Romidepsin (FK228) improves the survival of allogeneic skin grafts through downregulating the production of donor-specific antibody via suppressing the IRE1α-XBP1 pathway / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Antibody-mediated rejection (AMR) is one of the major causes of graft loss after transplantation. Recently, the regulation of B cell differentiation and the prevention of donor-specific antibody (DSA) production have gained increased attention in transplant research. Herein, we established a secondary allogeneic in vivo skin transplant model to study the effects of romidepsin (FK228) on DSA. The survival of grafted skins was monitored daily. The serum levels of DSA and the number of relevant immunocytes in the recipient spleens were evaluated by flow cytometry. Then, we isolated and purified B cells from B6 mouse spleens in vitro by magnetic bead sorting. The B cells were cultured with interleukin-4 (IL-4) and anti-clusters of differentiation 40 (CD40) antibody with or without FK228 treatment. The immunoglobulin G1 (IgG1) and IgM levels in the supernatant were evaluated by enzyme-linked immunosorbent assay (ELISA). Quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and western blotting were conducted to determine the corresponding levels of messenger RNA (mRNA) and protein expression in cultured cells and the recipient spleens. The results showed that FK228 significantly improved the survival of allogeneic skin grafts. Moreover, FK228 inhibited DSA production in the serum along with the suppression of histone deacetylase 1 (HADC1) and HDAC2 and the upregulation of the acetylation of histones H2A and H3. It also inhibited the differentiation of B cells to plasma cells, decreased the transcription of positive regulatory domain-containing 1 (Prdm1) and X-box-binding protein 1 (Xbp1), and decreased the expression of phosphorylated inositol-requiring enzyme 1 α (p-IRE1α), XBP1, and B lymphocyte-induced maturation protein-1 (Blimp-1). In conclusion, FK228 could decrease the production of antibodies by B cells via inhibition of the IRE1α-XBP1 signaling pathway. Thus, FK228 is considered as a promising therapeutic agent for the clinical treatment of AMR.

Identification and Production of Beauvericin by Fusarium subglutinansand F. sacchari from Sugarcane

Abstract Fusarium is producing several important mycotoxins including beauvericin (BEA). Two species of Fusarium viz. F. subglutinans and F. sacchari cause the Pokkahboeng disease of sugarcane. The studies on the occurrence and toxicity of BEA are scarce. Therefore, this study aimed to identify the isolates of Fusarium and detect their ability to produce BEA. The toxicity of BEA was also tested on brine shrimp Artemia salina. Many isolates of Fusarium were isolated from the infected plants of sugarcane in Malaysia. We identified the species of Fusarium according to their morphological characteristics. The capability of Fusarium isolates for producing the BEA was estimated by using a thin layer chromatography. The toxicity bioassay of BEA was conducted on the brine shrimp larvae. The results were identified on F. subglutinans and F. sacchari in 55 isolates of Fusarium. All isolates demonstrated the ability to produce BEA. Interestingly, BEA exhibited variation in toxicity between low toxic to very higher toxicity 100%. F. subglutinans and F. sacchari were able to produce BEA and possibly BEA may be causing toxicity in the host tissue and may be acting as a potential pathogenicity factor. Therefore, we consider BEA as an interesting factor in determining the virulence of fusarium isolate.

Romipeptides A and B, two new romidepsin derivatives isolated from Chromobacterium violaceum No.968 and their antitumor activities in vitro / 中国天然药物

Romipeptides A and B (1 and 2), two new romidepsin derivatives, and three known compounds, chromopeptide A (3), romidepsin (4) and valine-leucine dipeptide (5) were isolated from the fermentation broth of Chromobacterium violaceum No. 968. Their structures were elucidated by interpretation of their UV, HR-ESI-MS and NMR spectra. The absolute configuration of compound 1 and 2 were established by single crystal X-ray diffraction analysis. Compounds 1-5 were evaluated for their anti-proliferative activities against three human cancer cell lines, SW620, HL60, and A549. The results showed most of these compounds exhibited antitumor activities in vitro, in which compound 2 displayed potent cytotoxicity to SW620, HL60 and A549 cell lines, with IC of 12.5, 6.7 and 5.7 nmol·L, respectively.

