Biological pretreatment of sugarcane trash for its conversion to fermentable sugars

Pretreatment of lignocellulosic biomasses, the first step in their conversion to utilizable molecules requires very high energy (steam and electricity), corrosion resistant high-pressure reactors and high temperatures. These severe conditions not only add to the cost component of the entire process but also lead to the loss of sugars to the side reactions. Microbial pretreatments have been reported to be associated with reducing the cost factors as well as the severities of the reactions. Eight bioagents, including fungi and bacteria, were screened for their pretreatment effects on sugarcane trash. They narrowed down the C:N ratio of trash from 108:1 to a varying range of approximately 42:1 to 60:1.The maximum drop in C:N ratio of 61% was observed using Aspergillus terreus followed by Cellulomonas uda (52%) and Trichoderma reesei and Zymomonas mobilis (49%). The bioagents helped in degradation of sugarcane trash by production of cellulases, the maximum being produced by A. terreus, (12 fold) followed by C. uda (10 fold), Cellulomonas cartae (9 fold) and Bacillus macerans (8 fold). The microbial pretreatment of trash rendered the easy accessibility of sugars for enzymatic hydrolysis, which can be directed for production of alcohol.     

Immobilization of Xylan-Degrading Enzymes from Scytalidium thermophilum on Eudragit L-100

Summary Xylanase from Scytalidium thermophilum was immobilized on Eudragit L-100, a pH sensitive copolymer of methacrylic acid and methyl methacrylate. The enzyme was non-covalently immobilized and the system expressed 70% xylanase activity. The immobilized preparation had broader optimum temperature of activity between 55 and 65 °C as compared to 65 °C in case of free enzyme and broader optimum pH between 6.0 and 7.0 as compared to 6.5 in case of free enzyme. Immobilization increased the t_1/2 of enzyme at 60 °C from 15 to 30 min with a stabilization factor of 2. The K_m and V_max values for the immobilized and free xylanase were 0.5% xylan and 0.89 μmol/ml/min and 0.35% xylan and 1.01 μmol/ml/min respectively. An Arrhenius plot showed an increased value of activation energy for immobilized xylanase (227 kcal/mol) as compared to free xylanase (210 kcal/mol) confirming the higher temperature stability of the free enzyme. Enzymatic saccharification of xylan was also improved by xylanase immobilization.     

Cytodiagnosis of granulocytic sarcoma of liver arising in a milieu of myeloid metaplasia: a case report

BACKGROUND: Granulocytic sarcoma is an extramedullary tumor that is composed of granulocytic precursor cells. We report an unusual case of granulocytic sarcoma of the liver that arose in the background of myeloid metaplasia. Fine needle aspiration cytology (FNAC) was instrumental in making the diagnosis in absence of previously known Hematlogic abnormality. CASE: A 65-year-old woman presented with multiple nodules in the liver. USG-guided FNAC was performed on them. The aspirates showed many myeloid blasts, myelocytes, metamyelocytes, erytbroid precursors and lympboglandular bodies. We considered a differential diagnosis of granulocytic sarcoma and myeloid metaplasia. The presence of erytbroid precursors prompted us to consider myeloid metaplasia as a differential diagnorsis of granulocytic sarcoma. Peripberal smear showed a leukoerytbroblastic reaction. The patient died, and necropsy from the liver revealed extensive infiltration by undfferentiated blast cells with areas of myeloid metaplasia showing maturing erytbroid, myeloid and megakaryocytic elements. CONCLUSION: When a dual population of predominant myeloid blasts and normoblarts is encountered, a suspicion of granulocytic sarcoma arising in a background of myeloid metaplasia must be kept. Cells of all 3 lineages may not be always seen in myeloid metaplasia, and 1 cell line may predominate, causing a diagnostic dilemma.     

ERK activation is only one role of PKC in TCR-independent cytotoxic T cell granule exocytosis

Cytotoxic T cells (CTLs) kill target cells by releasing lytic agents via regulated exocytosis. Three signals are known to be required for exocytosis: an increase in intracellular Ca²⁺, activation of protein kinase C (PKC) and activation of extracellular signal regulated signal kinase (ERK). ERK activation required for exocytosis depends on activity of PKC. The simplest possibility is that the sole effect of PKC required for exocytosis is ERK activation. Testing this requires dissociating ERK and PKC activation. We did this using TCR-independent stimulation of TALL-104 human leukemic CTLs. When cells are stimulated with thapsigargin and PMA, agents that increase intracellular Ca²⁺ and activate PKC, respectively, PKC-dependent ERK activation is required for lytic granule exocytosis. Expressing a constitutively active mutant MAP kinase kinase activates ERK independent of PKC. However, activating ERK without PKC does not support lytic granule exocytosis, indicating that there are multiple effects of PKC required for granule exocytosis.     

