Progress in protein extraction methods for fungal identification with MALDI-TOF MS / 中华微生物学和免疫学杂志

With the development of techniques for rapid microbial identification, MALDI-TOF MS has become an important tool for clinical identification of fungi. Problems such as the applicability and standardization of protein extraction methods have hindered the development of MALDI-TOF MS technology in the fungal field. This paper analyzed the complex structure of fungal cell walls, introduced the protein extraction methods recommended by MALDI-TOF MS commercial mass spectrometry systems, discussed the protein extraction methods for the identification of various genera of yeast-like fungi and filamentous fungi by MALDI-TOF MS, such as direct smear method, formic acid acetonitrile extraction method and magnetic bead grinding method, and summarized the current status and drawbacks of protein extraction methods in fungal identification by MALDI-TOF MS with a view to providing theoretical reference for subsequent research.

Exosomes derived from Nr-CWS pretreated MSCs facilitate diabetic wound healing by promoting angiogenesis via the circIARS1/miR-4782-5p/VEGFA axis / 中国天然药物

Mesenchymal stem cell (MSC)-derived exosomes (Exos) were reported to a prospective candidate in accelerating diabetic wound healing due to their pro-angiogenic effect. MSCs pretreated with chemistry or biology factors were reported to advance the biological activities of MSC-derived exosomes. Hence, this study was designed to explore whether exosomes derived from human umbilical cord MSCs (hucMSCs) preconditioned with Nocardia rubra cell wall skeleton (Nr-CWS) exhibited superior proangiogenic effect on diabetic wound repair and its underlying molecular mechanisms. The results showed that Nr-CWS-Exos facilitated the proliferation, migration and tube formation of endothelial cells in vitro. In vivo, Nr-CWS-Exos exerted great effect on advancing wound healing by facilitating the angiogenesis of wound tissues compared with Exos. Furthermore, the expression of circIARS1 increased after HUVECs were treated with Nr-CWS-Exos. CircIARS1 promoted the pro-angiogenic effects of Nr-CWS-Exos on endothelial cellsvia the miR-4782-5p/VEGFA axis. Taken together, those data reveal that exosomes derived from Nr-CWS-pretreated MSCs might serve as an underlying strategy for diabetic wound treatment through advancing the biological function of endothelial cells via the circIARS1/miR-4782-5p/VEGFA axis.

Chemical Composition of Palm Oil Mill Sludge-Biodegraded Sweet Orange Peel Meal Mixture and Its Effect as Dietary Energy Source on Nutrient Digestibility by Broiler Chickens

A feeding trial, which lasted for seventy days, was conducted in which palm oil mill sludge and biodegraded sweet orange peel mixture was fed to substitute maize in broiler chicken diet at 0%, 5%, 10%, 15%, 20% and 25%. Sweet orange (Citrus sinensis) fruit peel was fermented by soaking for 48 h in retted cassava waste water (CWW) and sundried, to obtain biodegraded sweet orange peel (BSOP). Palm oil mill effluent was filtered with a 0.30 mm pore plastic mesh sieve, poured into a 0.75 ?m pore fine cheesecloth bag and allowed to stand for five hours to produce a paste of palm oil mill sludge (POMS). The POMS was mixed with BSOP in ratio 1:1, sundried, milled to produce a POMS-BSOP mixture. One hundred and eighty day-old Cobb 700 broilers divided into six equal parts, and three replicates of 10 birds each were used. Each part was assigned to one of 6 diets compounded with 0% (T1), 5% (T2), 10% (T3), 15% (T4), 20% (T5) and 25% (T6) of POMS-BSOP mixture. The microbial composition of retted CWW, chemical composition of the POMS-BSOP mixture, and the digestibility of nutrients by the broiler chickens were determined. Isolated from CWW were; Staphylococcus aureus, Streptococcus spp., Salmonella spp., Escherichia coli (bacteria), Aspergillus spp. (fungus) and Candidia spp. (yeast). POMS-BSOP was high in energy (4415.69 kcalME/kg), ether extract (41.50%), crude fibre (25.63%) and dry matter (92.28%), moderate in crude protein (6.83%), low in indigestible lignin (4.90% ADL), alkaloid (0.01%), tannin (0.02%), saponin (0.03%), phytate (0.05%), oxalate (0.15%) and flavonoid (0.17%). Dietary treatments significantly (P<0.05) affected digestibility of ether extract and metabolisable energy and crude protein digestibility by broiler chickens. Dietary maize can be replaced at up to 25% with a POMS-BSOP mixture to improve energy digestibility by broiler chickens.

