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Ocular repeated air blast injuries occur from low overpressure blast wave exposure, which are often repeated and in quick succession. We have shown that caspase-2 caused the death of retinal ganglion cells (RGC) after blunt ocular trauma. Here, we investigated if caspase-2 also mediates RGC apoptosis in a mouse model of air blast induced indirect traumatic optic neuropathy (b-ITON). C57BL/6 mice were exposed to repeated blasts of overpressure air (3 × 2 × 15 psi) and intravitreal injections of siRNA against caspase-2 (siCASP2) or against a control enhanced green fluorescent protein (siEGFP) at either 5 h after the first 2 × 15 psi (“post-blast”) or 48 h before the first blast exposure (“pre-blast”) and repeated every 7 days. RGC counts were unaffected by the b-ITON or intravitreal injections, despite increased degenerating ON axons, even in siCASP2 “post-blast” injection groups. Degenerating ON axons remained at sham levels after b-ITON and intravitreal siCASP2 “pre-blast” injections, but with less degenerating axons in siCASP2 compared to siEGFP-treated eyes. Intravitreal injections “post-blast” caused greater vitreous inflammation, potentiated by siCASP2, with less in “pre-blast” injected eyes, which was abrogated by siCASP2. We conclude that intravitreal injection timing after ocular trauma induced variable retinal and ON pathology, undermining our candidate neuroprotective therapy, siCASP2.

Wild-type erythropoietin (EPO) is promising for neuroprotection, but its therapeutic use is limited because it causes a systemic rise in hematocrit. We have developed an EPO-R76E derivative that maintains neuroprotective function without effects on hematocrit, but this protein has a short half-life in vivo. Here, we compare the efficacy and carrier-induced inflammatory response of two polymeric microparticle (MP) EPO-R76E sustained release formulations based on conventional hydrolytically degradable poly(lactic-co-glycolic acid) (PLGA) and reactive oxygen species (ROS)-degradable poly(propylene sulfide) (PPS). Both MP types effectively loaded EPO-R76E and achieved sustained release, providing detectable levels of EPO-R76E at the injection site in the eye in vivo for at least 28 days. Testing in an in vitro oxidative stress assay and a mouse model of blast-induced indirect traumatic optic neuropathy (bITON) showed that PPS and PLGA MP-mediated delivery of EPO-R76E provided therapeutic protection. While unloaded PLGA MPs inherently increase levels of pro-inflammatory cytokines in the bITON model, drug-free PPS MPs have innate antioxidant properties that provide therapeutic benefit both in vitro and in vivo. Both PLGA and PPS MPs enabled sustained release of EPO-R76E, providing therapeutic benefits including reduction in inflammation and axon degeneration, and preservation of visual function as measured by electroretinogram. The PPS-based MP platform is especially promising for further development, as the delivery system provides inherent antioxidant benefits that can be harnessed to work in complement with EPO-R76E or other drugs for neuroprotection in the setting of traumatic eye injury.

The power of CRISPRi to decrease targeted gene expression for clinical applications has been inhibited by delivery challenges. Existing constructs are too large to fit within the ∼4.7 kb packaging size limitation of adeno-associated virus (AAV), the only FDA approved viral vector for clinical use. Therefore, we optimized CRISPRi components to generate a single AAV vector that contains all functional elements and effectively knocks down expression of an endogenous gene in vivo. First, we increased nuclear targeting of Staphylococcus aureus deactivated Cas9 (SadCas9) 4-fold by using a helical linker and the c-Myc nuclear localization signal. Second, we identified an amino-terminal Krüppel associated box (KRAB) construct as the most effective in decreasing expression of target genes in vitro. Third, we optimized promoters for guide RNA and evaluated mini-promoters for expression of KRAB-SadCas9 in liver cells. Our final construct decreased protein convertase subtilisin/kexin type 9 (Pcsk9) mRNA and secreted protein 5-fold in vitro. The corresponding AAV2/8 vector was localized in nuclei of liver cells and decreased Pcsk9 mRNA and serum protein levels by 30% in vivo. This single AAV approach provides a potential clinically translatable method for decreasing targeted gene transcription by CRISPRi in vivo.

In this work, a series of 2-(1H-benzo[d]thiazole-2-yl)-N-arylbenzamides is synthesized by using diethyl phthalate, anilines and 2-amino-benzenethiol by one-pot three component synthesis in glycerol medium. Phosphoric acid is used as an effective reagent for this one-pot three component reaction. This reaction got completed in a short time, easy workup and gave an excellent yield in glycerol medium. The N-arylbenzamides was found to have significant cytotoxic potentials against various cancer cells viz., A549 (lung cancer), HeLa (cervical cancer) and MCF-7 (breast cancer) using MTT assay. The molecular docking study is also employed to understand the binding mechanism of N-arylbenzamides against the antibacterial target. The docking result shows the binding energy of compound 4a is -8.6 kcal/mol. The binding affinity is a major concern and it shows that Asn and Thr residues have an interaction with compound 4a.

Titanium(IV)-complex of chemical composition [{(NNO)2Ti}3O3] (2) bearing bidentate heteroditopic Schiff base [(C5H4OH)-N=CH-C4H3-NH] (L1) ligand in titanium coordination sphere was reported and its biological significance was evaluated. The in vitro cytotoxicity of L1 and 2 were evaluated by using MTT assay on cancer cell lines (MCF-7 & A549) and observed significant cytotoxicity. Further, the LDH and NO assay studies on both L1 and 2 on cancer cell lines revealed that the enhanced cytotoxicity compared to standard anticancer drug i.e. cisplatin. The DNA binding studies of tested compounds with Ct-DNA molecule by using UV-visible and fluorescence spectra and molecular docking studies revealed that moderate to good binding interactions with test molecules. Thus, the present work contributes and implies the biological significance of Ti-complex (2) and the correlation between the structure and the biological activities of such Ti-complexes supported by Schiff base systems opens up opportunities for further exploitation of similar biologically active titanium systems.

Economic nebulizer spray pyrolysis (NSP) technique has been employed to deposit Neodymium (Nd) doped Tin disulfide (SnS2) thin films at 325 °C by varying the doping concentration of Nd (0%, 2%, 4% and 6%). The impact of Nd concentration on crystalline structure, morphology and opto-electronic properties of films were studied using X-ray diffraction (XRD), FT-Raman spectrophotometer, atomic force microscope (AFM), energy-dispersive X-ray spectroscope, scanning electron microscope (SEM), UV–Visible spectrometer and Hall Effect measurements. Structural parameters such as lattice constants, texture orientation factor, micro strain, dislocation density and crystallite size were estimated using XRD data. The SEM and AFM images of films displayed an exceptional morphology. The surface roughness of the films was found to increase with the increase of dopant concentration. Optical analysis done on the films revealed variation in band gap from 2.75 to 2.92 eV as the Nd doping concentration is increased from 2 to 6%. SnS2 thin film exhibited n-type conductivity as, confirmed from Hall Effect studies. The resistivity and concentration of the carrier in the film were found to be 4.17 × 10−2 Ω cm and 4.06 × 1017 cm−3 respectively which were then correlated to the deposition parameters. Furthermore, FTO/n-Nd-SnS2/p-SnS hetero-junction based solar cells were prepared and their current voltage curve in dark condition was obtained.

Many neurodegenerations, including those of the visual system, have complex etiologies that include roles for both neurons and glia. In the retina there is evidence that retinal astrocytes play an important role in neurodegeneration. There are several approaches for isolating and growing primary retinal astrocytes, however, they often lead to different results. In this study, we examined the influence of culture conditions on phenotypic maturation of primary, purified retinal glia. We compared retinal astrocytes and Müller glia purified by immunomagnetic separation, as differentiation between these astrocyte subtypes is critical and immuno-based methods are the standard practice of purification. We found that while time in culture impacts the health and phenotype of both astrocytes and Müller glia, the phenotypic maturation of retinal astrocytes was most impacted by serum factors. These factors appeared to actively regulate intermediate filament phenotypes in a manner consistent with the induction of astrocyte-mesenchymal transition (AMT). This propensity for retinal astrocytes to shift along an AMT continuum should be considered when interpreting resulting data. Our goal is that this study will help standardize the field so that studies are replicable, comparable, and as accurate as possible for subsequent interpretation of findings.

