SDR Family Research Articles

New insights into the mechanism of azo dye biodegradation by Lactococcus lactis

Azo dye reduction by syntrophic microbial communities is still unclear, with conflicting observations reported in the literature. In this study, the biodegradation mechanism of a model azo dye by Lactococcus lactis strain LLSP-01 isolated from an acidogenic reactor system was investigated. Proteomics and RT-PCR analysis results showed that LLSP-01 azoreductase (AzoL) was not expressed by the isolate during exposure to Direct Black 22. In contrast, the mechanism appears to involve biosorption by glycoconjugates, particularly exopolysaccharides (EPS) and rhamnolipids, as proteins from the LPS O-antigen metabolism were statistically higher expressed in cells challenged with the target compound. Based on the proteomic observations, it is hypothesized that Direct Black 22 is adsorbed into the biofilm matrix and indirectly reduced by an SDR family oxidoreductase, in a mechanism mediated by riboflavin carriers. Induction of a ring-cleaving dioxygenase in the presence of azo dye indicates further degradation of the resulting aromatic amines. The collected results show that azo dye reduction by L. lactis is mediated by enzymes with broad range specificity, and not the typical azoreductases. This information can assist in the design of new strategies for the bioremediation of textile azo dyes.

Low mutation rate of spontaneous mutants enables detection of causative genes by comparing whole genome sequences.

In the early 1900s, mutation breeding to select varieties with desirable traits using spontaneous mutation was actively conducted around the world, including Japan. In rice, the number of fixed mutations per generation was estimated to be 1.38-2.25. Although this low mutation rate was a major problem for breeding in those days, in the modern era with the development of next-generation sequencing (NGS) technology, it was conversely considered to be an advantage for efficient gene identification. In this paper, we proposed an in silico approach using NGS to compare the whole genome sequence of a spontaneous mutant with that of a closely related strain with a nearly identical genome, to find polymorphisms that differ between them, and to identify the causal gene by predicting the functional variation of the gene caused by the polymorphism. Using this approach, we found four causal genes for the dwarf mutation, the round shape grain mutation and the awnless mutation. Three of these genes were the same as those previously reported, but one was a novel gene involved in awn formation. The novel gene was isolated from Bozu-Aikoku, a mutant of Aikoku with the awnless trait, in which nine polymorphisms were predicted to alter gene function by their whole-genome comparison. Based on the information on gene function and tissue-specific expression patterns of these candidate genes, Os03g0115700/LOC_Os03g02460, annotated as a short-chain dehydrogenase/reductase SDR family protein, is most likely to be involved in the awnless mutation. Indeed, complementation tests by transformation showed that it is involved in awn formation. Thus, this method is an effective way to accelerate genome breeding of various crop species by enabling the identification of useful genes that can be used for crop breeding with minimal effort for NGS analysis.

Identification of Potential Factors for the Promotion of Fucoxanthin Synthesis by Methyl Jasmonic Acid Treatment of Phaeodactylum tricornutum.

Fucoxanthin, a vital secondary metabolite produced by marine diatoms, has great economic value and research potential. However, its popularization and application have been greatly restricted due to its low content, difficult extraction, and high production cost. Methyl jasmonic acid (MeJA) exerts similar inductive hormones in the growth and development as well as metabolic processes of plants. In Phaeodactylum tricornutum (P. tricornutum), MeJA treatment can increase fucoxanthin content. In this study, the effects of different concentrations of MeJA on the cell growth and the fucoxanthin content of P. tricornutum were explored. Meanwhile, this study used high-throughput sequencing technology for transcriptome sequencing of P. tricornutum and subsequently performed differential gene expression analysis, gene ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, and weighted gene co-expression network analysis (WGCNA) for screening the hub genes for the promotion of fucoxanthin synthesis with MeJA-treated P. tricornutum. On this basis, the functions of the hub genes for the promotion of fucoxanthin synthesis with MeJA-treated P. tricornutum were further analyzed. The results revealed that the carotenoid synthesis-related genes PHATRDRAFT_54800 and PHATRDRAFT_20677 were the hub genes for the promotion of fucoxanthin synthesis with MeJA-treated P. tricornutum. PHATRDRAFT_54800 may be a carotenoid isomerase, while PHATRDRAFT_20677 may be involved in the MeJA-stimulated synthesis of fucoxanthin by exerting the role of SDR family NAD(P)-dependent oxidoreductases.