Systems biology applied to the study of papaya fruit ripening: the influence of ethylene / Biologia de sistemas aplicada ao estudo do amadurecimento de mamões: a influência do etileno

Fruit ripening is a biochemical process that results in flavor, odor, texture, and color suitable for human consumption, in addition to providing access to important nutrients. Although ripening promotes sensory and nutritional increases in fruits, there is also an increased susceptibility to physical damage, as is the case with papaya. These transformations occur due to changes in gene expression patterns at different stages of maturity, whose control and coordination result from the combined action of plant hormones, especially ethylene. As the action of this hormone in the regulation of gene expression is still elusive, this dissertation sought to address the global analysis of the transcriptome in an overview study of molecular processes involved in the ripening of ethylene-treated and non-treated papaya. Transcription factors related to ethylene synthesis and signaling had increased activity towards exogenous-ethylene treatment. Consequently, ethylene-induced enzymes had their coding genes differentially expressed, like genes related to the synthesis of carotenoids, linalool, and vitamins, which increase color, aroma, and antioxidant activity, respectively. Metabolic pathways related to the synthesis of sugars were suppressed while genes encoding the enzyme responsible for sucrose synthesis maintained a basal expression, showing that the accumulation of sugars occurs before the ripening process. The firmness of the peel and pulp of the fruits were strongly influenced by the treatment with ethylene and by the time of the experiment, suffering the action of numerous enzymes related to the degradation of the cell wall. The main enzyme responsible for softening the pulp was polygalacturonase, together with the activity of other pectinases and cellulases. In contrast to the need for the pre-climacteric action of pectate lyase and pectinesterase reported in other fleshy fruits, such as tomatoes and strawberries, papaya did not show a significant difference in their expression. The meta-analysis of several papaya ripening transcriptomes confirmed the expression profile observed in the previous RNA-seq, besides providing statistical enrichment to the biological narratives. Finally, the present study gathered a range of robust information on the gene regulation of the papaya ripening process, which opens possibilities for future approaches to transcriptomic analysis and validates the use of papaya as a model for such studies

O amadurecimento de frutos é um processo bioquímico que resulta em sabor, odor, textura e cor adequados para o consumo humano, além de propiciar o acesso a nutrientes importantes. Apesar do amadurecimento promover incrementos sensoriais e nutricionais nos frutos, ocorre também um aumento da suscetibilidade a danos físicos, como é o caso do mamão. Essas transformações ocorrem devido às alterações nos padrões de expressão gênica nos diferentes estádios de amadurecimento, cujo controle e coordenação decorrem da ação combinada de hormônios vegetais, principalmente do etileno. Como a ação deste hormônio na regulação da expressão gênica ainda é elusiva, a presente dissertação buscou abordar a análise global do transcriptoma em um amplo estudo dos processos moleculares envolvidos no amadurecimento de mamões tratados e não tratados com etileno. Os fatores de transcrição relacionados com a síntese e a sinalização do etileno tiveram sua atividade aumentada perante o tratamento exógeno com etileno. Consequentemente, as enzimas reguladas por esse hormônio tiveram seus genes de codificação expressos diferencialmente, como foi o caso de genes relacionados à síntese de carotenoides, linalool e vitaminas, que atuam no aumento da cor, aroma e atividade antioxidante, respectivamente. Vias metabólicas relacionadas com à síntese de açúcares foram reprimidas enquanto genes codificantes da enzima responsável pela síntese de sacarose mantiveram uma expressão basal, evidenciando que o acúmulo de açúcares ocorre antes do processo de amadurecimento. A firmeza da casca e da polpa dos frutos foram fortemente influenciadas pelo tratamento com etileno e pelo tempo de experimento, sofrendo ação de inúmeras enzimas relacionadas com a degradação da parede celular. A principal enzima responsável pelo amolecimento da polpa foi a poligalacturonase, em conjunto com a atividade de outras pectinases e celulases. Em contraste com a necessidade da ação pré-climatérica da pectato liase e da pectinesterase relatada em outras frutas carnosas, como tomates e morangos, o mamão não apresentou uma diferença significativa na expressão das mesmas. A meta-análise de diversos transcriptomas do amadurecimento do mamão reafirmaram o perfil de expressão observado no RNA-seq, além de prover enriquecimento estatístico às narrativas biológicas. Por fim, o presente estudo reuniu uma gama de informações robustas sobre a regulação gênica do processo de amadurecimento do mamão papaia, o que abrange a possibilidade para futuras abordagens de análise transcriptomica e valida o uso do mamão como modelo para tais estudos

