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Skin pigmentation is a highly heterogeneous trait with diverse consequences worldwide. SLC24A5, encoding a potent K+‐dependent Na+/Ca2+ exchanger, is among the known color‐coding genes that participate in melanogenesis by maintaining pH in melanosomes. Deficient SLC24A5 activity results in oculocutaneous albinism (OCA) type 6 in humans. In this study, by utilizing a exome sequencing (ES) approach, we identified two new variants [p. (Gly110Arg) and p. (IIe189Ilefs*1)] of SLC24A5 cosegregating with the OCA phenotype, including nystagmus, strabismus, foveal hypoplasia, albinotic fundus, and vision impairment, in three large consanguineous Pakistani families. Both of these variants failed to rescue the pigmentation in zebrafish slc24a5 morphants, confirming the pathogenic effects of the variants. We also phenotypically characterized a commercially available zebrafish mutant line (slc24a5ko) that harbors a nonsense (p.Tyr208*) allele of slc24a5. Similar to morphants, homozygous slc24a5ko mutants had significantly reduced melanin content and pigmentation. Next, we used these slc24a5ko zebrafish mutants to test the efficacy of nitisinone, a compound known to increase ocular and fur pigmentation in OCA1 (TYR) mutant mice. Treatment of slc24a5ko mutant zebrafish embryos with varying doses of nitisinone did not improve melanin production and pigmentation, suggesting that treatment with nitisinone is unlikely to be therapeutic in OCA6 patients. [ABSTRACT FROM AUTHOR]

Although the prevalence of migraine tends to decrease in the fifth to sixth decades of life, there are still a significant number of patients > 65 years of age who experience migraine or have new-onset migraine. Because these older patients are often excluded from clinical trials, there are fewer evidence-based treatment guidelines for them. Migraine treatment in the older population requires careful consideration of changes in medication metabolism and increased medical comorbidities. Furthermore, older patients can present with an atypical migraine phenotype and have a higher rate of secondary headache, which may lead to a delay in diagnosis and subsequent treatment. Classic preventive treatments for migraine, including tricyclic antidepressants, antiepileptic drugs, and beta blockers, often have intolerable side effects. In addition, the presence of coronary artery disease, stroke, and peripheral arterial disease precludes the use of typical rescue medications such as triptans. As such, there has been a dire need for novel acute and preventive treatments for older adults. The purpose of this review is to provide an update on novel acute and preventive treatments for migraine in the older population. The advantages of these therapies include their efficacy, favorable side-effect profile, particularly in patients with atherosclerotic disease, as well as their tolerability. [ABSTRACT FROM AUTHOR]

Background: Mycosis fungoides (MF) is the most common primary cutaneous lymphoma. It affects usually the covered areas of the body in elderly males in 6th and 7th decades of life. Atypical dermal lymphoid infiltrate is seen along with epidermotropism. Nuclei of neoplastic cells are convoluted. The neoplastic cells demonstrate positivity for CD3 (Pan T) immunohistochemical stain. Majority show increased CD4 to CD8 ratio. The present study was done to study the clinicopathological features, which might be of help in reaching a correct diagnosis in these cases. Materials and Methods: A retrospective descriptive study was conducted on 60 reported cases of MF. The retrieved slides were reviewed for clinical and histopathological features and immunohistochemical profile. Results: The ages ranged from 20–84 years, mean age was 47 years. Majority (75%) of patients were male. Trunk and extremities were the sites most commonly affected. There was significant inverse correlation between epidermal thickness and tumor stage (P = 0.02). Thickened epidermis was seen in patch stage and thickness reduced with progressing stage. The intensity of dermal infiltrate and cell size was also statistically significantly linked to stage progression (P < 0.001 each). In addition, proliferation index also correlated significantly with tumor stage (P = 0.002). Conclusion: Clinical information and histological features are equally important in the accurate diagnosis of MF. Papillary dermal fibrosis is a useful diagnostic clue. CD4:CD8 ratio is not increased in all cases; it may be decreased or remain unchanged. [ABSTRACT FROM AUTHOR]

Drought stress restricts plant growth and development by altering metabolic activity and biological functions. However, plants have evolved several cellular and molecular mechanisms to overcome drought stress. Drought tolerance is a multiplex trait involving the activation of signaling mechanisms and differentially expressed molecular responses. Broadly, drought tolerance comprises two steps: stress sensing/signaling and activation of various parallel stress responses (including physiological, molecular, and biochemical mechanisms) in plants. At the cellular level, drought induces oxidative stress by overproduction of reactive oxygen species (ROS), ultimately causing the cell membrane to rupture and stimulating various stress signaling pathways (ROS, mitogen-activated-protein-kinase, Ca2+, and hormone-mediated signaling). Drought-induced transcription factors activation and abscisic acid concentration co-ordinate the stress signaling and responses in cotton. The key responses against drought stress, are root development, stomatal closure, photosynthesis, hormone production, and ROS scavenging. The genetic basis, quantitative trait loci and genes of cotton drought tolerance are presented as examples of genetic resources in plants. Sustainable genetic improvements could be achieved through functional genomic approaches and genome modification techniques such as the CRISPR/Cas9 system aid the characterization of genes, sorted out from stress-related candidate single nucleotide polymorphisms, quantitative trait loci, and genes. Exploration of the genetic basis for superior candidate genes linked to stress physiology can be facilitated by integrated functional genomic approaches. We propose a third-generation sequencing approach coupled with genome-wide studies and functional genomic tools, including a comparative sequenced data (transcriptomics, proteomics, and epigenomic) analysis, which offer a platform to identify and characterize novel genes. This will provide information for better understanding the complex stress cellular biology of plants. [ABSTRACT FROM AUTHOR]

Intellectual disability (ID) is a genetically and clinically heterogeneous disorder, characterized by limited cognitive abilities and impaired adaptive behaviors. In recent years, exome sequencing (ES) has been instrumental in deciphering the genetic etiology of ID. Here, through ES of a large cohort of individuals with ID, we identified two bi-allelic frameshift variants in METTL5 , c.344_345delGA (p.Arg115Asnfs∗19) and c.571_572delAA (p.Lys191Valfs∗10), in families of Pakistani and Yemenite origin. Both of these variants were segregating with moderate to severe ID, microcephaly, and various facial dysmorphisms, in an autosomal-recessive fashion. METTL5 is a member of the methyltransferase-like protein family, which encompasses proteins with a seven-beta-strand methyltransferase domain. We found METTL5 expression in various substructures of rodent and human brains and METTL5 protein to be enriched in the nucleus and synapses of the hippocampal neurons. Functional studies of these truncating variants in transiently transfected orthologous cells and cultured hippocampal rat neurons revealed no effect on the localization of METTL5 but alter its level of expression. Our in silico analysis and 3D modeling simulation predict disruption of METTL5 function by both variants. Finally, mettl5 knockdown in zebrafish resulted in microcephaly, recapitulating the human phenotype. This study provides evidence that biallelic variants in METTL5 cause ID and microcephaly in humans and highlights the essential role of METTL5 in brain development and neuronal function. [ABSTRACT FROM AUTHOR]

For eliminating organic pollutants from wastewater, Fe 0 or Fe 0 -composites as catalyst for peroxymonosulfate (PMS) activation have attracted increasing attention in advanced oxidation processes. In this work, a novel Fe 0 -montmorillonite composites (Fe-Mt-C-H 2 ) have been prepared by thermal reduction and used as an activator for PMS to remove bisphenol A (BPA). The removal efficiency of BPA (25 mg·L −1 ) and total organic carbon (TOC) were 99.3% and 70.6%, respectively, in the presence of Fe-Mt-C-H 2 (0.4 g·L −1 ) and PMS (1 mM) at pH of 3. Electron paramagnetic resonance (EPR) spectroscopy and radical scavenger studies demonstrated that superoxide radical (O 2 − ) and singlet oxygen ( 1 O 2 ) were the crucial reactive oxygen species (ROS) in the Fe-Mt-C-H 2 /PMS system rather than sulfate (SO 4 − ) and hydroxyl radical (·OH). Besides, it was found that pH had a great influence on the catalytic oxidation rate of BPA and the Fe-Mt-C-H 2 exhibited best catalytic performance and reusability at initial pH of 3. Furthermore, the constituents of the iron surface oxidation layer at different initial pH were investigated with X-ray photoelectron spectroscopy studies, which indicated that the major constituents were Fe 3 O 4 in acidic solution and contained α-Fe 2 O 3 , FeO and FeSO 4 (s) in alkaline or neutral solution. The findings of this study provide new insights into the activation mechanism of PMS by Fe 0 -composites and exhibit a promising application of Fe-Mt-C-H 2 in wastewater treatment. [ABSTRACT FROM AUTHOR]

