Autosomal recessive non-syndromic hearing loss genes in Pakistan during the previous three decades.

Hearing loss is a clinically and genetically heterogeneous disorder, with over 148 genes and 170 loci associated with its pathogenesis. The spectrum and frequency of causal variants vary across different genetic ancestries and are more prevalent in populations that practice consanguineous marriages. Pakistan has a rich history of autosomal recessive gene discovery related to non-syndromic hearing loss. Since the first linkage analysis with a Pakistani family that led to the mapping of the DFNB1 locus on chromosome 13, 51 genes associated with this disorder have been identified in this population. Among these, 13 of the most prevalent genes, namely CDH23, CIB2, CLDN14, GJB2, HGF, MARVELD2, MYO7A, MYO15A, MSRB3, OTOF, SLC26A4, TMC1 and TMPRSS3, account for more than half of all cases of profound hearing loss, while the prevalence of other genes is less than 2% individually. In this review, we discuss the most common autosomal recessive non-syndromic hearing loss genes in Pakistani individuals as well as the genetic mapping and sequencing approaches used to discover them. Furthermore, we identified enriched gene ontology terms and common pathways involved in these 51 autosomal recessive non-syndromic hearing loss genes to gain a better understanding of the underlying mechanisms. Establishing a molecular understanding of the disorder may aid in reducing its future prevalence by enabling timely diagnostics and genetic counselling, leading to more effective clinical management and treatments of hearing loss.

A circuit for secretion-coupled cellular autonomy in multicellular eukaryotic cells.

Cancers represent complex autonomous systems, displaying self-sufficiency in growth signaling. Autonomous growth is fueled by a cancer cell's ability to "secrete-and-sense" growth factors (GFs): a poorly understood phenomenon. Using an integrated computational and experimental approach, here we dissect the impact of a feedback-coupled GTPase circuit within the secretory pathway that imparts secretion-coupled autonomy. The circuit is assembled when the Ras-superfamily monomeric GTPase Arf1, and the heterotrimeric GTPase Giαßγ and their corresponding GAPs and GEFs are coupled by GIV/Girdin, a protein that is known to fuel aggressive traits in diverse cancers. One forward and two key negative feedback loops within the circuit create closed-loop control, allow the two GTPases to coregulate each other, and convert the expected switch-like behavior of Arf1-dependent secretion into an unexpected dose-response alignment behavior of sensing and secretion. Such behavior translates into cell survival that is self-sustained by stimulus-proportionate secretion. Proteomic studies and protein-protein interaction network analyses pinpoint GFs (e.g., the epidermal GF) as key stimuli for such self-sustenance. Findings highlight how the enhanced coupling of two biological switches in cancer cells is critical for multiscale feedback control to achieve secretion-coupled autonomy of growth factors.

Multinuclear Zinc-Magnesium Hydroxide Carboxylates: A Predesigned Model System for Copolymerization of CO2 with Epoxides.

Among numerous catalysts in the ring-opening copolymerization of epoxides with carbon dioxide (CO2), zinc dicarboxylate complexes are the most common type, and in the family of metal-based homogeneous catalysts, zinc and magnesium complexes have attracted widespread attention. We report on the synthesis and structural characterization of a zinc-magnesium benzoate framework templated by the central hydroxide anion with µ3-κ2:κ2:κ2 coordination mode, [ZnMg2(µ3-OH)(O2CPh)5]n (n = 1 or 2). The resulting heterometallic system forms stable Lewis acid-base adducts with tetrahydrofuran (THF) and cyclohexene oxide (CHO), which crystallize as the hexanuclear zinc-magnesium hydroxide carboxylate cluster [ZnMg2(µ3-OH)(O2CPh)5(L)2]2 (L = THF or CHO). Their X-ray crystal structure analysis revealed that the Zn center prefers 4-fold coordination and the Mg centers demonstrated the ability to accommodate higher coordination numbers, and as a result, the heterocyclic molecules are exclusively bonded to 6-fold Mg atoms. The heteronuclear carboxylate aggregates appeared active in the copolymerization reaction at elevated temperatures to produce an alternating poly(cyclohexene carbonate).