Four binuclear Ni(II) complexes [[Ni2(H-DEAsal-tsc)2(μ-dppm)]·2Cl (1), [Ni2(DEAsal-mtsc)2(μ-dppm)] (2), [Ni2(DEAsal-etsc)2(μ-dppm)] (3) and [Ni2(DEAsal-ptsc)2(μ-dppm)] (4)] were synthesized from the ligands namely 4(N,N)-diethylaminosalicylaldehyde-4(N)-thiosemicarbazone [H2-DEAsal-tsc] H2L1/4(N,N)-diethylaminosalicylaldehyde-4(N)-methyl thiosemicarbazone [H2-DEAsal-mtsc] H2L2/4(N,N)-diethylaminosalicylaldehyde-4(N)-ethyl thiosemicarbazone [H2-DEAsal-etsc] H2L3/4(N,N)diethylaminosalicylaldehyde-4(N)-phenyl thiosemicarbazone [H2-DEAsal-ptsc] H2L4 and 1,1'-bis(diphenylphosphino)methane (dppm) and characterized by a number of spectro analytical techniques. The molecular structure of complexes [Ni2(H-DEAsal-tsc)2(μ-dppm)]·2Cl (1) and [Ni2(DEAsal-ptsc)2(μ-dppm)] (4) have been confirmed by single crystal X-ray diffraction studies. The analysis indicated that in complex 1, the ligand [H2-DEAsal-tsc] coordinated as monobasic tridentate donor through phenolic oxygen, azomethine nitrogen and thione sulfur atoms. However, in complex 4, the ligand [H2-DEAsal-ptsc] behaved as dibasic tridentate donor with thiolate sulfur coordination. Their ability to bind with Calf Thymus Deoxyribonucleic acid (CT-DNA) and Bovine Serum Albumin (BSA) were analysed spectrometrically. Intercalative interaction of the complexes with DNA was confirmed by ethidium bromide (EB) displacement studies and DNA viscosity measurements. The interaction mechanism of the complexes with BSA was found as static. In vitro antiproliferative studies of the ligands and complexes in A549 (human lung carcinoma cancer), MCF-7 (human breast cancer) and HeLa (human cervical cancer) cell lines witnessed significant cytotoxic nature of the complexes with low IC50 values (in μM) than the standard metallo-drug cisplatin. Further, the results of Lactate Dehydrogenase (LDH) and Nitric oxide (NO) release assays supported the effectiveness of the complexes on the above said cancer cells.

A series of four new, cyclometallated ruthenium (II) complexes was synthesized from 3-acetyl-8-methoxy-2H-chromen-2-one functionalized 4(N)- substituted thiosemicarbazones and characterized through various spectral and analytical methods. The molecular structures of the complexes 1, 2 and 4 were determined by single-crystal X-ray diffraction analysis, which confirmed that the complexes possess a distorted octahedral geometry with the ligands coordinating in a dibasic tridentate fashion via C, N and S atoms. DNA [Calf Thymus DNA (CT-DNA)] and protein [Bovine Serum Albumin (BSA) and Human Serum Albumin (HSA)] binding studies indicated an intercalative mode of binding with DNA and static quenching mechanism with proteins. The compounds cleaved plasmid DNA (pBR322) without application of any external agent and acted well as free radical scavengers. A good spectrum of antimicrobial activity was observed against four bacterial and five fungal pathogens. Two cancerous cell lines, MCF-7 (human breast cancer) and A549 (human lung carcinoma) were employed to test their in vitro cytotoxic activity using MTT assay. The complexes (1–4) showed better activity with lower IC50 values over cisplatin. Further non toxic nature of the complexes have been examined with human normal keratinocyte cell line HaCaT. Further, the results of Lactate dehydrogenase release (LDH) and Nitric Oxide (NO) release supported the cytotoxic nature of the compounds. The results of all the biological studies carried out implied that the complex 3 bearing an ethyl substituent was observed to be the best.

A pyrene scaffold bearing a phenol molecule has been designed and synthesized as a chemosensor L1 by an aldol type of condensation reaction. The sensor showed high sensitivity and selectivity for Al3+ ions by the enhancement of fluorescene intensity and distinguishes from the other potentially competing metal ions. The effect of pH, time and the reversibility of the sensor are studied. The fluorescence experiments suggested that the proposed binding of L1 and Al3+ ion is in a 1:1 stoichiometry. The chemosensor’s direct application in the electrosorptive removal of Al3+ ion and several other biological applications including the bio-imaging of bacterial cells were examined.

Four new water soluble chromone appended copper(II) complexes of the type [Cu(L)Cl] were synthesized from CuCl2·2H2O and 3-formylchromone-4N-substituted thiosemicarbazones (HL1–HL4). Characterization of the compounds was done by using analytical and spectral techniques such as elemental analyses, IR, UV–Visible, EPR and Mass spectrometry, which confirmed their formation. Single crystals suitable for X-ray diffraction were obtained for complex [Cu(L4)Cl], in which the ligand coordinated in a tridentate monobasic ONS donor fashion. The compounds strongly bound to CT-DNA (Calf Thymus DNA) through intercalation. BSA (Bovine Serum Albumin) and HSA (Human Serum Albumin) binding studies were carried out to check the binding ability of the compounds with protein and the mechanism of quenching was found to be static. The occurrence of microenvironmental change in protein was further confirmed by three dimensional (3D) fluorescence experiments. DNA cleavage experiments of the complexes showed that the complexes cleaved supercoiled DNA pBR322 without any external agent. The complexes have shown significant growth inhibitory activity against selected types of bacteria namely S. aureus, S. pneumonia, P. auroginosa, S. paratyphi and fungi namely C. albicans, T. rubrum, A. niger, A. fumigatus and C. tropicalis. Tests on cell proliferation of human lung cancer cell line (A549) and human breast cancer cell line (MCF-7) were performed for all the compounds. The new water soluble complexes overcome cisplatin resistance in the MCF-7 and A549 cell lines. All the compounds were found to be non-toxic against human normal keratinocyte cells (HaCaT). The biological studies indicated that the complex [Cu(L3)Cl] (3) exhibited better activity among the compounds and the complexes exhibited biological activity in the following order [Cu(L3)Cl] (3) > [Cu(L2)Cl] (2) > [Cu(L1)Cl] (1) > [Cu(L4)Cl] (4).

Four new bivalent organoruthenium complexes incorporated with 3-acetylcoumarin-4(N)-substituted thiosemicarbazones were synthesized. Comprehensive spectral and analytical techniques such as elemental analyses, IR, UV-vis, 1H NMR and 13C NMR provided proof of the formation of the complexes (1–4). Single crystals obtained for two ligands and the four complexes were characterized by X-ray crystallographic studies, which indicated CNS tridentate coordination of the ligands through C–H activation at the C(4) carbon of the pyrone ring of coumarin. The ligands and complexes intercalated with calf thymus DNA (CT DNA), supported by EB displacement studies and DNA viscosity measurements. The compounds bound well with two serum albumins (bovine serum albumin (BSA) and human serum albumin (HSA)) and the quenching mechanism was found to be static. Antioxidant studies revealed the radical scavenging proficiency of the compounds. The compounds showed better antimicrobial activity against two Gram-positive bacteria (Staphylococcus aureus and Streptococcus pneumonie), two Gram-negative bacteria (Pseudomonas aeruginosa and Salmonella paratyphi) and five fungi (Candida albicans, Trichophyton rubrum, Aspergillus niger, Aspergillus fumigates and Candida tropicalis). The in vitro cytotoxic abilities of the compounds were analysed using the MTT assay on MCF-7 (human breast cancer) and A549 (human lung carcinoma) cell lines and compared with standard cisplatin. Activities greater than cisplatin were observed for the ligands as well as the complexes. Assays on the human normal keratinocyte cell line HaCaT showed that the compounds were non-toxic to those cells. In all the biological studies done, the complexes displayed better activity than the ligands and among them, complex 3 stood out as the best, owing to the greater electron donating ability of the N-terminal ethyl group on the thiosemicarbazone. The results implied that N-terminal substitution played a significant role in bringing out the bioactivity of the complexes.

