Assessment of Quality of Life of Primary Caregivers of Egyptian Asthmatic Children and Adolescents.

BACKGROUND: Asthma as a serious public health problem worldwide exerts a serious load on children's health-related quality of life (HRQOL) and their families. OBJECTIVE: We assess the HRQOL of the primary caregivers of Egyptian asthmatic children and adolescents and its relation to HRQOL of their children and asthma severity. MATERIALS AND METHODS: A cross-sectional study was conducted on 128 pairs of asthmatic children (7-16 years) and their primary caregivers. Pediatric asthma quality of life (QOL) questionnaire, pediatric asthma caregiver's QOL questionnaire, and asthma control questionnaire were used. RESULTS: Uncontrolled asthmatic patients had statistically significantly lower mean caregiver score compared to controlled asthmatic (P < 0.005). There was a statistically significant positive correlation between caregiver's individual and overall QOL scores and their children (individual and overall QOL scores) (P < 0.05). A statistically significant negative correlation between asthma severity and QOL scores of the caregivers of asthmatic children and adolescents was found (P < 0.05). CONCLUSION: The QOL of the primary caregivers of asthmatic children is significantly adversely affected by their children's illness severity.

In silico predicted transcriptional regulatory control of steroidogenesis in spawning female fathead minnows (Pimephales promelas).

Oocyte development and maturation (or oogenesis) in spawning female fish is mediated by interrelated transcriptional regulatory and steroidogenesis networks. This study integrates a transcriptional regulatory network (TRN) model of steroidogenic enzyme gene expressions with a flux balance analysis (FBA) model of steroidogenesis. The two models were functionally related. Output from the TRN model (as magnitude gene expression simulated using extreme pathway (ExPa) analysis) was used to re-constrain linear inequality bounds for reactions in the FBA model. This allowed TRN model predictions to impact the steroidogenesis FBA model. These two interrelated models were tested as follows: First, in silico targeted steroidogenic enzyme gene activations in the TRN model showed high co-regulation (67-83%) for genes involved with oocyte growth and development (cyp11a1, cyp17-17,20-lyase, 3ß-HSD and cyp19a1a). Whereas, no or low co-regulation corresponded with genes concertedly involved with oocyte final maturation prior to spawning (cyp17-17α-hydroxylase (0%) and 20ß-HSD (33%)). Analysis (using FBA) of accompanying steroidogenesis fluxes showed high overlap for enzymes involved with oocyte growth and development versus those involved with final maturation and spawning. Second, the TRN model was parameterized with in vivo changes in the presence/absence of transcription factors (TFs) during oogenesis in female fathead minnows (Pimephales promelas). Oogenesis stages studied included: PreVitellogenic-Vitellogenic, Vitellogenic-Mature, Mature-Ovulated and Ovulated-Atretic stages. Predictions of TRN genes active during oogenesis showed overall elevated expressions for most genes during early oocyte development (PreVitellogenic-Vitellogenic, Vitellogenic-Mature) and post-ovulation (Ovulated-Atretic). Whereas ovulation (Mature-Ovulated) showed highest expression for cyp17-17α-hydroxylase only. FBA showed steroid hormone productions to also follow trends concomitant with steroidogenic enzyme gene expressions. General trends predicted by in silico modeling were similar to those observed in vivo. The integrated computational framework presented was capable of mechanistically representing aspects of reproductive function in fish. This approach can be extended to study reproductive effects under exposure to adverse environmental or anthropogenic stressors.

Effects of chronic exposure to 12‰ saltwater on the endocrine physiology of juvenile American alligator (Alligator mississippiensis).

