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Fusarium head blight (FHB), primarily caused by Fusarium graminearum, is one of the economically significant diseases in small grains. FHB causes severe damage to wheat production and grain quality. Several management strategies have been developed to control FHB, and chemical control through fungicides plays a significant role. Although fungicides have effectively controlled F. graminearum in the field, the continuous exposure causes a selection pressure in the pathogen population towards fungicide resistance. Several studies have identified fungicide-resistant F. graminearum isolates and fungicide-resistance mechanisms. Although new fungicides with a new mode of action can be introduced into the market, developing a new fungicide is time-consuming, and extra efforts are needed for testing, approvals, and registrations. Therefore, it is essential to strategize the methods to delay the fungicide resistance. This review focuses on the impact of several fungicide applications currently used on FHB, focusing on Fusarium graminearum, the status of the fungicide sensitivity for fungicide classes, the resistance mechanisms against fungicides, and the mitigation strategies to delay the development of fungicide resistance in the pathogen population. Studying the fungicide resistance mechanisms and the mitigation strategies will be helpful in the future to use the available fungicides against F. graminearum without losing its effectiveness. [ABSTRACT FROM AUTHOR]

Fusarium head blight (FHB), primarily caused by Fusarium graminearum, is one of the economically significant diseases in small grains. FHB causes severe damage to wheat production and grain quality. Several management strategies have been developed to control FHB, and chemical control through fungicides plays a significant role. Although fungicides have effectively controlled F. graminearum in the field, the continuous exposure causes a selection pressure in the pathogen population towards fungicide resistance. Several studies have identified fungicide-resistant F. graminearum isolates and fungicide-resistance mechanisms. Although new fungicides with a new mode of action can be introduced into the market, developing a new fungicide is time-consuming, and extra efforts are needed for testing, approvals, and registrations. Therefore, it is essential to strategize the methods to delay the fungicide resistance. This review focuses on the impact of several fungicide applications currently used on FHB, focusing on Fusarium graminearum, the status of the fungicide sensitivity for fungicide classes, the resistance mechanisms against fungicides, and the mitigation strategies to delay the development of fungicide resistance in the pathogen population. Studying the fungicide resistance mechanisms and the mitigation strategies will be helpful in the future to use the available fungicides against F. graminearum without losing its effectiveness.

Cotton is one of the important economic crops in the world, but the yield and quality of cotton are often affected by salt stress. Developing salt-resistant cotton varieties by genetic engineering technology is an effective method to solve this problem. ST6-40 gene was isolated from Thellungiella halophila by a functional gene mining method based on the construction of transgenic seed libraries composed of numerous independent lines and the screening of salt-tolerant lines by high-throughput genetic screening, and the obtained objective genes were directly associated with salt-tolerant function. In this study, the pCB2004-ST6-40 recombinant vector was transformed into an upland cotton variety R15 by Agrobacterium-mediated method, and the regenerated lines of transgenic cotton were obtained. To identify the salt tolerance of the transgenic cotton lines, physiological index assay and direct salt tolerance identification were taken. T1 and T2 generations of transgenic cotton lines were identified by measuring free proline content of leaves, while T3 generation of transgenic cotton lines were directly identified by saline irrigation, and the salt tolerance was determined by calculating the salt damage index according to the salt damage series. The results showed that 350 transformation events were performed, and the transformation rate was 19%, 65 positive T0generation lines were obtained, and 90% of the regenerated plants were positive plants. PCR detection of T1, T2 and T3 for three consecutive generations at the genome level and expression identification at the RNA level showed that the target gene could be inherited stably. Through the salt-tolerance detection of T1, T2 and T3 generation of transgenic positive plants, six transgenic homozygous lines with significantly enhanced salt tolerance compared with the receptor cotton R15 were obtained, and the salt tolerance of four transgenic lines was better than that of the salt tolerant control variety 99807, indicating that transgenic ST6-40 gene could effectively improve salt tolerance of cotton varieties. [ABSTRACT FROM AUTHOR]