The Molecular Basis of Catalysis by SDR Family Members Ketoacyl‐ACP Reductase FabG and Enoyl‐ACP Reductase FabI in Type‐II Fatty Acid Biosynthesis

AbstractThe short‐chain dehydrogenase/reductase (SDR) superfamily members acyl‐ACP reductases FabG and FabI are indispensable core enzymatic modules and catalytic orientation controllers in type‐II fatty acid biosynthesis. Herein, we report their distinct substrate allosteric recognition and enantioselective reduction mechanisms. FabG achieves allosteric regulation of ACP and NADPH through ACP binding across two adjacent FabG monomers, while FabI follows an irreversible compulsory order of substrate binding in that NADH binding must precede that of ACP on a discrete FabI monomer. Moreover, FabG and FabI utilize a backdoor residue Phe187 or a “rheostat” α8 helix for acyl chain length selection, and their corresponding triad residues Ser142 or Tyr145 recognize the keto‐ or enoyl‐acyl substrates, respectively, facilitating initiation of nucleophilic attack by NAD(P)H. The other two triad residues (Tyr and Lys) mediate subsequent proton transfer and (R)‐3‐hydroxyacyl‐ or saturated acyl‐ACP production.

The brown planthopper NlDHRS11 is involved in the detoxification of rice secondary compounds.

The brown planthopper (Nilaparvata lugens, BPH) is the most destructive serious pest for rice production. Resistant varieties are effective means to defend against BPH, but the impact of the ingestion of resistant rice on the BPH transcriptional regulation is still unclear. Here, we explored the molecular basis of the regulation by BPH feeding on resistant rice. BPH nymphs preferentially selected susceptible rice TN1 at 24 h after release in a choice test. Feeding on resistant rice IR56 under a non-selective condition increased mortality, decreased growth rate, and prolonged molting time of BPH. Transcriptomic sequencing revealed 38 dysregulated genes, including 31 down-regulated and 7 up-regulated genes in BPH feeding on resistant rice for 7 days compared with feeding susceptible rice TN1. These genes were mainly involved in the pathways of growth and development, metabolism, energy synthesis, and transport. Finally, we showed that the toxicities of rice defensive compounds to BPH were dose-dependent, and silencing of BPH gene dehydrogenase/reductase SDR family member 11 (NlDHRS11) increased sensibility to rice secondary compounds, ferulic acid, and resorcinol. The adaption of BPH feeding on resistant rice is orchestrated by dynamically regulating gene expressions, and NlDHRS11 is a gene involved in the detoxification of plant defensive chemicals. The current work provides new insights into the interaction between insects and plants, and helps to develop novel BPH control strategies. This article is protected by copyright. All rights reserved.

Transcriptomic Response of Clonostachys rosea Mycoparasitizing Rhizoctonia solani.

Clonostachys rosea is an important mycoparasitism biocontrol agent that exhibits excellent control efficacy against numerous fungal plant pathogens. Transcriptomic sequencing may be used to preliminarily screen mycoparasitism-related genes of C. rosea against fungal pathogens. The present study sequenced and analyzed the transcriptome of C. rosea mycoparasitizing a Basidiomycota (phylum) fungal pathogen, Rhizoctonia solani, under three touch stages: the pre-touch stage, touch stage and after-touch stage. The results showed that a number of genes were differentially expressed during C. rosea mycoparasitization of R. solani. At the pre-touch stage, 154 and 315 genes were up- and down-regulated, respectively. At the touch stage, the numbers of up- and down-regulated differentially expressed genes (DEGs) were 163 and 188, respectively. The after-touch stage obtained the highest number of DEGs, with 412 and 326 DEGs being up- and down-regulated, respectively. Among these DEGs, ABC transporter-, glucanase- and chitinase-encoding genes were selected as potential mycoparasitic genes according to a phylogenetic analysis. A comparative transcriptomic analysis between C. rosea mycoparasitizing R. solani and Sclerotinia sclerotiorum showed that several DEGs, including the tartrate transporter, SDR family oxidoreductase, metallophosphoesterase, gluconate 5-dehydrogenase and pyruvate carboxylase, were uniquely expressed in C. rosea mycoparasitizing R. solani. These results significantly expand our knowledge of mycoparasitism-related genes in C. rosea and elucidate the mycoparasitism mechanism of C. rosea.

