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Titanium aluminum nitride (TiAlN) is proposed as a coating to decrease the reflectivity of surgical instrumentation, thereby improving the field of vision of surgeons during procedures, although the corrosion resistance of the material against repeated sterilization processes must be guaranteed. The influence of aluminum (Al) on the microstructural and electrochemical properties of low-reflectivity titanium aluminum nitride coatings deposited by direct-current unbalanced magnetron sputtering was evaluated. The optical properties, chemical phases and composition were determined using the ultraviolet–visible spectroscopy, energy-dispersive X-ray spectroscopy, X-ray photoelectron spectroscopy and X-ray diffraction techniques. The microstructures were analyzed by atomic force microscopy, scanning electron microscopy and transmission electron microscopy–selected area electron diffraction. The corrosion resistance was evaluated by electrochemical impedance spectroscopy and potentiodynamic polarization. It was found that an increase in the aluminum content reduced the coating reflectivity by 60%, showing a dense columnar crystalline structure with a grain size of 200–100 nm and roughness of 6.7–4.0 nm and the coexistence of titanium nitride (TiN), aluminum nitride (AlN), titanium aluminum nitride and aluminum oxide (Al2O3) phases. In comparison with AISI 420 steel, the TiAlN-1300W aluminum coating showed a more positive corrosion potential, as well as a current density two orders of magnitude lower, thus guaranteeing corrosion resistance.

Introduction Alterations in the expression of the Angiotensin II type 1 receptors (AT1R) have been demonstrated in the development of several heart and renal diseases. The aim of this study was to evaluate the novel compound [18F]fluoropyridine-candesartan as a PET imaging tracer of AT1R in rat kidneys. Methods Competition binding assays were carried out with membranes from CHO-K1 cells expressing human AT1R. Binding to plasma proteins was assessed by ultrafiltration. Radiolabeled metabolites in rat plasma and kidneys of control and pretreated animals (candesartan 10 mg/kg or losartan 30 mg/kg) were analyzed by column-switch HPLC. Dynamic PET/CT images of [18F]fluoropyridine-candesartan in male Sprague-Dawley rats were acquired for 60 min at baseline, pre-treatment with the AT1R antagonist losartan (30 mg/kg) or the AT2R antagonist PD123,319 (5 mg/kg). Results Fluoropyridine-candesartan bound with a high affinity for AT1R (Ki = 5.9 ± 1.1 nM), comparable to fluoropyridine-losartan but lower than the parent compound candesartan (Ki = 0.4 ± 0.1 nM). [18F]Fluoropyridine-candesartan bound strongly to plasma proteins (99.3%) and was mainly metabolized to radiolabeled hydrophilic compounds, displaying minimal interference on renal AT1R binding with 82% of unchanged tracer in the kidneys at 20 min post-injection. PET imaging displayed high renal and liver accumulations and slow clearances, with maximum tissue-to-blood ratios of 14 ± 3 and 54 ± 12 in kidney cortex and liver, respectively, at 10 min post-injection. Binding specificity for AT1R was demonstrated with marked reductions in kidney cortex (−84%) and liver (−93%) tissue-to-blood ratios at 20 min post-injection, when blocking with AT1R antagonist losartan (30 mg/kg). No change was observed in kidney cortex of rats pre-treated with AT2R antagonist PD 123,319 (5 mg/kg), confirming binding selectivity for AT1 over AT2 receptors. Conclusion High kidney-to-blood ratios and binding selectivity to renal AT1R combined with tracer in vivo stability displaying minimal interference from labeled metabolites support further PET imaging studies with [18F]fluoropyridine-candesartan.

Algae are known to produce biomass faster as compared with lignocellulosic biomass. BC was produced by using reducing sugar from algae as a cheaper carbon source. The effect of immobilization on the production of biocellulose(BC) by Gluconacetobacter xylinus ATCC 10245 was studied. The BC production of all entrapped cultures within different gel materials was lower than that of free cell cultures but the entrapped G. xylinus cells in alginate beads recorded the highest BC productivity (12.5 g/l). The adsorption of G. xylinus cells on luffa pulp (LP) and ceramics particles (CP) gave the highest BC production (14.5 g/l), which represents increasing of 1.14 and 1.05- fold, respectively from the free cells. This attempt clarified that LP was the best immobilizing support as it gave the highest BC production. The effect of repeated reuse of LP was confirmed. Different physical properties of BC including; pH, water absorption capacity, dissolving in several solvents, and chemicals were determined. Observation of BC through SEM analysis showed the fine cellulose ribbons (fibrils). The crystallinity for the BC produced from X-ray diffraction (XRD) spectra was 84%. Upon the applicable level, the BC aerogel was formed to be applied in rising up the water and oil absorption capability by developing its hydrophilic properties. Data introduced strong affinities to the motor, olive, and cooking oils with high oil uptake contents of 7.5, 5.2, and 3.7 gg−1, respectively. Moreover, the BC aerogel exhibited potential adsorption for different heavy metal ions Pb2+, Cd2+, and Cu2+. The highest capability of Cd2+ removal was 95.7% at pH 6. The biodegrability of our produced BC and then its safety was proven.

Two novel PHA producing bacterial strains were chosen out of 12 strains collected from Mariout salt lakes. Analysis of 16srRNA gene sequence of the two new strains revealed 95.38% and 98.78% similarity to that of Halomonas pacifica and Halomonas salifodiane, respectively. A maximum polymer productivity of 6.9 g/l and 7.1 g/l was recorded by ASL10 and ASL11, respectively. Furthermore, a pH of 7 contributed to the highest polymer production for both strains. Interestingly, both ASL10 and ASL11showed a great ability to tolerate salinity up to 17 g/l NaCL. Moreover, both promising isolates were able to degrade crude oil efficiently by degradation percentages of 69.2% and 67.3% for ASL10 and ASL11, respectively. GCMS, FTIR, NMR, XRD and thermal properties were performed for poly (3 HV-co-3HB) characterization.