Objective：To construct the shRNA interference vector and over-expression vector of N-myc downstream regulatory gene 1（NDRG1)and transfect the human cerebral microvascular endothelial cells (hCMECs), and to verify the interference and over-expression effects and to obtain the cells stably overexpressing NDRG1. Methods：The pGPU6-GFP plasmid was double digested by BamH Ⅰ and Bbs Ⅰ, and then ligated with eight designed shRNAs to construct the pGPU6-GFP-NDRG1 shRNA interference vectors respectively, and the hCMECs were transfected by Lipofectamine 2000 after sequencing and identification, and the transfection efficiency of the vectors was observed under fluorescence microscope; the sequence of NDRG1 coding region amplified by PCR was ligated with the pMD19-T, after correct sequencing, the recombinant plasmid and the empty plasmid pcDNA3. 1 were digested with restriction endonucleases BamH Ⅰ and Hind Ⅲ, and the pcDNA3. 1-NDRG1 over-expression vector was constructed after ligation; the hCMECs were transfected with the correctly sequenced over-expression vector by using Lipofectamine 2000; the expression levels of NDRG1 mRNA in the hCMECs were detected by real-time fluorescence quantitative PCR（RT-qPCR)method, and the expression levels of NDRG1 protein in the hCMECs were detected by Western blotting method; the positive clone hCMECs were screened by neomycin. Results：After the pGPU6-GFP vector was doubly digested, the electrophoresis results showed that it was completely linear, and the sequencing results showed that the shRNAs were all successfully connected to the pGPU6-GFP vector; the transfection efficiency of vector was about 70 % observed under fluorescence microscope; the RT-qPCR results showed that there were three shRNA interference vectors with good interference effects, and the expression levels of NDRG1 mRNA in the hCMECs after transfection were about 26%, 21%, and 13% of those in control group, respectively; the corresponding specific protein bands were obtained by Western blotting method, the recombinant plasmid pcDNA3. 1- NDRG1 was identified by enzymatic digestion, and two DNA bands of 1 185 and 5 428 bp were identified by electrophoresis, and the sequencing results showed that the NDRG1 gene was successfully inserted into the pcDNA3. 1 vector, and the positive clone hCMECs were successfully constructed. The RT-qPCR results showed that the expression level of NDRG1 in the positive clone hCMECs was about 25. 96 times higher than that in control group; the Western blotting results showed that the expression level of NDRG1 protein in the hCMECs was increased compared with control group. Conclusion：The pGPU6-GFPNDRG1 shRNA interference vector is successfully constructed and the interference effect is verified; the pcDNA3. 1-NDRG1 over-expression vector and the hCMECs stably over-expressing NDRG1 are successfully constructed. [ABSTRACT FROM AUTHOR]

Introduction: Prostatic adenocarcinoma is the second leading cause of cancer related death in men in the western world and its incidence is increasing in Asian countries. Hence, it is of diagnostic challenge for pathologists to report in tissue biopsies especially in small focus of suspicious glands in radical prostatectomies and needle biopsies. Aim: To evaluate the role of Alpha-Methyl Acyl CoA Racemase (AMACR) immunohistochemical marker in diagnosing prostatic adenocarcinomas and in benign conditions of prostate. Materials and Methods: This was a cross-sectional study of 26 cases of prostatic adenocarcinomas, two cases of High Grade Prostatic Intraepithelial Neoplasias (HGPIN), 139 cases of Benign Prostatic Hyperplasia (BPH) and two cases of atypical adenomatous diagnosed on routine Haemotoxylin and Eosin (H&E) stained sections during a period of four years from January 2016 to December 2019 at ASRAM Medical College, Eluru. Immunohistochemistry with AMACR marker was done in all cases. Membranous staining pattern was accepted for prostatic adenocarcinomas. Semiquantitative scoring method was employed and tissue sections were examined at high power magnification (X400) by Olympus light microscope to evaluate AMACR expression. Results: Out of the 26 cases of prostatic adenocarcinomas in the study, majority were Grade group IV according to Gleason scoring system. Majority of the prostatic carcinomas showed strong and diffuse AMACR positivity. All BPH and Atypical Adenomatous Hyperplasia (AAH) were negative to AMACR immunohistochemical marker. Conclusion: The AMACR was found to be important diagnostic immune marker in prostatic adenocarcinomas especially in problematic situations where quantity and quality of tissue is limited.