P01-002-23 Effect of Dehydrogenase/Reductase SDR Family Member 9 (DHRS9) on Skin Responses to Acute Ultraviolet Light B Exposure in Mice

P01-002-23 Effect of Dehydrogenase/Reductase SDR Family Member 9 (DHRS9) on Skin Responses to Acute Ultraviolet Light B Exposure in Mice

Hif1α/Dhrs3a Pathway Participates in Lipid Droplet Accumulation via Retinol and Ppar-γ in Fish Hepatocytes.

Excessive hepatic lipid accumulation is a common phenomenon in cultured fish; however, its underlying mechanisms are poorly understood. Lipid droplet (LD)-related proteins play vital roles in LD accumulation. Herein, using a zebrafish liver cell line (ZFL), we show that LD accumulation is accompanied by differential expression of seven LD-annotated genes, among which the expression of dehydrogenase/reductase (SDR family) member 3 a/b (dhrs3a/b) increased synchronously. RNAi-mediated knockdown of dhrs3a delayed LD accumulation and downregulated the mRNA expression of peroxisome proliferator-activated receptor gamma (pparg) in cells incubated with fatty acids. Notably, Dhrs3 catalyzed retinene to retinol, the content of which increased in LD-enriched cells. The addition of exogenous retinyl acetate maintained LD accumulation only in cells incubated in a lipid-rich medium. Correspondingly, exogenous retinyl acetate significantly increased pparg mRNA expression levels and altered the lipidome of the cells by increasing the phosphatidylcholine and triacylglycerol contents and decreasing the cardiolipin, phosphatidylinositol, and phosphatidylserine contents. Administration of LW6, an hypoxia-inducible factor 1α (HIF1α) inhibitor, reduced the size and number of LDs in ZFL cells and attenuated hif1αa, hif1αb, dhrs3a, and pparg mRNA expression levels. We propose that the Hif-1α/Dhrs3a pathway participates in LD accumulation in hepatocytes, which induces retinol formation and the Ppar-γ pathway.

Antiandrogenic Effects of a Polyphenol in Carex kobomugi through Inhibition of Androgen Synthetic Pathway and Downregulation of Androgen Receptor in Prostate Cancer Cell Lines.

Prostate cancer (PC) represents the most common cancer disease in men. Since high levels of androgens increase the risk of PC, androgen deprivation therapy is the primary treatment; however this leads to castration-resistant PC (CRPC) with a poor prognosis. The progression to CRPC involves ectopic androgen production in the adrenal glands and abnormal activation of androgen signaling due to mutations and/or amplification of the androgen receptor (AR) as well as activation of androgen-independent proliferative pathways. Recent studies have shown that adrenal-derived 11-oxygenated androgens (11-ketotestosterone and 11-ketodihydrotestosterone) with potencies equivalent to those of traditional androgens (testosterone and dihydrotestosterone) are biomarkers of CRPC. Additionally, dehydrogenase/reductase SDR family member 11 (DHRS11) has been reported to be a 17β-hydroxysteroid dehydrogenase that catalyzes the production of the 11-oxygenated and traditional androgens. This study was conducted to evaluate the pathophysiological roles of DHRS11 in PC using three LNCaP, C4-2 and 22Rv1 cell lines. DHRS11 silencing and inhibition resulted in suppression of the androgen-induced expression of AR downstream genes and decreases in the expression of nuclear AR and the proliferation marker Ki67, suggesting that DHRS11 is involved in androgen-dependent PC cell proliferation. We found that 5,7-dihydroxy-8-methyl-2-[2-(4-hydroxyphenyl)ethenyl]-4H-1-benzopyran-4-one (Kobochromone A, KC-A), an ingredient in the flowers of Carex kobomugi, is a novel potent DHRS11 inhibitor (IC50 = 0.35 μM). Additionally, KC-A itself decreased the AR expression in PC cells. Therefore, KC-A suppresses the androgen signaling in PC cells through both DHRS11 inhibition and AR downregulation. Furthermore, KC-A enhanced the anticancer activity of abiraterone, a CRPC drug, suggesting that it may be a potential candidate for the development of drugs for the prevention and treatment of CRPC.