A novel 7-((4-(3-((2-[18F]fluoropyridin-3-yl)oxy)propyl)-1H-1,2,3-triazol-1-yl)methyl)-1H-benzo[d]imidazole derivative of the angiotensin II type-1 receptor (AT1R) blocker candesartan, [18F]fluoropyridine–candesartan, was synthesized via the copper-catalyzed azide–alkyne cycloaddition click reaction between 2-[18F]fluoro-3-(pent-4-yn-1-yloxy)pyridine ([18F]FPyKYNE) and the tetrazole-protected azido-candesartan derivative, followed by acid deprotection. This three-step, two-pot, and two-step purification synthesis was done within 2 h. The use of tris[(1-hydroxypropyl-1H-1,2,3-triazol-4-yl)methyl]amine (THPTA) as a Cu(I) stabilizing agent increased the overall radiochemical yield by 4-fold (10 ± 2%, n = 13) compared to the reaction without THPTA (2.4 ± 0.2%, n = 3; decay-corrected from 18F produced at the end-of-beam). Complete separation of [18F]FPyKYNE from its nitro precursor and [18F]fluoropyridine–candesartan from the deprotected azido-candesartan allowed for high molar activities (>380 GBq/μmol) of the tracer. The use of 0.1% trifluoroacetic acid in water for reformulation and the addition of sodium ascorbate to the final formulation (1.6 ± 0.2 GBq/mL, n = 3) prevented tracer radiolysis with >97% radiochemical purity for a period of up to 10 h after the end-of-synthesis. A significant reduction in the uptake (86 ± 3%, n = 8) of the tracer was observed ex vivo in rats (at 20 min postinjection) in the AT1R-rich kidney cortex following pretreatment with saturating doses of the AT1R antagonist candesartan or losartan. This specific binding to AT1R was confirmed in vitro in the rat renal cortex (autoradiography) by a reduction of 26 ± 5% (n = 12) with losartan coincubation (10 μM). These favorable binding properties support further studies to assess the potential of [18F]fluoropyridine–candesartan as a tracer for the positron emission tomography imaging of renal AT1R.

Coenzyme Q10 (CoQ10) deficiency syndrome is a rare disease included in the family of mitochondrial diseases, which is a heterogeneous group of genetic disorders characterized by defective energy production. CoQ10 biosynthesis in humans requires at least 11 gene products acting in a multiprotein complex within mitochondria. The high-throughput screening (HTS) method based on the stabilization of the CoQ biosynthesis complex (Q-synthome) produced by the COQ8 gene overexpression is proven here to be a successful method for identifying new molecules from natural extracts that are able to bypass the CoQ6 deficiency in yeast mutant cells. The main features of the new approach are the combination of two yeast targets defective in genes with different functions on CoQ6 biosynthesis to secure the versatility of the molecule identified, the use of glycerol as a nonfermentable carbon source providing a wide growth window, and the stringent conditions required to mark an extract as positive. The application of this pilot approach to a representative subset of 1200 samples of the Library of Natural Products of Fundación MEDINA resulted in the finding of nine positive extracts. The fractionation of three of the nine extracts allowed the identification of five molecules; two of them are present in molecule databases of natural extracts and three are nondescribed molecules. The use of this screening method opens the possibility of discovering molecules with CoQ10-bypassing action useful as therapeutic agents to fight against mitochondrial diseases in human patients., Theresearch group is funded by the Andalusian Government as the BIO177 group, through FEDER funds (European Commission), from the Junta de Andalucía Proyectos de Excelencia P12 CTS943. The MEDINA authors disclosed receipt of financial support from Fundación MEDINA, a public–private partnership of Merck Sharp & Dohme de España S.A./Universidad de Granada/Junta de Andalucía.

This study was done to indicate the activity of secondary metabolites of cyanophyta algal extracts for Anabaena Azolla which live in Azolla plant in Egypt and to study the effective of secondary metabolites in An.Azolla extracts (cold water and methanol extracts) to inhibition species of fungi (Fusarium oxysporum, penicillium expansum, Aspergillus niger and candida albicans) also used of HPLC analysis to detected some of active secondary metabolites was done only for methanol extracts also to preliminary statements that used of two extracts of blue green alga Anabaena Azolla. The role of An.Azolla extracts to inhibition growth of these fungi was detected through measured diameter of inhibition zone diameters were, 16.58, 11.42, 11.0 & 11.0 mm in case of Fusarium oxysporum, Penicillium expansum, Aspergillus niger and candida albicans respectively, but aqueous extract give inhibition zone were 18.15, 14.13, 14.0 & 13.0mm respectively. and the results showed these extracts had high correlation ratio and fungi shows high sensitivity to these extract. The mode of mechanism action to inhibition was on fungi cell wall by inhibition active enzymes, excelled treatment for cold water extract . The most important active compounds had been detected in HPLC were phenolic compounds and flavonoids compounds these all compounds detected specially in methanol extract of An .Azolla. The experiment has been conducted under temperature and artificial illumination at growth room of Soilless Culture Laboratory, Sadat city University, Egypt. The results showed that green fodder can be produced in 8 days using hydroponic technique. Trays treated with Azolla gave more green fresh yield (11.8 kg/tray) than untreated ones (7.2 kg/tray). Adding Azolla increased protein content and the percentages of N, P, K by 28.7% and 0.98, 0.82, 0.70 %, respectively, as compared to no addition of Azolla. Zero disease incidences were achieved with treated by Azolla as compared to 26.0 % with those untreated by Azolla. It could be concluded from this study that adding Azolla to hydroponic barley forage production improved green forage yield and its chemical analysis and free from root rot fungi.

This study represents a detailed investigation of the influence of post-deposition annealing on structural, morphological and optical properties of 1, 4, 5, 8-naphthalene tetracarboxylic dianhydride, NTCDA, thin films. NTCDA thin films were prepared at room temperature by the thermal evaporation technique. The prepared films were annealed at 373 K and 473 K for 2 h in a vacuum. The results of TGA proved that NTCDA is thermally stable up to 519 K. The X-ray diffraction technique (XRD) was used to investigate the crystal structure of NTCDA powder and thin films. Polycrystalline films were obtained with a preferred orientation in the [100] direction along (h00) zone axis. The negation of the decomposition of NTCDA molecular structure during preparation and/or annealing required employing Fourier transformation infrared (FTIR) technique. The topological properties characterization of as-deposited and annealed NTCDA thin films surfaces concluded that the annealing process induced an enhancement in the film smoothness. The annealing process resulted in significant variations in the optical constants of the NTCDA films. The indirect energy gap values decreased from 3.52 eV to 3.48 eV as a result of annealing up to 473 K. The nonlinear optical effects represented by the nonlinear refractive index, n2 and third-order nonlinear optical susceptibility χ (3) were determined. The nonlinear properties of NTCDA showed a magnificent switching behavior, which suggested the ability to use NTCDA in optical switching devices.

