Structural characterization of Ca2+/CaM in complex with the phosphorylase kinase PhK5 peptide

Phosphorylase kinase (PhK) is a large hexadecameric enzyme consisting of four copies of four subunits: (alphabetagammadelta)4. An intrinsic calmodulin (CaM, the delta subunit) binds directly to the gamma protein kinase chain. The interaction site of CaM on gamma has been localized to a C-terminal extension of the kinase domain. Two 25-mer peptides derived from this region, PhK5 and PhK13, were identified previously as potential CaM-binding sites. Complex formation between Ca2+/CaM with these two peptides was characterized using analytical gel filtration and NMR methods. NMR chemical shift perturbation studies showed that while PhK5 forms a robust complex with Ca2+/CaM, no interactions with PhK13 were observed. 15N relaxation characteristics of Ca2+/CaM and Ca2+/CaM/PhK5 complexes were compared with the experimentally determined structures of several Ca2+/CaM/peptide complexes. Good fits were observed between Ca2+/CaM/PhK5 and three structures: Ca2+/CaM complexes with peptides from endothelial nitric oxide synthase, with smooth muscle myosin light chain kinase and CaM kinase I. We conclude that the PhK5 site is likely to have a direct role in Ca2+-regulated control of PhK activity through the formation of a classical 'compact' CaM complex.     

Response of Java citronella (Cymbopogon winterianus Jowitt) to toxic heavy metal cadmium.

Cadmium at 200 mg kg-1 soil and above concentrations was fatal as growth was inhibited ultimately leading to death of Java citronella (Cymbopogon winterianus Jowitt.). The surviving plants at 50 and 100 mg kg-1 treatments also exhibited pronounced retardation of growth and biomass yield. There was considerable reduction in the level of essential oil in herbage and oil quality deteriorated. Cadmium accumulation profile showed that highest accumulation was in root, followed by stem, leaf sheath and leaf. Very high accumulation in root for higher doses appeared to be the reason for fatality.     

Flood plain wetland fisheries of India: with special reference to impact of climate change

Floodplain wetlands of India are biologically rich sensitive ecosystem that support unique aquatic biodiversity and play vital role in providing livelihood and nutritional security to a large section of the population of the country. Besides contributing to the environmental sustainability through Carbon sequestration, flood plain wetlands also serve as source for harvesting flood and rain water. Covering around 5.5 Lakh ha area, the flood plain wetlands of India are one of the major sources for fish production of the country, offering vast potential for capture as well as culture based fisheries. Degradation and shrinkage of the floodplain wetlands have been recorded due to several natural and anthropogenic reasons. In addition to that, the change in the climatic condition may have a far more devastating impact on these natural resources. Substantial change in climate with increasing temperature trend (0.60 °C during last 112 years) as well as changing pattern and intensity of rainfall have been reported in India. It is projected that extreme climate changes may have profound impact on wetlands, mediated through several direct or indirect pathways. Attention is urgently required at different levels for conservation and revamping of these resources along with coping up and mitigation strategies to address the impending challenges. However, there is dearth of scientific information specific to the flood plain wetlands of India. This paper reviews the present status and importance of the flood plain wetlands of India with special reference to impact of climate change along with coping-up and mitigation measures.     

Piroxicam confer neuroprotection in Cerebral Ischemia by inhibiting Cyclooxygenases,Acid- Sensing Ion Channel-1a and Aquaporin-4: an in silico comparison with Aspirin and Nimesulide

Cerebral ischemia (CI), caused by the deprivation of oxygen and glucose to the brain, is the leading cause of permanent disability.Neuronal demise in CI has been linked to several pathways which include cyclooxygenases (COX) − mediated production ofprostaglandins (PGs) and subsequently reactive oxygen species (ROS), aquaporin-4 (AQ-4) − mediated brain edema and acidsensingion channel-1a (ASIC-1a) − mediated acidotoxicity, matrix remodeling, in addition to others. Several non-steroidal antiinflammatorydrugs (NSAIDs) are presently in use to prevent these pathways. However, owing to the large number of processesinvolved, there is high drug load. So, identifying drugs with multimodal role has always been a frequently sought venture. Thepresent in silico study has been performed to find out the relative efficacy of three different NSAIDs (Piroxicam, Aspirin andNimesulide) in preventing neurodegeneration in CI, with respect to their inhibitory potential on COXs, AQ-4 and ASIC-1a. We findthat piroxicam is the most potent inhibitor of these receptors as compared to the NSAIDs under investigation. Since piroxicam hasalready been reported to inhibit N-methyl-D-aspartate (NMDA) receptor and matrix metalloproteinases (MMPs), which are alsolinked to CI-induced neurodegeneration, we hereby propose piroxicam to be a gold-standard drug in preventingneurodegeneration in CI.     