Background :To determine the efficacy of Daclatasvir and Sofosbuvir plus Ribavirin in achieving Rapid Virological Response (RVR) in patients of Hepatitis C genotype 3 who are noncirrhotic and have not received any previous treatment. Methods: In this descriptive case series study, 100 non cirrhotic, treatment-naïve patients of HCV genotype 3 were enrolled. Each patient was given oral tab Daclatasvir 60 mg once daily, tab Sofosbuvir once daily 400 mg and capsule Ribavirin 400 mg twice daily. HCV RNA PCR quantitative was obtained before treatment and repeated after completion of one month treatment with above mentioned drugs. Results: Mean age of included patients was 42.7 ± 10.68 years (mean ± SD). Females were dominant(59%). The BMI was 26.07 ± 3.00 kg/m2. Out of 100 patients 87% patients achieved Rapid Virological Response (RVR).The data was stratified according to age, gender and BMI. There was no effect of these parameters on the final results. Conclusion: Rapid Virological Response (RVR) of Daclatasvir plus Sofosbuvir and Ribavirin is impressive. However, more studies are required to ascertain the results. [ABSTRACT FROM AUTHOR]

Drought stress ‘particularly at seedling stage’ causes morpho-physiological differences in wheat which are crucial for its survival and adaptability. In the present study, 209 recombinant inbred lines (RILs) from synthetic wheat (W7984)× ‘Opata’ (also known as SynOpRIL) population were investigated under well-watered and water-limited conditions to identify quantitative trait loci (QTL) for morphological traits at seedling stage. Analysis of variance revealed significant differences (P < 0.01) among RILs, and water treatments for all traits with moderate to high broad sense heritability. Pearson's coefficient of correlation revealed positive correlation among all traits except dry root weight that showed poor correlation with fresh shoot weight (FSW) under water-limited conditions. A high-density linkage map was constructed with 2639 genotyping-by-sequencing markers and covering 5047 cM with an average marker density of 2 markers/cM. Composite interval mapping identified 16 QTL distributed over nine chromosomes, of which six were identified under well-watered and 10 in water-limited conditions. These QTL explained from 4 to 59% of the phenotypic variance. Six QTL were identified on chromosome 7B; three for shoot length under water-limited conditions (QSL.nust-7B) at 64, 104 and 221 cM, two for fresh root weight (QFRW.nust-7B) at 124 and 128 cM, and one for root length (QRL.nust-7B) at 122 cM positions. QFSW.nust-7B appeared to be the most significant QTL explaining 59% of the phenotypic variance and also associated with FSW at well-watered conditions. These QTL could serve as target regions for candidate gene discovery and marker-assisted selection in wheat breeding. [ABSTRACT FROM AUTHOR]

Objectives: This case of Klippel-Trénaunay syndrome (KTS) is being shared as a clinical experience for future reference regarding the presentation and management of bleeding per rectum in KTS. Bleeding per rectum is a potentially lethal complication due to the colorectal hemangiomas in rare cases of KTS. Decision making regarding the choice of treatment in these patients is subject to many variables and different methods of medical and surgical treatments have been attempted and published in the literature. We are sharing our experience of surgical treatment for bleeding per rectum in KTS with satisfactory early postoperative outcome. [ABSTRACT FROM AUTHOR]

Purpose. To present new clinical features, multimodal and ultrawide-field imaging characteristics of peripheral cone dystrophy (PCD), and results of laboratory and genetic investigation to decipher the etiology. Methods. Retrospective observational case-series. Results. Three patients with PCD presented with bilateral paracentral scotomas and a mean visual acuity of 20/25. All exhibited confluent macular hyperautofluorescence with a central bull’s eye lesion. Spectral-domain optical coherence tomography revealed loss of outer retinal elements, particularly the inner segment ellipsoid band and external limiting membrane, within the area of macular hyperautofluorescence. This area corresponded with a lightened fundus appearance and variable retinal pigment epithelium (RPE) abnormalities. Full field and multifocal electroretinography distinguished PCD from other photoreceptor dystrophies. Ultrawide-field imaging revealed irregular peripheral retinal lesions in a distribution greater nasally than temporally and not contiguous with the macular lesion. Functional and anatomic testing remained stable over a mean follow-up of 3 years. Laboratory investigation for causes of uveitis was negative. Whole exome sequencing identified rare variants in genes associated with macular or cone dystrophy or degeneration. Conclusions. In contrast to the original description, the funduscopic and fluorescein angiographic appearance of PCD is abnormal, although the defects are subtle. Peripheral lesions may be observed in some patients. Bilateral, symmetric, macular hyperautofluorescence associated with outer retinal atrophy that spares the fovea is a characteristic of PCD. Pathogenic variants in the same gene were not shared across the cohort, suggesting genetic heterogeneity. Further evaluation is warranted. [ABSTRACT FROM AUTHOR]

We report unusually high charge carrier mobilities for regioregular poly(3-hexyltiophene) (rrP3HT) in organic field-effect transistors (OFETs) using polyurethane (PU) as dielectric layer. Our devices display hole mobilities up to 1.37 cm 2 /V in the saturation regime and an ON/OFF current ratio higher than 10 3 , operating at voltages as low as −10 V, with a high I DS current of 1.5 μA. We assign the measured high mobilities mainly to the low density of randomly-oriented electric dipoles at the semiconductor/dielectric interface, which leads to a narrow energy distribution of the electronic levels available for charge transport in rrP3HT. This is confirmed by experimental and theoretical techniques: (1) temperature-dependent transport measurements for extraction of disorder-induced distribution of electronic levels; (2) density functional theory (DFT) calculations of electric dipole moments of PU; and (3) liquid contact-angle measurements for the dipolar component of dielectric surface tension. [ABSTRACT FROM AUTHOR]

The chitosan/vermiculite biocomposite (CTS-VMT) was synthesized successfully with epichlorohydrin (ECH) cross-linking agent and used to remove cadmium and lead from the aqueous solution. CTS-VMT was characterized by FTIR, SEM, BET, TG-DTG, zeta potential and XPS. The effects of critical parameters including solution pH, contact time, initial heavy-metal concentration and adsorbent regeneration were investigated. Besides, adsorption mechanisms were also researched. The results indicated that chitosan molecule cannot intercalate into the interlayer space but cross link on the external surface of VMT. The maximum adsorption capacities for Cd(II) and Pb(II) were 58.48 mg g − 1 and 166.67 mg g − 1 at pH 4, respectively. The adsorption process fitted well the pseudo-second-order model and the adsorption isotherms could be correctly simulated by the Langmuir isotherm model. The zeta potential analyses put forward that electrostatic attraction existed in the adsorption process between the metal cations and CTS-VMT. However, the principal mechanism for adsorption on CTS-VMT was chelation according to the kinetic study and XPS analyses. Moreover, the desorption experiments revealed the prepared adsorbent tended to be regenerated by using HCl and the removal rations for four cycles were all > 90%. Therefore, the synthesized CTS-VMT in this study has the potential to be utilized as an eco-friendly adsorbent for removing Cd(II) and Pb(II). [ABSTRACT FROM AUTHOR]

