RNA-Seq analysis in an avian model of maternal phenylketonuria

Cardiac malformations (CVMs) are a leading cause of infant morbidity and mortality. CVMs are particularly prevalent when the developing fetus is exposed to high levels of phenylalanine in-utero in mothers with Phenylketonuria. Yet, elucidating the underlying molecular mechanism leading to CVMs has proven difficult. In this study we used RNA-Seq to investigate an avian model of MPKU and establish differential gene expression (DEG) characteristics of the early developmental stages HH10, 12, and 14. In total, we identified 633 significantly differentially expressed genes across stages HH10, 12, and 14. As expected, functional annotation of significant DEGs identified associations seen in clinical phenotypes of MPKU including CVMs, congenital heart defects, craniofacial anomalies, central nervous system defects, and growth anomalies. Additionally, there was an overrepresentation of genes involved in cardiac muscle contraction, adrenergic signaling in cardiomyocytes, migration, proliferation, metabolism, and cell survival. Strikingly, we identified significant changes in expression with multiple genes involved in Retinoic Acid (RA) metabolism and downstream targets. Using qRTPCR, we validated these findings and identified a total of 42 genes within the RA pathway that are differentially expressed. Here, we report the first elucidation of the molecular mechanisms of cardiovascular malformations in MPKU conducted at early developmental timepoints. We provide evidence suggesting a link between PHE exposure and the alteration of RA pathway. These results are promising and offer novel findings associated with congenital heart defects in MPKU.     

Poly (ethylene glycol) hydrogel scaffolds with multiscale porosity for culture of human adipose-derived stem cells

Three-dimensional (3?D) hydrogel scaffolds are an attractive option for tissue regeneration applications because they allow for cell migration, fluid exchange, and can be synthesized to closely mimic the physical properties of the extracellular matrix environment. The material properties of hydrogels play a vital role in cellular migration and differentiation. In light of this, in-depth understanding of material properties is required before such scaffolds can be used to study their influence on cells. Herein, various blends and thicknesses of poly (ethylene glycol) dimethacrylate (PEGDMA) hydrogels were synthesized, flash frozen, and dried by lyophilization to create scaffolds with multiscale porosity. Environmental scanning electron microscopy (ESEM) images demonstrated that lyophilization induced microporous voids in the PEGDMA hydrogels while swelling studies show the hydrogels retain their innate swelling properties. Change in pore size was observed between drying methods, polymer blend, and thickness when imaged in the hydrated state. Human adipose-derived stem cells (hASCs) were seeded on lyophilized and non-lyophilized hydrogels to determine if the scaffolds would support cell attachment and proliferation of a clinically relevant cell type. Cell attachment and morphology of the hASCs were evaluated using fluorescence imaging. Qualitative observations in cell attachment and morphology of hASCs on the surface of the different hydrogel spatial configurations indicate these multiscale porosity hydrogels create a suitable scaffold for hASC culture. These findings offer another factor of tunability in creating biomimetic hydrogels for various tissue engineering applications including tissue repair, regeneration, wound healing, and controlled release of growth factors.     

Individual,household, and community level risk factors of stunting in children younger than 5 years: Findings from a national surveillance system in Nepal

Despite substantial reductions in recent years in Nepal, stunting prevalence in children younger than 5 years remains high and represents a leading public health concern. To identify factors contributing to the stunting burden, we report multilevel risk factors associated with stunting in 4,853 children aged 6–59 months in a nationally and agroecologically representative random sample from the first year of the Policy and Science for Health, Agriculture, and Nutrition Community Studies, a community‐based observational, mixed‐panel study. Mixed effects logistic regressions controlling for multilevel clustering in the study design were used to examine the association of individual‐, household‐, and community‐level factors associated with stunting. Stunting prevalence was 38% in our sample. After adjustment for potential confounding variables, maternal factors, including maternal height and education, were generally the strongest individual‐level risk factors for stunting, adjusted odds ratio (AOR) = 2.52, 95% CI [1.96, 3.25], short (<145 cm) versus not short mothers; AOR = 2.09, 95% CI [1.48, 2.96], uneducated mothers versus secondary school graduates. Among the household‐ and community‐level factors, household expenditure and community infrastructure (presence of paved roads, markets, or hospitals) were strongly, inversely associated with increased stunting risk, AOR = 1.68, 95% CI [1.27, 2.24], lowest versus highest household expenditure quintile; AOR = 2.38, 95% CI [1.36, 4.14], less developed (lacking paved roads, markets, or hospitals) versus more developed communities. Although most factors associated with stunting are not rapidly modifiable, areas for future research and possible interventions emerged.     