New cyclometallated ruthenium(ii) complexes of 3-acetyl-7-methoxycoumarin-4N-substituted thiosemicarbazones were synthesized and characterized by analytical and spectral techniques. The crystal structures of the ligands H2L1-3 and complexes (1, 2 and 4) were confirmed by X-ray crystallography. The analysis showed that the ligands have undergone C-H activation at the C(4) carbon of the pyrone ring and acted in a tridentate fashion by binding through C, N and S atoms. CT-DNA and protein (BSA/HSA) binding studies were carried out to analyze their interaction with biomolecules. Good binding affinity with DNA was observed with intercalative binding mode, which was further confirmed by EB displacement and viscosity measurement studies. The quenching mechanism with BSA/HSA was found to be static. Three dimensional (3D) fluorescence measurements were carried out to validate the micro environmental changes in the serum albumins. Their antioxidant propensity and antimicrobial study insisted that the compounds displayed good spectrum of activity. Evaluation of their anticancer potential against MCF-7 (human breast cancer) and A549 (human lung carcinoma) cell lines revealed that the complexes exhibited better activity than the ligands and cisplatin. Further, the results of LDH and NO release assays supported the cytotoxic nature of the compounds. The non-toxic nature of the compounds was established by testing against the non-cancerous cell line HaCaT (human normal keratinocyte). © 2018 The Royal Society of Chemistry.

Four Ru(II) carbonyl complexes of 4-oxo-4H-chromene-3-carbaldehyde thiosemicarbazones (C1-C4) were synthesized and characterized through spectroscopic techniques. The coordination behaviour of the ligands to the metal ion was confirmed by solving the single crystal X-ray structure of a representative complex C4, in which the ligand showed coordination via the C2 carbon of the chromone ring, the azomethine nitrogen and thiolate sulfur. The interaction with DNA and protein showed a good binding profile of the complexes. Antimicrobial effects of the complexes were tested against bacterial and fungal strains, for which a significant activity was observed. Furthermore, MTT assay on A549 and MCF-7 cell lines indicated the cytotoxic potential of the synthesized complexes, overcoming the cisplatin activity under similar experimental conditions.

Nanosized manganese ferrite (MnFe2O4) particles were prepared by sol–gel method using natural polymers like wheat flour (WF) and potato flour (PF) as surfactants and its structural, morphological, optical and magnetic characteristics were studied by X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FT-IR), scanning electron microscope (SEM), photoluminescence spectroscopy (PL) and vibration sample magnetometer (VSM). Brunauer–Emmett–Teller (BET) surface area test also performed and the results obtained were discussed. The average crystallite size was found to be 23 and 16 nm for WF/MnFe2O4 and PF/MnFe2O4 samples, respectively. Magnetic hysteresis loops confirmed the super-paramagnetic behavior for both the samples. For oxidation of benzyl alcohol to benzaldehyde, the catalytic activity of MnFe2O4 nanoparticles (NPs) was carried out. Antimicrobial and antifungal activity of WF/MnFe2O4 and PF/MnFe2O4 samples were investigated against two Gram-positive bacteria (Staphylococcus aureus, Streptococcus pneumoniae), two Gram-negative bacteria (Pseudomonas aeruginosa, Salmonella paratyphi) and fungus (Candida albicans) using inhibition zone method. Minimum Inhibitory Concentration (MIC) values also calculated to determine susceptibilities of bacteria to drugs and also to evaluate the activity of new antimicrobial agents. The in vitro cytotoxicity of newly synthesized samples were analyzed by MTT assay against MCF-7, A549 and HaCaT cell lines in a dose-dependent fashion. Among these two samples, sample B (using potato flour) shows better response than sample A (using wheat flour) and both the samples were non-toxic to normal cell line. The concentration required to kill 50% of the cell (IC50) was also calculated. MnFe2O4 nanoparticles were synthesized by sol–gel method using natural polymers, wheat flour and potato flour, as surfactant. The as-prepared MnFe2O4 was characterized by XRD, FT-IR, SEM, EDX, PL and VSM analysis. The average crystallite size was found to be 23 and 16 nm for WF/MnFe2O4 and PF/MnFe2O4 samples, respectively. Magnetic hysteresis loops confirmed the super-paramagnetic behavior for both the samples. The catalytic activity of MnFe2O4 nanoparticles (NPs) were carried out for oxidation of benzyl alcohol to benzaldehyde. Biological activities like antimicrobial, antifungal and anticancer activities of the samples were investigated. Among these two samples, PF/MnFe2O4 shows better response than WF/MnFe2O4 and both the samples were non-toxic to normal cell line. IC50 values of WF/MnFe2O4 (a) and PF/MnFe2O4 (b) against A549 and MCF-7 cell lines

Summary Sperm DNA Fragmentation has been extensively studied for more than a decade. In the 1940s the uniqueness of the spermatozoa protein complex which stabilizes the DNA was discovered. In the fifties and sixties, the association between unstable chromatin structure and subfertility was investigated. In the seventies, the impact of induced DNA damage was investigated. In the 1980s the concept of sperm DNA fragmentation as related to infertility was introduced as well as the first DNA fragmentation test: the Sperm Chromatin Structure Assay (SCSA). The terminal deoxynucleotidyl transferase nick end labelling (TUNEL) test followed by others was introduced in the nineties. The association between DNA fragmentation in spermatozoa and pregnancy loss has been extensively investigated spurring the need for a therapeutic tool for these patients. This gave rise to an increased interest in the aetiology of DNA damage. The present decade continues within this research area. Some of the more novel methods recently submerging are sorting of cells with increased DNA fragmentation and hyaluronic acid (HA) binding techniques. The clinical value of these tests remains to be elucidated. In spite of half a century of research within the area, this analysis is not routinely implemented into the fertility clinics. The underlying causes are multiple. The abundance of methods has impeded the need for a clinical significant threshold. One of the most promising methods was commercialized in 2005 and has been reserved for larger licensed laboratories. Myriads of reviews and meta‐analyses on studies using different assays for analysis of DNA fragmentation, different clinical Artificial Reproductive Treatments (ART), different definitions of successful ART outcome and small patient cohorts have been published. Although the area of DNA fragmentation in spermatozoa is highly relevant in the fertility clinics, the need for further studies focusing on standardization of the methods and clinical implementation persists.

The present work deals with the synthesis and characterization of four Co(III) complexes derived from 7-hydroxy-4-oxo-4H-chromene-3-carbaldehyde-4(N)-substituted thiosemicarbazones. Structural confirmation of the complexes 2 and 3 were aided by X-ray crystallographic analysis, which revealed the coordination of azomethine nitrogen, ring carbonyl oxygen and thiol sulfur atoms of the ligands. DNA and Bovine serum albumin (BSA) binding studies brought about the binding ability of the complexes to biomolecules. The binding constant values calculated for the complexes were in the range (0.30–1.19) × 106 M−1 and the mode of interaction was found to be partially intercalative. The mechanism of quenching with BSA was static with quenching constants in the order 1012 M−1s−1. Their antimicrobial profile has been established by testing against four pathogenic bacterial and fungal species. The cytotoxic potential of the complexes were examined against MCF-7 and A549 cells by MTT assay. The IC50 values of the complexes were less than cisplatin, which indicated their greater activity over the standard drug. In addition, testing on human normal keratinocyte cells HaCaT indicated their non toxic nature. Lactate dehydrogenase (LDH) and Nitric oxide (NO) release assays supported the cytotoxic ability and suggested apoptotic mode of cell death induced by the complexes.

