The Potential effect of G915C polymorphism in regulating TGF-β1 transport into Endoplasmic Reticulum for cytokine production

The TGF-β1 cytokine concentration is known to be higher in nephritis with implied Lupus Nephritis severity. The production of TGF-β1 cytokine is associated with G915C polymorphism. Therefore, it is of interest to study G915C polymorphism. The G915C polymorphism changes codon 25 which encodes arginine into proline in the signal peptide of TGF-β1. The amino acid substitution affects signal peptide properties that may inhibit the transport of TGF-β1 into the endoplasmic reticulum and eventually decline the cytokine production. Hence, the effect of G915C polymorphism on the properties of the signal peptide, the ability of TGF-β1 transport into the endoplasmic reticulum and the concentrations of urinary TGF-β1 in Lupus Nephritis patients was studied. The arginine substitution into proline decreased the polarity of the signal peptide for TGF-β1. The increased hydrophobicity with increased binding energy of the signal peptide for TGF-β1 to Signal Recognition Particle (SRP) and translocon is shown. This implies decreased protein complex stability in potentially blocking the transport of TGF-β1 into the endoplasmic reticulum. This transport retention possibly hampers the synthesis and maturation of TGF-β1 leading to decreased cytokine production.     

Free omental flap for partial breast reconstruction after breast-conserving surgery

Recent advances in endoscopic surgery have allowed laparoscopic harvesting of the omental flap with reduced donor-site morbidity and deformity. Between August of 2004 and May of 2010, the authors performed immediate breast reconstruction with the free omental flap in 10 breast cancer patients, employing it either as a pedicle flap or as a free flap for volume replacement. Only one complication occurred during the follow-up. Cosmetic results were mostly satisfactory. The free omental flap is a safe procedure with minimal donor-site morbidity and deformity and can be an additional option for immediate partial breast reconstruction after breast-conserving surgery. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, IV.     

GS143, an IkappaB ubiquitination inhibitor, inhibits allergic airway inflammation in mice

Asthma is characterized by airway inflammation with intense eosinophil infiltration and mucus hyper-production, in which antigen-specific Th2 cells play critical roles. Nuclear factor-κB (NF-κB) pathway has been demonstrated to be essential for the production of Th2 cytokines and chemokines in the airways in murine asthma models. In the present study, we examined the effect of GS143, a novel small-molecule inhibitor of IκB ubiquitination, on antigen-induced airway inflammation and Th2 cytokine production in mice. Intranasal administration of GS143 prior to antigen challenge suppressed antigen-induced NF-κB activation in the lung of sensitized mice. Intranasal administration of GS143 also inhibited antigen-induced eosinophil and lymphocyte recruitment into the airways as well as the expression of Th2 cytokines and eotaxin in the airways. Moreover, GS143 inhibited antigen-induced differentiation of Th2 cells but not of Th1 cells in vitro. Taken together, these results suggest that IκB ubiquitination inhibitor may have therapeutic potential against asthma.     

Molecular Identification of Hemolymph-Associated Symbiotic Bacteria in Red Imported Fire Ant Larvae

The red imported fire ant (RIFA) (Solenopsis invicta Buren), an exotic insect pest in Texas, has become well established throughout the eastern part of the state. More aggressive than native ant species, RIFA gradually have enlarged their range and spread north and west despite intense efforts to stop them. Symbiotic bacteria have an important relationship in the midgut of fourth instar RIFA larvae. However, the presence of symbiotic bacteria in hemolymph has not been explored. In this study, symbiotic bacteria isolated from the hemolymph of fourth instar larvae of RIFA were genetically identified in terms of genus using a partial sequence of the 16S rRNA gene. Using three different primer sets to amplify regions of the gyrA, gyrB, and SG850 genes, multiple species of the genus Bacillus were identified as inhabitants of fire ant hemolymph. Analysis of gyrA gene identified Bacillus cereus with a percentage match of 94.13–99.20% with DNA sequences from GenBank BLAST (). Using the gyrB gene, Bacillus species were identified with a percentage match of 95.48% to 100% using DNA sequences from GenBank. Finally, analysis of the SG850 gene identified Bacillus cereus with a percentage match of 96.20% to 99.83% using DNA sequences from GenBank.     

Initial Genetic Analysis of Xylella fastidiosa in Texas

Xylella fastidiosa is the causative agent of Pierce’s Disease of grape. No published record of X. fastidiosa genetics in Texas exists despite growing financial risk to the U.S. grape industry, a Texas population of the glassy-winged sharpshooter insect vector (Homalodisca vitripennis) now spreading in California, and evidence that the bacterium is ubiquitous to southern states. Using sequences of conserved gyrB and mopB genes, we have established at least two strains in Texas, grape strain and ragweed strain, corresponding genetically with subsp. piercei and multiplex, respectively. The grape strain in Texas is found in Vitis vinifera varieties, hybrid vines, and wild Vitis near vineyards, whereas the ragweed strain in Texas is found in annuals, shrubs, and trees near vineyards or other areas. RFLP and QRT PCR techniques were used to differentiate grape and ragweed strains with greater efficiency than sequencing and are practical for screening numerous X. fastidiosa isolates for clade identity.     