Sphingomyelinases generate ceramide from sphingomyelin as a second messenger in intracellular signaling pathways involved in cell proliferation, differentiation, or apoptosis. Children from 12 unrelated families presented with microcephaly, simplified gyral pattern of the cortex, hypomyelination, cerebellar hypoplasia, congenital arthrogryposis, and early fetal/postnatal demise. Genomic analysis revealed bi-allelic loss-of-function variants in SMPD4, coding for the neutral sphingomyelinase-3 (nSMase-3/SMPD4). Overexpression of human Myc-tagged SMPD4 showed localization both to the outer nuclear envelope and the ER and additionally revealed interactions with several nuclear pore complex proteins by proteomics analysis. Fibroblasts from affected individuals showed ER cisternae abnormalities, suspected for increased autophagy, and were more susceptible to apoptosis under stress conditions, while treatment with siSMPD4 caused delayed cell cycle progression. Our data show that SMPD4 links homeostasis of membrane sphingolipids to cell fate by regulating the cross-talk between the ER and the outer nuclear envelope, while its loss reveals a pathogenic mechanism in microcephaly.

Tannase enzyme (EC 3.1.1.20) is an enzyme used in many biotechnological applications as in chemical, beverage, pharmaceutical and food industries. was isolated and purified from marine Aspergillus nomius GWA5 by 75% acetone fractional precipitation, followed by gel filtration in Sephadex G-100 and ion exchange chromatography on DEAE-Sephadex A-50 yielding 4.48-fold purification. Estimation of tannase molecular weight was carried out using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) showing a molecular weight of 30 kDa. The highest activity (291 U/mg protein) were at pH 6.0 and 50 °C, respectively. Tannase stability was observed in acidic range (4-6) and was stable to heat treatment. In absence of its substrate it retained about 84.5% of its activity at 80 °C for 15 min. Effect of some metal ions and chelator on tannase activity was investigated. Mg2+ activated as activator of the pure enzyme while EDTA, Cd2+, Pb2+ and Hg2+ inhibited its activity and retained about 40.78, 51.55, 30.24 and 24.55% of its activity, respectively. Promising activity of Tannase was shown in removing tannin stains of tea from clothes.

L-asparaginase (E.C.3.5.1.1) is an enzyme responsible for hydrolysis of L-asparagine into aspartic acid and ammonia, and has its significant applications in the therapeutics and food technology. It was produced by the marine Aspergillus terreus and precipitated by 65% ammonium sulphate, followed by purification using gel filtration on Sephadex G-100 and DEAE-cellulose ion exchange chromatography, which yielded 11.96 fold purification. The molecular weight of the purified L-asparaginase was approximately 85 kDa, determined by a sodium dodecyl sulphate polyacrylamide gel electrophoresis. L-asparaginase showed high affinity for L-asparagine with a Km of 31.5 mM and Vmax of 500 U/ml. The optimum pH and temperature of the purified enzyme were 5.8 and 40 o C, respectively. The L-asparaginase enzyme was stable from pH 4 to 5.8 and stable up to 70 o C. The effect of activators and inhibitors was studied providing that CdCl2 , Pb Cl2 , and Hg Cl2 strongly inhibited the enzyme activity, while Na Cl highly enhanced activity. Anticancer activity of the purified L-asparaginase was detected against HCT-116, Hep-G2 and MCF-7 cell lines with IC50 ranged from 3.79-12.6 µg/ml.

Algae are omnipresent in all seas and oceans, which make thema target for many applications such as bio-fertilizers, fish feeding and removal of heavy metals. In the present study, different algal species were examined as sustainable alternatives substrates for PHA production by Halomonas sp. Several media simulations were utilized to achieve high polymer productivity. The maximum poly(3-hydroxybutyrate) (PHB) concentrations were determined by using Corallina mediterranea hydrolysates as a carbon and nitrogen source. The isolates Halomonas pacifica ASL10 and Halomonas salifodiane ASL11 were found to be able to produce PHA by 67 % wt and 63 % wt CDW, respectively. PHB nanoparticles (NPs) had high zeta potential values and small particle sizes. These properties make it suitable for several drug delivery and pharmaceutical applications. Interestingly, NPs showed a potent antibacterial activity against several reference strains. The antibacterial efficacy of PHA-NPs has not been previously studied, thus this study opens a promising use of PHA-NPs.

The light-induced functioning of photosynthetic pigment-protein complex of photosystem II (PSII) is linked to the vectorial translocation of charges across the membrane, which results in the formation of voltage. Direct measurement of the light-induced voltage (∆V) generated by spinach oxygen-evolving PSII core complexes adsorbed onto a Millipore membrane filter (MF) on an indium tin oxide (ITO) electrode under continuous illumination has been performed. PSII was shown to participate in electron transfer from water to the ITO electrode, resulting in ∆V generation. No photovoltage was detected in PSII deprived of the water-oxidizing complex. The maximal and stable photoelectric signal was observed in the presence of disaccharide trehalose and 2,6-dichloro-1,4-benzoquinone, acting as a redox mediator between the primary quinone acceptor QA of PSII and electrode surface. Long time preservation of the steady-state photoactivity at room temperature in a simple in design ITO|PSII-MF|ITO system may be related to the retention of water molecules attached to the PSII surface in the presence of trehalose.

Tumor invasion requires efficient cell migration, which is achieved by the generation of persistent and polarized lamellipodia. The generation of lamellipodia is supported by actin dynamics at the leading edge where a complex of proteins known as the WAVE regulatory complex (WRC) promotes the required assembly of actin filaments to push the front of the cell ahead. By using an U2OS osteosarcoma cell line with high metastatic potential, proven by a xenotransplant in zebrafish larvae, we have studied the role of the plasma membrane Ca2+ channel ORAI1 in this process. We have found that epidermal growth factor (EGF) triggered an enrichment of ORAI1 at the leading edge, where colocalized with cortactin (CTTN) and other members of the WRC, such as CYFIP1 and ARP2/3. ORAI1-CTTN co-precipitation was sensitive to the inhibition of the small GTPase RAC1, an upstream activator of the WRC. RAC1 potentiated ORAI1 translocation to the leading edge, increasing the availability of surface ORAI1 and increasing the plasma membrane ruffling. The role of ORAI1 at the leading edge was studied in genetically engineered U2OS cells lacking ORAI1 expression that helped us to prove the key role of this Ca2+ channel on lamellipodia formation, lamellipodial persistence, and cell directness, which are required for tumor cell invasiveness in vivo.

