Promoter polymorphism MMP-1 (-1607 2G/1G) and MMP-3 (-1612 5A/6A) in development of HAND and modulation of pathogenesis of HAND

The pathogenesis of HIV-associated neurocognitive disorder (HAND) is modulated by host genetic susceptibility factors such as Matrix metalloproteinases (MMPs). Promoter polymorphism of MMP-1 and MMP-3 may modify the expression of the gene. Hence, we evaluated the association of MMP-1-16072G/1G and MMP-3-1612 5A/6A polymorphisms with development of HAND and the modulation of pathogenesis of HAND. We enrolled a total of 180 individuals, 50 HIV-infected individuals with HAND, 130 without HAND, and 150 healthy controls. Polymorphism of MMP-1 and MMP-3 were genotyped by PCR-RFLP. MMP-1-1607 2G1G, -16071G/2G-1G/1G genotypes and -1607 1G allele were associated with the development of HAND (OR = 1.64, P = 0.05; OR = 1.45, P = 0.04; OR = 1.69, P = 0.05). MMP-1-16071G1G, MMP-3-16125A5A genotypes increased the risk for the development of HAND (OR = 1.78, P = 0.25; OR = 2.39, P = 0.13). MMP-3-1612 5A5A, -1612 6A/5A-5A/5A genotypes and -1612 5A allele were associated with the reduced risk of HAND (OR = 0.40, P = 0.05; OR = 0.53, P = 0.04; OR = 0.40, P = 0.01). Haplotype 5A1G increased the risk of development of HAND (OR = 1.93, P = 0.05). As observed in advanced HIV disease stage, MMP-1-1607 1G1G genotype enhance the risk for advancement of HIV disease (OR = 1.69, P = 0.89). MMP-3-1612 6A5A genotype showed higher risk for development of HAND in alcohol users (0R = 1.65, P = 0.44). MMP-1 genotype may have an influence on development of HAND whereas MMP3-1612 5A5A genotype may reduce risk for pathogenesis of HAND.     

Effects of ionizing radiation (60Co gamma rays) on growth and morphogenesis ofHaworthia mirabilis,haw. Callus tissue

Summary The effects of γ-radiation on growth and morphogenesis ofHaworthia callus in vitro were determined. The doses ranged from 100 to 5000 rads. Survival, growth pattern, growth rate, and differentiation of vegetative buds and roots in both irradiated and nonirradiated callus were compared. Growth data up to 24 weeks for irradiated and control cultures were analyzed. The dose range between 800 to 2500 rads produced compact callus as compared to the controls which were friable. After 12 weeks all control cultures differentiated vegetative buds with roots, whereas callus exposed to 800 to 2500 rads continued to grow with little or no organogenesis. However, it was observed that the wet and dry weights of callus receiving 1000 to 1500 rads ultimately exceeded those of nonirradiated controls.     

Inhibition of galectin-3 mediated cellular interactions by pectic polysaccharides from dietary sources

Pectic polysaccharides from dietary sources such as Decalepis hamiltonii—swallow root (SRPP), Hemidesmus indicus (HPP), Nigella sativa—black cumin (BCPP), Andrographis serpyllifolia—(APP), Zingiber officinale—ginger (GRPP) and, citrus pectin (CPP) were examined for galectin inhibitory activity. Inhibition of (a) galectin-3 of MDA-MB-231 cells induced hemagglutination of red blood cells; (b) galectin-3 mediated interaction between normal/metastatic human buccal cells (NBC)/(MBC) and; (c) invasion of MDA-MB-231 and MBC in the invasive chamber was assessed. Results indicated that SRPP inhibited hemagglutination at Minimum Inhibitory Concentration (MIC) of 1.86 μg ml⁻¹ equivalent of carbohydrate as apposed to those of BCPP (130 μg ml⁻¹), APP (40 μg ml⁻¹), HPP (40 μg ml⁻¹) and CPP (25 μg ml⁻¹). GRPP even at concentration >1–6 mg ml⁻¹ did not inhibit agglutination. Also SRPP showed ∼15 and 2 fold potent anti hemagglutination activity relative to that of galectin-3 specific sugars—galactose (MIC-27.1 μg ml⁻¹) and lactose (MIC-4.16 μg ml⁻¹) respectively. Further, SRPP at 10 μg ml⁻¹ inhibited agglutination of NBC by galectin-3 of MDA-MB-231 cells. Modified swallow root pectic polysaccharide (MSRPP) of 50 kDa retained anti hemagglutination activity (MIC of 1.03 μg ml⁻¹) and inhibited MDA-MB-231 and MBC invasion by 73 and 50% with an IC₅₀ of 136 and 200 μg ml⁻¹ respectively. Both SRPP and MSRPP induced apoptosis up to 80% at 100 μg ml⁻¹ concentration by activating ∼2 and 8 folds of Caspase-3 activity. Sugar composition analysis and its correlation with the galectin inhibitory property indicated that pectic polysaccharides with higher arabinose and galactose content—arabinogalactan inhibited hemagglutination significantly.     