Effect of Unsaturated Metal Site Modulation in Highly Stable Microporous Materials on CO2 Capture and Fixation.

We have designed and synthesized two unprecedented microporous three-dimensional metal-organic frameworks, {[Cd6(TPOM)3(L)6]·12DMF·3H2O}n (1) and {[Zn2(TPOM)(L)2]·2DMF·H2O}n (2), based on a flexible quadritopic ligand, tetrakis(4-pyridyloxymethylene)methane (TPOM), and a bent dicarboxylic acid, 4,4'-(dimethylsilanediyl)bis-benzoic acid (H2L). The networks of 1 and 2 share a 4-c uninodal net NbO topology but exhibit different metal environments due to coordination preferences of Cd(II) and Zn(II). The Cd(II) center in 1 is six-coordinated, whereas the Zn(II) center in 2 is only four-coordinated, making the latter an unsaturated metal center. Such modulation of coordination atmosphere of metal centers in MOFs with the same topology is possible due to diverse binding of the carboxylate groups of L2-. Both 1 and 2 have relatively high thermal stability and exhibit permanent porosity after the removal of guest solvent molecules based on variable temperature powder X-ray diffraction and gas adsorption analysis. These materials exhibit similar gas adsorption properties, especially highly selective CO2 uptake/capture over other gases (N2 and CH4). However, because of the presence of an unsaturated Lewis acidic metal site, 2 acts as a very efficient heterogeneous catalyst toward the chemical conversion of CO2 to cyclic carbonates under mild conditions, whereas 1 shows very less activity. This work provides experimental evidence for the postulate that an unsaturated metal site in MOFs enhances adsoprtion of CO2 and promotes its conversion via the Lewis-acid catalysis.

Structural and Biophysical Characterization of Stable Alpha-Synuclein Oligomers.

The aggregation of α-synuclein (α-syn) into neurotoxic oligomers and fibrils is an important pathogenic feature of synucleinopatheis, including Parkinson's disease (PD). A further characteristic of PD is the oxidative stress that results in the formation of aldehydes by lipid peroxidation. It has been reported that the brains of deceased patients with PD contain high levels of protein oligomers that are cross-linked to these aldehydes. Increasing evidence also suggests that prefibrillar oligomeric species are more toxic than the mature amyloid fibrils. However, due to the heterogenous and metastable nature, characterization of the α-syn oligomeric species has been challenging. Here, we generated and characterized distinct α-syn oligomers in vitro in the presence of DA and lipid peroxidation products 4-hydroxy-2-nonenal (HNE) and 4-oxo-2-nonenal (ONE). HNE and ONE oligomer were stable towards the treatment with SDS, urea, and temperature. The secondary structure analysis revealed that only HNE and ONE oligomers contain ß-sheet content. In the seeding assay, both DA and ONE oligomers significantly accelerated the aggregation. Furthermore, all oligomeric preparations were found to seed the aggregation of α-syn monomers in vitro and found to be cytotoxic when added to SH-SY5Y cells. Finally, both HNE and ONE α-syn oligomers can be used as a calibrator in an α-syn oligomers-specific ELISA.

Outcome of surgical condition of neonates who underwent surgery: A prospective study from a tertiary care center.

Background: The neonatal period is a highly vulnerable time for an infant, who is completing many of the physiologic adjustments required for extra- uterine survival. If the neonate has a coexisting pathology which needs surgery, this challenge is magnified. Neonatal surgical conditions are unique in their type because some require early diagnosis, prompt surgery and postoperative care to improve the survival and outcome. Objective: The aim of this study was to know the clinical profile of congenital surgical conditions and to estimate the burden and outcome in special new born care unit. Method: The study population include 138 surgical neonate admitted in special new born care unit, department of paediatrics, Kamla Raja Hospital, Gajra Raja Medical College, Gwalior (M.P.) from April 2017 to April 2018 including six month follow up period. Results: Total admission in special new born care unit were 5378 out of which 138 (2.5%) neonates of surgical condition were admitted in the study period. Incidence of neonatal surgical condition was found to be 8.48%. Surgery was performed in 57 (41.30%) neonates. The Commonest neonatal surgical condition was constituted by gastrointestinal system (39.13%). Among gastrointestinal system anomalies, tracheoesophageal fistula were 28.6% of total gastrointestinal system cases. The most common surgical condition encountered was meningomyelocele, 23.36% of total cases. The survival of neonatal surgical condition in hospital was 52.89% and after six month follow up was 26.08%. The overall mortality with neonatal surgical condition in this study was 73.91%. Maternal age, antenatal care, history of congenital malformation, socioeconomic status, mode of delivery, prematurity, type of admission, single or multiple surgical condition, inotropic and ventilation support, post operative complication were significantly associated with final outcome of neonatal surgical condition. Conclusion: High mortality was found in neonates suffering from surgical conditions. Commonest anomaly includes conditions of gastrointestinal tract. Maternal age more than 35 year, poor antenatal care, prematurity, vaginal delivery, extra mural neonate, multiple surgical condition, inotropic and ventilation support and post operative complications were associated with increased mortality.