ZnAlTi layered double oxide (ZnAlTi-LDO) supported C 60 @AgCl nanoparticles were synthesized by the coprecipitation-light-induced method and their photocatalytic activity for photo-degradation of Bisphenol A (BPA) was analyzed quantitatively by high performance liquid chromatography (HPLC). The morphology, structure and composition of the nanocomposites were analyzed by Scanning Electron Microscopy, Transmission electron microscopy, X-ray diffraction patterns, Fourier Transform Infrared spectra and Specific Surface Area. According to the analysis, ZnAlTi-LDO supported C 60 @AgCl nanoparticles (C 60 @AgCl-LDO) was characteristic of mesoporous materials. Pore size distribution plots of all the catalysts were well-distributed, ranging between 10 and 20 nm. Because the heterostructure of Ag@AgCl enhanced near-field for light scattering, reflection and absorption. The Ag@AgCl-LDO nanoparticles and C 60 @AgCl-LDO nanoparticles showed a very intense absorption band in the near-UV region. The C 60 was used as support materials to enhance the stability of Ag-based photocatalysts. The C 60 @AgCl can reduce the recombination of photo-induced electron–hole pairs and therefore increase the efficiency of the photocatalysis. The photo-degradation experiment was conducted by using a series of the nanoparticles to degrade BPA under simulated visible light irradiation. Test results show that, photogenerated holes, superoxide radical species, OH bulk and singlet oxygen are responsible for the photo-degradation, among which the superoxide radical species plays an indispensable role in the photocatalytic reaction system. The degradation rate of the C 60 @AgCl-LDO in photocatalytic degradation of BPA under simulated visible light irradiation is for 90%. The C 60 @AgCl-LDO nanoparticles with remarkable photocatalytic activity can be used for large-scale environmental remediation. [ABSTRACT FROM AUTHOR]

Purpose To describe a novel macular phenotype that is associated with normal visual function. Design Retrospective, observational case series. Participants Thirty-six affected individuals from 23 unrelated families. Methods This was a retrospective study of patients who had a characteristic macular phenotype. Subjects underwent a full ocular examination, electrophysiologic studies, spectral-domain optical coherence tomography (OCT), and fundus autofluorescence imaging. Genomic analyses were performed using haplotype sharing analysis and whole-exome sequencing. Main Outcome Measures Visual acuity, retinal features, electroretinography, and whole-exome sequencing. Results Twenty-six of 36 subjects were female. The median age of subjects at presentation was 15 years (range, 5–59 years). The majority of subjects were asymptomatic and presented after a routine eye examination (22/36 subjects) or after screening because of a positive family history (13/36 subjects) or by another ophthalmologist (1/36 subjects). Of the 3 symptomatic subjects, 2 had reduced visual acuity secondary to nonorganic visual loss and bilateral ametropic amblyopia with strabismus. Visual acuity was 0.18 logarithm of the minimum angle of resolution (logMAR) or better in 30 of 33 subjects. Color vision was normal in all subjects tested, except for the subject with nonorganic visual loss. All subjects had bilateral symmetric multiple yellow dots at the macula. In the majority of subjects, these were evenly distributed throughout the fovea, but in 9 subjects they were concentrated in the nasal parafoveal area. The dots were hyperautofluorescent on fundus autofluorescence imaging. The OCT imaging was generally normal, but in 6 subjects subtle irregularities at the inner segment ellipsoid band were seen. Electrophysiologic studies identified normal macular function in 17 of 19 subjects and normal full-field retinal function in all subjects. Whole-exome analysis across 3 unrelated families found no pathogenic variants in known macular dystrophy genes. Haplotype sharing analysis in 1 family excluded linkage with the North Carolina macular dystrophy (MCDR1) locus. Conclusions A new retinal phenotype is described, which is characterized by bilateral multiple early-onset yellow dots at the macula. Visual function is normal, and the condition is nonprogressive. In familial cases, the phenotype seems to be inherited in an autosomal dominant manner, but a causative gene is yet to be ascertained. [ABSTRACT FROM AUTHOR]

Background: Deferring percutaneous coronary intervention in nonischemic lesions by fractional flow reserve (FFR) is associated with excellent long-term prognosis in patients with stable ischemic heart disease (SIHD). Although FFR is increasingly used for clinical decision making in acute coronary syndrome (ACS) patients with intermediate lesions, its effect on long-term prognosis has not been well established.Objectives: This study investigated the clinical and prognostic utility of FFR in ACS patients with percutaneous coronary intervention deferred on the basis of nonischemic FFR.Methods: We studied 206 consecutive ACS patients with 262 intermediate lesions and 370 patients with SIHD (528 lesions) in whom revascularization was deferred on the basis of a nonischemic FFR (>0.75). The primary outcome measure was a composite of myocardial infarction and target vessel failure (major adverse cardiovascular events [MACE]).Results: In the entire cohort, the long-term (3.4 ± 1.6 years) MACE rate was higher in the ACS group than in the SIHD group (23% vs. 11%, p < 0.0001). After propensity score matching (200 patients/group), MACE remained significantly higher (ACS 25% vs. SIHD 12%; p < 0.0001). On Cox proportional hazards analysis for MACE, ACS had a hazard ratio of 2.8 (95% confidence interval: 1.9 to 4.0; p < 0.0001). In both the matched and unmatched cohorts, across all FFR categories, ACS patients had a significantly higher annualized myocardial infarction/target vessel revascularization rate compared with SIHD (p < 0.05). Receiver-operating characteristic analysis identified FFR cutoffs (best predictive accuracy for MACE) of <0.84 for ACS (MACE 21% vs. 36%; p = 0.007) and <0.81 for SIHD (MACE 17% vs. 9%; p = 0.01).Conclusions: Deferring percutaneous coronary intervention on the basis of nonischemic FFR in patients with an initial presentation of ACS is associated with significantly worse outcomes than SIHD. Caution is warranted in using FFR values derived from patients with SIHD for clinical decision making in ACS patients. [ABSTRACT FROM AUTHOR]

A major cause of blindness worldwide is disease and damage in the posterior ocular segment. Due to poor penetration of therapeutic molecules, topical eye drops tend to be the least effective. We demonstrate a potential approach of penetration enhancement in topical drug formulations for such diseases using three penetration enhancing agents (PEAs) — an in‐house synthesized novel helical polymer and two commonly used cell penetrating peptides (CPPs), TAT and Penetratin, to enable increased penetration of the most commonly used anti‐inflammatory drug, dexamethasone sodium phosphate (DSP). A rapid and cost‐effective ex‐vivo screening model of excised porcine cornea and sclera is used to assay trans‐corneal/scleral penetration of the three PEAs and results quantified using UV & fluorescence spectroscopy for PEAs and by high performance liquid chromatography (HPLC) for DSP. All three PEAs effectively cross both the cornea and sclera individually. A significant enhancement of DSP trans‐corneal/scleral permeation after topical application with PEAs is observed. ZO‐1 immunostaining of the tight junctions in corneal epithelium demonstrated intact tight junctions and transepithelial electrical resistance (TEER) measurements of the corneal epithelium before and 60 minutes after application show very negligible to no difference, thus proving no damage has been done to the integrity of corneal barrier and barrier function. Finally, in‐vitro cytotoxicity of PEAs was determined using MTT assays in ARPE‐19 and HCE‐2 cells, whilst immunostaining with β‐III tubulin was used in assays involving adult rat primary retinal neurons. None of the PEAs exhibit any trace of cytotoxicity. We conclude that CPPs are able to increases the efficiency of drug delivery through the cornea and improve the chances of drug delivery to the posterior ocular segment. [ABSTRACT FROM AUTHOR]

