Detection of polymorphism and sequence characterization of toll-like receptor 7 gene of Indian goat revealing close relationship between ruminant species

In this study, approximately 3.4?kb nucleotide sequence of caprine TLR7 (Toll-like receptor 7) gene was generated from twelve different Indian goat breeds belonging to different geographical regions. Goat TLR7 gene ORF (Open Reading Frame) was found to be 3141 nucleotides long coding for 1046?amino acids similar to sheep. The sequence analysis at nucleotide level revealed goat TLR7 having 99.5% homology with sheep, followed by other livestock species. Simple Modular Architecture Research Tool (SMART) was used for the structural analysis of goat TLR7 that showed the presence of 22 leucine rich repeats (LRRs) along with single Toll/interleukin-1 receptor (TIR) domains. TIR domain, when compared, was found to be similar in ruminant species, goat, sheep, cattle, and buffalo. The phylogenetic analysis also revealed grouping of all ruminant species together, goat being closer to sheep followed by cattle and buffalo. A total of 22 polymorphic sites were observed in TLR7 gene of 24 goats representing 12 different breeds, out of which 19 were present within the coding region and three in 3'UTR. Out of the seven nonsynonymous SNPs, two were in ectodomains and one in TIR domain. Overall our results indicate substantial variation within goat TLR7 gene, which could be exploited for association with disease susceptibility.     

Prevention of cardiac hypertrophy and heart failure by silencing of NF-kappaB

Activation of the nuclear factor (NF)-κB signaling pathway may be associated with the development of cardiac hypertrophy and its transition to heart failure (HF). The transgenic Myo-Tg mouse develops hypertrophy and HF as a result of overexpression of myotrophin in the heart associated with an elevated level of NF-κB activity. Using this mouse model and an NF-κB-targeted gene array, we first determined the components of NF-κB signaling cascade and the NF-κB-linked genes that are expressed during the progression to cardiac hypertrophy and HF. Second, we explored the effects of inhibition of NF-κB signaling events by using a gene knockdown approach: RNA interference through delivery of a short hairpin RNA against NF-κB p65 using a lentiviral vector (L-sh-p65). When the short hairpin RNA was delivered directly into the hearts of 10-week-old Myo-Tg mice, there was a significant regression of cardiac hypertrophy, associated with a significant reduction in NF-κB activation and atrial natriuretic factor expression. Our data suggest, for the first time, that inhibition of NF-κB using direct gene delivery of sh-p65 RNA results in regression of cardiac hypertrophy. These data validate NF-κB as a therapeutic target to prevent hypertrophy/HF.     

SDS-capped 1-pyrenecarboxaldehyde nanoprobe for selective detection of Cu2+ ion from water samples: Spectroscopic approach

Sodium dodecyl sulfate (SDS)-capped 1-pyrenecarboxaldehyde nanoparticles (PyalNPs) were prepared using a reprecipitation method in an aqueous medium and exhibited red-shifted aggregation-induced enhanced emission (AIEE). The dynamic light scattering (DLS) examination showed narrower particle size distribution with an average particle size of 41 nm, whereas −34.5 mV zeta potential value indicate the negative surface charge and good stability of nanoparticles (NPs) in an aqueous medium. The AIEE was seen at λ_max = 473 nm in a fluorescence spectrum of a PyalNP suspension. In the presence of Cu²⁺ ions, the fluorescence of PyalNPs quenches very significantly, even in the presence of other metal ions like Ba²⁺, Ca²⁺, Cd²⁺, Co²⁺, Al³⁺, Fe²⁺, Hg²⁺, Ni²⁺ and Mg²⁺. The changes in the fluorescence lifetime of PyalNPs in the presence of Cu²⁺ ions suggested that the type of quenching was dynamic. The fluorescence quenching data for the NPs suspension fitted well into a typical Stern–Volmer relationship in the concentration range 1.0–25 μg/ml of Cu²⁺ ions. The estimated value of the correlation coefficient R² = 0.9877 was close to 1 and showed the linear relationship between quenching data and Cu²⁺ ion concentration. The limit of detection (LOD) was found to be 0.94 ng/ml and is far below the tolerable intake limit value of 1.3 μg/ml accepted by the World Health Organization for Cu²⁺ ions in drinking water. The fluorescence quenching approach for a SDS-capped Pyal nanosuspension for copper ion quantification is of high specificity and coexisting ions were found to interfere very negligibly. The developed method was successfully applied for the estimation of copper ions in river water samples.     

Plant regeneration from a cryopreserved embryogenic cell suspension of a commercial sugarcane hybrid (Saccharum sp.)

Efficient plant regeneration was obtained from a cryopreserved embryogenic cell suspension of sugarcane established from leaf derived callus. Pregrowing the cells for three days in MS basal medium supplemented with 0.33 M sorbitol was essential to the process. The cells were cooled at a rate of 0.5°C/min to –40°C and then stored in liquid nitrogen. Thawing was carried out rapidly in water at +40°C, and the cells were then plated without washing onto filter paper discs placed on a semi-solid regeneration medium (MS basal + 3% sucrose + 0.13 mg/1 2,4-D +0.25 mg/1 BAP + 0.25 mg/1 kinetin + 0.25 mg/1 zeatin). The filter paper discs, along with the cells, were transferred to the same, fresh medium after five hours. After 24 hours the cells were scraped off, placed on fresh semi-solid medium and incubated at 28°C in the dark for two weeks before transfer to light. A regeneration efficiency of 92% was obtained (regenerated plants, expressed as a percent of unfrozen control). Plants regenerated from cryopreserved cells, and grown to maturity in the greenhouse, were morphologically identical to regenerated control plants.Abbreviations DMSO dimethyl sulfoxide - PEG polyethylene glycol - 2,4-D 2,4-dichlorophenoxyacetic acid - BAP benzyl aminopurine - TTC 2,3,5-triphenyl tetrazolium chloride     

Haematological and haematopoietic responses to starvation in an air-breathing fish Channapunctatus Bloch

Changes in haematological values (RBC numbers, haemoglobin content, haematocrit value, MCV, MCH, MCHC, TLC and DLC) based on weekly samples from a group of starved fish were investigated. After 8 weeks of starvation, the effects of restoration to a normal diet was evaluated. Parallel studies on haematopoietic tissues were also made. Changes in some biochemical values such as blood glucose, liver and muscle glycogen were also examined to correlate biochemical effects with those of haematological changes. Erythrocytes, thrombocytes and neutrophils were found to be most sensitive to starvation. The initial response to deprivation of food was an increase in RBCs and related values and in total leukocyte population. However, from week 5 onwards a sharp decline in these cell populations was noted. The leukocytes and thrombocytes showed a change parallel to RBC and the total leukocyte counts. However, neutrophils were observed to show a consistent increase throughout the starvation period. A blood glucose level below SOmglOOmh¹ appeared critical in relation to blood cell population. Haematopoietic studies revealed that reticulocytes and mesomyelocytes were unable to keep pace with the changing peripheral blood picture. Other stages in development responded to the changes in the peripheral blood.