The redox state of the neural progenitors regulates physiological processes such as neuronal differentiation and dendritic and axonal growth. The relevance of endoplasmic reticulum (ER)-associated oxidoreductases in these processes is largely unexplored. We describe a severe neurological disorder caused by bi-allelic loss-of-function variants in thioredoxin (TRX)-related transmembrane-2 (TMX2); these variants were detected by exome sequencing in 14 affected individuals from ten unrelated families presenting with congenital microcephaly, cortical polymicrogyria, and other migration disorders. TMX2 encodes one of the five TMX proteins of the protein disulfide isomerase family, hitherto not linked to human developmental brain disease. Our mechanistic studies on protein function show that TMX2 localizes to the ER mitochondria-associated membranes (MAMs), is involved in posttranslational modification and protein folding, and undergoes physical interaction with the MAM-associated and ER folding chaperone calnexin and ER calcium pump SERCA2. These interactions are functionally relevant because TMX2-deficient fibroblasts show decreased mitochondrial respiratory reserve capacity and compensatory increased glycolytic activity. Intriguingly, under basal conditions TMX2 occurs in both reduced and oxidized monomeric form, while it forms a stable dimer under treatment with hydrogen peroxide, recently recognized as a signaling molecule in neural morphogenesis and axonal pathfinding. Exogenous expression of the pathogenic TMX2 variants or of variants with an in vitro mutagenized TRX domain induces a constitutive TMX2 polymerization, mimicking an increased oxidative state. Altogether these data uncover TMX2 as a sensor in the MAM-regulated redox signaling pathway and identify it as a key adaptive regulator of neuronal proliferation, migration, and organization in the developing brain.

Recombinant l.asparaginase, L.ASNase, from Pseudomonas aeruginosa was purified using nickel affinity chromatography. The affinity purified L.ASNase exhibited a protein band with a molecular weight of 72.4 kDa on a native polyacrylamide gel and 36.276 kDa using SDS–PAGE. The activity of the purified L.ASNase was enhanced by Mg2+ and inhibited by Zn2+ at a concentration of 5 mM. The specificity of the recombinant L.ASNase towards different substrates was examined, and it was found that the enzyme showed the highest activity towards l.asparagine. Moreover, the enzyme showed lower activity towards other substrates such as L.glutamine, urea and acrylamide. The in vitro hemolysis assay revealed that the purified L.ASNase did not show hemolysis effect on blood erythrocytes. Serum and trypsin half-life of L.ASNase suggested that the recombinant L.ASNase retained 50% of its initial activity after 90 and 60 min incubation period in serum and trypsin separately.

l-Asparaginase (EC 3.5.1.1) is an important medical enzyme that catalysis the hydrolysis of l-asparagine to aspartic acid and ammonium. For over four decades l. asparaginase utic agent for the treatment of a variety of lymphoproliferative disorders and lymphoma such as acute lymphoblastic leukemia. In the present study A. terreus full length l. asparaginase gene, 1179bp was optimized for expression in Escherichia coli BL21 (DE3) pLysS. The full length A. terreusl. asparaginase gene encoding a protein of 376 amino acids with estimated molecular weight of 42.0kDa and a theoretical isoelectric point (pI) of 5.0. BLAST and phylogeny analysis revealed that the A. terreusl. asparaginase shared high similarity with other known fungal l. asparaginase (75% homology with A. nomius and 71% with A. nidulans). The recombinant protein was overexpressed in the form of amorphous submicron proteinaceous inclusion bodies upon induction with 1mM IPTG at 37°C for 18h.

Silk fibroin (SF) is a non-pathological amyloidogenic protein prone, in solution, to the formation of amyloid-like aggregated species, displaying similarities in fibrillation kinetics with pathological amyloids, as widely reported in the literature. We show here, on the basis of different biophysical approaches (turbidity, Congo Red assays, CD, DLS and fluorescence), that fusidic acid (FA), a well-known antibiotic, acts on SF as an anti-aggregating agent in a dose-dependent manner, being also able to revert SF aggregation. FA binds to SF inducing changes in the environment of SF aromatic residues. We further provide the proof of principle that FA, already approved as drug on humans and used in ophthalmic preparations, displays its anti-aggregation properties also on lens material derived from cataract surgery and is capable of reducing aggregation. Thus it is suggested that FA can be foreseen as a therapeutic treatment for cataract and other protein aggregation disorders.

Calcium phosphate coatings (CaPO4) are widely used in the manufacture of diverse types of osseous implants. However, the CaPO4 deposited by most manufacturing methods presents low adhesion and low crystallinity, which makes it difficult to apply immediately without further treatments. In this research, adherent and crystalline TiN/TiO2/HA coatings were deposited onto Ti6Al4V alloy substrates without the necessity of carrying out additional treatments of the coated samples. The reactive magnetron sputtering technique was used for the deposition of the coatings applying a 13.56 MHz rf power of 650 W to the HA target and a DC power of 1.2 kW to Ti target (99,9% of purity). The compositional and microstructural analysis of synthetized coatings were performed using FTIR, microraman, XRD, SEM/EDS and AFM characterization techniques. The hardness, elastic modulus and adhesion of individual coatings were studied and analyzed using nanoindentation and scratch tests in order to elucidate the effects of these on the adhesion of the composite coatings. It was found that the deposited HA coatings have a crystalline structure obtained by appropriate adjustment of the deposition parameters without requiring a posterior heat treatment. Also the inclusion of the TiN/TiO2- bilayer improved the adhesion strength of the whole coating system.