Background: Intravitreal injections to the eye are the standard treatment for wet age‐related macular degeneration (AMD) but are associated with several drawbacks. Intravitreal injections not only cause structural damage but the invasive procedure also causes discomfort and pain, resulting in decreased patient adherence to the therapy. Our work aims to develop a topical liposomal formulation, functionalised with a cell penetrating peptide (CPP) that has the potential to reach the posterior segment of the eye and inhibit neovascularisation and inflammation, which are the hallmarks of wet AMD and Diabetic Retinopathy. The proposed formulation will offer non‐invasive administration (topical) with decreased dosing frequency (owing to its sustained release action), facilitating increased patient compliance. Methods: The liposomes were prepared by the thin film hydration method, followed by the extrusion of obtained multilamellar vesicles into single unilamellar vesicles. The formulations were characterized for their size, PDI and % encapsulation efficiency (%EE). Initially, a Design of Experiments approach was adopted to optimize the encapsulation efficiency of dexamethasone in liposomes, with the amounts of DPPC, cholesterol and dexamethasone being the factors and the size, PDI and %EE as the responses. HPLC was used to quantify the amount of encapsulated dexamethasone. Bevacizumab was then co‐loaded with optimized dexamethasone liposome formulation and bicinchoninic acid (BCA) assay was used to quantify the amount of encapsulated bevacizumab in the liposomes. Results: Liposomes with an average size of 137 ± 3 nm, PDI of 0.07 ± 0.03 and a zeta potential of −3.09 ± 0.42 were synthesized. Dexamethasone encapsulation efficiency was improved from initially observed value of 13.01 ± 0.06 to the optimized value of 52.8 ± 2.5. This optimized formulation has a %EE 5.8 time greater than the previously reported values for DPPC‐dexamethasone liposomes (1). Bevacizumab was successfully co‐encapsulated into the liposomes along with dexamethasone. Bevacizumab demonstrated %EE of 91.3 ± 4.07 in the dual encapsulated liposomes, whereas %EE of dexamethasone was 75.1 ± 4.4. Further studies: The liposome formulation will be functionalised with the CPP and characterized for drug release, transport through ocular layers, cytotoxicity, anti‐angiogenesis and anti‐inflammatory activities using in vitro, ex vivo and in vivo models. References 1. Bhardwaj U, Burgess DJ. Physicochemical properties of extruded and non‐extruded liposomes containing the hydrophobic drug dexamethasone. Int J Pharm. 2010;388(1–2):181–9. This project is funded by the European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska‐Curie Actions (grant agreement – No 813440). [ABSTRACT FROM AUTHOR]

The article reports 7 pathogenic variants in five Hermansky-Pudlak syndrome (HPS) genes segregating with the phenotype in seven large consanguineous Pakistani families using whole-exome sequencing (WES). Detailed hematological examinations and chest computerized tomography (CT) scans were performed on the affective individuals. Data suggests a genotype-phenotype correlation where the missense alleles are hypomorphic and associated with a milder phenotype.

Radiotherapy of individuals suffering with head & neck or brain tumors subserve the risk of sensorineural hearing loss. Here, we evaluated the protective effect of Aminothiol PrC-210 (3-(methyl-amino)-2-((methylamino)methyl)propane-1-thiol) on the irradiated inner ear of guinea pigs. An intra-peritoneal or intra-tympanic dose of PrC-210 was administered prior to receiving a dose of gamma radiation (3000 cGy) to each ear. Auditory Brainstem Responses (ABRs) were recorded one week and two weeks after the radiation and compared with the sham animal group. ABR thresholds of guinea pigs that received an intra-peritoneal dose of PrC-210 were significantly better compared to the non-treated, control animals at one week post-radiation. Morphologic analysis of the inner ear revealed significant inflammation and degeneration of the spiral ganglion in the irradiated animals not treated with PrC-210. In contrast, when treated with PrC-210 the radiation effect and injury to the spiral ganglion was significantly alleviated. PrC-210 had no apparent cytotoxic effect in vivo and did not affect the morphology or count of cochlear hair cells. These findings suggest that aminothiol PrC-210 attenuated radiation-induced cochlea damage for at least one week and protected hearing. [ABSTRACT FROM AUTHOR]

Melanocortin 1 receptor ( MC1R), a Gs protein-coupled receptor of the melanocyte's plasma membrane, is a major determinant of skin pigmentation and phototype. Upon activation by α-melanocyte stimulating hormone, MC1R triggers the cAMP cascade to stimulate eumelanogenesis. We used whole-exome sequencing to identify causative alleles in Pakistani families with skin and hair hypopigmentation. Six MC1R mutations segregated with the phenotype in seven families, including a p.Val174del in-frame deletion and a p.Tyr298* nonsense mutation, that were analyzed for function in heterologous HEK293 cells. p.Tyr298* MC1R showed no agonist-induced signaling to the cAMP or ERK pathways, nor detectable agonist binding. Conversely, signaling was comparable for p.Val174del and wild-type in HEK cells overexpressing the proteins, but binding analysis suggested impaired cell surface expression. Flow cytometry and confocal imaging studies revealed reduced plasma membrane expression of p.Val174del and p.Tyr298*. Therefore, p.Tyr298* was a total loss-of-function ( LOF) allele, while p.Val174del displayed a partial LOF attribute. [ABSTRACT FROM AUTHOR]

Hepatitis C virus (HCV) infection is thought to be associated with an increased risk of coronary heart disease (CHD) events, perhaps secondary to increased inflammation. We sought to examine the angiographic burden of coronary artery disease (CAD) in patients with HCV compared to HCV-negative patients. All consecutive HCV RNA-positive patients (n = 61) who underwent coronary angiography at the University of Arkansas for Medical Sciences from 2001 to 2013 were identified. A parallel group of HCV-negative controls (n = 61), matched for age, gender, and indication for coronary angiography served as control. Angiographic burden of CAD was assessed by computing Gensini scores. Statistical analysis was performed using SPSS 21.0. Patients with HCV had significantly lower levels of total and low-density lipoprotein cholesterol. Preangiographic use of aspirin and statin was significantly lower in the HCV cohort. Number of patients with obstructive CAD was less in HCV group (23% vs 39%, p <0.05). However, angiographic Gensini score was similar in both groups. There was no correlation between HCV RNA titers and Gensini score (p = 0.9, analysis of variance). In conclusion, patients with active HCV infection have similar angiographic CAD burden as HCV-negative patients. Furthermore, viral load does not appear to correlate with atherosclerosis burden. Patients with HCV have less-obstructive CAD and less-frequent use of aspirin and statins. [ABSTRACT FROM AUTHOR]

Hearing loss is a complex disorder caused by both genetic and environmental factors. Previously, mutations in CIB2 have been identified as a common cause of genetic hearing loss in Pakistani and Turkish populations. Here we report a novel (c.556C>T; p.(Arg186Trp)) transition mutation in the CIB2 gene identified through whole exome sequencing (WES) in a Caribbean Hispanic family with non-syndromic hearing loss. CIB2 belongs to the family of calcium-and integrin-binding (CIB) proteins. The carboxy-termini of CIB proteins are associated with calcium binding and intracellular signaling. The p.(Arg186Trp) mutation is localized within predicted type II PDZ binding ligand at the carboxy terminus. Our ex vivo studies revealed that the mutation did not alter the interactions of CIB2 with Whirlin, nor its targeting to the tips of hair cell stereocilia. However, we found that the mutation disrupts inhibition of ATP-induced Ca2+ responses by CIB2 in a heterologous expression system. Our findings support p.(Arg186Trp) mutation as a cause for hearing loss in this Hispanic family. In addition, it further highlights the necessity of the calcium binding property of CIB2 for normal hearing. [ABSTRACT FROM AUTHOR]

Achromatopsia (ACHM) is an early-onset retinal dystrophy characterized by photophobia, nystagmus, color blindness and severely reduced visual acuity. Currently mutations in five genes CNGA3, CNGB3, GNAT2, PDE6C and PDE6H have been implicated in ACHM. We performed homozygosity mapping and linkage analysis in a consanguineous Pakistani ACHM family and mapped the locus to a 15.12-Mb region on chromosome 1q23.1-q24.3 with a maximum LOD score of 3.6. A DNA sample from an affected family member underwent exome sequencing. Within the ATF6 gene, a single-base insertion variant c.355_356dupG (p.Glu119Glyfs*8) was identified, which completely segregates with the ACHM phenotype within the family. The frameshift variant was absent in public variant databases, in 130 exomes from unrelated Pakistani individuals, and in 235 ethnically matched controls. The variant is predicted to result in a truncated protein that lacks the DNA binding and transmembrane domains and therefore affects the function of ATF6 as a transcription factor that initiates the unfolded protein response during endoplasmic reticulum (ER) stress. Immunolabeling with anti-ATF6 antibodies showed localization throughout the mouse neuronal retina, including retinal pigment epithelium, photoreceptor cells, inner nuclear layer, inner and outer plexiform layers, with a more prominent signal in retinal ganglion cells. In contrast to cytoplasmic expression of wild-type protein, in heterologous cells ATF6 protein with the p.Glu119Glyfs*8 variant is mainly confined to the nucleus. Our results imply that response to ER stress as mediated by the ATF6 pathway is essential for color vision in humans. [ABSTRACT FROM AUTHOR]