Proteomics study of mitochondrial proteins in tilapia red meat and their effect on color change during storage

The underlying mechanism of the role of mitochondria in color changing of tilapia fillet during 0–4 d storage is not completely clear. A total of 209 differentially significant expressed proteins (DSEPs) were identified by using label-free mitochondrial proteomics, with 56 proteins up-regulated in T2 and 61 proteins (up-regulated) in T3. Protein-Protein interaction reveled proteins which participate in TCA cycles (Citrate synthase (cs)), Oxidoreductase (Malate dehydrogenase (mdh1, mdh2), Succinyl-CoA (Oxct1), Hydroxyacyl-coenzyme a dehydrogenase (hadh), Dehydrogenase/reductase (SDR family) member 1 (dhrs1)) interacted strongly with each other. In turn, they can increase the level of mitochondrial respiration and mitochondrial function, leading to color changing of tilapia fillet. The heat shock 60kD protein 1 (chaperonin, hspd1) interacted with metabolic enzymes (cs and mdh2) and had important effects on color. These results could help researchers better understand the color changing mechanism on the surface of tilapia fillet during the storage.

The functional emulsifying component of SL-bioemulsifier is an SDR family oxidoreductase protein

HypothesisBioemulsifiers play an important role in microbial enhanced oil recovery (MEOR) due to their excellent emulsion stability. Different from biosurfactants, bioemulsifiers cannot significantly reduce the oil-water interfacial tension, and the mechanism through which they improve oil recovery remains poorly understood. It is helpful to study the mechanism by analyzing its functional emulsifying components of bioemulsifier. ExperimentsIn our previous work, the thermophilic strain Aeribacillus pallidus SL-1 efficiently degraded crude oil and insoluble polycyclic aromatic hydrocarbons (naphthalene and phenanthrene) with the help of a bioemulsifier. In this study, the glycoprotein SL-bioemulsifier was studied. FindingsThe main functional emulsifying component was identified as a 26 kDa SDR family oxidoreductase protein, called E26. The recombinant E26 protein exhibited excellent emulsifying activity with different hydrophobic substrates, and stabilized the emulsions for a long time even at extreme pH, high salt concentrations and high temperatures. Notably, the emulsion formed by E26 protein was found to have a gel-like structure, facilitating the formation of an oil-water barrier to stabilize the emulsion. In addition, E26 protein effectively improved the surface wettability of calcite minerals in the oil-water system at a low concentration, which may play a role in the enhancement of oil recovery by bioemulsifiers.

Burkholderia Pseudomallei Short-Chain Dehydrogenase/ Oxidoreductase: Potential Urine Biomarker Candidate for Acute Melioidosis