Background: With a global prevalence of about 10%, gastric cancer is among the most prevalent cancers. Currently, there has been an ongoing trend toward investigating genetic disruptions in different cancers because they can be used as a target-specific therapy. We aimed to systemically review some gene expression patterns in gastric cancer. Methods: The current systematic review was designed and executed in 2020. Scopus, PubMed, Cochrane Library, Google Scholar, web of knowledge, and Science Direct were searched for relevant studies. A manual search of articles (hand searching), reference exploring, checking for grey literature, and seeking expert opinion were also done. Results: In this review, 65 studies were included, and the expression pattern of HER2/ ERBB2, ER1/Erb1/EGFR, PIK3CA, APC, KRAS, ARID1A, TP53, FGFR2 and MET was investigated. TP53, APC, KRAS, and PIK3CA mutation cumulative frequency were 24.8 (I² = 95.05, Q value = 525.53, df = 26, P < 0.001), 7.2 (I² = 89.79, Q value = 48.99, df = 5, P < 0.001), 7.8 (I² = 93.60, Q value = 140.71, df = 9, P = 0.001) and 8.6 (I² = 80.78, Q value = 525.53, df = 9, P < 0.001) percent, respectively. Overexpression was investigated for HER1/ Erb1/EGFR, PIK3CA, APC, KRAS, ARID1A, TP53, CCND1, FGFR2, MET and MYC. The frequency of TP53 and HER2/ERBB2 were 43.1 (I² = 84.06, Q value = 58.09, df = 9, P < 0.001) and 20.8 (I² = 93.61, Q value = 234.89, df = 15, P < 0.001) percent, respectively. Conclusion: More research is encouraged to investigate the genes for which we could not perform a meta-analysis. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Hydrobiology is the property of Editorial Department of Journal of Hydrobiology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Genetic studies including the quest, cloning and expression of genes encoding proteins responsible for various vital physiological processes and beneficial characteristics of economic perspective have made the biotechnology research progressively auspicious. Due to its great zootechnical and industrial importance somatotropin gene have been cloned from various animal species. Current study was designed to clone mature ovine growth hormone complementary DNA (oGH cDNA) of a sheep breed, Kajli and carry out over expression studies of cloned GH cDNA in a suitable prokaryotic expression system. Sheep GH cDNA was cloned in T/A (thymine / adenine) vector with signal peptide and confirmed by nested polymerase chain reaction (PCR) and restriction digestion. The gene was then ligated in pLEX expression vector and restricted plasmids showed a fragment insert of ~ 600 bps. Restriction analysis confirmed positive clones, were induced for protein expression analysis. The pET vectors (plasmid for expression by T7 RNA polymerase) have an isopropylthio-ß-galactoside (IPTG) inducible strong T7 promoter and Escherichia coli expression strain of BL21 (DE3) pLysS contains DNA fragment from T7 phage which harbors RNA polymerase. Therefore, for expressing recombinant proteins, cells were induced with various IPTG concentrations to optimize expression levels. Cells were induced with different IPTG concentrations (0.1 to 0.8 mM) followed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Results indicated maximum expression level of oGH at 5 hrs after induction of cells with 0.3 mM IPTG concentration with a molecular weight of 22 kDa. As for as cellular localization of protein is concerned accumulation of expressed oGH is observed in inclusion bodies. The successful expression of the cloned GH cDNA of sheep confirmed the functional viability of the clone. The above mentioned technique of genetic engineering has provided to boost the dairy industry by the production of large quantities of recombinant bovine somatotropin (rbST). [ABSTRACT FROM AUTHOR]