American alligator (Alligator mississippiensis) habitats are prone to saltwater intrusion following major storms, hurricanes or droughts. Anthropogenic impacts affecting hydrology of freshwater systems may exacerbate saltwater intrusion into freshwater habitats. The endocrine system of alligators is susceptible to changes in the environment but it is currently not known how the crocodilian physiological system responds to environmental stressors such as salinity. Juvenile alligators were exposed to 12‰ saltwater for 5 weeks to determine the effects of chronic exposure to saline environments. Following 5 weeks, plasma levels of hormones [e.g. progesterone, testosterone, estradiol, corticosterone, aldosterone (ALDO), angiotensin II (ANG II)] were quantified using liquid chromatography and tandem mass spectrometry. Compared with freshwater-kept subjects, saltwater-exposed alligators had significantly elevated plasma levels of corticosterone, 11-deoxycortisol, 17α-hydroxyprogesterone, testosterone, 17ß-estradiol, estrone and estriol whereas pregnenolone and ANG II were significantly depressed and ALDO levels were unchanged (slightly depressed). On the one hand, saltwater exposure did not affect gene expression of renal mineralocorticoid and glucorticoid and angiotensin type 1 (AT-1) receptors or morphology of lingual glands. On the other hand, saltwater exposure significantly reduced plasma glucose concentrations whereas parameters diagnostic of perturbed liver function (aspartate aminotransferase and alanine aminotransferase) and kidney function (creatinine and creatine kinase) were significantly elevated. Except for plasma potassium levels (K+), plasma ions Na+ and Cl- were significantly elevated in saltwater alligators. Overall, this study demonstrated significant endocrine and physiological effects in juvenile alligators chronically exposed to a saline environment. Results provide novel insights into the effects of a natural environmental stressor (salinity) on the renin-angiotensin-aldosterone system and steroidogenesis of alligators.

The Effects of Sertraline on Fathead Minnow (Pimephales promelas) Growth and Steroidogenesis.

The purpose of this study was to evaluate the steroidogenic effects of sertraline, a popular selective serotonin reuptake inhibitor, on larval fathead minnow (FHM; Pimephales promelas) and adult FHM. Larvae were exposed to 0.1, 1, and 10 µg/L sertraline for 28 days and analyzed for differential mRNA expression of 11ß-hydroxysteroid dehydrogenase (11ß-HSD), 20ß-hydroxysteroid dehydrogenase (20ß-HSD), aromatase (CYP19a), nuclear thyroid receptor alpha (TRα), and normalized to RP-L8. Adult FHM were exposed to 3 or 10 µg/L sertraline for 7 days and analyzed for differential expression of the same genes with the addition of thyroid receptor beta (TRß). Larval FHM exposed to 0.1 µg/L had a significant upregulation of both 20ß-HSD and TRα while adult FHM exposed to 10 µg/L had a significant upregulation of 11ß-HSD expression in brain tissue. The significance of these findings with respect to survival, growth and reproduction are currently unknown, but represent areas for future research.

In silico predicted reproductive endocrine transcriptional regulatory networks during zebrafish (Danio rerio) development.

The interconnected topology of transcriptional regulatory networks (TRNs) readily lends to mathematical (or in silico) representation and analysis as a stoichiometric matrix. Such a matrix can be 'solved' using the mathematical method of extreme pathway (ExPa) analysis, which identifies uniquely activated genes subject to transcription factor (TF) availability. In this manuscript, in silico multi-tissue TRN models of brain, liver and gonad were used to study reproductive endocrine developmental programming in zebrafish (Danio rerio) from 0.25h post fertilization (hpf; zygote) to 90 days post fertilization (dpf; adult life stage). First, properties of TRN models were studied by sequentially activating all genes in multi-tissue models. This analysis showed the brain to exhibit lowest proportion of co-regulated genes (19%) relative to liver (23%) and gonad (32%). This was surprising given that the brain comprised 75% and 25% more TFs than liver and gonad respectively. Such 'hierarchy' of co-regulatory capability (brain

Environmental stressors and the epigenome.

Epigenetic modification and transgenerational transfer of phenotype at the individual or population level, particularly in response to environmental change, is at the forefront of biological investigation. The plasticity of this process allows an organism to respond to changes in environmental conditions, potentially conferring a survival advantage. In this review, we discuss epigenetic transgenerational phenomena in the specific context of environmental stressors including hypoxia and environmental toxicants.:

In silico predicted structural and functional robustness of piscine steroidogenesis.