Differential proteomics on synthetic antimicrobial decapeptide against Streptococcus mutans / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To compare the protein profiles between decapeptide-treated and untreated planktonic cells of Streptococcus mutans (S. mutans) by differential proteomic analysis to determine and identify the key proteins.</p><p><b>METHODS</b>In our previous study, we investigated decapeptide (KKVVFKVKFK-NH2), which was a novel adenosine monophosphate. Compared with other oral pathogens tested, decapeptide had a preferential antibacterial activity against S. mutans. It also inhibited S. mutans biofilm formation and reduced the one-day developed biofilm. In the present study, we first synthesized decapeptide, and then compared the protein profiles between decapeptide-treated and untreated planktonic cells of S. mutans by two-dimensional gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. We also verified different expressions of key protein enolase in the protein level.</p><p><b>RESULTS</b>The results showed that decapeptide altered the protein expression of planktonic S. mutans. These proteins were functionally involved in carbohydrate degradation by glycolysis, protein folding, conjunction, transport, translation, adenosine triphosphate binding, protein binding, sequence-specific DNA binding, transcription factor activity, and two-component response regulator activity. Western blot results showed that enolase protein expression decreased obviously in decapeptide-treated cells of S. mutans.</p><p><b>CONCLUSION</b>The protein expression of S. mutans significantly changed after synthetic antimicrobial decapeptide treatment, suggesting that decapeptide may present a preferential effect on oral caries by changing the expression of certain key proteins, such as enolase protein.</p>

Evaluation of emerging Fusarium mycotoxins beauvericin, Enniatins, Fusaproliferin and Moniliformin in domestic rice in Iran

The occurrence of emerging Fusarium mycotoxins beauvericin [BEA], enniatins [ENNs] [A, A1, B, B1], Fusaproliferin and moniliformin was evaluated by a liquid chromatography/ electrospray ionization-tandem mass spectrometric [LC/ESI-MS/MS] technique in 65 domestic rice samples produced in Gilan and Mazandaran Provinces in Iran. The results showed that 46% of the samples were contaminated with at least one of the emerging mycotoxins. BEA was the most prevalent mycotoxin, which was found in 26 out of 65 rice samples at the concentrations up to 0.47 microg/Kg. Enniatin A1 which was the only member of ENNs was detected in the samples, occurred in 7.7% of samples with an average level of 0.06 microg/Kg. No detectable level of Fusaproliferin and moniliformin was found. This is the first report concerning the contamination of Iranian domestic rice samples with the emerging Fusarium mycotoxins

Construction, expression and enzymatic activity analysis of AUR1 eukaryotic expression vector of Botrytis cinerea / 生物工程学报

In order to study the expression and the activity of inositol phosphorylceramide synthase (BcAUR1 gene) in Botrytis cinerea, we amplified BcAUR1 by RT-PCR from Botrytis cinerea, using the special primers with FLAG and BamH I/Xho I restriction sites. Recombinant pYES2-BcAUR1 was constructed to transform into Saccharomyces cerevisae deltayorl by LiAC. The expression of inositol phosphorylceramide (IPC) synthase and its activity were detected by Western blotting and HPLC, respectively. The results show that pYES2-BcAUR1 could express in uracil mutant deltayorl of Saccharomyces cerevisae. IPC synthase enzyme activity of pYES2-BcAUR1 transformants significantly increased and was approximately double than no-load BcAUR1 transformants. The low concentration of Aureobasidin A could inhibit growth of no-load BcAUR1 transformants, but pYES2-BcAUR1 transformants could resist fungal growth inhibition which was induced by Aureobasidin A.