Correction of Niemann-Pick type C1 trafficking and activity with the histone deacetylase inhibitor valproic acid.

Niemann-Pick type C (NPC) disease is primarily caused by mutations in the NPC1 gene and is characterized by the accumulation of unesterified cholesterol and lipids in the late endosomal (LE) and lysosomal (Ly) compartments. The most prevalent disease-linked mutation is the I1061T variant of NPC1, which exhibits defective folding and trafficking from the endoplasmic reticulum to the LE/Ly compartments. We now show that the FDA-approved histone deacetylase inhibitor (HDACi) valproic acid (VPA) corrects the folding and trafficking defect associated with I1061T-NPC1 leading to restoration of cholesterol homeostasis, an effect that is largely driven by a reduction in HDAC7 expression. The VPA-mediated trafficking correction is in part associated with an increase in the acetylation of lysine residues in the cysteine-rich domain of NPC1. The HDACi-mediated correction is synergistically improved by combining it with the FDA-approved anti-malarial, chloroquine, a known lysosomotropic compound, which improved the stability of the LE/Ly-localized fraction of the I1061T variant. We posit that combining the activity of VPA, to modulate epigenetically the cellular acetylome, with chloroquine, to alter the lysosomal environment to favor stability of the trafficked I1061T variant protein can have a significant therapeutic benefit in patients carrying at least one copy of the I1061T variant of NPC1, the most common disease-associated mutation leading to NPC disease. Given its ability to cross the blood-brain barrier, we posit VPA provides a potential mechanism to improve the response to 2-hydroxypropyl-ß-cyclodextrin, by restoring a functional NPC1 to the cholesterol managing compartment as an adjunct therapy.

Spatial modeling of mid-infrared spectral data with thermal compensation using integrated nested Laplace approximation.

The problem of analyzing substances using low-cost sensors with a low signal-to-noise ratio (SNR) remains challenging. Using accurate models for the spectral data is paramount for the success of any classification task. We demonstrate that the thermal compensation of sample heating and spatial variability analysis yield lower modeling errors than non-spatial modeling. Then, we obtain the inference of the spectral data probability density functions using the integrated nested Laplace approximation (INLA) on a Bayesian hierarchical model. To achieve this goal, we use the fast and user-friendly R-INLA package in R for the computation. This approach allows affordable and real-time substance identification with fewer SNR sensor measurements, thereby potentially increasing throughput and lowering costs.

Production and characterization of cow milk based low-protein milk protein concentrate (MPC) powders.

Ultrafiltration and Diafiltration processes are used to concentrate proteins present in defatted milk in order to manufacture milk protein concentrate (MPC) powders. Selective passage of the water-soluble components causes retention as well as concentration of colloidal milk components in these processes. Increase in calcium and casein contents decreases the stability of milk proteins present in ultrafiltered retentates and negatively influence properties of manufactured MPC powders. Homogenization, diafiltration and disodium phosphate induced changes in properties of low-protein MPC powders were targeted in this study. Applied treatments significantly (P < 0.05) improved foaming and emulsification, solubility, viscosity, heat stability, dispersibility, specific surface area and buffer index of resultant MPC powders over control. Fresh, treated low-protein MPC powders showed significantly higher (< 0.05) solubility values over control sample, which remains higher even after 60 days of storage at 25 ± 1 °C. The rheological behaviour of reconstituted low-protein MPC solutions was also studied. It was best explained as Herschel-Bulkley rheological behaviour. Low-protein MPC powders with improved functional properties may find better use as a protein ingredient in different dairy and food applications.