Human C2orf69 is an evolutionarily conserved gene whose function is unknown. Here, we report eight unrelated families from which 20 children presented with a fatal syndrome consisting of severe autoinflammation and progredient leukoencephalopathy with recurrent seizures; 12 of these subjects, whose DNA was available, segregated homozygous loss-of-function C2orf69 variants. C2ORF69 bears homology to esterase enzymes, and orthologs can be found in most eukaryotic genomes, including that of unicellular phytoplankton. We found that endogenous C2ORF69 (1) is loosely bound to mitochondria, (2) affects mitochondrial membrane potential and oxidative respiration in cultured neurons, and (3) controls the levels of the glycogen branching enzyme 1 (GBE1) consistent with a glycogen-storage-associated mitochondriopathy. We show that CRISPR-Cas9-mediated inactivation of zebrafish C2orf69 results in lethality by 8 months of age due to spontaneous epileptic seizures, which is preceded by persistent brain inflammation. Collectively, our results delineate an autoinflammatory Mendelian disorder of C2orf69 deficiency that disrupts the development/homeostasis of the immune and central nervous systems.

This article provides a new series of semiconducting quantum dots based on the doping of gallium arsenide with protactinium rare earth element. The Ga1-xPaxAs QDs were prepared using organometallic pyrolysis approach. The effect of protactinium rare earth element on the phase of the GaAs crystal lattice was emphasized. It was found that the GaAs cubic lattice is expanded and its size was increased. The synthetic recipe disclosed the formation of uniform nanocrystals with average size in the range 2.5 nm to 8.1 nm as Pa atoms increased. The optical properties of the Ga1-xPaxAs QDs inspected a redshift of the absorption spectrum and reduction of the bandgaps from 2.4 eV to 1.81 eV. The Ga1-xPaxAs QDs exhibited multicolors in the wavelength range 560 nm to 690 nm. The emerged light photons spectrum from these Ga1-xPaxAs QDs was very sharp and intense. The Pa-dopant suppressed the defects of the GaAs QDs and increased the quantum yield to 73 %. These marvel behaviors may enable the developed Ga1-xPaxAs QDs to be used for the manufacture of highly luminescent QLEDs.

Shortcomings encountered in commercial L-Asparaginases greatly restrict their therapeutic potential and hence, searching for novel L-Asparaginases has become a mandatory issue. Present work highlights cloning, expression and characterization of L-Asparaginase open reading frame (ORF) from Pseudomonas aeruginosa strain EGYII DSM 101801 in E.coli BL21 (DE3) pLysS. A DNA fragment of 984 bp encoding L-Asparaginase ORF was cloned on pGEM-T easy vector via polymerase chain reaction. Successful expression of ORF fragment, harbored in frame on pET28-a(+) expression vector, was achieved in E.coli DE3 (BL21) pLysS as a Six His-Tag fusion protein after 18 h of induction with 1 mM IPTG at 37 °C. The recombinant enzyme (aeruginosa EGYII L-Asparaginase) was purified to homogeneity using Ni2+ chelated Fast Flow Sepharose resin with 8.567 purification fold and 14.42% recovery. It exhibited an approximate molecular mass of 36 kDa as deduced from SDS-PAGE. The recombinant enzyme exhibited its maximal activity at pH 8.5 and 45 °C. It retained almost 80% and 60% of its activity at 37 °C after 20 min and 30 min, respectively. Two fold enhancement in enzyme activity was retained in presence of 5 mM MgCl2. EDTA, β-mercaptoethanol and ZnCl2 at a concentration of 1 mM each slightly enhanced enzyme activity. Kinetics parameters of the recombinant enzyme were Km (63.11 mM) and Vmax (1121 μmol min−1 mg−1) for L-Asparagine as calculated from Lineweaver–Burk plot. Present data would encourage testing the therapeutic potential of the recombinant aeruginosa EGYII L-Asparaginase.

Twenty one moderately halophilic bacterial strains were isolated from seawater and sediment in Alexandria Eastern Harbour, Egypt. The isolates were screened for the production of four extracellular degradative enzymes. The majority of isolates (57.1%) possessed significant enzyme activities, 43% of them have potentiality to produce amylase enzyme. The most active isolate for the production of amylase enzyme was identified by using a 16S rRNA sequence analysis as Bacillus lichineformis AH214. Optimization of the fermentation medium components and environmental factors using One Variable at a Time Approach and Plackett-Burman design was applied to enhance the amylase production by Bacillus lichineformis AH214. The maximum microbial amylase production could be achieved using an optimized medium of the following composition (g/l): 1.0 g yeast extract, 0.05 g K2HPO4, 0.25 g FeCl3, 15.0 g starch, 30.0 g NaCl, 0.75 g MgSO4.7H2O and inoculums size of 1.5 ml/50 ml and incubated at optimum conditions of pH 7, agitation speed 160 rpm, time 30 h and temperature 40°C. On applying optimized medium in the fermentation process, an enzyme productivity of 13.44 U/mg protein was achieved with two fold increase compared to the basal one. The crude amylase produced by Bacillus lichineformis was stable up to 40°C, pH 7.5 and 1.5 M NaCl. Key words: Halophiles, amylase, Bacillus lichineformis, Plackett-Burman, optimization.

Since the use of organotin as antifouling additives was prohibited in 2003, many researchers have endeavored to design and develop novel economic environment-friendly marine antifouling additives. This work reports the successful functionalization of biopolymeric chitosan, isolated from shrimp shells, with salicylidene ionic liquid (IL-Sal) brushes, (ILCSB1–6). These designed architectures were structurally and morphologically characterized. Marine biofouling-inducing bacterial strains (S. aureus, E. coli, A. hydrophila and Vibrio) were selected as microfoulants for a laboratory antibacterial and biofilm susceptibility assay investigation. Our outcomes unveiled a novel promising ecofriendly biocidal agent with excellent and broad antibacterial efficacy compared to parent chitosan and the standard antifoulant, Diuron®. The fabricated poly-IL-brushes chitosan architectures were subjected to a rigorous test in a field trial in Red Sea water. Our findings provide new insights into eco-friendly antifouling additives as an alternative to traditional antifouling agents. Novel IL-functionalized chitosan-based coatings exhibited long-term durability, surface inertness and promising antifouling performance.

l -asparaginase plays a vital role in medical application, particularly in treatment of acute lymphoblastic leukemia as an effective anti-tumor agent. In the present study, twenty-one fungal strains were isolated from marine environment of the Red Sea coasts of Egypt. Screening for fungal l -asparaginase production was done, and only five fungal strains were selected and identified as Aspergillus , Penicillium and Fusarium . The most potent fungal isolate was Aspergillus terreus which yielded the highest l -asparaginase specific activity (4.81 U/mg protein). The highest enzyme productivity was observed on the 5th day and the optimized fermentation parameters were pH 6.0, temperature 35 °C. The yield was also high up on using dextrose and asparagine (8.26 U/mg protein) as carbon and nitrogen sources. The cultural conditions were studied using the Plackett–Burman experimental design. Immobilization using A. terreus adsorbed on sponge enhanced the enzyme production by 1.33-fold compared to the conventional free-cells. Repeated reuse of the adsorbed cells achieved the maximum enzyme specific activity after three cycles (33.86 U/mg protein).