Nickel ferrite NiFe2O4 nanoparticles (NPs) were successfully synthesized by using nickel nitrate, ferric nitrate, citric acid, and ethyl cellulose as a surfactant by a simple sol–gel-assisted combustion method (SACM) and microwave-assisted combustion method (MACM). Structural, morphological, optical, and magnetic properties of the obtained powder were characterized by powder X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), photoluminescence (PL) spectroscopy, and vibrating sample magnetometry (VSM). XRD results show that the resultant powder was pure crystalline with cubic structure. The average crystalline size was found to be 18.8 and 10.2 nm synthesized by SACM and MACM, respectively. FT-IR spectra indicate the type of bonds between Ni–O and Fe–O (metal and oxygen). SEM images show that the morphology of the powder consists of well-defined structure. VSM results showed a ferromagnetic behavior of the sample. Antimicrobial activity of NiFe2O4 nanoparticles was performed. Both sample 1 (SACM) and sample 2 (MACM) show good inhibition in the zone 100 μg/ml. While comparing, sample 2 shows high inhibition than sample 1.

Early life nutrition plays an important role in brain development. Emerging research in rodents, piglets and humans suggest that prebiotics, milk fat globule membrane and lactoferrin may each play unique roles in brain development and cognitive functions. However, knowledge of their combined impact is lacking. We show here that providing weanling rats with a diet containing milk fat globule membrane, lactoferrin and a polydextrose/galactooligosaccharide prebiotic blend led to a significant increase in total dendritic spine density in hippocampal dentate gyrus neurons. Region-specific alterations in dendritic spine density and morphology could provide a mechanistic basis underlying broader cognitive benefits, but further research is required to demonstrate functional consequences of these observations.

Thin films of PbS doped with tin (2 ≤ Sn ≤ 8 wt.%) were deposited on glass substrates by cost- effective nebulizer spray method. X-ray diffraction studies confirm the polycrystalline nature with a face centered cubic structure along with the orientation of (200) plane. SEM/AFM studies reveal that the nano-sized spherical and granular shaped grains roofed the whole film surface and the grain shapes were changed with the change in Sn doping percentage. Bandgap was found to decrease from 1.98 eV to 1.66 eV with an increase in the doping concentration from 2 to 6 wt%. Hall Effect measurement confirmed p-type conducting nature for all the doped PbS films. The resistivity and carrier concentration values are found to be about 0.32 × 103 Ωcm and 6.78 × 1013 cm−3, respectively, for the optimized PbS:Sn film (6 wt%). The response of the heterostructured (FTO/n-CdS/p-PbS:Sn/Ag) solar cell with the optimal Sn doping (6 wt% Sn) was studied under dark and illuminated conditions.

This paper fortifies the idea of smart drug vehicles, by exploring nickel ferrite nanoparticles. The material is made smart by covering with a novel biocompatible dextran fastened with a hydrophobic cavity containing molecule, β-cyclodextrin (β-CD). The approximately 6 nm MNPs, embodied with the β-CD-dextran conjugate, permit a sustained in vitro release of camptothecin. Cytotoxicity studies are done on HaCaT, 3T3, HeLa, MDA-MB-231, and A549 cell lines. It is fascinating the stacked camptothecin indicates upgraded viability against cancer cells, rendering the nanocarrier viable for transporting the anticancer drug.

This study investigated the wound healing properties of the aqueous extract of Morinda tinctoria Roxb leaves in rats. The study also provides information on the purification, qualitative, and quantitative analysis of phytochemical components present in M. tinctoria Roxb. Wound contraction and period of epithelialization was determined and topical application of M. tinctoria Roxb aqueous leaf extract showed better healing than orally treated and control groups. The results suggest that the efficacy of M. tinctoria Roxb aqueous leaf extract as a wound healing agent and can also be used as a therapeutic agent for internal as well as external wounds.

We investigated the role of oxidative stress and the inflammasome in trauma-induced axon degeneration and vision loss using a mouse model. The left eyes of male mice were exposed to over-pressure air waves. Wild-type C57Bl/6 mice were fed normal, high-vitamin-E (VitE), ketogenic or ketogenic-control diets. Mice lacking the ability to produce vitamin C (VitC) were maintained on a low-VitC diet. Visual evoked potentials (VEPs) and retinal superoxide levels were measured in vivo. Tissue was collected for biochemical and histological analysis. Injury increased retinal superoxide, decreased SOD2, and increased cleaved caspase-1, IL-1α, IL-1β, and IL-18 levels. Low-VitC exacerbated the changes and the high-VitE diet mitigated them, suggesting that oxidative stress led to the increase in IL-1α and activation of the inflammasome. The injury caused loss of nearly 50% of optic nerve axons at 2 weeks and astrocyte hypertrophy in mice on normal diet, both of which were prevented by the high-VitE diet. The VEP amplitude was decreased after injury in both control-diet and low-VitC mice, but not in the high-VitE-diet mice. The ketogenic diet also prevented the increase in superoxide levels and IL-1α, but had no effect on IL-1β. Despite this, the ketogenic diet preserved optic nerve axons, prevented astrocyte hypertrophy, and preserved the VEP amplitude. These data suggest that oxidative stress induces priming and activation of the inflammasome pathway after neurotrauma of the visual system. Further, blocking the activation of the inflammasome pathway may be an effective post-injury intervention.

The retrosplenial cortex (RSC) plays a critical role in episodic memory, but the molecular mechanisms governing plasticity in this structure are poorly understood. Diverse studies have demonstrated a role for RSC in acquisition, early consolidation and retrieval similar to the hippocampus (HC), as well as in systems consolidation similar to the anterior cingulate cortex. Here, we asked whether established molecular and structural substrates of memory consolidation in the HC also engage in RSC shortly after learning. We show striking parallels in training induced gene-activation in HC and RSC following contextual conditioning, which is blocked by systemic administration of an NMDA receptor antagonist. Long-term memory is enhanced by retrosplenial and hippocampal knockdown (KD) of the cAMP specific phosphodiesterase Pde4d. However, while training per se induces lasting spine changes in HC, this does not occur in RSC. Instead, increases in the number of mature dendritic spines are found in the RSC only if cAMP signaling is augmented by Pde4d KD, and spine changes are at least partially independent of training. This research highlights parallels and differences in spine plasticity mechanisms between HC and RSC, and provides evidence for a functional dissociation of the two.

A series of 3-acetyl-8-methoxycoumarin appended thiosemicarbazones (1-4) was prepared from the reaction of 3-acetyl-8-methoxycoumarin with 4(N)-substituted thiosemicarbazides in a view of ascertaining their biological properties with the change of N-terminal substitution in the thiosemicarbazide moiety. Comprehensive characterization was brought about by various spectral and analytical methods. The molecular structures of all the compounds were determined by single crystal X-ray diffraction analysis. Binding studies with Calf thymus DNA (CT-DNA) and proteins such as Bovine Serum Albumin (BSA) and Human Serum Albumin (HSA) indicated an intercalative mode of binding with DNA and static quenching mechanism with proteins. The compounds cleaved plasmid DNA (pBR322) and acted well as free radical scavengers. A good spectrum of antimicrobial activity was observed against four bacterial and five fungal pathogens. The compounds exhibited profound antiproliferative activity on MCF-7 (human breast cancer) and A549 (human lung carcinoma) cell lines. Assay on human normal keratinocyte cell line HaCaT showed that the compounds were non-toxic to normal cells.

