��-Catenin Is Genetically and Biologically Associated with Cortical Cataract and Future Alzheimer-Related Structural and Functional Brain Changes

Multiple lines of evidence suggest that specific subtypes of age-related cataract (ARC) and Alzheimer disease (AD) are related etiologically. To identify shared genetic factors for ARC and AD, we estimated co-heritability of quantitative measures of cataract subtypes with AD-related brain MRI traits among 1,249 members of the Framingham Eye Study who had a brain MRI scan approximately ten years after the eye exam. Cortical cataract (CC) was found to be co-heritable with future development of AD and with several MRI traits, especially temporal horn volume (THV, ρ = 0.24, P<10⁻⁴). A genome-wide association study using 187,657 single nucleotide polymorphisms (SNPs) for the bivariate outcome of CC and THV identified genome-wide significant association with CTNND2 SNPs rs17183619, rs13155993 and rs13170756 (P<2.6×10⁻⁷). These SNPs were also significantly associated with bivariate outcomes of CC and scores on several highly heritable neuropsychological tests (5.7×10⁻⁹≤P<3.7×10⁻⁶). Statistical interaction was demonstrated between rs17183619 and APP SNP rs2096488 on CC (P = 0.0015) and CC-THV (P = 0.038). A rare CTNND2 missense mutation (G810R) 249 base pairs from rs17183619 altered δ-catenin localization and increased secreted amyloid-β_1–42 in neuronal cell culture. Immunohistopathological analysis of lens tissue obtained from two autopsy-confirmed AD subjects and two non-AD controls revealed elevated expression of δ-catenin in epithelial and cortical regions of lenses from AD subjects compared to controls. Our findings suggest that genetic variation in delta catenin may underlie both cortical lens opacities in mid-life and subsequent MRI and cognitive changes that presage the development of AD.     

Lymphocytic microparticles inhibit angiogenesis by stimulating oxidative stress and negatively regulating VEGF-induced pathways

Recent studies have demonstrated that lymphocyte-derived microparticles (LMPs) impair endothelial cell function. However, no data currently exist regarding the contribution of LMPs in the regulation of angiogenesis. In the present study, we investigated the effects of LMPs on angiogenesis in vivo and in vitro and demonstrated that LMPs strongly suppressed aortic ring microvessel sprouting and in vivo corneal neovascularization. In vitro, LMPs considerably diminished human umbilical vein endothelial cell survival and proliferation in a concentration-dependent manner. Mechanistically, the antioxidants U-74389G and U-83836E were partially protective against the antiproliferative effects of LMPs, whereas the NADPH oxidase (NOX) inhibitors apocynin and diphenyleneiodonium significantly abrogated these effects. Moreover, LMPs increased not only the expression of the NOX subunits gp91(phox), p22(phox), and p47(phox), but also the production of ROS and NOX-derived superoxide (O(2)(-)). Importantly, LMPs caused a pronounced augmentation in the protein expression of the CD36 antiangiogenic receptor while significantly downregulating the protein levels of VEGF receptor type 2 and its downstream signaling mediator, phosphorylated ERK1/2. In summary, LMPs potently suppress neovascularization in vivo and in vitro by augmenting ROS generation via NOX and interfering with the VEGF signaling pathway.     

Plantlet regeneration from leaf derived callus of <Emphasis Type="Italic">Vanilla planifolia</Emphasis> Andr.