STIM1 is an endoplasmic reticulum protein with a role in Ca2+mobilization and signaling. As a sensor of intraluminal Ca2+levels, STIM1 modulates plasma membrane Ca2+channels to regulate Ca2+entry. In neuroblastoma SH-SY5Y cells and in familial Alzheimer’s disease patient skin fibroblasts, STIM1 is cleaved at the transmembrane domain by the presenilin-1-associated γ-secretase, leading to dysregulation of Ca2+homeostasis. In this report, we investigated expression levels of STIM1 in brain tissues (medium frontal gyrus) of pathologically confirmed Alzheimer’s disease patients, and observed that STIM1 protein expression level decreased with the progression of neurodegeneration. To study the role of STIM1 in neurodegeneration, a strategy was designed to knock-out the expression ofSTIM1gene in the SH-SY5Y neuroblastoma cell line by CRISPR/Cas9-mediated genome editing, as an in vitro model to examine the phenotype of STIM1-deficient neuronal cells. It was proved that, while STIM1 is not required for the differentiation of SH-SY5Y cells, it is absolutely essential for cell survival in differentiating cells. Differentiated STIM1-KO cells showed a significant decrease of mitochondrial respiratory chain complex I activity, mitochondrial inner membrane depolarization, reduced mitochondrial free Ca2+concentration, and higher levels of senescence as compared with wild-type cells. In parallel, STIM1-KO cells showed a potentiated Ca2+entry in response to depolarization, which was sensitive to nifedipine, pointing to L-type voltage-operated Ca2+channels as mediators of the upregulated Ca2+entry. The stable knocking-down ofCACNA1Ctranscripts restored mitochondrial function, increased mitochondrial Ca2+levels, and dropped senescence to basal levels, demonstrating the essential role of the upregulation of voltage-operated Ca2+entry through Cav1.2 channels in STIM1-deficient SH-SY5Y cell death.Key messagesSTIM1 protein expression decreases with the progression of neurodegeneration in Alzheimer’s disease.STIM1 is essential for cell viability in differentiated SH-SY5Y cells.STIM1 deficiency triggers voltage-regulated Ca2+entry-dependent cell death.Mitochondrial dysfunction and senescence are features of STIM1-deficient differentiated cells.

Some coordination biopolymeric metal–alginate ionotropic hydrogel complexes were prepared by the replacement of the Na+ counter ions of alginate sol polysaccharide by monovalent silver(I) or polyvalent metal ions forming their corresponding complexes in either granule or hydrogel phases. The type of such phase and the capillary or non-capillary structures property were found to be dependent on the method of preparation and the direction of diffusion of the metal ion toward the alginate sol matrix whether is of upward or downward direction. The net process of exchange leads to the so-called sol-gel transformation to give its respective hydrogel complexes. This process takes place through formation of partially ionic and partially coordinate bonds between the metal ion and the carboxylate and hydroxyl functional groups of alginate, respectively. This kind of chelation forms a sort of bridges in an egg box-like structure. The anisotropic property of the hydrogels is owing to the orientation of the solvent molecules and macromolecular chains toward the chelated metal ions. The geometrical configuration and physicochemical properties of the hydrogel complexes depend on the nature of the metal ions such as the valence and its coordination number as well as on the strength of chelating of the bonds formed. The kinetics and mechanism of sol-gel transformation, electrical conductivity, and thermal decomposition with their evaluated kinetic parameters along with the other physicochemical properties such as FTIR, XRD, morphology, configuration geometry, and rheological properties have been investigated and discussed.

Obstructive sleep apnea syndrome (OSAS) is a risk factor for arterial thrombosis and cardiovascular morbidity. Activated platelets play key roles in the development of atherothrombosis, thus may be involved in these complications of OSAS. Herein, we evaluated the relationship between severity of OSAS and the degree of platelet aggregates as a marker of activated platelets in 64 patients with OSAS. Platelet aggregations were determined by means of optical aggregometry, using adenosine diphosphate (ADP) as an agonist. Compared with the control group, ADP-induced platelet aggregability was increased in patients with total OSAS, severe OSAS, and in mild to moderate OSAS. Moreover, ADP-induced platelet aggregation was correlated with the Epworth sleepiness scale (ESS) in patients with severe OSAS. Obstructive sleep apnea syndrome is associated with enhanced platelets aggregations, which may predispose the cardiovascular sequels. The ESS may be important in predicting platelet activation and thus atherothrombotic complications in those with OSAS.

Eruption of blue-green algal blooms occurs frequently in eutrophic lakes and fish ponds, with associated unpleasant odor and horrid scums. In the present study, we conducted a pre-test experiment in 3 m(3) outdoor concrete ponds to determine the optimum concentration of aluminum sulfate (alum) required for reduction of the cyanobacterial blooms without negative effect on fish growth. As a consequence, 10 mg L(-1) alum was named as the optimum concentration that was applied in 1000 m(3) earthen fish ponds. Obtained results showed that Secchi disc values significantly increased from 10 to 24 cm after 14 days of alum application. Alum-treated ponds showed a reduction in total phytoplankton counts by 94 and 96% compared to the corresponding controls after 10 and 14 days, respectively. Abundance of blue-green algae in the treated ponds was decreased by 98% compared to the corresponding control after 14 days of alum application. Consequently, dissolved oxygen, pH, total phosphorus, orthophosphate, and chlorophyll "a" content declined significantly. Our study revealed that using 10 mg L(-1) of alum is an effective way to control cyanobacterial blooms in eutrophic waters, especially in fish ponds, without negative effect in water quality.