New Schiff base ligands are prepared by the condensation of 7-hydroxy-3-formylchromone with semicarbazone and phenyl semicarbazone. The complexation of these ligands with Cu(II) ion is proposed in the light of spectral studies (IR, UV-Vis, 1H NMR, 13C NMR, Mass and ESR). In the complexes 1 and 2, the ligands coordinate to the Cu(II) ion in a neutral fashion via ONO donor atoms. The single crystal XRD studies reveal a slightly distorted square-pyramidal geometry for cationic complex (1) and an octahedral geometry for neutral complex (2). Preliminary biological studies such as DNA and Protein binding are carried out by using absorption and emission titration methods. Observation of intercalative mode of binding with Calf Thymus DNA (CT-DNA) is confirmed by means of viscosity measurements. The micro-environmental changes occurring in Bovine Serum Albumin (BSA) and Human Serum Albumin (HSA) are monitored via three dimensional (3D) fluorescence studies. The compounds ability in inhibiting microbial growth is tested against different pathogens. MCF-7 (human breast cancer) and A549 (human lung carcinoma) cell lines are utilized to check the anticancer potential of the synthesized compounds by using MTT, LDH and NO assays. The results show that complexes 1 and 2 exhibited potent cytotoxic activity over standard drug cisplatin.

Erythropoietin (EPO) is critical for red blood cell production and is also an effective neuroprotective agent. However, it may also contribute to pathological angiogenesis. Here we investigate the angiogenic potential of EPO and a mutant form with attenuated erythropoietic activity, EPO-R76E, on primary human retinal microvascular endothelial cells (HRMEC) and in the adult retina. Assays of death, proliferation, and tube-formation were performed on HRMECs exposed to EPO, EPO-R76E, or media alone. Postnatal day 9 wild-type mice were injected intramuscularly with adeno-associated virus vectors expressing either enhanced green fluorescent protein or EpoR76E. At 3 months, levels of EPO-R76E in the eye were quantified, and the health of the retinal vasculature was assessed by fluorescein angiography and isolectin immunolabeling. Immunohistochemistry, histology, and electroretinogram assessments were performed as measures of retinal health. Neither EPO nor EPO-R76E induced proliferation or tube-formation in HRMEC under the conditions used. EPO-R76E decreased HRMEC death in a dose-dependent manner. Long-term systemic gene delivery of EPO-R76E was safe in terms of retinal vasculature, histology, and the electroretinogram in vivo. Our results show that EPO-R76E can block HRMEC death, consistent with its role in erythropoiesis and neuroprotection. In addition, long-term gene delivery of EPO-R76E is safe in the adult retina.

The eutectic spacing (λ) was investigated in three alloys of succinonitrile–neopentylglycol–(D)camphor–aminomethylpropanediol (SCN–NPG–DC–AMPD) system over a wide range of temperature gradients (G) and growth velocities (V), both in thin ⩽5 μm (two-dimensional (2-D)) samples and in bulk (three-dimensional (3-D)) samples. The compositions were selected such that fraction of rod-forming phase (f) was equal to 11.5 vol.% for the invariant SCN–12.2 wt.% NPG alloy, 23.3 vol.% for the univariant SCN–24.2 wt.% DC–0.5 wt.% NPG alloy and 29.8 vol.% for the bivariant SCN–33.0 wt.% DC–6.0 wt.% NPG–3.0 wt.% AMPD alloy. For high G/V conditions significant deviations from growth at minimum undercooling were found with spacing selection closer to the pinch-off limit. The experimental results show that confining the 3-D eutectics to 2-D growth conditions invariably leads to larger values of the mean rod spacing, but to smaller values of the mean rod diameter. The 2-D/3-D ratio of the square of mean rod diameters η = d 2 D R / d 3 D R 2 was found to be independent of the phase fraction of the rod-forming phase f and equal to η = 0.80 ± 0.02. The 2-D/3-D ratio of square of rod spacing k = λ 2 D R / λ 3 D R 2 was found to dependent on f following k = ( 2 f ) - 1 or else k = η 3 8 π f - 1 . These findings are discussed in terms of the Jackson–Hunt model of eutectic growth and simple geometrical considerations.

7-Hydroxy-4-oxo-4H-chromene Schiff bases and their corresponding water soluble copper(II) complexes were synthesized and characterized by various spectral techniques. The structural confirmation of the complexes 2 and 3 has been provided by X-ray crystallographic analysis. The binding affinities of the ligands and complexes with Calf Thymus DNA (CT-DNA) and proteins (Bovine Serum Albumin (BSA) and Human Serum Albumin (HSA)) were studied by using absorption and emission titration methods. An intercalative mode of binding with CT-DNA and a static quenching mechanism with albumin (BSA and HSA) was confirmed from the obtained results. The efficiency of the compounds in inhibiting the microbial growth was tested against various pathogenic bacteria Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pneumoniae and Acinetobacter baumannii by using Gentamicin as standard and fungi Candida albicans, Aspergillus niger, Trichophyton rubrum, Candida tropicalis and Aspergillus fumigatus by using ketoconazole as standard and a significant activity has been found for the complexes. The anticancer potential of the ligands and complexes were tested against A549 and MCF-7 cells by MTT assay using cisplatin as a standard. The complexes 1–4 showed better cytotoxic profile than cisplatin, when incubated for a period of 48 h and 72 h. The results of all the biological analyses indicated the higher activity of the complexes over the ligands. Out of the complexes, complex 3 with the more electron donating ethyl group exhibited the higher activity.

This work deals with the study on the key properties of PbS thin films prepared by simple nebulizer spray method. The structural study done by XRD exhibited cubic crystal structure with polycrystalline nature for the deposited PbS films. The obtained crystallite size was varied between 41 to 55 nm for various molar concentrations. The morphology study done by SEM displays uniform distribution of the grains all over the surface of the prepared PbS thin films. The observed band gap value was decreased from 1.8 eV to 1.51 eV for various molar concentrations. The emitted PL intensity was strong at about 580 nm which is related to the near band edge (NBE) emission. Electrical studies performed by Hall Effect measurement confirm the p-type conductivity of prepared films. Resistivity and carrier concentrations are 2.92 × 103 Ωcm and 4.83 × 1013cm−3 respectively for the deposited films. In addition, a heterojunction solar cell of structure FTO/n-CdS/p-PbS/Ag was produced and its conversion efficiency was ~0.39%. The nebulizer spray technique is very useful and novel method for the preparation of device quality PbS thin film.

Alloys from the succinonitrile–(D)camphor (SCN–DC) system are widely used as model alloys for the in situ investigation of solidification using light optical microscopy, but literature on the binary phase diagram is contradictory with respect to the solubility limit of DC in (SCN). Phase equilibria of the system were therefore revisited experimentally and critically assessed in the present work. The results prove that the maximum solubility of DC in the succinonitrile solid solution (SCN) is far less 1 wt% and the volume fraction of the (DC) phase in the eutectic solid is 23.3%. On this basis and on recently reported experimental data the CALPHAD description of the SCN–DC system was re-optimized.

Characterization of the response of the Balb/c mouse to an eye-directed overpressure airwave, with the hypothesis that this mouse strain and model is useful for testing potential therapeutics for the treatment of traumatic eye injury.The left eyes of adult Balb/c mice were exposed to an eye-directed overpressure airwave. Intraocular pressure (IOP) was measured and eyes were inspected for gross pathology changes. Optical coherence tomography and histology were used to examine the structural integrity of the retina and optic nerve. Immunohistochemistry, in vivo molecular fluorophores, and a multiplex enzyme-linked immunosorbent assay were utilized to identify changes in cell death, neuroinflammation, and oxidative stress.This model induced a transient increase in IOP, corneal injuries, infrequent large retinal detachments, retinal pigment epithelium (RPE) vacuolization, glial reactivity, and retinal cell death. Both the corneal damage and RPE vacuolization persisted with time. Optic nerve degeneration occurred as early as 7 days postinjury and persisted out to 60 days. Retinal cell death, increased levels of reactive oxygen species, and neuroinflammation were detected at 7 days postinjury.The injury profile of the Balb/c mouse is consistent with commonly observed pathologies in blast-exposed patients. The damage is throughout the eye and persistent, making this mouse model useful for testing cell-based therapies.