This study describes the anthelmintic efficacy of an organic fraction (EtOAc-F) from Guazuma ulmifolia leaves and the evaluation of its reactive oxidative stress on Haemonchus contortus. The first step was to assess the anthelmintic effect of EtOAc-F at 0.0, 3.5, 7.0 and 14 mg kg of body weight (BW) in gerbil's (Meriones unguiculatus) artificially infected with H. contortus infective larvae (L3). The second step was to evaluate the preliminary toxicity after oral administration of the EtOAc-F in gerbils. Finally, the third step was to determine the relative expression of biomarkers such as glutathione (GPx), catalase (CAT), and superoxide dismutase (SOD) against H. contortus L3 post-exposition to EtOAc-F. Additionally, the less-polar compounds of EtOAc-F were identified by gas mass spectrophotometry (GC-MS). The highest anthelmintic efficacy (97.34%) of the organic fraction was found in the gerbils treated with the 14 mg/kg of BW. Histopathological analysis did not reveal changes in tissues. The relative expression reflects overexpression of GPx (p<0.05, fold change: 14.35) and over expression of SOD (p≤0.05, fold change: 0.18) in H. contortus L3 exposed to 97.44 mg/mL of EtOAc-F compared with negative control. The GC-MS analysis revealed the presence of 4-hydroxybenzaldehyde (1), leucoanthocyanidin derivative (2), coniferyl alcohol (3), ferulic acid methyl ester acetate (4), 2,3,4-trimethoxycinnamic acid (5) and epiyangambin (6) as major compounds. According to these results, the EtOAc-F from G. ulmifolia leaves exhibit anthelmintic effect and increased the stress biomarkers on H. contortus. [Display omitted] • Ethyl acetate fraction reduces the larvae count in the in vivo gerbil test. • There were no histopathological lesions attributable to the treatments in gerbils. • Ethyl acetate fraction increases stress biomarkers on Haemonchus contortus larvae. [ABSTRACT FROM AUTHOR]

The Iranian poppy by the scientific name of Papaver bracteatum Lindl. has been long propounded as a source for thebaine. Thebaine is among secondary metabolites that due to its wide spread and unique medicinal properties play a valuable role in production of herbal medicines and pharmaceutical industries on the whole. The objective of the present study is to investigate the enhanced production of thebaine by over expression of salutaridinol 7-o-acetyltransferase (SalAT) gene, a key gene in thebaine biosynthesis pathway, in Agorobacterium-mediated transformed cell suspension culture of P. bracteatum. In this study, the cDNA of SalAT with P. bracteatum origin was cloned in pBI121 expression vector under CaMV35s promoter. In order to transfer the cloned gene Agrobacterium tumefaciens of LBA4404 strain and hypocotyl-cotyledon explants were used. For recognition of transformed calli obtained, PCR and Southern blot analysis were employed and high performance liquid chromatography analysis (HPLC) was applied for determination of thebaine content. The comparison of callus induction and thebaine content between transformed and non-transformed cell culture was studied on completely randomized design with three replications. According to HPLC analysis which revealed a significant 1.7 fold (0.58 mg/g) enhancement in thebaine content in transformed suspension cell culture comparing to nontransformed cell culture (0.34 mg/g), it can be concluded that the over expression of key genes such as SalAT gene in the cellular suspension culture of P. bracteatum could be a promising potential for mass production of this valuable alkaloid. [ABSTRACT FROM AUTHOR]