STIM1 is a key regulator of store-operated calcium entry (SOCE), and therefore a mediator of Ca 2 + entry-dependent cellular events. Phosphorylation of STIM1 at ERK1/2 target sites has been described as enhancing STIM1 activation during intracellular Ca 2 + emptying triggered by the inhibition of the sarco(endo)plasmic Ca 2 + -ATPase with thapsigargin. However, no physiological function is known for this specific phosphorylation. The present study examined the role of STIM1 phosphorylation in cell signaling triggered by EGF. Using a human endometrial adenocarcinoma cell line (Ishikawa cells) EGF or H-Ras(G12V), an active mutant of H-Ras, was found to trigger STIM1 phosphorylation at residues Ser575, Ser608, and Ser621, and this process was sensitive to PD0325901, an inhibitor of ERK1/2. Both, ERK1/2 activation and STIM1 phosphorylation took place in the absence of extracellular Ca 2 + , indicating that both events are upstream steps for Ca 2 + entry activation. Also, EGF triggered the dissociation of STIM1 from EB1 (a regulator of microtubule plus-ends) in a manner similar to that reported for the activation of STIM1 by thapsigargin. Migration of the Ishikawa cells was impaired when STIM1 phosphorylation was targeted by Ser-to-Ala substitution mutation of ERK1/2 target sites. This effect was also observed with the Ca 2 + channel blocker SKF96365. Phosphomimetic mutation of STIM1 restored the migration to levels similar to that found for STIM1-wild type. Finally, the increased vimentin expression and relocalization of E-cadherin triggered by EGF were largely inhibited by targeting STIM1 phosphorylation, while STIM1-S575E/S608E/S621E normalized the profiles of these two EMT markers.

STIM1, the endoplasmic reticulum Ca2+ sensor that modulates the activity of plasma membrane Ca2+ channels, becomes phosphorylated at ERK1/2 target sites during Ca2+ store depletion triggered by thapsigargin or epidermal growth factor (EGF). This ERK1/2-dependent phosphorylation regulates STIM1 localization and dissociation from microtubules, and it is known that enhances the binding to ORAI1, a store-operated Ca2+ entry (SOCE) channel, leading to the activation of this Ca2+ influx pathway. However, there remained some evidence of a role for SOCE in the activation of ERK1/2, and here we assessed the contribution of SOCE to ERK1/2 activation by generating a STIM1-deficient cell line by CRISPR/Cas9 genome editing of the STIM1 locus in prostate cancer PC3 cells. The genomic modification consisted of a 16 base-pair insertion in exon 5 of both alleles, therefore abrogating STIM1 synthesis. STIM1-KO cells did show a striking decrease in Ca2+ influx in response to thapsigargin or EGF, a result that demonstrates that SOCE mediates Ca2+ entry in PC3 cells during stimulation with EGF. Moreover, identical levels of total ERK1/2 were found in STIM1-KO cells and the parental cell line, and ERK1/2 activation was fully activated in KO cells, both in the presence and in the absence of extracellular Ca2+, a result that supports that STIM1 and SOCE are not required for ERK1/2 activation. This activation was sensitive to Src kinase inhibition, but not to CAMKII nor PKC inhibition, a result that sets STIM1 and SOCE as downstream targets of the axis Src-Raf-MEK-ERK, rather than upstream regulators.

A novel marine fungus was isolated and classified as Aspergillus flavus strain EGY11. The heat-inactivated form of isolated Aspergillus flavus was investigated and evaluated as a new eco-friendly and highly efficient biosorbent for removal of toxic heavy metals such as Cd(II), Hg(II), and Pb(II) from aqueous solutions. The SEM morphological studies of biosorbent-loaded metal ions confirmed their direct binding on the surface of heat-inactivated Aspergillus flavus. The metal biosorption capacity values were determined and optimized by the batch technique in the presence of various experimental controlling factors such as pH, contact time, biosorbent dosage, initial metal ion concentration, and coexisting species. The maximum metal capacity values of Cd(II), Hg(II), and Pb(II) were cauterized as 1550 (pH 7.0), 950 (pH 7.0), and 1000 μmol g−1 (pH 6.0), respectively. The equilibrium time for removal of metal ions was identified as 40 min. The maximum sorption capacity values (1200.0–4000.0 μmol g−1) were established by 5.0 mg as the optimum mass of biosorbent. The collected biosorption data obtained from the equilibrium studies using the initial metal ion concentration were described by the Langmuir, Freundlich, Brunauer–Emmett–Teller (BET), and Dubinin–Radushkevich isotherm (D–R) isotherm models. The potential implementation of heat-inactivated Aspergillus flavus biosorbent for heavy metal removal from different water samples was successfully accomplished using multistage microcolumn technique. The results refer to excellent percentage recovery values in the ranges 92.7–99.0, 91.3–95.6, and 95.3–98.2% for the biosorptive removal of Cd(II), Hg(II), and Pb(II), respectively, from the examined environmental samples.

In this work, two dispiro[3H-indole-3,2′-pyrrolidine-3′,3″-piperidines], C31H29Cl2N3O3 (4a) and C32H31Cl2N3O3 (4b), have been synthesized and characterized by IR, 1H NMR, 13C NMR, 1H 1H COSY, 1H 13C HSQC and single crystal X-ray diffraction. Both compounds were found to be crystallized in the triclinic space group P 1 ¯ with Z = 2. Molecules of the crystalline structure of 4a are linked with intermolecular C H⋯O and N H⋯O hydrogen bonding. Meanwhile, an intermolecular N H⋯O hydrogen bonding was recognized in the crystalline structure of 4b. Molecular mechanics force field (MM+), followed by either semi-empirical AM1 or PM3, has been used to calculate the optimized geometrical parameters for the two compounds. The determined theoretical geometry parameters were found to be in a good agreement with the parameters obtained using X-ray studies. Moreover, the Molecular Electrostatic Potential (MEP) of both compounds have been calculated at B3LYP/6-31G(d,p) level of theory exhibiting that, the most electrophilic site of the synthesized compounds, is the piperidone O3 atom however, the most nucleophilic site is the indolyl N2. Additionally, the Frontier Molecular Orbitals (FMOs) 4a and 4b have been determined by the same technique exhibiting large energy gap between HOMO and LUMO orbitals (3.40, 3.43 eV for 4a and 4b, respectively), inferring good stability, high excitation energies and large chemical hardness of these molecules.