Inhibitory effect of a novel histone deacetylases inhibitor FK228 on human colon cancer HCT-116 cells in vitro and in vivo / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the inhibitory effects of a novel histone deacetylases inhibitor FK228 on human colon cancer HCT-116 cells in vitro and in vivo, and evaluate its toxicity and side effects.</p><p><b>METHODS</b>The in vitro growth inhibitions of HCT-116 cells by different concentrations of FK228 and 5-Fu for 24, 48 and 72 h were assessed by CCK-8 assay. BALB/c nude mouse models of tumor xenografts were prepared by subcutaneous implantation of tumor tissue, and 4 mg/kg FK228 and 50 mg/kg 5-Fu were i.p. injected, respectively. The inhibitory effects on tumor growth, hematology, and liver and kidney function were evaluated.</p><p><b>RESULTS</b>CCK-8 assay indicated that FK228 had an obvious growth inhibitory effect on HCT-116 cells in a dose- and time-dependent manner. The IC50 of FK228 in HCT-116 cells was 12.05 ng/ml for 48 h, while the IC50 of 5-Fu was 18.92 µg/ml. At 20 days after FK228 and 5-Fu treatment, the tumor volume of the FK228 group was (139.71 ± 44.54)mm(3), significantly lower than that of the 5-Fu group [(282.28 ± 58.81)mm(3)] and that of the model group [(520.65 ± 39.73)mm(3), P < 0.01 for both]. The average tumor weight was (0.07 ± 0.02)g in the FK228 group, significantly lower than that of the 5-Fu group [(0.20 ± 0.08)g, P < 0.01]. The tumor growth inhibition rate of the FK228 group was 73.2%, significantly higher than that of the 5-Fu group (45.8%, P < 0.01). The ALT levels of the FK228 and 5-Fu groups were significantly higher than that of the model group (P < 0.01). The BUN of 5-Fu group was significantly higher than that of the model group (P < 0.01), but the BUN of FK228 group was not significantly different from that of the blank and control groups (P > 0.05 for both). Routine blood test showed that WBC, RBC, Hb and PLT of the 5-Fu group were significantly lower than those of the model group (P < 0.05 for all), but only WBC of the FK228 group was significantly lower than that of the model group (P < 0.05). The pathological examination using HE staining revealed that in the FK228 group, there were fibrosis and inflammatory cell infiltration in the liver tissue, and mild edema of the renal tubules in the kidney. However, in the 5-Fu group there were extensive hepatocyte edema and necrosis in the liver, and evident deformation and necrosis of glomeruli and tubules, and tubular wall thinning in the kidney.</p><p><b>CONCLUSIONS</b>The results of this study indicate that FK228 can more effectively than 5-Fu inhibit the growth of HCT-116 cells in vitro and vivo, and without obvious toxic effect on the kidney and hematology. Its clinical value in colon cancer treatment deserves further investigation.</p>

Progress in the study of some important natural bioactive cyclopeptides / 药学学报

Natural cyclopeptides are hot spots in chemical and pharmaceutical fields because of the wide spreading bio-resources, complex molecular structures and various bioactivities. Bio-producers of cyclopeptides distribute over almost every kingdom from bacteria to plants and animals. Many cyclopeptides contain non-coded amino acids and non-pepditic bonds. Most exciting characteristic of cyclopeptides is a range of interesting bioactivities such as antibiotics gramicidin-S (2), vancomycin (3) and daptomycin (4), immunosuppressive cyclosporin-A (1) and astin-C (8), and anti-tumor aplidine (5), RA-V (6) and RA-VII (7). Compounds 1-4 are being used in clinics; compounds 5-8 are in the stages of clinical trial or as a candidate for drug research. In this review, the progress in chemical and bioactive studies on these important natural bioactive cyclopeptides 1-8 are introduced, mainly including discovery, bioactivity, mechanism, QSAR and synthesis.

Effects of destruxin A on Rhipicephalus (Boophilus) microplus ticks (Acari: Ixodidae) / Efeitos da destruxina A no carrapato Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

This study evaluated the effects of destruxin A on Rhipicephalus (Boophilus) microplus females, since this toxin is one of the likely causes of high mortality induced by the entomopathogenic fungus Metarhizium anisopliae in arthropods. Ticks were immersed or inoculated with different concentrations of destruxin A. Despite the doses applied, there were no deaths or significant alterations in oviposition between the groups treated with destruxin A and the control groups. No other external effect caused by destruxin, such as tetanic paralysis, was observed in these engorged female ticks after the treatment.