Development and characterization of novel microsatellite markers in the Asiatic lion (Panthera leo persica)

All cat species, except domestic cats, are either threatened or endangered. The Asiatic lion (Panthera leo persica), a critically endangered species, is driven to a single habitat zone in Gir forests in India for its survival. The present status of this big cat compels us to acquire deeper knowledge of its genetic variability and phylogenetic relationships. Here we report the isolation of highly polymorphic microsatellite loci from a partial genomic library of the Asiatic lion. These loci were characterized and have shown high levels of variation ranging from six to 11 alleles per locus in the population studied.     

Identification, structure, and differential expression of members of a BURP domain containing protein family in soybean.

Expressed sequence tags (ESTs) exhibiting homology to a BURP domain containing gene family were identified from the Glycine max (L.) Merr. EST database. These ESTs were assembled into 16 contigs of variable sizes and lengths. Consistent with the structure of known BURP domain containing proteins, the translation products exhibit a modular structure consisting of a C-terminal BURP domain, an N-terminal signal sequence, and a variable internal region. The soybean family members exhibit 35-98% similarity in a -100-amino-acid C-terminal region, and a phylogenetic tree constructed using this region shows that some soybean family members group together in closely related pairs, triplets, and quartets, whereas others remain as singletons. The structure of these groups suggests that multiple gene duplication events occurred during the evolutionary history of this family. The depth and diversity of G. max EST libraries allowed tissue-specific expression patterns of the putative soybean BURPs to be examined. Consistent with known BURP proteins, the newly identified soybean BURPs have diverse expression patterns. Furthermore, putative paralogs can have both spatially and quantitatively distinct expression patterns. We discuss the functional and evolutionary implications of these findings, as well as the utility of EST-based analyses for identifying and characterizing gene families.     

A compilation of soybean ESTs: generation and analysis.

Whole-genome sequencing is fundamental to understanding the genetic composition of an organism. Given the size and complexity of the soybean genome, an alternative approach is targeted random-gene sequencing, which provides an immediate and productive method of gene discovery. In this study, more than 120000 soybean expressed sequence tags (ESTs) generated from more than 50 cDNA libraries were evaluated. These ESTs coalesced into 16928 contigs and 17336 singletons. On average, each contig was composed of 6 ESTs and spanned 788 bases. The average sequence length submitted to dbEST was 414 bases. Using only those libraries generating more than 800 ESTs each and only those contigs with 10 or more ESTs each, correlated patterns of gene expression among libraries and genes were discerned. Two-dimensional qualitative representations of contig and library similarities were generated based on expression profiles. Genes with similar expression patterns and, potentially, similar functions were identified. These studies provide a rich source of publicly available gene sequences as well as valuable insight into the structure, function, and evolution of a model crop legume genome.     

Isolation and characterization of a bacteriocin produced by Cicer-Rhizobium

A constitutively expressed bacteriocin from Cicer-Rhizobium was purified to homogeneity. The purified preparation yielded a homogenous protein with a molecular weight of about 29 kDa. This protein was heat stable, unaffected by nucleases and was found to have an iso-electric point (pI) of 4.6. The N-terminal sequence of the protein was found to be M-N-N-N-Y-R-E-L-L-P-I-I-G-P-P-W-A-E-I-E, sharing 78% homology with linocin M18. Bacteriocin bioactivity was correlated with the presence of a 29 kDa protein in the growth diffusates of the culture. A mutant strain unable to produce this bacteriocin was found to have a statistically significant reduction in nodule occupancy and competitiveness against the wild type and indigeneous populations under unsterile conditions. Bacteriocin production by the mutant carrying the complement clone pJNP365 was found to be stable even in an unsterile environment.     

Antagonists of the human A2A receptor. Part 5: Highly bio-available pyrimidine-4-carboxamides

A novel series of antagonists of the human A_2A receptor have been identified and have been shown to display good potency and high degrees of selectivity over other receptor sub-types. Displaying in vivo potency in commonly used disease models and high oral bio-availability, this class of compounds may serve as clinically useful treatments for the relief of the symptoms associated with Parkinson’s disease.