A Highly Stable Triazole-Functionalized Metal-Organic Framework Integrated with Exposed Metal Sites for Selective CO2 Capture and Conversion.

A new triazole-functionalized tetracarboxylic acid ligand (H4 L) has been synthesized and utilized for the fabrication of a 3D ZnII organic framework with a Zn4 (-COO)6 cluster as the secondary building unit. The framework exhibits very good thermal stability and consists of dual functionalities of exposed Lewis acidic metal sites and accessible nitrogen-donor Lewis basic sites. The Lewis basic nitrogen sites in the framework serve as CO2 binding sites for highly selective CO2 capture and the presence of exposed Lewis acidic metal sites in the framework make it an efficient heterogeneous catalyst for the chemical fixation of CO2 into value-added cyclic carbonates under ambient conditions.

A Microporous Metal-Organic Framework Catalyst for Solvent-free Strecker Reaction and CO2 Fixation at Ambient Conditions.

The self-assembly of zinc(II) acetate tetrahydrate, a flexible tetrapyridyl ligand, tetrakis(3-pyridyloxymethylene)methane (3-tpom), a bent dicarboxylic acid, and 4,4'-(dimethylsilanediyl)bis- benzoic acid (H2L) under solvothermal conditions has resulted in the formation of a microporous zinc(II)-organic framework, {[Zn2(3-tpom)(L)2]·2H2O}n (1). The framework exhibits very good thermal stability as evident from the thermogravimetric analysis, which is further supported by variable temperature powder X-ray diffraction analysis. The microporous nature of the framework has been established by the gas adsorption analysis. The framework exhibits exceptionally selective carbon dioxide adsorption in contrast with other gases having comparatively larger kinetic diameters (3.64 Å for N2 and 3.8 Å for CH4) under ambient conditions (298 K and 1 bar pressure). Further, the framework decorated with catalytically active unsaturated metal sites acts as a good catalyst toward the cycloaddition reaction of CO2 with epoxides and the three-component Strecker reaction at ambient conditions and without the requirement of any solvent. The heterogeneous nature along with good catalytic activity at ambient and solvent-free conditions entitles 1 as an excellent catalyst for these organic transformations.

Microplastics from lagooning sludge to composts as revealed by fluorescent staining- image analysis, Raman spectroscopy and pyrolysis-GC/MS.

Lagooning sludge (LS), which is used as soil amendment in Morocco, may contain microplastics (MPs). The aim of this study was to examine the effect of dewatering and co-composting of LS with green waste (GW) on the MPs' evolution. In this context the present study proposes fast-preliminary steps to detect plastics in lagooning sewage sludge before the extraction and identification process. We used pyrolysis GC/MS spectrometry to investigate the presence of chemical compounds possibly derived from plastics, and fluorescence staining by Nile Red to detect fluorescent particles suspected as plastics. Thereafter, we quantified the MPs particles after density fractionation and investigated their nature by Raman spectroscopy. RESULTS: indicated the presence of an average of 40.5 ± 11.9 × 103 MPs particles/kg (dry matter) and 36 ± 9.7 × 103 MPs particles/kg (dry matter) in fresh sludge and dewatered sludge respectively. Sludge dewatering in drying beds resulted a loss of small MPs (<500 µm). In co-composts, the quantity of MPs varied with the proportion of sewage sludge. The distribution of MPs types differentiated by colour and types (polypropylene, polyethylene, polyamide and polyester) evolved differently. Conventional co-composting did not have any effect on MPs quantity, indicating that they are not biodegradable under these temperature conditions, but it influenced their particle size. The risks of these pollutants after repeated field application and the possibility of their reduction through others co-composting procedures and techniques would be further investigated.

Antibodies against alpha-synuclein: tools and therapies.

