity of the assay in the euploid were 100％ （27/27）.Conclusions Trisomy 21 can be prenatally diagnosed by the QF-PCR method in AF supernatant.This karyotype analysis method greatly reduces the requirement for the specimen size.It will be a benefit for early amniocentesis and could avoid pregnancy complications.The method may become an ancillary method for prenatal diagnosis of trisomy 21.     

Association of Sleep Duration and Quality With Subclinical Atherosclerosis

Background

Sleep duration and quality have been associated with increased cardiovascular risk. However, large studies linking objectively measured sleep and subclinical atherosclerosis assessed in multiple vascular sites are lacking.