Next-generation sequencing (NGS) of exomes and genomes has accelerated the identification of genes involved in Mendelian phenotypes. However, many NGS studies fall short of identifying causal variants, with estimates for success rates as low as 25% for uncovering the pathological variant underlying disease etiology. An important reason for such failures is familial locus heterogeneity, where within a single pedigree causal variants in two or more genes underlie Mendelian trait etiology. As examples of intra- and inter-sibship familial locus heterogeneity, we present 10 consanguineous Pakistani families segregating hearing impairment due to homozygous variants in two different hearing impairment genes and a European-American pedigree in which hearing impairment is caused by four variants in three different genes. We have identified 41 additional pedigrees with syndromic and nonsyndromic hearing impairment for which a single previously reported hearing impairment gene has been identified but only segregates with the phenotype in a subset of affected pedigree members. We estimate that locus heterogeneity occurs in 15.3% (95% confidence interval: 11.9%, 19.9%) of the families in our collection. We demonstrate novel approaches to apply linkage analysis and homozygosity mapping (for autosomal recessive consanguineous pedigrees), which can be used to detect locus heterogeneity using either NGS or SNP array data. Results from linkage analysis and homozygosity mapping can also be used to group sibships or individuals most likely to be segregating the same causal variants and thereby increase the success rate of gene identification. [ABSTRACT FROM AUTHOR]

Axotomised retinal ganglion cells (RGCs) die rapidly by apoptosis and fail to regenerate because of the limited availability of neurotrophic factors and a lack of axogenic stimuli. However, we have recently showed that pigment epithelium-derived factor (PEDF) promotes RGC survival and axon regeneration after optic nerve crush injury. PEDF has multiple fragments of the native peptide that are neuroprotective, anti-angiogenic and anti-inflammatory. Here we investigated the neuroprotective and axogenic properties of a fragment of PEDF, PEDF-34, in retinal neurons in vitro and when delivered by intravitreal injection and eye drops in vivo . We found that PEDF-34 was 43% more neuroprotective and 52% more neuritogenic than PEDF-44 in vitro . Moreover, in vivo , intravitreal delivery of 1.88 nM PEDF-34 was 71% RGC neuroprotective at 21 days after optic nerve crush compared to intact controls, whilst daily eye drops containing 1.88 nM PEDF-34 promoted 87% RGC survival. After topical eye drop delivery, PEDF-34 was detected in the vitreous body within 30 min and attained physiologically relevant concentrations in the retina by 4 h peaking at 1.4 ± 0.05 nM by 14 days. In eye drop- compared to intravitreal-treated PEDF-34 animals, 55% more RGC axons regenerated 250 μm beyond the optic nerve lesion. We conclude that daily topical eye drop application of PEDF-34 is superior to weekly intravitreal injections in promoting RGC survival and axon regeneration through both direct effects on retinal neurons and indirect effects on other retinal cells. [ABSTRACT FROM AUTHOR]

A duplication variant within the middle ear-specific gene A2ML1 cosegregates with otitis media in an indigenous Filipino pedigree (LOD score = 7.5 at reduced penetrance) and lies within a founder haplotype that is also shared by 3 otitis-prone European-American and Hispanic-American children but is absent in non-otitis-prone children and >62,000 next-generation sequences. We identified seven additional A2ML1 variants in six otitis-prone children. Collectively, our studies support a role for A2ML1 in the pathophysiology of otitis media. [ABSTRACT FROM AUTHOR]

Saline soils are the source of severe abiotic stress, which hinders crop productivity. To compete the challenges of food security for rapidly growing population of the world, it is necessary to utilize marginal lands for cultivation by developing saline adapted varieties. Total six elite maize lines were used as parents following diallel mating design. Resultantly 30 Fjs were generated. Data were recorded for different adaptability indicators under saline environments in maize. Differences among mean squares for general combining ability (GcA), specific combining ability (ScA) and reciprocal effects for all the indicators were highly significant. Root length, shoot length, Na+ and K+ ion contents were governed by non-additive type of gene action whereas; leaf area was controlled by additive gene action. Under different saline environments, better adaptability level regarding GCA, SCA and their reciprocal effects were indicated in maize genotypes L7-2, OH-41 and WFTMS. The cross Q67 × L7-2 exhibited comparatively highest level of adaptability regarding SCA effects under saline environments. For reciprocal effects maize genotypes L7-2, Q67, OH-41 and WFTMS could be used in different crossing combinations for the development of maize hybrids adapted to different saline environments. © 2015 Friends Science Publishers [ABSTRACT FROM AUTHOR]

Pathogenic mutations of MARVELD2, encoding tricellulin, a tricelluar tight junction protein, cause autosomal recessive non-syndromic hearing loss (DFNB49) in families of Pakistan and Czech Roma origin. In fact, they are a significant cause of prelingual hearing loss in the Czech Roma, second only to GJB2 variants. Previously, we reported that mice homozygous for p.Arg497* variant of Marveld2 had a broad phenotypic spectrum, where defects were observed in the inner ear, heart, mandibular salivary gland, thyroid gland and olfactory epithelium. The current study describes the types and frequencies of MARVELD2 alleles and clinically reexamines members of DFNB49 families. We found that MARVELD2 variants are responsible for about 1.5 % (95 % CI 0.8-2.6) of non-syndromic hearing loss in our cohort of 800 Pakistani families. The c.1331+2T>C allele is recurrent. In addition, we identified a novel large deletion in a single family, which appears to have resulted from non-allelic homologous recombination between two similar Alu short interspersed elements. Finally, we observed no other clinical manifestations co-segregating with hearing loss in DFNB49 human families, and hypothesize that the additional abnormalities in the Marveld2 mutant mouse indicates a critical non-redundant function for tricellulin in other organ systems. [ABSTRACT FROM AUTHOR]

We assessed a large consanguineous Pakistani family (PKAB157) segregating early onset low vision problems. Funduscopic and electroretinographic evaluation of affected individuals revealed juvenile cone-rod dystrophy (CRD) with maculopathy. Other clinical symptoms included loss of color discrimination, photophobia and nystagmus. Whole-exome sequencing, segregation and haplotype analyses demonstrated that a transition variant (c.955T>C; p.(Cys319Arg)) in CNGA3 co-segregated with the CRD phenotype in family PKAB157. The ability of CNGA3 channel to influx calcium in response to agonist, when expressed either alone or together with the wild-type CNGB3 subunit in HEK293 cells, was completely abolished due to p.Cys319Arg variant. Western blotting and immunolocalization studies suggest that a decreased channel density in the HEK293 cell membrane due to impaired folding and/or trafficking of the CNGA3 protein is the main pathogenic effect of the p.Cys319Arg variant. Mutant alleles of the human cone photoreceptor cyclic nucleotide-gated channel (CNGA3) are frequently associated with achromatopsia. In rare cases, variants in CNGA3 are also associated with cone dystrophy, Leber's congenital amaurosis and oligo cone trichromacy. The identification of predicted p.(Cys319Arg) missense variant in CNGA3 expands the repertoire of the known genetic causes of CRD and phenotypic spectrum of CNGA3 alleles. [ABSTRACT FROM AUTHOR]

Background: In the present study we aimed: 1) To establish the prevalence and clinical impact of DFNB49 mutations in deaf Roma from 2 Central European countries (Slovakia and Hungary), and 2) to analyze a possible common origin of the c.1331+2T>C mutation among Roma and Pakistani mutation carriers identified in the present and previous studies. Methods: We sequenced 6 exons of the MARVELD2 gene in a group of 143 unrelated hearing impaired Slovak Roma patients. Simultaneously, we used RFLP to detect the c.1331+2T>C mutation in 85 Hungarian deaf Roma patients, control groups of 702 normal hearing Romanies from both countries and 375 hearing impaired Slovak Caucasians. We analyzed the haplotype using 21 SNPs spanning a 5.34Mb around the mutation c.1331+2T>C. Results: One pathogenic mutation (c.1331+2T>C) was identified in 12 homozygous hearing impaired Roma patients. Allele frequency of this mutation was higher in Hungarian (10%) than in Slovak (3.85%) Roma patients. The identified common haplotype in Roma patients was defined by 18 SNP markers (3.89 Mb). Fourteen common SNPs were also shared among Pakistani and Roma homozygotes. Biallelic mutation carriers suffered from prelingual bilateral moderate to profound sensorineural hearing loss. Conclusions: We demonstrate different frequencies of the c.1331+2T>C mutation in hearing impaired Romanies from 3 Central European countries. In addition, our results provide support for the hypothesis of a possible common ancestor of the Slovak, Hungarian and Czech Roma as well as Pakistani deaf patients. Testing for the c.1331+2T>C mutation may be recommended in GJB2 negative Roma cases with early-onset sensorineural hearing loss. [ABSTRACT FROM AUTHOR]