Synucleinopathies including Parkinson's disease, dementia with Lewy bodies and multiple system atrophy are characterized by the abnormal accumulation and propagation of α-synuclein (α-syn) pathology in the central and peripheral nervous system as Lewy bodies or glial cytoplasmic inclusions. Several antibodies against α-syn have been developed since it was first detected as the major component of Lewy bodies and glial cytoplasmic inclusions. Over the years, researchers have generated specific antibodies that alleviate the accumulation of intracellular aggregated α-syn and associated pathology in cellular and preclinical models of synucleinopathies. So far, antibodies have been the first choice as tools for research and diagnosis and currently, a wide variety of antibody fragments have been developed as an alternative to full-length antibodies for increasing its therapeutic usefulness. Recently, conformation specific antibody-based approaches have been found to be promising as therapeutic strategies, both to block α-syn aggregation and ameliorate the resultant cytotoxicity, and as diagnostic tools. In this review, we summarize different α-syn specific antibodies and provide their usefulness in tackling synucleinopathies. This article is part of the Special Issue "Synuclein".

Design and Construction of a Chiral Cd(II)-MOF from Achiral Precursors: Synthesis, Crystal Structure and Catalytic Activity toward C-C and C-N Bond Forming Reactions.

Using achiral components, a V-shaped dicarboxylic acid (H2L) and a conformationally flexible bidentate linker (bpp), a thermally stable chiral metal organic framework {[Cd(bpp)(L)(H2O)]·DMF} n (1), where H2L = 4,4'-(dimethylsilanediyl)bis-benzoic acid, bpp = 1,3-bis(4-pyridyl)propane and DMF = N,N-dimethylformamide, has been solvothermally synthesized and crystallographically characterized. It consists of 1D helical chains linked at the cadmium centers resulting in an overall 2D framework. Its microporous nature was confirmed by gas-sorption measurements. Upon thermal activation of 1, where both guest DMF molecules present in the 1D open channels and the coordinated H2O molecules are removed, its active metal site shows Lewis acid character to be an excellent heterogeneous catalyst for the C-C (Knoevenagel condensation reaction) and C-N (Strecker) bond forming reactions.

Diastereoselective multi-component tandem condensation: synthesis of 2-amino-4-(2-furanone)-4H-chromene-3-carbonitriles.

A general strategy for a one-pot stereoselective synthesis of 2-amino-4-(2-furanone)-4H-chromene-3-carbonitriles by reaction of salicylaldehyde, malononitrile and butenolides via a tandem Knoevenagel/Pinner/vinylogous Michael condensation is presented. The ß,γ-butenolides gave a syn-selective MCR adduct with a dr up to 11.5 : 1. The mechanistic insight into the MCR was obtained by DFT calculations.

GIV/Girdin activates Gαi and inhibits Gαs via the same motif.

We previously showed that guanine nucleotide-binding (G) protein α subunit (Gα)-interacting vesicle-associated protein (GIV), a guanine-nucleotide exchange factor (GEF), transactivates Gα activity-inhibiting polypeptide 1 (Gαi) proteins in response to growth factors, such as EGF, using a short C-terminal motif. Subsequent work demonstrated that GIV also binds Gαs and that inactive Gαs promotes maturation of endosomes and shuts down mitogenic MAPK-ERK1/2 signals from endosomes. However, the mechanism and consequences of dual coupling of GIV to two G proteins, Gαi and Gαs, remained unknown. Here we report that GIV is a bifunctional modulator of G proteins; it serves as a guanine nucleotide dissociation inhibitor (GDI) for Gαs using the same motif that allows it to serve as a GEF for Gαi. Upon EGF stimulation, GIV modulates Gαi and Gαs sequentially: first, a key phosphomodification favors the assembly of GIV-Gαi complexes and activates GIV's GEF function; then a second phosphomodification terminates GIV's GEF function, triggers the assembly of GIV-Gαs complexes, and activates GIV's GDI function. By comparing WT and GIV mutants, we demonstrate that GIV inhibits Gαs activity in cells responding to EGF. Consequently, the cAMP→PKA→cAMP response element-binding protein signaling axis is inhibited, the transit time of EGF receptor through early endosomes are accelerated, mitogenic MAPK-ERK1/2 signals are rapidly terminated, and proliferation is suppressed. These insights define a paradigm in G-protein signaling in which a pleiotropically acting modulator uses the same motif both to activate and to inhibit G proteins. Our findings also illuminate how such modulation of two opposing Gα proteins integrates downstream signals and cellular responses.