Trisomy for human chromosome 21 (Hsa21) results in Down syndrome (DS), one of the most genetically complex conditions compatible with human survival. Assessment of the physiological consequences of dosage-driven overexpression of individual Hsa21 genes during early embryogenesis and the resulting contributions to DS pathology in mammals are not tractable in a systematic way. A recent study looked at loss-of-function of a subset of Caenorhabditis elegans orthologs of Hsa21 genes and identified ten candidates with behavioral phenotypes, but the equivalent over-expression experiment has not been done. We turned to zebrafish as a developmental model and, using a number of surrogate phenotypes, we screened Hsa21 genes for effects on early embyrogenesis. We prepared a library of 164 cDNAs of conserved protein coding genes, injected mRNA into early embryos and evaluated up to 5 days post-fertilization (dpf). Twenty-four genes produced a gross morphological phenotype, 11 of which could be reproduced reliably. Seven of these gave a phenotype consistent with down regulation of the sonic hedgehog (Shh) pathway; two showed defects indicative of defective neural crest migration; one resulted consistently in pericardial edema; and one was embryonic lethal. Combinatorial injections of multiple Hsa21 genes revealed both additive and compensatory effects, supporting the notion that complex genetic relationships underlie end phenotypes of trisomy that produce DS. Together, our data suggest that this system is useful in the genetic dissection of dosage-sensitive gene effects on early development and can inform the contribution of both individual loci and their combinatorial effects to phenotypes relevant to the etiopathology of DS.

Membrane proteins play important functions, such as cellular communication and transferring materials in the cell. Many membrane proteins are involved in human diseases. However, little information is available on their structure, stability and folding. To study their structure, membrane proteins should primarily be produces in large amounts. However, production of these proteins is limited by various technical issues. Developing novel strategies to circumvent these issues seems to be highly important to proceed in membrane protein science. In this review, we have summarized the recent findings which can promote membrane protein expression and purification. [ABSTRACT FROM AUTHOR]

Background and Objective: Calcitonin is a small peptide hormone including 32 amino acids and 3.4 KD molecular weight which is produced by the parafollicular cells of the thyroid gland in respond to increasing calcium ions in serum. This peptide is used for adjuvant therapy of osteoporosis, Paget's disease and hypercalcemic shock. In this study, the heterologuse expression of calcitonin was done in Escherichia coli. Methods: In this experimental study, the thioredoxin fusion partner was added to n-terminal of the Salmon calciton in order to increase its stability by the synthetic biology. The recombinant construct was transformed and over expressed into Escherichia coli BL21 (DE3) host cell. Results: SDS-PAGE analysis showed the over expression of recombinant protein after IPTG induction. Conclusion: In this study, the construct including fused Salmon calcitonin gene with thioredoxin was cloned. The SDS-PAGE result showed the stable expression of fused calcitonin.

To explore the potential genes from the industrially polluted Amlakhadi canal, located in Ankleshwar, Gujarat, India, its community genome was extracted and cloned into E. coli EPI300™-T1 R using a fosmid vector (pCC2 FOS™) generating a library of 3,92,000 clones with average size of 40 kb of DNA-insert. From this library, the clone DM1 producing brown colored melanin-like pigment was isolated and characterized. For over expression of the pigment, further sub-cloning of the clone DM1 was done. Sub-clone containing 10 kb of the insert was sequenced for gene identification. The amino acids sequence of a protein 4-Hydroxyphenylpyruvate dioxygenase (HPPD), which is know to be involved in melanin biosynthesis was obtained from the gene sequence. The sequence-homology based 3D structure model of HPPD was constructed and analyzed. The physico-chemical nature of pigment was further analysed using 1 H and 13 C NMR, LC-MS, FTIR and UV–visible spectroscopy. The pigment was readily soluble in DMSO with an absorption maximum around 290 nm. Based on the genetic and chemical characterization, the compound was confirmed as melanin-like pigment. The present results indicate that the metagenomic library from industrially polluted environment generated a microbial tool for the production of melanin-like pigment. [ABSTRACT FROM AUTHOR]