INTRODUCTION: In the current climate of urgency in identifying biomarkers for the development of rapid diagnostic kits, the use of urine samples to diagnose acute melioidosis was evaluated, comparing urine samples from Burkholderia pseudomallei culture-positive and culture-negative patients, and comparing pneumonic and septicemic melioidosis. MATERIAL AND METHODS: Eleven urine samples from clinically suspected melioidosis patients from a tertiary referral center, Hospital Tengku Ampuan Afzan, Pahang was used. An in-solution method for the detection of bacterial proteins using liquid chromatography-mass spectrometry quadrupole time-of-flight (LCMS QTOF) was used. RESULTS: Three bacterial proteins were consistently detected among all the culture[1]positive and PCR-positive cases tested, namely SDR family NAD(P)-dependent oxidoreductase protein (32kDa), 3-hydroxyacyl-CoA dehydrogenase Burkholderia sp. (32kDa), and NAD(P)-dependent dehydrogenase (short-subunit alcohol dehydrogenase family) Burkholderia sp. (33kDa). CONCLUSIONS: Short-chain dehydrogenase (SDO) proteins could potentially be a urine biomarker candidate as these have shown to aid in the ability of Burkholderia spp. to invade host cells as this action is important for the initial intracellular survival of the organism.

Monosaccharide biosynthesis pathways database.

A distinctive feature of glycans vis-à-vis proteins and nucleic acids is its structural complexity, which arises from the huge repertoire of monosaccharides, isomeric linkages and branching. A very large number of monosaccharides have so far been discovered in natural glycans. Experimentally, pathways for the biosynthesis have been characterized completely for 55 monosaccharides and partially for a few more. However, there is no single platform, which provides information about monosaccharide biosynthesis pathways and associated enzymes We have gathered 572 experimentally characterized enzymes of 66 biosynthesis pathways from literature and set up a first of its kind database called the Monosaccharide Biosynthesis Pathways Database http://www.bio.iitb.ac.in/mbpd/). Annotations such as the reaction catalyzed, substrate specificity, biosynthesis pathway and PubMed IDs are provided for all the enzymes in the database. Sequence homologs of the experimentally characterized enzymes found in nearly 13,000 completely sequenced genomes from Bacteria and Archaea have also been included in the database. This platform will help in the deduction of evolutionary relationships among enzymes such as aminotransferases, nucleotidyltransferases, acetyltransferases and SDR family enzymes. It can also facilitate experimental studies such as direct enzyme assays to validate putative annotations, establish structure-function relationship, expression profiling to determine the function, determine the phenotypic consequences of gene knock-out/knock-in and complementation studies.

Increasing cellulosic ethanol production by enhancing phenolic tolerance of Zymomonas mobilis in adaptive evolution

Cellulosic ethanol fermentability of ethanologenic strain Zymomonas mobilis is severely inhibited by phenolic aldehydes generated from lignocellulose pretreatment. Here, a 198 days’ laboratory adaptive evolution of Z. mobilis 8b in corn stover hydrolysate was conducted to increase its phenolic aldehydes tolerance and ethanol fermentability. The obtained Z. mobilis Z198 demonstrated a significantly improved conversion of the most toxic phenolic aldehyde (vanillin) by 6.3-fold and cellulosic ethanol production by 21.6%. The transcriptional analysis using qRT-PCR revealed that the gene ZMO3_RS07160 encoding SDR family oxidoreductase in Z. mobilis Z198 was significantly up-regulated by 11.7-fold. The overexpression of ZMO3_RS07160 in the parental Z. mobilis increased the ethanol fermentability to that of the adaptively evolved strain Z. mobilis Z198. This study provided a practical method to obtain a robust cellulosic ethanol fermenting strain, and a candidate gene for synthetic biology of biorefinery strains with strong phenolic aldehydes tolerance.

Chemical Proteomic Profiling of Protein 4′‐Phosphopantetheinylation in Mammalian Cells

AbstractProtein 4′‐phosphopantetheinylation is an essential post‐translational modification (PTM) in prokaryotes and eukaryotes. So far, only five protein substrates of this specific PTM have been discovered in mammalian cells. These proteins are known to perform important functions, including fatty acid biosynthesis and folate metabolism, as well as β‐alanine activation. To explore existing and new substrates of 4′‐phosphopantetheinylation in mammalian proteomes, we designed and synthesized a series of new pantetheine analogue probes, enabling effective metabolic labelling of 4′‐phosphopantetheinylated proteins in HepG2 cells. In combination with a quantitative chemical proteomic platform, we enriched and identified all the currently known 4′‐phosphopantetheinylated proteins with high confidence, and unambiguously determined their exact sites of modification. More encouragingly, we discovered, using targeted chemical proteomics, a potential 4′‐phosphopantetheinylation site in the protein of mitochondrial dehydrogenase/reductase SDR family member 2 (DHRS2).