Cloning and expression of three formate oxidase genes from Debaryomyces vanrijiae MH201

Debaryomyces vanrijiae MH201 produces formate oxidase (FOD) at estimated pI values by native isoelectric focusing of 5.1, 5.4, and 5.9. We cloned and expressed three formate oxidase cDNAs, FOD1, FOD2, and FDO3, of the yeast using Escherichia coli. The open reading frames of FOD1, FOD2, and FDO3 were 1,731 bp long, and encoded 576-amino acid polypeptides with molecular masses of 64,142, 63,794, and 63,836 Da respectively. Expression of FOD1, FOD2, and FOD3 resulted in the production of three isozymes, with pI values of 5.1, 5.9, and 5.9 respectively. Co-expression of FOD1 and FOD2 and of FOD1 and FOD3 resulted in the production of additional isozymes with pI values, of 5.4. The three amino acid sequences of FOD1, FOD2, and FOD3 contained a consensus motif of a flavin adenine dinucleotide binding site in their N-terminal parts and a glucose-methanol-choline oxidoreductase signature pattern, suggesting that formate oxidase ought to be classified in the glucose-methanol-choline oxidoreductase family.     

Intraspecific variations of phosphorus absorption and remobilization, P forms, and their internal buffering in Brassica cultivars exposed to a P-stressed environment

Translocation of absorbed phosphorus (P) from metabolically inactive sites to active sites in plants growing under P deprivation may increase its P utilization efficiency (PUE). Acclimation to phosphate (Pi) starvation may be caused by a differential storage pool of vacuolar P, its release, and the intensity of re-translocation of absorbed P as P starvation inducible environmental cues (PSIEC) from ambient environment. Biomass assay and three P forms, namely inorganic (Pi),organic (Po), and acid-soluble total (Ptas) were estimated in Brassica cultivars exposed to 10 d P deprivation in the culture media. Considering that -δPi/δt denotes the rate of Pi release, Pi release velocity (RSPi) was determined as the tangent to the equations obtained for Pi f(t) at the mean point in the period of greatest Pi decrease, whereas the inverse of the RSPi was art estimate of the internal Pi buffering capacity (IBCPi). Inter cultivar variations in size of the non-metabolic Pi pool,RSPi, re-translocation of Pi from less to more active metabolic sites, and preferential Pi source and sink compartments were evaluated under P starvation. The cultivar 'Brown Raya' showed the highest Pi storage ability under adequate external P supply, and a more intensive release than 'Rain Bow' and 'Dunkled' under P stress. Cultivar 'B.S.A' was inferior to 'Con-1'in its ability to store and use Pi. Roots and upper leaves were the main sink of Pi stored in the lower and middle leaves of all cultivars and showed lower IBCPi and larger RSPi values than lower and middle leaves. In another trial, six cultivars were exposed to P-free nutrition for 29 d after initial feeding on optimum nutrition for 15 d. With variable magnitude, all of the cultivars re-translocated P from the above ground parts to their roots under P starvation, and [P] at 44 d after transplanting was higher in developing leaves compared with developed leaves. Under P deprivation, translocation of absorbed P from metabolically inactive to active sites may have helped the tolerant cultivars to establish a better rooting system, which provided a basis for tolerance against P starvation and increased PUE. A better understanding of the extent to which changes in the flux of P absorption and re-translocation under PSlEC will help to scavenge Pi from bound P reserves and will bring more sparingly soluble P into cropping systems and obtain capitalization of P reserves.     

Rapid synthesis and turnover of brain microsomal ether phospholipids in the adult rat.

The rates of synthesis, turnover, and half-lives were determined for brain microsomal ether phospholipids in the awake adult unanesthetized rat. A multicompartmental kinetic model of phospholipid metabolism, based on known pathways of synthesis, was applied to data generated by a 5 min intravenous infusion of [1,1-(3)H]hexadecanol. At 2 h post-infusion, 29%, 33%, and 31% of the total labeled brain phospholipid was found in the 1-O-alkyl-2-acyl-sn-glycero-3-phosphate, ethanolamine, and choline ether phospholipid fractions, respectively. Autoradiography and membrane fractionation showed that 3% of the net incorporated radiotracer was in myelin at 2 h, compared to 97% in gray matter microsomal and synaptosomal fractions. Based on evidence that ether phospholipid synthesis occurs in the microsomal membrane fraction, we calculated the synthesis rates of plasmanylcholine, plasmanylethanolamine, plasmenylethanolamine, and plasmenylcholine equal to 1.2, 9.3, 27.6, and 21.5 nmol. g(-1). min(-1), respectively. Therefore, 8% of the total brain ether phospholipids have half-lives of about 36.5, 26.7, 23.1, and 15.1 min, respectively. Furthermore, we clearly demonstrate that there are at least two pools of ether phospholipids in the adult rat brain. One is the static myelin pool with a slow rate of tracer incorporation and the other is a dynamic pool found in gray matter.The short half-lives of microsomal ether phospholipids and the rapid transfer to synaptosomes are consistent with evidence of the marked involvement of these lipids in brain signal transduction and synaptic function.