Assessments of metabolic robustness or susceptibility are inherently dependent on quantitative descriptions of network structure and associated function. In this paper a stoichiometric model of piscine steroidogenesis was constructed and constrained with productions of selected steroid hormones. Structural and flux metrics of this in silico model were quantified by calculating extreme pathways and optimal flux distributions (using linear programming). Extreme pathway analysis showed progestin and corticosteroid synthesis reactions to be highly participant in extreme pathways. Furthermore, reaction participation in extreme pathways also fitted a power law distribution (degree exponent γ=2.3), which suggested that progestin and corticosteroid reactions act as 'hubs' capable of generating other functionally relevant pathways required to maintain steady-state functionality of the network. Analysis of cofactor usage (O2 and NADPH) showed progestin synthesis reactions to exhibit high robustness, whereas estrogen productions showed highest energetic demands with low associated robustness to maintain such demands. Linear programming calculated optimal flux distributions showed high heterogeneity of flux values with a near-random power law distribution (degree exponent γ≥2.7). Subsequently, network robustness was tested by assessing maintenance of metabolite flux-sum subject to targeted deletions of rank-ordered (low to high metric) extreme pathway participant and optimal flux reactions. Network robustness was susceptible to deletions of extreme pathway participant reactions, whereas minimal impact of high flux reaction deletion was observed. This analysis shows that the steroid network is susceptible to perturbation of structurally relevant (extreme pathway) reactions rather than those carrying high flux.

Relationship between interleukin-10 polymorphism and maternal serum leptin level in preeclampsia.

The objective of this study is to show that pregnancy is a unique immune phenomenon because the feto-placental unit can develop without being attacked by the maternal immune system despite the admixing of maternal and fetal cells. There is a growing body of evidence that regulatory T cells (Tregs) act as modulators of vascular homeostasis. The aim of this study was to evaluate the role of interleukin-10 polymorphism along with leptin in the pathogenesis of preeclampsia. The study was carried out on 20 primigravida pregnant women with preeclampsia and 20 normal primigravida pregnant women. Blood samples sent for laboratory tests showed the presence of serum leptin, which was determined by DRGLeptin sandwich (Enzyme-linked immunosorbent assay (ELISA) technique) (EIA-2395; DRG International, Inc., Mountainside, NJ, USA) and serum IL-10 was determined by ELISA (Genzyme, Cambridge, MA, USA). The detection of IL-10 polymorphism was done by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) techniques. The results of this study show that there was a significant difference between the frequency of genotype in patients and control group with regard to GG genotype, AA genotype, A allele, and G allele (P < .05) However, it was not significant with regard to the frequency of AG genotype (P > .05). The serum leptin was found to be significantly higher in preeclampsia group and with a more significant increase in AA genotype.

Early life-stage toxicity of eight pharmaceuticals to the fathead minnow, Pimephales promelas.

Human pharmaceuticals are routinely being detected in the environment, and there is growing concern about whether these drugs could elicit effects on aquatic organisms. Regulatory paradigms have shifted accordingly, with a greater emphasis on chronic toxicity data compared with acute data. The Organisation for Economic Co-operation and Development 210 Early Life Stage Test has been proposed as a good measure of the potential for pharmaceuticals to elicit chronic toxicity. To begin building a data set regarding the early life-stage toxicity of pharmaceuticals to fish, fathead minnows (FHM) were exposed to amiodarone, carbamazepine, clozapine, dexamethasone, fenofibrate, ibuprofen, norethindrone, or verapamil. Survival and growth were used to assess chronic toxicity in FHM at 28 days posthatch. Exposure of FHM to carbamazepine, fenofibrate, and ibuprofen resulted in no significant adverse effects at the concentrations tested. FHM survival was not impacted by verapamil exposure; however, growth was significantly decreased at 600 µg/L. Dexamethasone-exposed FHM showed a significant decrease in survival at a concentration of 577 µg/L; however, growth was not impacted at the concentration tested. Norethindrone exposure resulted in a significant decrease in survival and dry weight at 14.8 and 0.74 µg/L, respectively. Exposure to amiodarone and clozapine resulted in a significant decrease in survival and a significant increase in growth at concentrations of 1020 and 30.8 µg/L, respectively. Although the effect levels derived in this study are greater then concentrations observed in the environment, these data suggest that synthetic progestins may require additional research.