Erythropoietin (Epo) is neuroprotective in a number of preparations, but can lead to unacceptably high and even lethal hematocrit levels. Recent reports show that modified Epo variants confer neuroprotection in models of glaucoma and retinal degeneration without raising hematocrit. In this study, neuroprotective effects of two Epo variants (EpoR76E and EpoS71E) were assessed in a model of Parkinson's disease. The constructs were packaged in recombinant adeno-associated viral (rAAV) vectors and injected intramuscularly. After 3 weeks, mice received five daily injections of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and were killed 5 weeks later. The MPTP-lesioned mice pretreated with rAAV.eGFP (negative control) exhibited a 7- to 9-Hz tremor and slower latencies to move on a grid test (akinesia). Both of these symptomatic features were absent in mice pretreated with either modified Epo construct. The rAAV.eGFP-treated mice lesioned with MPTP exhibited a 41% reduction in tyrosine hydroxylase (TH)-positive neurons in the substantia nigra. The rAAV.EpoS71E construct did not protect nigral neurons, but neuronal loss in mice pretreated with rAAV.EpoR76E was only half that of rAAV.eGFP controls. Although dopamine levels were normal in all groups, 3,4-dihydroxyphenylacetic acid (DOPAC) was significantly reduced only in MPTP-lesioned mice pretreated with rAAV.eGFP, indicating reduced dopamine turnover. Analysis of TH-positive fibers in the striatum showed normalized density in MPTP-lesioned mice pretreated with rAAV.EpoS71E, suggesting that enhanced sprouting induced by EpoS71E may have been responsible for normal behavior and dopaminergic tone in these mice. These results show that systemically administered rAAV-generated non-erythropoietic Epo may protect against MPTP-induced parkinsonism by a combination of neuroprotection and enhanced axonal sprouting.

A new experimental method for in-situ observation of microstructure formation in top view during unidirectional solidification of bulk, transparent, organic alloys is presented. This method allows observing growth patterns over an extended interface area with high resolution and minimal optical aberrations. With (D)camphor – neopentylglycol – succinonitrile (DC–NPG–SCN) alloys a series of unidirectional solidification experiments were performed in order to validate the set-up. By means of multi-focus exposition eutectic cells were observed over a depth of several millimeters, followed by 3D reconstruction of their shape. The method also allows capturing the integral interface dynamics and measuring its relative undercooling.

The retinal degeneration slow (rds/rds) mouse was used to test photoreceptor protection by systemic gene delivery of non-erythropoietic forms of erythropoietin (EPO). Two Epo mutants were generated and packaged into recombinant adeno-associated virus (rAAV) serotype 2/5, controls included rAAV2/5.Epo and rAAV2/5.enhanced green fluorescent protein (eGFP). Mice were injected in the quadriceps at postnatal day seven and analyses were performed at postnatal day 90. Hematocrit, serum EPO levels, and outer nuclear layer (ONL) thickness were quantified. Hematocrit and serum EPO levels in rAAV2/5.eGFP, rAAV2/5.Epo, and rAAV2/5.EpoR103E treated mice were: 46%, 8 mU/ml; 63%, 117 mU/ml; and 52%, 332 mU/ml, respectively. The ONL from rds/rds mice treated with the Epo vectors were approximately twice as thick as the negative controls. This demonstrates that the photoreceptors can be protected without performing an intraocular injection and without increasing the hematocrit to unsafe levels. Intramuscular delivery of rAAV.EpoR103E is an attractive treatment for retinal degenerative diseases.

Learning-induced trophic activity is thought to be critical for maintaining health of the aging brain. We report here that learning, acting through an unexpected pathway, activates synaptic receptors for one of the brain's primary trophic factors. Unsupervised learning, but not exploratory activity alone, robustly increased the number of postsynaptic densities associated with activated (phosphorylated) forms of BDNF's TrkB receptor in adult rat hippocampus; these increases were blocked by an NMDA receptor antagonist. Similarly, stimulation of hippocampal slices at the learning-related theta frequency increased synaptic TrkB phosphorylation in an NMDA receptor–dependent fashion. Theta burst stimulation, which was more effective in this regard than other stimulation patterns, preferentially engaged NMDA receptors that, in turn, activated Src kinases. Blocking the latter, or scavenging extracellular TrkB ligands, prevented theta-induced TrkB phosphorylation. Thus, synaptic TrkB activation was dependent upon both ligand presentation and postsynaptic signaling cascades. These results show that afferent activity patterns and cellular events involved in memory encoding initiate BDNF signaling through synaptic TrkB, thereby ensuring that learning will trigger neurotrophic support.

The thermodynamic description of the ternary system B–Nb–Ti is obtained by modelling the Gibbs energy of all individual phases in the system using the CALPHAD approach. The model parameters have been evaluated by means of a computer optimization technique based on the available descriptions of the constituent binary systems B–Nb, B–Ti and Nb–Ti and relevant experimental information on phase equilibria for the ternary system. The thermodynamic descriptions of both systems with boron, B–Nb and B–Ti, were published in the first part of the work. For the ternary system B–Nb–Ti several vertical and isothermal sections as well as the projection of the liquidus and solidus surface are calculated using the elaborated thermodynamic description. A tolerable agreement between the calculations and experimental data is achieved.

The temperature of phase transformations, as well as the enthalpy of melting of 29 different alloys from the Bi–In–Sn system were measured by DSC using a Perkin–Elmer DSC-7 instrument. The following tie-triangles were characterized in terms of analysing the chemical composition of the phases in equilibrium, using energy dispersive X-ray analysis (EDX): (Bi)–BiIn–(Sn) at 77 °C, BiIn 2 -β-γ at 59 °C including the composition of the liquid phase at 59 °C and the tie-line BiIn 2 -γ at 59 °C. A new thermodynamic description is presented for the ternary Bi–In–Sn system in the entire composition range. The parameters for the thermodynamic models of the constituent binary systems Bi–In, Bi–Sn and In–Sn are adopted from earlier assessments, and those for the Bi–In–Sn system are optimized in this study using experimental data on phase equilibria and data on the enthalpy of melting of different alloys available in the literature and measured in the present work. Several vertical and isothermal sections as well as the liquidus surface and selected thermodynamic properties are calculated using the thermodynamic description. They show reasonably good agreement with reported experimental data.