In vitro neoplastic transformation of plant callus tissue by γ-radiation

Tumors have been induced by γ-radiation in callus tissue derived from a monocotyledonous flowering plant, Haworthia mirabilis Haw. The transformed tissue exhibited compact texture, excessive cell proliferation and loss of capacity for organogenesis. Tumors were characterized by their ability to undergo continuous autonomous growth on minimal media in the subsequent 4 generations of subculture. In contrast, the nonirradiated control tissue grew with friable texture, required inositol or growth hormones and showed prolific differentiation of vegetative buds.     

Interspecific hybridization inBrassica: Application of flow cytometry for analysis of ploidy and genome composition in hybrid plants

Interspecific hybrids from the crosses betweenBrassica campestris, B. carinata, B. juncea andB. napus were obtained throughin vitro ovary and ovule culture. F₁ hybrids were studied morphologically and flow cytometry was used to estimate 2C nuclear DNA content both in parentalBrassica species and their hybrids. It was found that in comparison with the A genome, the B and the C genomes ofBrassica contained 26.9 % and 43.9 % more DNA, respectively. This finding may be used to distinguish interspecific hybrids containing various genome combinations. It was concluded that flow cytometric analysis of nuclear DNA content might be useful tool inBrassica breeding.     

Topographic mapping and compression elasticity analysis of skinned cardiac muscle fibers in vitro with atomic force microscopy and nanoindentation

Surface topography and compression elasticity of bovine cardiac muscle fibers in rigor and relaxing state have been studied with atomic force microscopy. Characteristic sarcomere patterns running along the longitudinal axis of the fibers were clearly observed, and Z-lines, M-lines, I-bands, and A-bands can be distinguished through comparing with TEM images and force curves. AFM height images of fibers had shown a sarcomere length of 1.22±0.02 μm (n=5) in rigor with a significant 9% increase in sarcomere length in relaxing state (1.33±0.03 μm, n=5), indicating that overlap moves with the changing physiological conditions. Compression elasticity curves along with sarcomere locations have been taken by AFM compression processing. Coefficient of Z-line, I-band, Overlap, and M-line are 25±2, 8±1, 10±1, and 17±1.5 pN/nm respectively in rigor state, and 18±2.5, 4±0.5, 6±1, and 11±0.5 pN/nm respectively in relaxing state. Young's Modulus in Z-line, I-band, Overlap, and M-line are 115±12, 48±9, 52±8, and 90±12 kPa respectively in rigor, and 98±10, 23±4, 42±4, and 65±7 kPa respectively in relaxing state. The elasticity curves have shown a similar appearance to the section analysis profile of AFM height images of sarcomere and the distance between adjacent largest coefficient and Young's Modulus is equal to the sarcomere length measured from the AFM height images using section analysis, indicating that mechanic properties of fibers have a similar periodicity to the topography of fibers.     

A Hump-Backed Trend in Bacterial Diversity with Elevation on Mount Fuji, Japan

Little is known of how bacterial diversity in soils varies with elevation. One previous study found a decline with elevation, whereas another found no trend. We chose Mount Fuji of Japan as a geologically and topographically simple mountain system. Samples were taken at elevational intervals, between the base of the mountain at 1,000 m and its summit at 3,700 m. Polymerase chain reaction-amplified soil DNA for the bacterial 16S gene targeting V1–V3 region was pyrosequenced using the 454 Roche machine, and taxonomically classified with reference to a bioinformatic database. There was a significant “peak” in total bacterial diversity at around 2,500 m above the tree line with a decline towards the highest elevations around 3,700 m near the summit. Individual bacterial phyla show distinct trends—increase, decrease, or a mid-elevational “bulge” in diversity. Bacterial diversity does not parallel woody plant or herbaceous plant diversity. We suggest that beyond the tree and vegetation line, the more extreme temperature fluctuations, stronger UV, lack of nutrients, and more frequent disturbance of the loose substrate of these slopes allows less competition and greater bacterial species diversity due to “lottery” recruitment. However, at the highest elevations, the physiological challenges are so extreme that fewer bacterial species are capable of surviving.