In this study, the physicochemical and photocatalytic properties of two kinds of stannate perovskite oxides (MgSnO 3 and CaSnO 3) were investigated under simulated sunlight, where dimethyl phthalate (DMP) and diethyl phthalate (DEP) were selected as the probe pollutants. The results of photochemical characterization showed that MgSnO 3 perovskite exhibited better photocatalytic performance than CaSnO 3 perovskite. MgSnO 3 perovskite could effectively degrade 75% of DMP and 79% of DEP through pseudo-first-order reaction kinetics, which remained good in pH 3.0 to 9.0. Quenching experiments and electron paramagnetic resonance (EPR) characterization indicated that photogenerated holes (h +), superoxide (O 2 −), and hydroxyl radicals (OH) worked in the photo-degradation, while O 2 − played the most important role. Furthermore, intermediates identification and density functional theory (DFT) calculations were used to explore the degradation mechanism. For both DMP and DEP, the reactive oxygen species (ROS, including O 2 − and OH) were responsible for the hydroxylation of benzene ring and the breaking of the aliphatic chain, while h + was prone to break the aliphatic chain. This work is expected to provide new insights on the photocatalytic mechanism of stannate perovskites for environmental remediation. [Display omitted] • MgSnO 3 and CaSnO 3 perovskites were firstly used for photocatalytic degradation of PAEs. • MgSnO 3 perovskite exhibited better photocatalytic performance. • h + and O 2 − were the predominant radicals in photo-degradation. • Experiments and DFT calculations were combined to elucidate the degradation mechanism. [ABSTRACT FROM AUTHOR]

Multiple sclerosis is an immune-mediated, demyelinating and neurodegenerative disease that currently lacks any neuroprotective treatments. Innovative neuroprotective trial designs are required to hasten the translational process of drug development. An ideal target to monitor the efficacy of strategies aimed at treating multiple sclerosis is the visual system, which is the most accessible part of the human central nervous system. A novel C57BL/6 mouse line was generated that expressed transgenes for a myelin oligodendrocyte glycoprotein-specific T cell receptor and a retinal ganglion cell restricted-Thy1 promoter-controlled cyan fluorescent protein. This model develops spontaneous or induced optic neuritis, in the absence of paralytic disease normally associated with most rodent autoimmune models of multiple sclerosis. Demyelination and neurodegeneration could be monitored longitudinally in the living animal using electrophysiology, visual sensitivity, confocal scanning laser ophthalmoscopy and optical coherence tomography all of which are relevant to human trials. This model offers many advantages, from a 3Rs, economic and scientific perspective, over classical experimental autoimmune encephalomyelitis models that are associated with substantial suffering of animals. Optic neuritis in this model led to inflammatory damage of axons in the optic nerve and subsequent loss of retinal ganglion cells in the retina. This was inhibited by the systemic administration of a sodium channel blocker (oxcarbazepine) or intraocular treatment with siRNA targeting caspase-2. These novel approaches have relevance to the future treatment of neurodegeneration of MS, which has so far evaded treatment. [ABSTRACT FROM AUTHOR]

Optic nerve injuries cause death of retinal ganglion cells and degeneration of their axons. Vigneswara et al. show that combined delivery of caspase-2 and caspase-6 inhibitors into the eye promotes retinal ganglion cell survival and axon regeneration in a rodent model, an effect that is mediated through the JAK/STAT pathway.We have previously shown that crushing the optic nerve induces death of retinal ganglion cells by apoptosis, but suppression of CASP2, which is predominantly activated in retinal ganglion cells, using a stably modified short interfering RNA CASP2, inhibits retinal ganglion cell apoptosis. Here, we report that combined delivery of short interfering CASP2 and inhibition of CASP6 using a dominant negative CASP6 mutant activates astrocytes and Müller cells, increases CNTF levels in the retina and leads to enhanced retinal ganglion cell axon regeneration. In dissociated adult rat mixed retinal cultures, dominant negative CASP6 mutant + short interfering CASP2 treatment also significantly increases GFAP+ glial activation, increases the expression of CNTF in culture, and subsequently increases the number of retinal ganglion cells with neurites and the mean retinal ganglion cell neurite length. These effects are abrogated by the addition of MAB228 (a monoclonal antibody targeted to the gp130 component of the CNTF receptor) and AG490 (an inhibitor of the JAK/STAT pathway downstream of CNTF signalling). Similarly, in the optic nerve crush injury model, MAB228 and AG490 neutralizes dominant negative CASP6 mutant + short interfering CASP2-mediated retinal ganglion cell axon regeneration, Müller cell activation and CNTF production in the retina without affecting retinal ganglion cell survival. We therefore conclude that axon regeneration promoted by suppression of CASP2 and CASP6 is CNTF-dependent and mediated through the JAK/STAT signalling pathway. This study offers insights for the development of effective therapeutics for promoting retinal ganglion cell survival and axon regeneration. [ABSTRACT FROM PUBLISHER]

Albinism is a rare genetic condition globally characterized by a number of specific deficits in the visual system, resulting in poor vision, in association with a variable hypopigmentation phenotype. This lack or reduction in pigment might affect the eyes, skin, and hair (oculocutaneous albinism, OCA), or only the eyes (ocular albinism, OA). In addition, there are several syndromic forms of albinism (e.g. Hermansky-Pudlak and Chediak-Higashi syndromes, HPS and CHS, respectively) in which the described hypopigmented and visual phenotypes coexist with more severe pathological alterations. Recently, a locus has been mapped to the 4q24 human chromosomal region and thus represents an additional genetic cause of OCA, termed OCA5, while the gene is eventually identified. In addition, two new genes have been identified as causing OCA when mutated: SLC24A5 and C10orf11, and hence designated as OCA6 and OCA7, respectively. This consensus review, involving all laboratories that have reported these new genes, aims to update and agree upon the current gene nomenclature and types of albinism, while providing additional insights from the function of these new genes in pigment cells. [ABSTRACT FROM AUTHOR]

Human X-linked blue-cone monochromacy (BCM), a disabling congenital visual disorder of cone photoreceptors, is a candidate disease for gene augmentation therapy. BCM is caused by either mutations in the red ( OPN1LW) and green ( OPN1MW) cone photoreceptor opsin gene array or large deletions encompassing portions of the gene array and upstream regulatory sequences that would predict a lack of red or green opsin expression. The fate of opsin-deficient cone cells is unknown. We know that rod opsin null mutant mice show rapid postnatal death of rod photoreceptors. Using in vivo histology with high-resolution retinal imaging, we studied a cohort of 20 BCM patients (age range 5-58) with large deletions in the red/green opsin gene array. Already in the first years of life, retinal structure was not normal: there was partial loss of photoreceptors across the central retina. Remaining cone cells had detectable outer segments that were abnormally shortened. Adaptive optics imaging confirmed the existence of inner segments at a spatial density greater than that expected for the residual blue cones. The evidence indicates that human cones in patients with deletions in the red/green opsin gene array can survive in reduced numbers with limited outer segment material, suggesting potential value of gene therapy for BCM. [ABSTRACT FROM AUTHOR]