Hypoxia and its effect on the cellular system

Eukaryotic cells utilize oxygen for different functions of cell organelles owing to cellular survival. A balanced oxygen homeostasis is an essential requirement to maintain the regulation of normal cellular systems. Any changes in the oxygen level are stressful and can alter the expression of different homeostasis regulatory genes and proteins. Lack of oxygen or hypoxia results in oxidative stress and formation of hypoxia inducible factors (HIF) and reactive oxygen species (ROS). Substantial cellular damages due to hypoxia have been reported to play a major role in various pathological conditions. There are different studies which demonstrated that the functions of cellular system are disrupted by hypoxia. Currently, study on cellular effects following hypoxia is an important field of research as it not only helps to decipher different signaling pathway modulation, but also helps to explore novel therapeutic strategies. On the basis of the beneficial effect of hypoxia preconditioning of cellular organelles, many therapeutic investigations are ongoing as a promising disease management strategy in near future. Hence, the present review discusses about the effects of hypoxia on different cellular organelles, mechanisms and their involvement in the progression of different diseases.     

流行性出血热病毒核酸的提取

本文报告流行性出血热R22和A9株病毒培养,同位素标记及核酸提取的初步研究,並获得了该病毒核酸的3个片段即L、M、S、分子量分别约为:3.8、1.9和0.86×10~5道尔顿,不论用20～70％蔗糖密度梯度离心提纯的病毒,或用30％蔗糖垫层离心的粗制病毒,均获同样结果,但多数情况下,用蔗糖密度梯度离心时,除病毒峰外,还发现主要由细胞组份(即线粒体和核糖体等)组成的另一峰,并经常影响病毒RNA的提取,对如何获得纯净病毒及其核酸进行了讨论。     

Nuclear Translocation and Regulation of Intranuclear Distribution of Cytoplasmic Poly(A)-Binding Protein Are Distinct Processes Mediated by Two Epstein Barr Virus Proteins

Many viruses target cytoplasmic polyA binding protein (PABPC) to effect widespread inhibition of host gene expression, a process termed viral host-shutoff (vhs). During lytic replication of Epstein Barr Virus (EBV) we observed that PABPC was efficiently translocated from the cytoplasm to the nucleus. Translocated PABPC was diffusely distributed but was excluded from viral replication compartments. Vhs during EBV infection is regulated by the viral alkaline nuclease, BGLF5. Transfection of BGLF5 alone into BGLF5-KO cells or uninfected 293 cells promoted translocation of PAPBC that was distributed in clumps in the nucleus. ZEBRA, a viral bZIP protein, performs essential functions in the lytic program of EBV, including activation or repression of downstream viral genes. ZEBRA is also an essential replication protein that binds to viral oriLyt and interacts with other viral replication proteins. We report that ZEBRA also functions as a regulator of vhs. ZEBRA translocated PABPC to the nucleus, controlled the intranuclear distribution of PABPC, and caused global shutoff of host gene expression. Transfection of ZEBRA alone into 293 cells caused nuclear translocation of PABPC in the majority of cells in which ZEBRA was expressed. Co-transfection of ZEBRA with BGLF5 into BGLF5-KO cells or uninfected 293 cells rescued the diffuse intranuclear pattern of PABPC seen during lytic replication. ZEBRA mutants defective for DNA-binding were capable of regulating the intranuclear distribution of PABPC, and caused PABPC to co-localize with ZEBRA. One ZEBRA mutant, Z(S186E), was deficient in translocation yet was capable of altering the intranuclear distribution of PABPC. Therefore ZEBRA-mediated nuclear translocation of PABPC and regulation of intranuclear PABPC distribution are distinct events. Using a click chemistry-based assay for new protein synthesis, we show that ZEBRA and BGLF5 each function as viral host shutoff factors.     

Conformation of double-stranded polydeoxynucleotides in solution by proton two-dimensional nuclear Overhauser enhancement spectroscopy

Proton 2D-NOE spectroscopy has been used to investigate the three-dimensional conformations of several sonicated polydeoxynucleotides in solution. The observed pattern of cross peaks indicate that poly(dA-dT) · poly(dA-dT) in all salt concentrations studied (up to 6.6M CsF), and poly(dG-m⁵dC) · poly(dG-m⁵dC) in low salt (0.1M NaCl) are righthanded B-structures. Poly(dG-m⁵dC) · poly(dG-m⁵dC) in Mg²⁺ (3 mM) solution exhibits a pattern characteristic of the left-handed Z-form. These results for poly(dA-dT) · poly(dA-dT) are in contrast to suggestions that this copolymer exists as a left-handed form, either in low or high salt. We present pure absorption-mode 2D-NOE spectra that enable us to compare several distances and define the conformations of these polydeoxynucleotides in solution.     

Island scalp flap for superior forehead reconstruction

An island scalp fasciocutaneous flap, based on the posterior superficial temporal vessels, is described for single-stage reconstruction of full-thickness forehead and scalp defects. The hairline can be precisely determined and tailored to restore symmetry. By removing the hair-bearing dermis of the forehead portion of the flap and placing a full-thickness skin graft, aesthetic reconstitution of the forehead skin is achieved. This flap is especially useful when exposed calvarium limits other techniques.