Quantification of 2-hydrazinopyridine derivatized steroid hormones in fathead minnow (Pimephales promelas) blood plasma using LC-ESI+/MS/MS.

Fathead minnows (Pimephales promelas) comprise a species-of-choice for the hazard assessments of various environmental contaminants, including compounds capable of disrupting endocrine function. Towards this end, the use of liquid chromatography coupled with mass spectrometry (LC-MS) and/or tandem mass spectrometry (MS/MS) is gaining common use for the quantification of steroid hormones as biomarkers of endocrine stress in small-fish toxicological studies. In this work, 2-hydrazinopyridine (2-HP) was used to derivatize and quantify the physiologically relevant steroid hormones of: 17α-hydroxypregnenolone, progesterone, 11-ketotestosterone, 11-deoxycortisol and 17α,20ß-dihydroxypregnenone, in the blood plasma of male and female fathead minnows. Liquid chromatographic separation was achieved using a Waters™ Sunfire C(18) column (2.1 mm×50 mm with a 3.5 µm particle size) and Milli-Q water:methanol (both with 0.1% formic acid) mobile phase over a gradient of 15 min. All mass analyses were conducted using electrospray ionization in the positive mode with tandem mass spectrometry (ESI+/MS/MS). This is the first such application of 2-HP derivatization for the quantifications of the structurally and functionally diverse C19 androgen of 11-ketotestosterone; C21 progestogens of 17α-hydroxypregnenolone, progesterone and17α,20ß-dihydroxypregnenone; and C21 corticosteroid of 11-deoxycortisol, in fathead minnow blood plasma. The limits of detection (LOD) were set to the lowest calibration standard that gave a signal-to-background response of ≥3, and were: 0.16 ng/ml for progesterone, 0.63 ng/ml for 17α-hydroxypregnenolone, 11-deoxycortisol and 17α,20ß-dihydroxypregnenone, and 1.25 ng/ml for 11-ketotestosterone. This study demonstrates the application of 2-HP derivatization for the analysis of a variety of steroid hormones representative of endocrine function in a species of fish commonly used in toxicological studies.

The unexpected sources of organotin contamination in aquatic toxicological laboratory studies.

Unaccounted sources of contamination can be problematic in toxicological studies and can range from the presence of impurities, breakdown products or isoforms of the parent compound to the unexpected compounds leaching from dosing apparatus. As these compounds are not being tested, they may not be measured in the dosed aquaria and hence go undetected, potentially contributing as confounding factors in toxicological assessments. In this paper we report the unexpected detection of butyltin compounds (mono, di and tributyltin) in flow-through aquaria waters of an aquatic toxicological set-up. High and variable leaching rates for dibutyltin of 2.0 and 6.6 microg/h were detected during the first week of each of two separate flow-through studies. Following this initial 'surge' of dibutyltin leachate, a decrease in leachate rate was seen with values of 0.9 and 1.2 microg/h by Day 14 (second week of study). The main source of the butyltin leachates was shown, to be the airline tubing used in the assembly of the air-supply into each flow-through tank. A 24h period of incubation of the airline tubing with clean water led to the leaching of concentrations of 63.8 ng/l TBT-Sn, 1638.8 ng/l DBT-Sn and 4054.6 ng/l MBT-Sn. The concentration of tributyltin detected was within its toxicologically effective range and as such could have potentially confounding effects on the toxicological bioassays being used. These accidental findings could be of enormous relevance to aquatic toxicologists and have an important bearing on the choice of materials used to construct experimental exposure aquaria.