[Display omitted] • LDH and kaolin would affect the stability of PSNPs in aquatic environment. • IS and pH played significant roles in both homoaggregation and heteroaggregation. • Electrostatic interaction was the dominant force of PSNPs-minerals heteroaggregation. • DLVO theory and DFT calculations were used to elucidate the interaction mechanism. The transport, fate, and toxic effects of nanoplastics (NPs) would be affected by various environmental factors. In this study, the aggregation behaviors of polystyrene nanoplastics (PSNPs) with or without the presence of oppositely charged minerals (Mg/Al layered double hydroxides (LDH) and kaolin) in aqueous phase were investigated through settling experiment, and the influencing factors including pH and ionic strength (IS) were also analyzed systematically. Our results revealed that pH had a negligible effect on the fate of individual PSNPs under natural aquatic conditions, while high IS would cause homoaggregation. The positively charged LDH decreased the stability of PSNPs, while negatively charged kaolin had a weak effect on it, suggesting that electrostatic interaction was directly related to the stability of PSNPs in aquatic environment. Further experiments of PSNPs with other metal oxides with different surface charges (Al 2 O 3 and SiO 2) confirmed the importance of electrostatic interaction for PSNPs-minerals heteroaggregation. Transmission electron microscopic (TEM), Derjaguin–Landau–Verwey–Overbeek (DLVO) theory, and Density function theory (DFT) calculations were combined for the first time to explain the heteroaggregation mechanism, demonstrating that there were expected to exist hydrogen bond and van der Waals interaction in addition to the dominant force of electrostatic interaction. Our findings are expected to shed light on the environmental behavior of PSNPs in a complex aquatic environment. [ABSTRACT FROM AUTHOR]

Congenital cataracts (CC) are responsible for approximately one-tenth of childhood blindness cases globally. Here, we report an African American family with a recessively inherited form of CC. The proband demonstrated decreased visual acuity and bilateral cataracts, with nuclear and cortical cataracts in the right and left eye, respectively. Exome sequencing revealed a novel homozygous variant (c.563A > G; p.(Asn188Ser)) in GJA3, which was predicted to be pathogenic by structural analysis. Dominantly inherited variants in GJA3 are known to cause numerous types of cataracts in various populations. Our study represents the second case of recessive GJA3 allele, and the first report in African Americans. These results validate GJA3 as a bona fide gene for recessively inherited CC in humans. [ABSTRACT FROM AUTHOR]

Hearing loss (HL) is the most common neurosensory defect in humans that affects the normal communication. Disease is clinically and genetically heterogeneous, rendering challenges for the molecular diagnosis of affected subjects. This study highlights the phenotypic and genetic complexity of inherited HL in a large consanguineous Pakistan kindred. Audiological evaluation of all affected individuals revealed varying degree of mild to profound sensorineural HL. Whole exome (WES) of four family members followed by Sanger sequencing revealed candidate disease-associated variants in five known deafness genes: GJB2 (c.231G>A; p.(Trp77 *)), SLC26A4 (c.1337A>G; p.(Gln446Arg)), CDH23 (c.2789C>T; p.(Pro930Leu)), KCNQ4 (c.1672G>A; p.(Val558Met)) and MPDZ (c.4124T>C; p.(Val1375Ala)). All identified variants replaced evolutionary conserved residues, were either absent or had low frequencies in the control databases. Our in silico and 3-Dimensional (3D) protein topology analyses support the damaging impact of identified variants on the encoded proteins. However, except for the previously established "pathogenic" and "likely pathogenic" categories for the c.231G>A (p.(Trp77 *)) allele of GJB2 and c.1377A>G (p.(Gln446Arg)) of SLC26A4, respectively, all the remaining identified variants were classified as "uncertain significance" based on the American College of Medical Genetics and Genomics/Association for Molecular Pathology (ACMG/AMP) variant pathogenicity guidelines. Our study highlights the complexity of genetic traits in consanguineous families, and the need of combining the functional studies even with the comprehensive profiling of multiple family members to improve the genetic diagnosis in complex inbred families. [ABSTRACT FROM AUTHOR]

Green method have been used for fabrication of Graphene oxide/ Ginger extractive based lamellar membranes. Such membranes showed exellence selectvity of dye molecules (˃99%) in both aqeous and organic medium with extraordinary stability. [Display omitted] • Graphene oxide / Ginger extractive (GO/GE) based lamellar membranes were prepared using green route. • GO/GE membranes exhibited ultrafast permeability for DI water (permeance ∼ 420 ± 10 Lm-2h-1bar−1). • Such membranes showed an excellent selectivity for several organic dyes up to ∼ 100% for methyl blue. • As-prepared laminates were used for organic separation and exhibits excellent rejection toward rhodamine B up to ˃98% • Those laminates are highly stable under aqueous environment up to ∼ 70 days. Graphene oxide (GO) has attracted an enormous attention for membrane-based technology in wastewater treatment due to its supreme properties such as two-dimensional (2D) structure, good chemical and mechanical stabilities, and high antifouling. However, due to presence of several functional groups on its basal plane and edge, which hinder its applications in wastewater treatment. Herein, we prepared GO-based lamellar membranes from GO and ginger plant extractive (GE) with various thicknesses (∼280 ± 20 nm, ∼500 ± 30 nm, ∼780 ± 30 nm and ∼ 1130 ± 30 nm) by using vacuum filtration method. The GO/GE lamellar membrane (∼280 ± 10 nm) showed an excellent separation efficiency for various organic dyes with ultrafast permeability i.e. rhodamine B (RB, Rejection 99 ± 1% and permeance 320 ± 10 Lm-2h-1bar−1) and methyl blue (MB, Rejection 99 ± 1% and permeance 210 ± 10 Lm-2h-1bar−1). Further, the GO/GE lamellar membrane also shows high water permeance ∼ 420 ± 10 Lm-2h-1bar−1. In addition, these membranes also used for organic separation and results show good separation efficiency of dye molecules in organic solvent (methanol) and exhibited high rejection (˃98%) for both RB and MB dye. This high separation efficiency against several dye molecules with good permeance and stability enlighten the great potential of our GO/GE lamellar membranes in water purification, desalination and other nanofiltration applications. [ABSTRACT FROM AUTHOR]

Backscatter scanning electron microscopy and conventional whole cell patch-clamp experiments reveal a two-step mechanism for the regeneration of tip links, the crucial element of mechanotransduction machinery in the hair cells of the inner ear. [ABSTRACT FROM AUTHOR]

A letter to the editor is presented about the identification of four Pakistani albinism mutations.

Sciatic nerve (SN) transection-induced apoptosis of dorsal root ganglion neurons (DRGN) is one factor determining the efficacy of peripheral axonal regeneration and the return of sensation. Here, we tested the hypothesis that caspase-2 (CASP2) orchestrates apoptosis of axotomised DRGN both in vivo and in vitro by disrupting the local neurotrophic supply to DRGN. We observed significantly elevated levels of cleaved CASP2 (C-CASP2), compared to cleaved caspase-3 (C-CASP3), within TUNEL+DRGN and DRG glia (satellite and Schwann cells) after SN transection. A serum withdrawal cell culture model, which induced 40% apoptotic death in DRGN and 60% in glia, was used to model DRGN loss after neurotrophic factor withdrawal. Elevated C-CASP2 and TUNEL were observed in both DRGN and DRG glia, with C-CASP2 localisation shifting from the cytosol to the nucleus, a required step for induction of direct CASP2-mediated apoptosis. Furthermore, siRNA-mediated downregulation of CASP2 protected 50% of DRGN from apoptosis after serum withdrawal, while downregulation of CASP3 had no effect on DRGN or DRG glia survival. We conclude that CASP2 orchestrates the death of SN-axotomised DRGN directly and also indirectly through loss of DRG glia and their local neurotrophic factor support. Accordingly, inhibiting CASP2 expression is a potential therapy for improving both the SN regeneration response and peripheral sensory recovery. [ABSTRACT FROM AUTHOR]