Efficacy and safety of Nocardia rubra cell wall skeleton for the treatment of erosive oral lichen planus / 北京大学学报(医学版)

OBJECTIVE@#To observe the efficacy and safety of Nocardia rubra cell wall skeleton (Nr-CWS) for the treatment of erosive oral lichen planus (EOLP).@*METHODS@#Sixty patients with clinically and pathologically diagnosed EOLP were randomly divided into the experimental group and control group according to the random number. Patients in the experimental group were treated with lyophilized powder containing Nr-CWS combined with normal saline. Patients in the control group received topical placebo without Nr-CWS combined with normal saline. Changes in the EOLP lesion area and the patient's pain level were recorded at the timepoints of weeks 1, 2, and 4 after the two different treatments, respectively. The changes of the patient's REU scoring system (reticulation, erythema, ulceration), the visual analogue scale and the oral health impact score (OHIP-14) were compared between the experimental group and control group after treatment, and the safety indicators of the two groups at the initial diagnosis and after 4 weeks' treatment were also observed, respectively.@*RESULTS@#Totally, 62 patients with clinically and pathologically diagnosed EOLP were enrolled, 2 of whom were lost to the follow-up, with 31 in the experimental group, and 29 in the control group. The mean age of the experimental group and control group were (52.9±12.4) years and (54.07±12.40) years, respectively. There was no significant difference in the oral periodontal index between the experimental group and control group. In the experimental group, the erosive area of oral lichen planus was significantly reduced 1, 2, and 4 weeks after the Nr-CWS's treatment (P < 0.05), the reduction rate was 81.75%, the patient's pain index was also decreased (P < 0.05), and in addition, the OHIP-14 was reduced (P < 0.05). The changes of the REU scoring system, the visual analogue scale and the OHIP-14 were significantly different between the experimental group and control group after treatment. There was no significant difference in the safety index between the two groups.@*CONCLUSION@#The priliminary data show that the Nr-CWS is effective and safe to treat EOLP.

Effect of berberine hydrochloride on cell wall integrity of Candida albicans hypha / 中国中药杂志

The aim of this paper was to investigate the effect of berberine hydrochloride on the cell wall integrity of Candida albicans hypha. The minimal inhibitory concentration(MIC) of berberine hydrochloride against clinical and standard C. albicans strains was detected by micro liquid-based dilution method; the effect of berberine hydrochloride on the colony formation of C. albicans SC5314 was investigated by spot assay; the effect of berberine hydrochloride on the metabolism of C. albicans SC5314 hypha was checked by XTT reduction assay, and the viability of C. albicans SC5314 hypha was tested by fluorescent staining assay. The effect of berberine hydrochloride on the morphology of C. albicans SC5314 hypha was examined by scanning electron microscope. The changes in the cell wall of C. albicans SC5314 hypha after berberine hydrochloride treatment were detected by transmission electron microscopy. The effect of berberine hydrochloride on β-glucan from C. albicans SC5314 was detected by flow cytometry. The effect of berberine hydrochloride on hypha-specific gene ECE1 and β-glucan synthase genes FKS1 and FKS2 in C. albicans was examined by qRT-PCR. The results showed that berberine hydrochloride showed a strong inhibitory effect on both clinical and standard strains of C. albicans, and the MIC was 64-128 μg·mL~(-1). Spot assay, XTT redunction assay and fluorescent staining assay showed that with the increase of berberine hydrochloride concentration, the viability of C. albicans SC5314 gradually decreased. The transmission electron microscopy scanning assay showed that this compound could cause cell wall damage of C. albicans. The flow cytometry analysis showed the exposure degree of C. albicans β-glucan. The qRT-PCR further showed that berberine hydrochloride could significantly down-regulate hypha-specific gene ECE1 and β-glucan synthase-related gene FKS1 and FKS2. In conclusion, this compound can down-regulate C. albicans and β-glucan synthase-related gene expressions, so as to destroy the cell wall structure of C. albicans, expose β-glucan and damage the integrity of the wall.