Vanilla planifolia is a tropical orchid mainly known for the aromatic flavor of its cured pods. Callus cultures were initiated from leaf and nodal explants of V. planifolia. Leaf explants showed better callus initiation than the nodal explants with callus biomass production maximal when cultured on Murashige and Skoog (MS) basal medium containing 4.52 mM 2,4-dichlorophenoxy acetic acid and 2.22 mM benzyladenine (BAP). Callus transferred to MS basal medium supplemented with 13.32 μM BAP, and 13.43 μM naphthaleneacetic acid (NAA) showed superior growth response and produced 14.0 ± 1.0 shoots with an average length of 3.6 ± 0.1 cm after 40 d. Subsequent transfer of the proliferated shootlets to MS basal medium supplemented with 8.88 μM BAP and 8.08 μM NAA produced 12.3 ± 0.14 plantlets with an average height of 3.6 cm ± 0.10 cm. All plantlets produced profuse rooting within 35–40 d after transfer to half-strength MS basal medium supplemented with NAA in combination with indole-3-acetic acid. Rooted plantlets were transferred for hardening, with 80% of the plantlets becoming successfully established in the field. Potentially, more than 100,000 plantlets could be produced within eight subcultures from callus obtained from leaf explant through the methods described here.     

Oxidation of isoeugenol by Nocardia iowensis

Isoeugenol is a starting material for both the synthetic and biotechnological production of vanillin and vanillic acid. Nocardia iowensis DSM 45197 (formerly Nocardia species NRRL 5646) resting cells catalyze the conversion of isoeugenol to vanillic acid, vanillin, vanillyl alcohol and guaiacol. The present study used a variety of chemical, microbial and enzymatic approaches to probe the pathways used by N. iowensis in the oxidation of isoeugenol to these products. Of three possible pathways considered, initial side-chain olefin epoxidation, epoxide hydrolysis to a vicinal diol, and diol cleavage to vanillin and subsequently further oxidation to vanillic acid appears as the most likely route. Isoeugenol was not oxidized to ferulic acid, a well-known microbial transformation precursor for vanillin and vanillic acid. ¹⁸O-Labeled oxygen (one atom) and water (two oxygen atoms) were incorporated into vanillic acid during the whole-cell biotransformation reaction with isoeugenol indicating the likely involvement of oxygenase and hydrolase systems in the bioconversion reaction. Vanillin was converted to singly labeled vanillic acid in the presence of H₂¹⁸O suggesting the presence of an aldehyde oxidase. Cell extracts achieved the conversion of isoeugenol to vanillic acid and vanillin without cofactors. Partial fractionation of two enzyme activities supported the presence of isoeugenol monooxygenase and vanillin oxidase activities in N. iowensis.     

Evolution of a multi‐step phosphorelay signal transduction system in Ensifer: recruitment of the sigma factor RpoN and a novel enhancer‐binding protein triggers acid‐activated gene expression

Most Ensifer strains are comparatively acid sensitive, compromising their persistence in low pH soils. In the acid‐tolerant strain Ensifer medicae WSM419, the acid‐activated expression of lpiA is essential for enhancing survival in lethal acidic conditions. Here we characterise a multi‐step phosphorelay signal transduction pathway consisting of TcsA, TcrA, FsrR, RpoN and its cognate enhancer‐binding protein EbpA, which is required for the induction of lpiA and the downstream acvB gene. The fsrR, tcrA, tcsA and rpoN genes were constitutively expressed, whereas lpiA and acvB were strongly acid‐induced. RACE mapping revealed that lpiA/acvB were co‐transcribed as an operon from an RpoN promoter. In most Ensifer species, lpiA/acvB is located on the chromosome and the sequence upstream of lpiA lacks an RpoN‐binding site. Nearly all Ensifer meliloti strains completely lack ebpA, tcrA, tcsA and fsrR regulatory loci. In contrast, E. medicae strains have lpiA/acvB and ebpA/tcrA/tcsA/fsrR co‐located on the pSymA megaplasmid, with lpiA/acvB expression coupled to an RpoN promoter. Here we provide a model for the expression of lpiA/acvB in E. medicae. This unique acid‐activated regulatory system provides insights into an evolutionary process which may assist the adaptation of E. medicae to acidic environmental niches.     