Severing the axons of retinal ganglion cells (RGC) by crushing the optic nerve (ONC) causes the majority of RGC to degenerate and die, primarily by apoptosis. We showed recently that after ONC in adult rats, caspase-2 activation occurred specifically in RGC while no localisation of caspase-3 was observed in ganglion cells but in cells of the inner nuclear layer. We further showed that inhibition of caspase-2 using a single injection of stably modified siRNA to caspase-2 protected almost all RGC from death at 7 days, offering significant protection for up to 1 month after ONC. In the present study, we confirmed that cleaved caspase-2 was localised and activated in RGC (and occasional neurons in the inner nuclear layer), while TUNEL+ RGC were also observed after ONC. We then investigated if suppression of caspase-2 using serial intravitreal injections of the pharmacological inhibitor z-VDVAD-fmk (z-VDVAD) protected RGC from death for 15 days after ONC. Treatment of eyes with z-VDVAD suppressed cleaved caspase-2 activation by =85% at 3-4 days after ONC. Increasing concentrations of z-VDVAD protected greater numbers of RGC from death at 15 days after ONC, up to a maximum of 60% using 4000 ng/ml of z-VDVAD, compared to PBS treated controls. The 15-day treatment with 4000 ng/ml of z-VDVAD after ONC suppressed levels of cleaved caspase-2 but no significant changes in levels of cleaved caspase-3, -6, -7 or -8 were detected. Although suppression of caspase-2 protected 60% of RGC from death, RGC axon regeneration was not promoted. These results suggest that caspase-2 specifically mediates death of RGC after ONC and that suppression of caspase-2 may be a useful therapeutic strategy to enhance RGC survival not only after axotomy but also in diseases where RGC death occurs such as glaucoma and optic neuritis. [ABSTRACT FROM AUTHOR]

We ascertained two large Pakistani consanguineous families (PKDF231 and PKDF608) segregating profound hearing loss, vestibular dysfunction, and retinitis pigmentosa; the defining features of Usher syndrome type 1 (USH1). To date, seven USH1 loci have been reported. Here, we map a novel locus, USH1K, on chromosome 10p11.21-q21.1. In family PKDF231, we performed a genome-wide linkage screen and found a region of homozygosity shared among the affected individuals at chromosome 10p11.21-q21.1. Meiotic recombination events in family PKDF231 define a critical interval of 11.74 cM (20.20 Mb) bounded by markers D10S1780 (63.83 cM) and D10S546 (75.57 cM). Affected individuals of family PKDF608 were also homozygous for chromosome 10p11.21-q21.1-linked STR markers. Of the 85 genes within the linkage interval, PCDH15, GJD4, FZD4, RET and LRRC18 were sequenced in both families, but no potential pathogenic mutation was identified. The USH1K locus overlaps the non-syndromic deafness locus DFNB33 raising the possibility that the two disorders may be caused by allelic mutations. [ABSTRACT FROM AUTHOR]

Abstract: Ethnopharmacological relevance: Viscum album L. is claimed in traditional medical practice, to be useful in the treatment of epilepsy and insomnia in Himachal Pradesh, India. Materials and methods: The effect of Viscum album L. on epilepsy, psychosis and sedative activity was evaluated in mice and rats using standard procedure. Results: The aqueous leaf extract of Viscum album L. prolonged the pentobarbital induced sleeping time and reduced the locomotor activity in actophotometer. This suggests that reduced locomotor activity facilitate GABAergic transmission. In addition the extract reduced MES, INH and PTZ-induced convulsions which suggest that there may be possibility of blocking Na+ channels, opening of Cl−channels or enhancing the GABAergic system. The extract decreased the apomorphine-induced stereotyped behavior and potentiates the HAL-induced cataleptic score which suggests the extract possess antidopaminergic activity. Conclusion: The results obtained in present study suggested that title plant exhibited sedative, antiepileptic and antipsychotic activity in mice and rats. [Copyright &y& Elsevier]

Background: Oculocutaneous albinism (OCA) is caused by a group of genetically heterogeneous inherited defects that result in the loss of pigmentation in the eyes, skin and hair. Mutations in the TYR, OCA2, TYRP1 and SLC45A2 genes have been shown to cause isolated OCA. No comprehensive analysis has been conducted to study the spectrum of OCA alleles prevailing in Pakistani albino populations. Methods: We enrolled 40 large Pakistani families and screened them for OCA genes and a candidate gene, SLC24A5. Protein function effects were evaluated using in silico prediction algorithms and ex vivo studies in human melanocytes. The effects of splice-site mutations were determined using an exon-trapping assay. Results: Screening of the TYR gene revealed four known (p.Arg299His, p.Pro406Leu, p.Gly419Arg, p.Arg278*) and three novel mutations (p.Pro21Leu, p.Cys35Arg, p.Tyr411His) in ten families. Ex vivo studies revealed the retention of an EGFP-tagged mutant (p.Pro21Leu, p.Cys35Arg or p.Tyr411His) tyrosinase in the endoplasmic reticulum (ER) at37°C, but a significant fraction of p.Cys35Arg and p.Tyr411His left the ER in cells grown at a permissive temperature (31°C). Three novel (p.Asp486Tyr, p.Leu527Arg, c.1045-15 T > G) and two known mutations (p.Pro743Leu, p. Ala787Thr) of OCA2 were found in fourteen families. Exon-trapping assays with a construct containing a novel c.1045-15 T > G mutation revealed an error in splicing. No mutation in TYRP1, SLC45A2, and SLC24A5 was found in the remaining 16 families. Clinical evaluation of the families segregating either TYR or OCA2 mutations showed nystagmus, photophobia, and loss of pigmentation in the skin or hair follicles. Most of the affected individuals had grayish-blue colored eyes. Conclusions: Our results show that ten and fourteen families harbored mutations in the TYR and OCA2 genes, respectively. Our findings, along with the results of previous studies, indicate that the p.Cys35Arg, p.Arg278* and p.Gly419Arg alleles of TYR and the p.Asp486Tyr and c.1045-15 T > G alleles of OCA2 are the most common causes of OCA in Pakistani families. To the best of our knowledge, this study represents the first documentation of OCA2 alleles in the Pakistani population. A significant proportion of our cohort did not have mutations in known OCA genes. Overall, our study contributes to the development of genetic testing protocols and genetic counseling for OCA in Pakistani families. [ABSTRACT FROM AUTHOR]

A missense mutation of Gipc3 was previously reported to cause age-related hearing loss in mice. Point mutations of human GIPC3 were found in two small families, but association with hearing loss was not statistically significant. Here, we describe one frameshift and six missense mutations in GIPC3 cosegregating with DFNB72 hearing loss in six large families that support statistically significant evidence for genetic linkage. However, GIPC3 is not the only nonsyndromic hearing impairment gene in this region; no GIPC3 mutations were found in a family cosegregating hearing loss with markers of chromosome 19p. Haplotype analysis excluded GIPC3 from the obligate linkage interval in this family and defined a novel locus spanning 4.08 Mb and 104 genes. This closely linked but distinct nonsyndromic hearing loss locus was designated DFNB81. [ABSTRACT FROM AUTHOR]

Warburg Micro syndrome and Martsolf syndrome are heterogenous autosomal-recessive developmental disorders characterized by brain, eye, and endocrine abnormalities. Previously, identification of mutations in RAB3GAP1 and RAB3GAP2 in both these syndromes implicated dysregulation of the RAB3 cycle (which controls calcium-mediated exocytosis of neurotransmitters and hormones) in disease pathogenesis. RAB3GAP1 and RAB3GAP2 encode the catalytic and noncatalytic subunits of the hetrodimeric enzyme RAB3GAP (RAB3GTPase-activating protein), a key regulator of the RAB3 cycle. We performed autozygosity mapping in five consanguineous families without RAB3GAP1/2 mutations and identified loss-of-function mutations in RAB18. A c.71T > A (p.Leu24Gln) founder mutation was identified in four Pakistani families, and a homozygous exon 2 deletion (predicted to result in a frameshift) was found in the fifth family. A single family whose members were compound heterozygotes for an anti-termination mutation of the stop codon c.619T > C (p.X207QextX20) and an inframe arginine deletion c.277_279 del (p.Arg93 del) were identified after direct gene sequencing and multiplex ligation-dependent probe amplification (MLPA) of a further 58 families. Nucleotide binding assays for RAB18(Leu24Gln) and RAB18(Arg93del) showed that these mutant proteins were functionally null in that they were unable to bind guanine. The clinical features of Warburg Micro syndrome patients with RAB3GAP1 or RAB3GAP2 mutations and RAB18 mutations are indistinguishable, although the role of RAB18 in trafficking is still emerging, and it has not been linked previously to the RAB3 pathway. Knockdown of rab18 in zebrafish suggests that it might have a conserved developmental role. Our findings imply that RAB18 has a critical role in human brain and eye development and neurodegeneration. [ABSTRACT FROM AUTHOR]