Chemical Proteomic Profiling of Protein 4'-Phosphopantetheinylation in Mammalian Cells.

Protein 4'-phosphopantetheinylation is an essential post-translational modification (PTM) in prokaryotes and eukaryotes. So far, only five protein substrates of this specific PTM have been discovered in mammalian cells. These proteins are known to perform important functions, including fatty acid biosynthesis and folate metabolism, as well as β-alanine activation. To explore existing and new substrates of 4'-phosphopantetheinylation in mammalian proteomes, we designed and synthesized a series of new pantetheine analogue probes, enabling effective metabolic labelling of 4'-phosphopantetheinylated proteins in HepG2 cells. In combination with a quantitative chemical proteomic platform, we enriched and identified all the currently known 4'-phosphopantetheinylated proteins with high confidence, and unambiguously determined their exact sites of modification. More encouragingly, we discovered, using targeted chemical proteomics, a potential 4'-phosphopantetheinylation site in the protein of mitochondrial dehydrogenase/reductase SDR family member 2 (DHRS2).

Identification of molecular markers for superior quantitative traits in a novel sea cucumber strain by comparative microRNA-mRNA expression profiling

To investigate the adaptability of Apostichopus japonicus (A. japonicus) strain “Anyuan No. 1” in the South China Sea, field monitoring and microRNA-mRNA integrated analyses were conducted between “Anyuan No. 1” and a regular A. japonicus population from Wendeng (Shandong Province, as a control) in the Xiapu farming area in Fujian Province, China. The results showed that “Anyuan No. 1” exhibited greater body weight increase and a higher number of papillae compared to the control during two and a half months of field monitoring. Comparative microRNA (miRNA) and mRNA transcriptome analyses identified 12 differentially expressed miRNAs (DEMs) and 165 differentially expressed genes (DEGs) in “Anyuan No. 1” compared to the control. Long-chain specific acyl-CoA dehydrogenase (ACADL), transmembrane protein 251 (TMEM251), dehydrogenase/reductase SDR family protein 7-like (Dhrs7), insulin-like growth factor-binding protein 7 (IGFBP-7), CDK5 regulatory subunit-associated protein 1 (CDK5RAP1), visual pigment-like receptor peropsin, 39S ribosomal protein, miR-10, miR-153, miR-7, and miR-3529 were identified as gene and miRNA candidates correlated with superior economic traits in “Anyuan No. 1”. Collectively, “Anyuan No. 1” is suitable for large-scale cultivation extension due to its better adaptability to the South China Sea area. Furthermore, we identified “miR10-ACADL” as a potential module for further molecular marker-assisted selective breeding of A. japonicus.

Human dehydrogenase/reductase SDR family member 11 (DHRS11) and aldo-keto reductase 1C isoforms in comparison: Substrate and reaction specificity in the reduction of 11-keto-C19-steroids