Trisomy for human chromosome 21 (Hsa21) results in Down syndrome (DS), one of the most genetically complex conditions compatible with human survival. Assessment of the physiological consequences of dosage-driven overexpression of individual Hsa21 genes during early embryogenesis and the resulting contributions to DS pathology in mammals are not tractable in a systematic way. A recent study looked at loss-of-function of a subset of Caenorhabditis elegans orthologs of Hsa21 genes and identified ten candidates with behavioral phenotypes, but the equivalent over-expression experiment has not been done. We turned to zebrafish as a developmental model and, using a number of surrogate phenotypes, we screened Hsa21 genes for effects on early embyrogenesis. We prepared a library of 164 cDNAs of conserved protein coding genes, injected mRNA into early embryos and evaluated up to 5 days post-fertilization (dpf). Twenty-four genes produced a gross morphological phenotype, 11 of which could be reproduced reliably. Seven of these gave a phenotype consistent with down regulation of the sonic hedgehog (Shh) pathway; two showed defects indicative of defective neural crest migration; one resulted consistently in pericardial edema; and one was embryonic lethal. Combinatorial injections of multiple Hsa21 genes revealed both additive and compensatory effects, supporting the notion that complex genetic relationships underlie end phenotypes of trisomy that produce DS. Together, our data suggest that this system is useful in the genetic dissection of dosage-sensitive gene effects on early development and can inform the contribution of both individual loci and their combinatorial effects to phenotypes relevant to the etiopathology of DS. [ABSTRACT FROM AUTHOR]

Natural rubber (cis-1, 4-polyisoprene) is being produced from bark laticifer cells of Hevea brasiliensis and the popular high latex yielding Indian rubber clones are easily prone to onset of tapping panel dryness syndrome (TPD) which is considered as a physiological syndrome affecting latex production either partially or completely. This report describes an efficient protocol for development of transgenic rubber plants by over-expression of 3-hydroxy 3-methylglutaryl Co-enzyme A reductase 1 ( hmgr1 ) gene which is considered as rate limiting factor for latex biosynthesis via Agrobacterium -mediated transformation. The pBIB plasmid vector containing hmgr1 gene cloned under the control of a super-promoter was used for genetic transformation using embryogenic callus. Putatively transgenic cell lines were obtained on selection medium and produced plantlets with 44% regeneration efficiency. Transgene integration was confirmed by PCR amplification of 1.8 kb hmgr1 and 0.6 kb hpt genes from all putatively transformed callus lines as well as transgenic plants. Southern blot analysis showed the stable integration and presence of transgene in the transgenic plants. Over expression of hmgr1 transgene was determined by Northern blot hybridization, semi-quantitative PCR and real-time PCR (qRT-PCR) analysis. Accumulation of hmgr1 mRNA transcripts was more abundant in transgenic plants than control. Increased level of photosynthetic pigments, protein contents and HMGR enzyme activity was also noticed in transgenic plants over control. Interestingly, the latex yield was significantly enhanced in all transgenic plants compared to the control. The qRT-PCR results exhibit that the hmgr1 mRNA transcript levels was 160-fold more abundance in transgenic plants over untransformed control. These results altogether suggest that there is a positive correlation between latex yield and accumulation of mRNA transcripts level as well as HMGR enzyme activity in transgenic rubber plants. It is presumed that there is a possibility for enhanced level of latex biosynthesis in transgenic plants as the level of mRNA transcripts and HMGR enzyme activity is directly correlated with latex yield in rubber tree. Further, the present results clearly suggest that the quantification of HMGR enzyme activity in young seedlings will be highly beneficial for early selection of high latex yielding plants in rubber breeding programs. [ABSTRACT FROM AUTHOR]