Concentration‐dependent studies of Nd3+‐doped zinc phosphate glasses for NIR photoluminescence at 1.05 μm

Nd³⁺‐doped lead‐free zinc phosphate glasses with the chemical compositions (60‐x) NH₄H₂PO₄ + 20ZnO + 10BaF₂ + 10NaF + xNd₂O₃ (where x = 0.5, 1.0, 1.5, 2.0 and 2.5 mol%) were prepared using a melt quenching technique. Vibrational bands were assigned and clearly elucidated by Raman spectral profiles for all the glass samples. Judd–Ofelt (J–O) intensity parameters (Ω_λ: λ = 2, 4, 6) were obtained from the spectral intensities of different absorption bands of Nd³⁺ ions. Radiative properties such as radiative transition probabilities (A_R), radiative lifetimes (τ_R) and branching ratios (β_R) for different excited states were calculated using J–O parameters. The near infrared (NIR) photoluminescence spectra exhibited three emission bands (⁴F_3/2 level to ⁴I_13/2, ⁴I_11/2 and ⁴I_9/2 states) for all the concentrations of Nd³⁺ ions. Various luminescence properties were studied by varying the Nd³⁺ concentration for the three spectral profiles. Fluorescence decay curves of the ⁴F_3/2 level were recorded. The energy transfer mechanism that leads to quenching of the ⁴F_3/2 state lifetimes was discussed at higher concentration of Nd³⁺ ions. These glasses are suggested as suitable hosts to produce efficient lasing action in NIR region at 1.05 μm.     

Efficient Regeneration of Curcuma amada Roxb. Plantlets from Rhizome and Leaf Sheath Explants

Efficient and simple protocols were developed for conversion of embryos derived from microspores in rape. The frequency of embryo conversion was higher than 80% on improved media without pretreatments using ABA, GA₃ or desiccation, which had been required before embryos were transferred to solid media for conversion. While on basal media such as 1/2MS, the conversion frequency was no more than 32%. Two groups of embryos at different developmental stages, 7 mm embryos and 3 mm embryos, were used as material to select the most suitable medium. Different components were required for successful conversion of the two groups of embryos. For 7 mm embryos 1/2MS medium with Ca²⁺ concentration of 900 mg l^–1 was optimal, while for 3 mm embryos, 1/2 MS_N+V+Ca (half strength MS with 450 mg l^–1 CaCl₂·2H₂O, 100 mg l^–1 NH₄NO₃ and enhanced vitamins) was a suitable medium. The results indicate that calcium may play an important role in the conversion of embryos derived from microspores, and can replace, to some extent, the practice of drying and growth supplements, which have been widely used for maturation, and desiccation of embryos.     

The time evolution of sequential enzyme reactions: A singular perturbation approach

A perturbative procedure is developed to describe the entire time course of a sequential enzyme reaction occurring in a closed system. The perturbation parameter is the ratio of the enzyme concentration to a reference concentration defined in analogy with the Michaelis-Menten constant. It is shown that damped oscillations may occur about the quasi-steady state for certain realistic values of the parameters. Even under the most favourable circumstances, the damping is exceptionally heavy. In contrast, the evolution to the equilibrium is always monotonic.     

Effect of multigeneration alcohol feeding on murine immune system

Mice belonging to F8, F12, F14 and F20 generation of a multigeneration study reared on 20% (v/v) ethanol in water as the sole drinking source were investigated for their immune competence using various parameters. The results indicated lack of any significant effect on delayed type hypersensitivity to dinitro fluorobenzene (DNFB) or sheep red blood cells (SRBC) in mice consuming ethanol. Further, alloskin graft and tumor graft response was similar in both ethanol and water fed mice. Humoral response to SRBC was also intact. However, NK cell activity was reduced significantly in ethanol fed mice. Phagocytic index as assessed by the carbon clearance test was also reduced considerably in mice consuming ethanol. The results clearly indicate that ethanol per se has a significant effect on the nonspecific limb of the immune system, in chronically fed mice.