Eighteen bacterial strains were isolated from Sea water collected from different sites of Red Sea, Egypt. They were screened for antagonistic activity against many bacterial pathogens. The most active antimicrobial producer (R6) which isolated from Safaga middle was identified as Bacillus badius strain AAUM using 16S rRNA phylogenetic analysis. The most control variables were selected from Plackett–Burman (PB) factorial design for the production of antimicrobial agent from Bacillus badius strain AAUM. It achieved 1.56-fold fold increase (against S. aureus) when grown in medium composed of g/l: peptone, 2.5; yeast extract, 5.0; sea water, 25ml; size of inoculum (3ml/50ml), adjusted to pH 9 and incubation period 20 hrs at 45oC. Immobilization using entrapment and adsorption techniques were applied to improve the productivity of cells. B. badius strain AAUM adsorbed on polyurethane foam realized 1.4-fold increase than the free cells. Reused of the adsorbed culture caused an increase of antimicrobial agent production by 1.14-fold than the free cells. The predicted crude antimicrobial spectra suggested that the B. badius strain AAUM can produce important and novel antimicrobial drugs.

Two field experiments were carried out at a Farm in El-Mataryia district, Dakahlia Governorate during the two growing seasons of 2011 and 2012 to study the effect of organic fertilization and ammonium thiosulfate (ATS) levels that reduce salinity effect on vegetative growth, yield and its components and chemical constituents of eggplant crop Black Beauty cultivar. The experiments were carried out by using split – plots system in a randomized complete blocks design with three replications. The main plots were arranged with organic fertilization treatments. The sub plots were assigned to ammonium thiosulfate levels. The obvious results of this investigation can be summarized as follows: Using farmyard manure (FYM) and potassium humate (KH) treatments significantly increased all studied characters as compared with other organic fertilization treatments in both seasons. Application rice residues (RS) caused significant increases on all studied characters as compared with control treatment (without organic fertilization) in both seasons. All studied characters were significantly influenced by different ATS levels in the both seasons. The highest values of these traits were resulted from using 30 L/fed and 40 L/fed of ATS, respectively in both seasons. Generally, it could be recommended that using FYM or KH, respectively along with 30 L ATS/fed to enhance vegetative growth characters, yield and its components and chemical constituents of eggplant fruits. Also, it could be recommended that using RS combined with 30 L ATS/fed to enhance studied characters as compared with control treatment (without organic fertilization) and also to reduce sources of environmental pollution and maintain human health.

Calcium handling is critical for the oocyte function, since the first steps of fertilization are dependent on the appropriate Ca(2+) mobilization to originate transient spikes of the cytosolic Ca(2+) concentration. It is well known that the Ca(2+) influx from the extracellular milieu is required to maintain this signaling in mammalian oocytes. However, the regulation of the Ca(2+) channels involved in this process is still unknown in oocytes. STIM1, a key regulator of store-operated Ca(2+) entry (SOCE), relocates in the mouse oocyte shortly after sperm stimulation, suggesting that SOCE is involved in the maintenance of cytosolic Ca(2+)-spiking in the fertilized oocyte. Here, we show that there is an up-regulation of the expression of STIM1 at the germinal vesicle breakdown stage, and this expression remains steady during following maturation stages. We found that oocytes express ORAI1, a store-operated Ca(2+) channel, and that ORAI1 expression level was stable during oocyte maturation. Immature oocytes showed no Ca(2+) entry and no increase in STIM1-ORAI1 colocalization in response to the store depletion induced by thapsigargin. On the contrary, in mature oocytes, STIM1-ORAI1 colocalization is enhanced 3-fold by depletion of Ca(2+) stores, enabling the activation of store-operated calcium channels and therefore Ca(2+) entry. Finally, the correlation between SOCE activation during the maturation of oocytes and STIM1-ORAI1 colocalization strongly suggests that ORAI1 is involved in the Ca(2+) entry pathway in the mature oocyte. SOCE up-regulation in the final stage of maturation is further evidence of a major role for SOCE in fully mature oocytes, and therefore in Ca(2+) signaling at fertilization.

© 2011 American Association of Pharmaceutical Scientists, The aim of this work is to prepare tretinoin/dimethyl-beta-cyclodextrin complexes and fully characterize them through various analytical techniques. According to the phase solubility studies performed, the equilibrium for maximum complexation is reached in about 8 days presenting an AL-type diagram (soluble complexes) corresponding mainly to 1:1 stoichiometry (Ks=13,600 M−1), although the possibility of the presence of 1:2 complexes was mathematically proven. Differential scanning calorimetry, X-ray diffraction and all the other analytical techniques have proven the presence of true complex formation in all the preparation methods tested. H-NMR and FTIR spectra allowed the selection of the best complexation method. The comparison between Raman spectra revealed that the more relevant feature is the band at 1,573 cm−1, which corresponds to the entire delocalization of the superconjugated system, and after inclusion is observed as a positive frequency shift. Based on these results and the data obtained by molecular modelling calculations, it is proposed that the structure of the drug included into the cyclodextrin corresponds to the side chain including the functional group COOH. The complex was also analysed by atomic force microscopy to determine its size distribution which was heterogeneous and polymodal. However, it could be observed that they all have the same phase constitution.

The kinetics and mechanism of the reaction of hydrogen peroxide with some Fe(II) Schiff base complexes were investigated spectrophotometrically in aqueous solution at pH 8 and 35 °C under pseudo-first-order conditions. The used ligands were derived from salicylaldehyde or o-hydroxynaphthaldehyde and some amino acids ( l -leucine, l -iso-leucine, l -serine, l -methionine and dl -tryptophan). It was found that the formation of the purple interaperoxo complex appears only above pH 7.5. The reaction consists of two steps. The first step involves reversible formation of the intraperoxo intermediate which renders irreversible at pH 8. The second step consists of inner-sphere electron transfer. The suggested scheme illustrated first-order kinetics at low [H2O2] and zero-order at high [H2O2]. Moreover, the activation parameters of the reaction were evaluated.