The Mouse GalR2 Galanin Receptor: Genomic Organization, cDNA Cloning, and Functional Characterization

Abstract: The diverse physiological actions of galanin are thought to be mediated through activation of galanin receptors (GalRs). We report the genomic and cDNA cloning of a mouse GalR that possesses a genomic structure distinct from that of GalR1 and encodes a functional galanin receptor. The mouse GalR gene consists of two exons separated by a single intron within the protein-coding region. The splicing site for the intron is located at the junction between the third transmembrane domain and the second intracellular loop. The cDNA encodes a 370-amino acid putative G protein-coupled receptor that is markedly different from human GalR1 and rat GalR3 (38 and 57%) but shares high homology with rat GalR2 (94%). In binding studies utilizing membranes from COS-7 cells transfected with mouse GalR2 cDNA, the receptor displayed high affinity ( K _D = 0.47 n M ) and saturable binding with ¹²⁵I-galanin ( B _max = 670 fmol/mg). The radioligand binding can be displaced by galanin and its analogues in a rank order: galanin ⋍ M40 ⋍ M15 ⋍ M35 ⋍ C7 ⋍ galanin (2–29) ⋍ galanin (1–16) ≫ galanin (10–29) ⋍ galanin (3–29), which resembles the pharmacological profile of the rat GalR2. Receptor activation by galanin in COS-7 cells stimulated phosphoinositide metabolism, which was not reversed by pertussis toxin. Thus, the galanin receptor encoded in the cloned mouse GalR gene is the type 2 galanin receptor and is active in both ligand binding and signaling assays.     

Minimotif Miner: a tool for investigating protein function

In addition to large domains, many short motifs mediate functional post-translational modification of proteins as well as protein-protein interactions and protein trafficking functions. We have constructed a motif database comprising 312 unique motifs and a web-based tool for identifying motifs in proteins. Functional motifs predicted by MnM can be ranked by several approaches, and we validated these scores by analyzing thousands of confirmed examples and by confirming prediction of previously unidentified 14-3-3 motifs in EFF-1.     

Near optimal manufacturing flow controller design

Flow control of flexible manufacturing systems (FMSs) addresses an important real-time scheduling requirement of modern manufacturing facilities, which are prone to failures and other controllable or stochastic discrete events affecting production capacity, such as change of setup and maintenance scheduling. Flow controllers are useful both in the coordination of interconnected flexible manufacturing cells through distributed scheduling policies and in the hierarchical decomposition of the planning and scheduling problem of complex manufacturing systems. Optimal flow-control policies are hedging-point policies characterized by a generally intractable system of stochastic partial differential equations. This article proposes a near optimal controller whose design is computationally feasible for realistic-size systems. The design exploits a decomposition of the multiple-part-type problem to many analytically tractable one-part-type problems. The decomposition is achieved by replacing the polyhedra production capacity sets with inscribed hypercubes. Stationary marginal densities of state variables are computed iteratively for successive trial controller designs until the best inscribed hypercubes and the associated optimal hedging points are determined. Computational results are presented for an illustrative example of a failureprone FMS.     

Crataegus grossidentata sp. nov. (Rosaceae–Pyreae), a new hawthorn from northern Iran

Crataegus grossidentata Sharifnia & K. I. Chr., found in northern Iran, is described and illustrated as a species new to science. Its ecology, distribution and taxonomic relationships are discussed. A key to C. grossidentata and other one‐styled taxa of Crataegus occurring in Iran is provided.     