Este estudo avaliou os efeitos da destruxina A em fêmeas de Rhipicephalus (Boophilus) microplus, uma vez que essa toxina é uma das prováveis causas da alta mortalidade induzida pelo fungo entomopatogênico Metarhizium anisopliae em artrópodes. Os carrapatos foram imersos ou inoculados com diferentes concentrações de destruxina A. Apesar das doses aplicadas, não houve mortes ou alterações significativas de postura entre os grupos tratados com destruxinas A e os grupos controle. Nenhum outro efeito externo provocado pela destruxina A, tal como paralisia tetânica, foi observado nas fêmeas ingurgitadas de carrapato após o tratamento.

Effect of fluorouracil combined with FK228 on the proliferation, apoptosis and Fas mRNA level in HepG2 hepatoma cell lines / 中南大学学报(医学版)

OBJECTIVE@#To explore the effect of fluorouracil (FU) combined with epigenetic drug FK228 (depsipeptide FR901228) on the proliferation, apoptosis and Fas mRNA level in HepG2 hepatoma cell lines.@*METHODS@#There were 4 groups in this experiment: an untreated group, an FK228 treated group, a FU treated group,and a FU combined with FK228 group.Tetrazolium salt colorimetry (MTT) assay was used to assess the cell inhibition rate. Q value of Kingos formula and multi-factor analysis of variance (ANOVA ) were used to judge the combination treatment effect,and flow cytometry was used to detect the apoptosis rate. Fas mRNA level was analyzed by RT-PCR.@*RESULTS@#FK228 or FU would inhibit the growth of HepG2 cells in a concentration-dependent and time-dependent manner. Cell inhibition rates of HepG2 were significantly enhanced in the FU combined with FK228 group, compared with that in the FU treated group alone (P<0.05). Both Q values were more than 1, the 2 drug combinations showed interaction,and FU combined with FK228 had synergistic effect.Compared with the FK228 treated group and the FU treated group, apoptosis rate of HepG2 cells was significantly increased (P<0.05), and the Fas mRNA level was up-regulated in HepG2 cells in FU combined FK228 group (P<0.05).@*CONCLUSION@#Combination of FK228 and FU can enhance the proliferation inhibition and apoptosis induction of FU in hepatoma cell lines, up-regulate the Fas mRNA level, and increase the sensitivity of hepatoma cell lines to FU.

A Case of Emetic Toxin Producing Bacillus cereusStrains Isolated from Outbreak / 대한임상미생물학회지

Bacillus cereus causes two types of gastrointestinal diseases: emesis and diarrhea. It produces one emetic toxin and nine different enterotoxins. In March 2008, eight of a family became sick after eating slices of raw fish. We isolated emetic toxin producing B. cereus from the stools of 6 patients and 2 subclincal humans. In this study, the presence of enterotoxin genes, such as those of haemolysin BL (Hbl), nonhemolytic enterotoxin (Nhe), B. cereus enterotoxin T (BceT), enterotoxin FM (EntFM), cytotoxin K (cytK) and cereulide were assayed by polymerase chain reaction (PCR) methods. Their enterotoxin activities were assayed using the BCET- RPLA, Tecra ELISA kit and Hep-2 vacuole activity. Bacterial isolates were subtyped by pulsed-field gel electrophoresis (PFGE). This study demonstrates the emetic toxin-producing stains of B. cereus in clinical specimens, for the first time in the Republic of Korea.

Advance of several types of important marine antitumor drugs / 药学学报

Marine antitumor drugs have been the research focus in the world. Recently, advancement has been made in the investigation of six types of compounds including bryostatin-1, ecteinascidin-743, dolastatin, didemnin B, psammaplin and halichondrin B. In this review, we summarized the recent research progress of the above mentioned marine antitumor drugs and their derivatives. Also, the development tendency of marine antitumor drugs was discussed.

Effect of alternaria pathotoxin(s) on expression of p53-like apoptotic protein in calli and leaves of Brassica campestris.

Possible involvement of apoptosis was investigated in pathotoxin-treated and nutritionally-depleted in vitro cultured calli by comparing levels of p53-like protein. Antibodies raised against human p53 were used to detect and quantify p53 in B. campestris. Expression of p53-like protein increased from proliferating to static growth stage and reached to constant level at decaying stage. Both ELISA and dot immuno-binding assay showed that p53-like protein was over expressed in toxin treated and nutritionally depleted calli. Almost similar changes were seen in senescent damage in Brassica species indicating involvement of p53 dependent pathways.