UV filter instability: consequences for the human lens

Human lenses appear to become coloured with age primarily due to the covalent binding of UV filter compounds to lens proteins. These crystallin modifications result from the inherent instability of the kynurenine UV filters. Here we investigate this decomposition, the role this may have in the formation of other primate UV filters, and the interaction of the intermediates (alpha,beta-ketoalkenes) with lens components. The UV filters kynurenine, 3-hydroxykynurenine and 3-hydroxykynurenine glucoside were incubated at neutral pH in the presence or absence of NADH or NADPH. The three UV filters underwent spontaneous deamination, such that at pH 7 less than half of the starting materials (kynurenine (42%), 3-hydroxykynurenine glucoside (30%) and 3-hydroxykynurenine (21%)) remained after 7 days. In the presence of NAD(P)H, the double bond of the UV filter-derived deamination compounds, were reduced. Deamination of 3-hydroxykynurenine glucoside, followed by reduction with NAD(P)H, could thus account for the formation of the major lens UV filter 4-(2-amino-3-hydroxyphenyl)-4-oxobutanoic acid glucoside. beta-Benzoylacrylic acid, which possesses the same alpha,beta-ketoalkene sidechain as the deaminated kynurenine UV filters, underwent Michael addition with glutathione, was reduced (hydrogenated) by NAD(P)H, however was unreactive with ascorbate. Surprisingly, at pH 7 the UV filter-derived alpha,beta-ketoalkene intermediates, do not readily undergo intramolecular cyclization. This feature makes the double bond more available for reaction with protein nucleophilic residues and other lens components such as glutathione. On the basis of these data it is likely that glutathione and NAD(P)H, but not ascorbate, protect proteins in the lens from modification by UV filters.     

Sorbents in acute renal failure and end-stage renal disease: middle molecule and cytokine removal

Renal replacement therapy in acute renal failure (ARF) and chronic renal failure (end-stage renal disease; ESRD) has been based on the use of modifications of dialysis (continuous arteriovenous hemofiltration and hemodiafiltration) to remove middle-molecular-weight toxins, consisting of low-molecular-weight proteins and peptides (LMWP) and cytokines involved in inflammation. High-flux dialyzers are not efficient at removing LMWP, and for this reason, sorbents have been studied to augment or replace dialysis. Removal of LMWP such as beta2-microglobulin, leptin, complement factor D, angiogenin and cytokines such as interleukin (IL)-1, IL-6, IL-10, IL-18 and tumor necrosis factor-alpha has been established in animal models of sepsis and in ESRD patients using sorbents. Sorbent devices added to hemodialysis, or the use of such devices alone in inflammatory states, including sepsis, ARF, cardiopulmonary bypass, pre-explantation of donor organs and ESRD, are being studied.     

The incidence of concha bullosa and its relationship to nasal septal deviation and paranasal sinus disease

BACKGROUND AND PURPOSE: The incidence of middle turbinate pneumatization, or concha bullosa, has been well described in the literature. However, to our knowledge, no study has evaluated concha bullosa in relation to nasal septal deviation. We sought to analyze the incidence of concha bullosa and any correlation with nasal septal deviation and paranasal sinus disease. METHODS: Three neuroradiologists retrospectively reviewed findings of 1095 consecutive paranasal sinus CT studies conducted between 2001 and 2002. All examinations were performed for evaluation of a symptom referable to the sinonasal region. Paranasal sinus inflammatory disease was identified and graded as mild, moderate, or severe. Sphenoid, ethmoid, maxillary, and frontal sinuses were each graded separately on both sides. If a concha bullosa was present, it was graded in size as small, moderate, or large. If bilateral concha were present, sizes were compared and when one was larger, it was identified as dominant. When nasal septal deviation was present, it was graded as mild, moderate, or severe. The direction of nasal septal deviation was identified as the face of the convex surface. RESULTS: There was a clear association between the presence of a unilateral concha, or a dominant concha (in the case of bilateral concha), and the presence of nasal septal deviation (P < .0001). Moreover, there was a significant relationship between the presence of concha bullosa and deviation of the nasal septal to the contralateral side (P < .0001). This inverse association was present regardless of the size of the concha bullosa or degree of septal deviation. In every case, there was some preservation of air channels between the dominant concha and the nasal septum. Seventy-three percent of patients with concha bullosa had paranasal sinus inflammatory disease; 78% of patients without concha bullosa also had some form of inflammatory disease. CONCLUSION: Concha bullosa is a common anatomic variant. There is a strong association between the presence of a concha bullosa and contralateral deviation of the nasal septum. Nasal septal deviation away from the dominant concha, with preserved adjacent air channels, suggests that the deviation is not a direct result of mass effect from the concha. No increased incidence of paranasal sinus disease exists in patients with concha bullosa.     

Further evidence of a right hand advantage in motor skill by chimpanzees (Pan troglodytes)

Asymmetries in motor skill when grasping small food items was evaluated in a sample of captive chimpanzees. In two experiments, error rates in grasping food were assessed when controlling for individual differences in grip morphology. In both experiments, chimpanzees were found to make more errors with the left compared to the right hand. Male chimpanzees were also found to make more errors compared to females. These results are discussed in the context of a presumed disassociation between hand preference and performance as indicators of cerebral dominance in motor functions.     

CASK and Dlg form a PDZ protein complex at the mammalian neuromuscular junction

Membrane-associated guanylate kinases (MAGUKs) are modular adapter proteins that serve as scaffolding molecules and anchor channels and receptors via their PDZ (PSD-95, Dlg, Zo-1) domains. Calcium, calmodulin-associated serine/threonine kinase (CASK) is a MAGUK that is critical at synapses in the central nervous system and at cell-cell junctions because of its interactions with channels, receptors, and structural proteins. We show via confocal microscopy that CASK and another MAGUK, Discs Large (Dlg), are present at the mammalian neuromuscular junction in skeletal muscle. Immunoprecipitation data from mouse muscle show that CASK associates with Dlg, providing evidence of a MAGUK protein complex at this synapse. These data indicate that CASK and Dlg may act as a scaffold for organizing receptors and channels at the postsynaptic membrane of the neuromuscular junction.