Water hyacinth pretreated with microwave-assisted dilute acid was used for fermentation to produce hydrogen using four kinds of genetically modified bacteria for the first time in this study. The genes of several hydrogenase were over-expressed in Enterobacter cloacae CICC10017 and Enterobacter aerogenes ATCC13408. Four recombinant strains including E. cloacae/HPP, E. aerogenes/HycE, E. aerogenes/HycG, and E. aerogenes/HoxEFUYH were obtained. Hydrogen yields from water hyacinth were investigated, which were enhanced using the recombinant strains. When water hyacinth was pretreated by microwave at 140°C for 15 min with 1% H2SO4 and then hydrolyzed by cellulase, the reducing sugar yield of 49.4 g/100 g hyacinth was 91.4% of the theoretical reducing sugar yield. Among the four genetically modified bacteria, E. cloacae/HPP gave the highest hydrogen yield of 74.9 mL/g- total volatile solids, which was 50% higher than that given by the wild strain from pretreated water hyacinth. © 2017 American Institute of Chemical Engineers Environ Prog, 36: 1296-1300, 2017 [ABSTRACT FROM AUTHOR]

Aims RAD51 participates in homologous recombination repair (HRR) of double-stranded DNA breaks (DSBs) which may cause genomic instability and cancer. The aim of this study was to investigate RAD51 gene expression at transcriptional and translational levels to measure mRNA and protein level and to correlate its relationship with proliferation marker, Ki67 in thyroid cancer patients. This study also explored correlation of these genes with different clinicopathological parameters of the study cohort by Spearman's rank correlation coefficient. Methods Quantitative real time polymerase chain reaction (qRT-PCR) and immunohistochemistry were used to detect mRNA transcript levels and protein expression of RAD51 and Ki67 in 102 cases of thyroid cancer tissues and equal number of uninvolved healthy thyroid tissue controls. Results Data showed that expression for both RAD51 and Ki67 was significantly increased in thyroid cancer (p < 0.001). High RAD51 and Ki67 expression was associated with later stages, poor tissue differentiation, large tumor size, positive lymph node metastasis and distant metastasis. The correlation analysis demonstrated a strong positive correlation (r = 0.461) between RAD51 and Ki67 on mRNA level and on protein level (r = 0.866). Strong correlation was observed between clinicopathological characteristics and selected molecules. Conclusion The present study concluded that upregulation of RAD51 and overexpression of Ki67 may be associated with the progression of thyroid cancer. [ABSTRACT FROM AUTHOR]

Background and Objective: Calcitonin is a small peptide hormone including 32 amino acids and 3.4 KD molecular weight which is produced by the parafollicular cells of the thyroid gland in respond to increasing calcium ions in serum. This peptide is used for adjuvant therapy of osteoporosis, Paget's disease and hypercalcemic shock. In this study, the heterologuse expression of calcitonin was done in Escherichia coli. Methods: In this experimental study, the thioredoxin fusion partner was added to n-terminal of the Salmon calciton in order to increase its stability by the synthetic biology. The recombinant construct was transformed and over expressed into Escherichia coli BL21 (DE3) host cell. Results: SDS-PAGE analysis showed the over expression of recombinant protein after IPTG induction. Conclusion: In this study, the construct including fused Salmon calcitonin gene with thioredoxin was cloned. The SDS-PAGE result showed the stable expression of fused calcitonin. [ABSTRACT FROM AUTHOR]

Protoporphyrin IX has been used as an efficient sensitizer in photodynamic diagnosis, photodynamic therapy, and sonodynamic therapy. The level of protoporphyrin IX is very important for diagnostic or therapy effects. 5-aminolevulinic acid synthase 2 (ALAS2) is the key enzyme upstream of protoporphyrin IX synthesis. To increase protoporphyrin IX accumulation, ALAS2 over-expression transgenic mice were generated. Plasmid containing alas2 gene was transfected in colonic carcinoma cell lines. Both in tissues of transgenic mice and in colonic carcinoma cells, the amount of protoporphyrin IX accumulation did increase. In addition, the level of heme, which is down stream of protoporphyrin IX did not change. Overexpression ALAS2 in nonerythriod cells may become a novel approach to cause protoporphyrin IX accumulation. [ABSTRACT FROM AUTHOR]