These are exciting times in the field of retinal gene therapy, as impressive successes have been achieved by recombinant adeno-associated viral (rAAV) gene delivery of Rpe65 or Rep1 to the retinal pigment epithelium of patients with Lebers congential amaurosis or choroideremia, respectively.1–5 Instead of being left with incurable blindness, patients are reporting restoration of vision. An advantage of these disease targets is that the affected cell type, the retinal pigment epithelium, is amenable to gene therapy since it is naturally phagocytic, resulting in efficient transduction, and is not adversely affected by overexpression of exogenous transgenes. In contrast, the majority of inherited retinal degenerations are due to mutations in photoreceptor-specific transcripts causing primary death of these cells and permanent vision loss.6 Novel studies published in Molecular Therapy–Methods & Clinical Development address such challenges for photoreceptor gene therapy, including rapid onset of cell death (resulting in pathological changes to the retinal environment) and inefficient transduction of photoreceptors by gene therapy vectors. Palfi et al.7 address the latter challenge by demonstrating that rAAV2/rh10 transduces photoreceptors as efficiently as the current gold standard, rAAV2/8.8,9 The number of cells transduced and level of transgene expression achieved by Palfi et al., with rAAV2/rh10 is sufficient to provide histological, electrophysiological, and behavioral improvements in a mouse model of autosomal recessive retinitis pigmentosa due to mutations in rhodopsin. Their studies are among the first to show success in the most common form of retinitis pigmentosa.10–12 While very exciting and full of promise, the effect is not long-lasting, consistent with results in other studies using rAAV-mediated gene addition to rescue photoreceptors.12–19 A decreased rate of degeneration is certainly clinically relevant as it could translate to the addition of years of precious sight in patients. At the same time we need to understand why the efficacy of gene therapy in photoreceptors decreases over time despite addressing the intrinsic deficit (lack of functional protein). This article will discuss two potential explanations: (i) insufficient gene expression within the cells causing intrinsic cell death pathways to still activate; and (ii) changes in the retinal milieu that initiate cell death by extrinsic signaling pathways. Palfi et al.11, both in the recent article in this journal7 and in their 2010 paper,12 achieved a fantastic feat—improvement of structure and function by gene delivery of rhodopsin into the rhodopsin knock-out mouse. Their studies also highlight the challenges of gene augmentation therapy for retinal degeneration. Despite a significant effort by Palfi et al. to increase expression of rhodopsin, the disorganization of the outer segment discs and lower visual function (spatial acuity threshold and electroretinogram) in the treated mice as compared to wild-type controls suggest that normal levels of rhodopsin were still not achieved. Further, the mice still exhibited progressive retinal degeneration. Studies by other groups suggest that increasing gene expression alone may not be sufficient to completely block progression of inherited retinal degenerations. First, increasing gene expression either through repeated injections of rAAV or by use of a more efficient rAAV vector does not overcome this progressive decline in photoreceptors.16,17 Second, studies in both dogs and mice show that the therapeutic effects of augmentation gene therapy are more sustained when it is given early, prior to onset of photoreceptor degeneration.13,15 Many inherited retinal degenerations due to deficits in photoreceptor-specific proteins have a very early onset. Therefore, the retinal environment is likely altered prior to intervention by gene therapy. For example, glial reactivity can be detected prior to significant retinal degeneration in multiple models of retinal degeneration.20–24 Glial cell reactivity can be beneficial acutely, but can cause oxidative stress and neuroinflammation, leading to neuronal death if the glia remain reactive long-term (for review, see ref. 25). In fact, Palfi et al., detected infiltrates in the rhodopsin knock-out mouse, although they did not assess the level of reactivity of the endogenous glial cells. However, robust and long-term benefit may be achieved by a combined approach targeting both the intrinsic gene defect and extrinsic signaling (i.e., neuroprotection). This approach has shown success in a mouse model of inherited retinal degeneration.26 Gene delivery of several neuroprotective agents has shown promise in models of inherited retinal degeneration.27–34 It is important to note that not all neuroprotective factors are created equal, and all inherited retinal degenerations are not the same. It is critical to have a good understanding of the molecular pathways both activated by the inherited retinal degeneration of interest and inhibited by each potential neuroprotective factor in order to properly pair them. For example, if the photoreceptor degeneration is driven entirely by intrinsic pathways that activate apoptotic cell death, then only those factors that block apoptosis will be effective. In contrast, if autophagy, pyroptosis, or necroptosis are critical, then one needs a factor that will target those pathways. In addition, late stage inherited retinal degenerations, as well as complex retinal degenerations such as glaucoma, diabetic retinopathy, and age-related macular degeneration, are affected by extrinsic factors such as glial reactivity (as mentioned above), oxidative stress, and/or neovascularization, thus requiring factors that will target these processes. Fortunately, most neuroprotective factors are pluripotent such that they can address both intrinsic and extrinsic pathways. Shanab et al.35 present a promising neuroprotective gene therapy approach that targets neovascularization in diabetic retinopathy. In the retina, neovascularization most commonly occurs during retinopathy of prematurity and as a common secondary complication to diabetic retinopathy. It can also occur in advanced stages of inherited retinal degenerative diseases, in which the lack of photoreceptors creates a more oxidative environment leading to neovascularization in the retina.36 This group previously demonstrated that an oxidative environment leads to decreased MMP-7 activity resulting in decreased processing of pro-nerve growth factor (NGF) into NGF in both experimental and clinical diabetes.37–39 While it is well known that NGF is neuroprotective through activation of TrkA (for review, see ref. 40), they showed that proNGF promoted neuroinflammation and cell death by activation of p75NTR.41–43 In their current paper, the authors used virus-mediated (lentivirus) gene delivery of a cleavage resistant proNGF to induce endothelial cell death and counteracted the damage by treatment with a lentivirus carrying shRNA targeting the p75NTR. This approach successfully protected the endothelial cells both in vivo and in vitro. These results are encouraging and further studies should be performed to test the general applicability of this approach for retinal neovascularization as a result of retinopathy of prematurity, and choroidal neovascularization as a result of age-related macular degeneration or end stage inherited retinal degenerations. There is a need for a new approach for blocking neovascularization since recent studies show that anti–vascular endothelial growth factor therapies are ineffective at blocking neovascularization in a subset of patients.44 In summary, two studies now published in Molecular Therapy–Methods & Clinical Development highlight the great strides being made in retinal gene therapy and the future of the field. Palfi et al. demonstrate great progress in the treatment of retinal degenerations due to lack of functional rhodopsin using a new rAAV serotype. Shanab et al. illustrate the utility of virus-mediated gene delivery for understanding the molecular events that underlie disease, in this case neovascularization, and demonstrate a new approach for blocking pathological angiogenesis. This and other neuroprotective approaches will be necessary for treating complex retinal degenerations and may improve outcomes in inherited retinal degenerations. The lack of long-term preservation of vision after gene addition into photoreceptors underscores the need for both gene augmentation and neuroprotection approaches in order to achieve great clinical success. The field is also propelled forward by the development of new serotypes of rAAV that have greater transduction efficiency, faster onset of transgene expression, and target different cell types (i.e., photoreceptors, glia, or ganglion cells). In conclusion, for the field of retinal gene therapy to reach the ultimate goal of treating all forms of blinding disease, multiple groups will need to work together, collaboratively, bringing their expertise on the biology of disease, vector development, and both gene targeted and neuroprotective strategies together to develop synergistic, multipronged gene therapies.

Several key experiments were performed to determine the temperature of transformations of organic alloys from the ternary systems 2-amino-2-methyl-1,3-propanediol–(D)camphor–succinonitrile (AMPD-DC-SCN) and 2-amino-2-methyl-1,3-propanediol–neopentylglycol–succinonitrile (AMPD-NPG-SCN) by means of differential scanning calorimetry (DSC). The phase diagrams of these ternary systems were assessed via the CALPHAD approach using Thermo-Calc and the experimental data on phase equilibria were determined by the above key experiments. Good agreement between the experimental and the calculated data was achieved for both ternary systems. Combining the obtained thermodynamic descriptions with those obtained for two other ternary systems, AMPD–DC–NPG and DC–NPG–SCN, published earlier, the thermodynamic description for the entire quaternary system was created. In order to identify the phases involved in solid–liquid equilibria and the nature of coupled eutectic growth in quaternary alloys, several unidirectional solidification experiments were performed: two quaternary alloys with composition 37.5SCN–44.5DC–16.5NPG–1.5AMPD (wt%) and 34SCN–44DC–19NPG–3AMPD (wt%) were used for the experiments on monovariant eutectic growth. We find that in the last alloy the monoclinic NPG phase is fully suppressed by the addition of AMPD and three solid phases show coupled eutectic growth at the solid/liquid interface: the hexagonal phase (DC solid solution), the FCC phase (NPG–AMPD solid solution) and the BCC phase (SCN solid solution). The quasi-2D eutectic patterns obtained in a thin sample show the stacking sequence –BCC–Hex–FCC–Hex–. The volume ratio of Hex:BCC:FCC phases equals 4:3:2. In the quaternary alloy with 1.5 wt% AMPD the monoclinic NPG phase is present at the solid/liquid interface and the eutectic growth is rather irregular.