wbeT sequence typing and IS1004 profiling of Vibrio cholerae isolates

Aims: To investigate the molecular basis for serotype variation in Vibrio cholerae O1 and the genetic relatedness amongst different serotypes isolated from 2004 to 2008 in Iran. Methods and Results: Despite the presence of all three serotypes of V. cholerae O1 (Ogawa, Inaba and Hikojima) in Iran in the last decade, the Inaba strains have been the dominated serotype. Sequence analysis of wbeT determined only a single substitution of G for A at position 295 in all Inaba strains resulting in a replacement of serine to proline. No difference was found in the copy numbers and profile of IS1004 between the classical and El Tor V. cholerae O1 strains, supporting the clonality amongst the isolates obtained over 5 years in Iran. In addition, Southern blots of HpaII‐digested chromosomal DNAs of our Ogawa and Inaba isolates showed the presence of an incomplete copy of IS1004 for all isolates. Conclusions: IS1004 profiling can be a reliable method for analysis of clonal dissemination of V. cholerae. The results indicated that specific point mutation at a particular position within the wbeT of V. cholerae O1 strains in Iran may occur which, in turn, may result in serotype switching. Significance and Impact of the Study: Understanding the molecular basis for serotype conversion of V. cholerae and their genetic relatedness could give insights for the incoming cholera epidemic prediction and control.     

Crataegus coriifolia sp. nov. (Rosaceae) from Iran

Crataegus coriifolia Sharifnia & Zarrinkolah is described as a new species from Iran. Its taxonomic relationships, ecology and distribution are discussed. An identification key to C. coriifolia and other two‐styled species of Crataegus occurring in Iran is provided.     

Improving the recovery of clarification process of recombinant hepatitis B surface antigen in large-scale by optimizing adsorption-desorption parameters on Aerosil-380

Abstract

In the downstream process of recombinant hepatitis B surface antigen (rHBsAg), nano-colloidal silica adsorbent (Aerosil-380) is one of the possible methods to separate the antigen from other main impurities partially. The current study aimed to maximize the adsorptive capacity of Aerosil-380 as well as rHBsAg recovery for large-scale production of recombinant hepatitis B vaccine. The experimental design methodology was used to optimize the eight critical parameters influencing the efficiency, rHBsAg recovery, of the adsorption-desorption process in the lab-scale. These examined parameters were the adsorption–desorption temperature, pH, contact time, agitation speed, antigen concentration, and desorption buffer. Under optimal condition, the maximum adsorption capacity of Aerosil-380 was equal to 3333?μg.g^?1 (rHBsAg/adsorbent), and we could recover about 95% of rHBsAg with purity of 54% (rHBsAg/total protein) in the lab scale. Using the optimum parameters for rHBsAg clarification process in large-scale by Aerosil-380, we recovered about 78% of rHBsAg with 43% purity. Based on the obtained experimental data, Langmuir adsorption isotherm and pseudo-first-order kinetic model provide the best correlations of experimental data for the adsorbent. Findings of this study significantly increase the recovery of clarification process of rHBsAg in large-scale compared to previous reports.     

Molecular diversity of CTX prophage in Vibrio cholerae

Aims: The objective of this study was to investigate the molecular diversity of CTX genetic element within toxigenic Vibrio cholerae genomes and to determine the genetic diversity of V. cholerae population collected in a 6‐year period (2004–2009) in Iran. Methods and Results: The results of mismatch amplification mutation assay (MAMA)‐PCR and sequencing showed cytosine nucleotide in positions 203 and 115 in all 50 El Tor V. cholerae strains, which is the same as classical ctxB sequence. One strain yielded amplicons with both El Tor and classical biotype primers in MAMA‐PCR indicative of presence of two copies of CTX phages with different genotypes (rstR^ET ctxB^class and rstR^ET ctxB^ET) integrated within the genome of this isolate, which suggested the integration of two different CTX phages at different occasions or point mutation in one copy of CTX. Sequencing and PCR analysis indicated the presence of hybrid CTX genotype (rstR^ET ctx^class) in 70·6% of the isolates; however, only El Tor RS1 phage has been integrated in flanking to the CTX phages with different genotypes. Conclusions: Enterobacterial repetitive intergenic consensus‐PCR (ERIC‐PCR) and ribosomal gene spacer‐PCR (RS‐PCR) showed a relatively homogenous population in different years. Our findings indicate that sequence analysis of RS and ctxB regions has more discriminative power than restriction‐based methods. Significance and Impact of the Study: Investigating the molecular diversity of CTX prophage among V. cholerae strains helps to establish a new valuable database of genetic information about isolates, which is of great importance for epidemiologic studies in Iran and other countries encountering cholera epidemics.