Toll receptors play an important role in the innate immunity of invertebrates. All reported Tolls have only one Toll/interleukin-1 receptor (TIR) domain at the C-terminal. In this study, numerous Tolls with tandem TIRs at the C-terminal were found in molluscs. Such Tolls presented an extra TIR (TIR-1) compared with Toll-I. Thus, Toll-I might be the ancestor of tandem TIRs containing Toll. To test this hypothesis, 83 Toll-I and Toll-2 (most have two TIRs, but others seem to be the evolutionary intermediates) genes from 29 shellfish species were identified. These Tolls were divided into nine groups based on phylogenetic analyses. A strong correlation between phylogeny and motif composition was found. All Toll proteins contained the TIR-2 domain, whereas the TIR-1 domain only existed in some Toll-2 protein, suggesting that TIR-1 domain insertion may play an important role in Toll protein evolution. Further analyses of functional divergence and adaptive evolution showed that some of the critical sites responsible for functional divergence may have been under positive selection. An additional intragenic recombination played an important role in the evolution of the Toll-I and Toll-2 genes. To investigate the functional difference of Toll-I and Toll-2, over expression of Hcu_Toll-I or Hcu_Toll-2-2 in Drosophila S2 cells was performed. Results showed that Hcu_Toll-2-2 had stronger antimicrobial peptide (AMP) activity than Hcu_Toll-I. Therefore, enhanced AMP-induced activity resulted from tandem TIRs in Toll-2s of molluscs during evolution history.

To further improve the expression level of recombinant xylanase in Pichia pastoris, the xynB gene, encoding the mature peptide from Aspergillus niger NL-1, was designed and synthesized based on the synonymous condon bias of P. pastoris and optimized G+C content. 155 nucleotides were changed, and the GC content decreased from 57.7% to 43.6%. The synthetic xynB was inserted into the pPICZaA and then integrated into P. pastoris GS115. The activity of the recombinant xylanase reached 1414.7 U/mL, induced with 0.8% methanol after 14-day cultivation at a temperature of 28oC in shake flasks, which was 267% higher than that of the native gene. Furthermore, the maximum xylanase activity of 20424.2 U/mL was obtained by high-density fermentation in a 5-L fermenter, which was the highest xylanase expression in P. pastoris yet reported. The recombinant xylanase had its optimal activity at a pH of 5.0 and temperature of 50oC. The recombinant xylanase was stable over a pH range of 4.5 to 8.0. Thus, this report provides an industrial means to produce the recombinant xylanase in P. pastoris.

Copyright of Chinese Journal of Applied Ecology / Yingyong Shengtai Xuebao is the property of Chinese Journal of Applied Ecology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Dark fermentative H 2 production is a promising approach since it requires minimal energy input as compared to other conventional methods of hydrogen production. Moreover, a wide variety of low cost-feedstock can be used that are released in the environment, thus providing an added advantage of waste management. This paper reviews the application of genomic and proteomic approaches to heterotrophic hydrogen production that could be useful in improvement of the same. Molecular techniques like polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) are conventionally used for microbial profiling of mixed consortia. An impending application of molecular techniques like terminal-restriction fragment length polymorphism (T-RFLP), ribosomal intergenic spacer analysis (RISA), quantitative PCR (qPCR), single strand conformation polymorphism (SSCP), fluorescence in situ hybridization (FISH) and fluorescence activated cell sorting (FACS) could be potentially used for advanced and rapid microbial characterization. Molecular techniques like gene overexpression, homologous recombination, gene knockout and antisense RNA are extensively used for improvement of hydrogen production processes. Challenges of applying above contemporary molecular techniques in thermophilic biohydrogen production processes were also studied. Use of proteomics helps in understanding the effect of genetic improvement leading to changes in entire proteome of the organism. Moreover, it is a useful tool for study the functional aspect of overexpression, site directed mutagenesis, random mutagenesis etc. towards improvement of H 2 production. Thus the study of genomic and proteomic techniques improve understanding of dark fermentative hydrogen production processes. [ABSTRACT FROM AUTHOR]