Enterogenous Cyst Presenting as Mediastinal Mass in a Newborn.

Duplication cyst is a rare congenital malformation. Enterogenous cyst are foregut duplication cyst with or without a vertebral defect. We report a case of a 36hr old neonate with mediastinal enterogenous cyst associated with vertebral defects. The embryology, differentials, and management of enterogenous cyst in the newborn have been discussed.

Alteration in physicochemical, functional, rheological and reconstitution properties of milk protein concentrate powder by pH, homogenization and diafiltration.

Concentration of milk proteins by ultrafiltration (UF) and diafiltration (DF) processes during manufacturing of milk protein concentrate (MPC) powders alter their natural milk protein stabilization system. Increasing calcium and protein contents often leads to poor functional properties in MPC powders. The pH adjustment using disodium phosphate (DSP, Na2HPO4) and DF with 150 mM NaCl solution of UF retentate were hypothesized to produce desirable changes in various properties of resulted MPC powders. Addition of Na2HPO4 followed by homogenization; DF of 5 × UF retentate with 150 mM NaCl solution resulted in significant improvement in the dispersibility, wettability, flowability, solubility, heat stability, buffer index, emulsification and foaming and water and oil binding capacities of the MPC powders. The solubility of developed MPC powders was significantly higher than MPC-C powder in fresh as well as even after 90 days of storage at 25 ± 1 °C. Rheological behaviour of reconstituted MPC was best explained by Herschel Bulkley model. Scanning electron microscopy micrograph indicated that MPC powders were having smooth surfaced, intact and separate smaller particles compared to rough, larger, infused aggregates with dents in MPC-C. Technological interventions applied are easier to adopt, cost-effective and efficient in producing excellent quality MPC powders that may find applications in wide range of novel food formulations.

Facile synthesis of triphenylenes and triphenylene/phenanthrene fused heteroaromatics.

In the recent past, development of milder, safer and comparatively less toxic methods in organic synthesis has received great attention. We report a simple and facile method for the synthesis of substituted triphenylenes and their heteroaryl analogues using ceric ammonium nitrate (CAN) via oxidative biaryl coupling. We describe the structural and photophysical properties of these unique heteroaryl fused triphenylenes.

Cyclin-dependent kinase 5 activates guanine nucleotide exchange factor GIV/Girdin to orchestrate migration-proliferation dichotomy.

Signals propagated by receptor tyrosine kinases (RTKs) can drive cell migration and proliferation, two cellular processes that do not occur simultaneously--a phenomenon called "migration-proliferation dichotomy." We previously showed that epidermal growth factor (EGF) signaling is skewed to favor migration over proliferation via noncanonical transactivation of Gαi proteins by the guanine exchange factor (GEF) GIV. However, what turns on GIV-GEF downstream of growth factor RTKs remained unknown. Here we reveal the molecular mechanism by which phosphorylation of GIV by cyclin-dependent kinase 5 (CDK5) triggers GIV's ability to bind and activate Gαi in response to growth factors and modulate downstream signals to establish a dichotomy between migration and proliferation. We show that CDK5 binds and phosphorylates GIV at Ser1674 near its GEF motif. When Ser1674 is phosphorylated, GIV activates Gαi and enhances promigratory Akt signals. Phosphorylated GIV also binds Gαs and enhances endosomal maturation, which shortens the transit time of EGFR through early endosomes, thereby limiting mitogenic MAPK signals. Consequently, this phosphoevent triggers cells to preferentially migrate during wound healing and transmigration of cancer cells. When Ser1674 cannot be phosphorylated, GIV cannot bind either Gαi or Gαs, Akt signaling is suppressed, mitogenic signals are enhanced due to delayed transit time of EGFR through early endosomes, and cells preferentially proliferate. These results illuminate how GIV-GEF is turned on upon receptor activation, adds GIV to the repertoire of CDK5 substrates, and defines a mechanism by which this unusual CDK orchestrates migration-proliferation dichotomy during cancer invasion, wound healing, and development.