Rex, Christopher S., Julie C. Lauterborn, Ching-Yi Lin, Eniko A. Kramár, Gary A. Rogers, Christine M. Gall, and Gary Lynch. Restoration of long-term potentiation in middle-aged hippocampus after induction of brain-derived neurotrophic factor. J Neurophysiol 96: 677-685, 2006. First published May 17, 2006; doi:10.1152/jn.00336.2006. Restoration of neuronal viability and synaptic plasticity through increased trophic support is widely regarded as a potential therapy for the cognitive declines that characterize aging. Previous studies have shown that in the hippocampal CA1 basal dendritic field deficits in the stabilization of long-term potentiation (LTP) are evident by middle age. The present study tested whether increasing endogenous brain-derived neurotrophic factor (BDNF) could reverse this age-related change. We report here that in middle-aged (8- to 10-mo-old) rats, in vivo treatments with a positive AMPA-type glutamate receptor modulator both increase BDNF protein levels in the cortical telencephalon and restore stabilization of basal dendritic LTP as assessed in acute hippocampal slices 18 h after the last drug treatment. These effects were not attributed to enhanced synaptic transmission or to facilitation of burst responses used to induce LTP. Increasing extracellular levels of BDNF by exogenous application to slices of middle-aged rats was also sufficient to rescue the stabilization of basal dendritic LTP. Finally, otherwise stable LTP in ampakine-treated middle-aged rats can be eliminated by infusion of the extracellular BDNF scavenger TrkB-Fc. Together these results indicate that increases in endogenous BDNF signaling can offset deficits in the postinduction processes that stabilize LTP.

The temperature and enthalpy of transformations of organic alloys from the ternary system 2-amino-2-methyl-1,3-propanediol–(D)camphor–neopentylglycol (AMPD–DC–NPG) were measured by means of differential scanning calorimetry (DSC). The phase diagram of this ternary system was assessed via the CALPHAD approach using Thermo-Calc based on the thermodynamic descriptions of the constituent binaries published earlier and on the thermodynamic and phase equilibrium data measured in the present work for ternary alloys. Good agreement between the experimental and calculated data was achieved. Experiments and calculations show that the AMPD–DC–NPG exhibits a nonvariant quasiperitectic reaction at 336.1 K and composition AMPD-17.5 mol% DC-42.56 mol% NPG, with all solid phases being nonfaceted. Unidirectional solidification experiments were performed with three different alloys selected such that each exhibits a small amount of the primary BCC (AMPD solid solution), the FCC (NPG solid solution) and the hexagonal (DC solid solution) phase, respectively, in order to verify the nature of the phases involved in solid–liquid equilibria and the nature of coupled growth in this ternary system. We find that in all these cases once steady state growth is reached two solid phases grow at the solid–liquid interface with a rod-like coupled growth pattern: The hexagonal phase (DC solid solution) and the FCC phase (NPG solid solution).

The temperature and enthalpy of transformations of organic alloys from the binary systems neopentylglycol–succinonitrile (NPG–SCN) and 2-amino-2-methyl-1,3-propanediol–succinonitrile (AMPD–SCN) were measured by means of differential scanning calorimetry (DSC). The phase diagrams of these binary systems were assessed via the CALPHAD approach using Thermo-Calc by simultaneously optimizing the thermodynamic and phase equilibrium data measured in the present work. Proper agreements between the experimental and calculated data for the phase diagrams as well as for the thermochemical properties were achieved. Experiments and calculations show that both the NPG–SCN and the AMPD–SCN systems exhibit a non-variant eutectic reaction with the eutectic point at 90.45 mol% SCN (318.0 K) and at 97.39 mol% SCN (325.7 K), respectively. In the NPG–SCN system the temperature of the eutectic reaction is about 3 K higher than the temperature of the transformation from the ordered crystals (OCs) to the orientationally disordered crystals (ODICs), whereas the eutectic reaction in the AMPD–SCN involves the OCs of AMPD and the ODICs of SCN. Unidirectional solidification experiments were performed with selected NPG–SCN and AMPD–SCN alloys in order to verify phases involved in solid–liquid equilibria and the nature of eutectic growth in these systems. We find that eutectic growth in NPG–SCN eutectic alloy occurs with both solid phases being non-facetted and with a rod-like eutectic structure. The eutectic as well as some hypo-eutectic alloys from the AMPD–SCN system show irregular eutectic growth with a non-facetted BCC_A2 phase of SCN and a facetted monoclinic phase of AMPD.

Aiming to obtain a reliable description of the ternary Ag–Al–Cu system the thermodynamic evaluation of the constituent binaries Ag–Al, Ag–Cu and Al–Cu are revised by modelling of the Gibbs energy of all individual phases using the CALPHAD approach. The model parameters have been evaluated using a computer optimisation technique based on the established descriptions of the systems and taking into account the data on thermodynamic properties and phase equilibria both reported in recent publications and obtained by own measurements. The phase diagrams and the thermodynamic properties calculated with the evaluated parameters are in good agreement with the corresponding experimental data.

Oxidative stress is involved in the pathogenesis of many diseases. Overexpression of antioxidant enzymes by gene therapy may protect tissues from oxidative damage. Because the reactive oxygen species hydrogen peroxide can diffuse across cell membranes, we hypothesized that overexpression of the antioxidant catalase within certain cells might protect neighboring cells. To test this hypothesis, we transduced retinal pigment epithelial (RPE) cells in vitro and in vivo with adenovirus carrying the catalase gene (Ad.CMV.catalase). After transduction of only a subset of RPE cells in vitro, all cells in the culture were protected from exogenous hydrogen peroxide. Similarly, in vivo, eyes injected with Ad. CMV. catalase had high catalase levels in the RPE, which protected the adjacent photoreceptors from light damage and reduced photoreceptor oxidative stress as measured by the markers 4-hydroxynonenal and nitrotyrosine. Both in vitro and in vivo, gene therapy with Ad. CMV. catalase protected neighboring cells from oxidative stress-induced terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) positivity. The data provide a paradigm for antioxidant gene therapy with catalase, designed to protect not only transduced cells, but also neighboring cells.

The temperature and enthalpy of transformations of organic alloys from the binary system succinonitrile-( d )camphor were measured by means of differential scanning calorimetry (DSC) within the entire composition range. The analytical description of the Gibbs energies of pure succinonitrile (SCN) and pure ( d )camphor (DC) were derived utilizing the data on temperature and enthalpy of transformations, and temperature dependencies of heat capacity available in the literature. The phase diagram for the binary SCN-DC system was assessed via the CALPHAD approach using Thermo-Calc by simultaneously optimizing the thermodynamic and phase equilibrium data available in the literature and measured in the present work. A good agreement between the experimental and calculated data for the phase diagram as well as for the thermochemical properties was achieved. Experiments and calculations show that the binary system SCN-DC has an eutectic reaction with the eutectic point at 311.5 K and 13.9 mol% DC. The enthalpy of mixing derived in the optimisation proves weak attractive interaction between dissimilar molecules. Unidirectional solidification of the eutectic alloy was performed in order to verify the nature of the eutectic: we find that eutectic growth occurs with both solid phases being nonfacetted and with a rod-like eutectic structure consisting of 23 vol% (DC) and 77 vol% (SCN). Due to the optical activity of DC its distribution in the solid sample is well detectible in polarised light.