Recent studies have shown that an adrenal steroid 11β-hydroxy-4-androstene-3,17-dione serves as the precursor to androgens, 11-ketotestosterone and 11-ketodihydrotestosterone (11KDHT). The biosynthetic pathways include the reduction of 3- and 17-keto groups of the androgen precursors 11-keto-C19-steroids, which has been reported to be mediated by three human enzymes; aldo-keto reductase (AKR)1C2, AKR1C3 and 17β-hydroxysteroid dehydrogenase (HSD) type-3. To explore the contribution of the enzymes in the reductive metabolism, we kinetically compared the substrate specificity for 11-keto-C19-steroids among purified recombinant preparations of four AKRs (1C1, 1C2,1C3 and 1C4) and DHRS11, which shows 17β-HSD activity. Although AKR1C1 did not reduce the 11-keto-C19-steroids, AKR1C3 and DHRS11 reduced 17-keto groups of 11-keto-4-androstene-3,17-dione, 11-keto-5α-androstane-3,17-dione (11K-Adione) and 11-ketoandrosterone with Km values of 5–28 μM. The 3-keto groups of 11KDHT and 11K-Adione were reduced by AKR1C4 (Km 1 μM) more efficiently than by AKR1C2 (Km 5 and 8 μM, respectively). GC/MS analysis of the products showed that DHRS11 acts as 17β-HSD, and that AKR1C2 and AKR1C4 are predominantly 3α-HSDs, but formed a minor 3β-metabolite from 11KDHT. Since DHRS11 was thus newly identified as 11-keto-C19-steroid reductase, we also investigated its substrate-binding mode by molecular docking and site-directed mutagenesis of Thr163 and Val200, and found the following structural features: 1). There is a space that accommodates the 11-keto group of the 11-keto-C19-steroids in the substrate-binding site. 2) Val200 is a critical determinant for exhibiting the strict 17β-HSD activity of the enzyme, because the Val200Leu mutation resulted in both significant impairment of the 17β-HSD activity and emergence of 3β-HSD activity towards 5α-androstanes including 11KDHT.

The preliminary studies on protein profile in retained and not retained foetal membranes in heavy draft mares.

Protein profile of the placenta expresses its function and maintenance. Any alterations can be reflected in qualitative and quantitative changes in this profile. The aim of the present study was the evaluation of protein profile in the placenta of mares suffering from the retention of foetal membranes (FMR) by two separation methods and the comparison with physiologically released tissues. Placentas from 14 healthy, heavy draft mares were collected immediately after the expulsion of newborn. Tissues after homogenization and staining with fluorescent dyes were subjected to electrophoretic as well as chromatographic separation. Electrophoretic gels were statistically analysed for the presence and abundance of examined proteins, while some proteins were identified in chromatographic fractions. Out of 248 spots detected in endometrium, 38 differed significantly between FMR and control animals, while in allantochorion, respective values reached 241 and 27 spots (p<.05). Among identified proteins that expressed higher abundance in endometrium of FMR mares than control animals were prostaglandin reductase, dehydrogenase/reductase SDR family, and placental growth factor. These proteins are involved in regulation of parturition. Additionally, the following proteins responsible for physiological activity of a cell-guanine methyl transferase, aspartyl/asparaginyl beta-hydroxylase and GTP-binding protein, were identified. These proteins expressed higher abundance in allantochorion of FMR mares than in controls. This preliminary study confirmed the disturbances in protein pattern between foetal membranes in FMR and healthy mares. Further qualitative and quantitative experiments are necessary to deepen our knowledge on the mechanisms of the retention of foetal membranes in mares.

Dehydrogenase/reductase SDR family member2 silencing sensitizes an oxaliplatin‑resistant cell line to oxaliplatin by inhibiting excision repair cross‑complementing group 1 protein expression.

Oxaliplatin (Oxa)-based chemotherapy is widely used as the first-line treatment for colorectal cancer (CRC). However, Oxa-resistance is common for many postoperative CRC patients. To explore drug resistance in CRC, an Oxa-resistant cell line, HCT116/Oxa, was established from parental HCT116 cells. These Oxa-resistant cells exhibited characteristics of epithelial-mesenchymal transition (EMT) and a higher migratory capacity than parental cells. Protein profiles of HCT116/Oxa and HCT116 cells were compared using a tandem mass tag-based quantitative proteomics technique. The protein dehydrogenase/reductase SDR family member 2 (DHRS2) was revealed to be highly expressed in HCT116/Oxa cells. Silencing of DHRS2 in HCT116/Oxa cells effectively restored Oxa-sensitivity by suppressing the expression of excision repair cross-complementing group 1 protein via a p53-dependent pathway, and reversed the EMT phenotype. Overall, the suppression of DHRS2 expression may be a promising strategy for the prevention of Oxa-resistance in CRC.