An efficient algae cell wall disruption methodology for recovery of intact chloroplasts from microalgae

The microalgal cell wall breakage has been identified as complex phenomenon which is highly dependent onthe nature and composition of cell wall. A detailed analysis of plastids and their function requires the breakingopen of cell without any damage to cellular components. To develop a rapid and universal methodology forcell wall breakage, liquid nitrogen crushing, sonication, enzymatic lysis, and homogenization procedures wereapplied to various microalgal species. Homogenization-based procedure for the isolation of intact chloroplastwas found to be universal for all algal species under the study. The isolated chloroplasts were subjected tochloroplast integrity analysis. The intact chloroplast exhibited a positive maximum quantum yield and Fv/Fm values ranging from 0.1 to 0.4 as measured by pulse amplitude modulation fluorometry and was found tobe suitable for further downstream applications such as isolation of protein–pigment complexes involved inphotosynthetic O2 evolution. The developed methodology is a quick and efficient technique for the isolation ofintact chloroplasts across different genera of microalgae by employing minor changes in the base protocol as aspecies-specific characteristic

Nutritive value and anatomical characterization from Pennisetum purpureum genotypes

The research submitted samples from stems and leaf blades from tree genotypes of Pennisetum purpureum called 93-32-02, 92-70-02, and 91-06-02 (EMBRAPA - Dairy cattle) and elephant grass cv. Napier (reference cultivar) to the chemical, anatomical evaluations, and in vitro dry matter digestibility (IVDMD) measurement. The anatomical characteristics of the stems and leaf blades, the chemical composition, and the IVDMD of these genotypes at 70 days of re-growth were correlated. Concerning IVDMD, the data highlighted differences, and the cultivar Napier presented the smallest value. Digital images obtained by light microscopy from cross-section reveal that all the stem and leaf blade have similar structural organization. Quantitative differences were verified mainly in the stem. The leaves displayed differences only in the mesophyll thickness. The genotypes showed higher potential in the rainy season since they had the largest IVDMD when compared to the cultivar Napier.

Identification of chemical components and monosaccharide assay in Nocardia rubra cell wall skeleton / 药学实践杂志

Objective To analyze and identify the chemical components in the Nocardia rubra cell wall skeleton (Nr-CWS), and to determine the contents of monosaccharides accurately. Methods The extract of Nr-CWS was separated and analyzed by UHPLC-Q-TOF/MS method. The chemical components were quickly identified by matching the data with the information in the Metlin database. The monosaccharide contents in the Nr-CWS extract were determined by UHPLC-MS/MS method after derivatization. Results A total of 64 chemical components were identified in the extract of Nr-CWS, including amino acids, monosaccharides and so on. A assay method for 8 monosaccharides by UHPLC-MS/MS was successfully established. The content of arabinose in Nr-CWS was the highest, followed by galactose, which indicated that the main polysaccharide components in Nr-CWS may be composed of these monosaccharides. Conclusion In this study, we analyzed the main chemical components of Nr-CWS, which are amino acids, fatty acids and so on. The content of monosaccharide after polysaccharide hydrolysis was determined by UHPLC-MS/MS. This will lay a foundation for the screening of the active components of Nr-CWS and the study of its pharmacological mechanism.

Cryo-EM snapshots of mycobacterial arabinosyltransferase complex EmbB-AcpM

Inhibition of Mycobacterium tuberculosis (Mtb) cell wall assembly is an established strategy for anti-TB chemotherapy. Arabinosyltransferase EmbB, which catalyzes the transfer of arabinose from the donor decaprenyl-phosphate-arabinose (DPA) to its arabinosyl acceptor is an essential enzyme for Mtb cell wall synthesis. Analysis of drug resistance mutations suggests that EmbB is the main target of the front-line anti-TB drug, ethambutol. Herein, we report the cryo-EM structures of Mycobacterium smegmatis EmbB in its "resting state" and DPA-bound "active state". EmbB is a fifteen-transmembrane-spanning protein, assembled as a dimer. Each protomer has an associated acyl-carrier-protein (AcpM) on their cytoplasmic surface. Conformational changes upon DPA binding indicate an asymmetric movement within the EmbB dimer during catalysis. Functional studies have identified critical residues in substrate recognition and catalysis, and demonstrated that ethambutol inhibits transferase activity of EmbB by competing with DPA. The structures represent the first step directed towards a rational approach for anti-TB drug discovery.

Cryo-EM snapshots of mycobacterial arabinosyltransferase complex EmbB-AcpM

Inhibition of Mycobacterium tuberculosis (Mtb) cell wall assembly is an established strategy for anti-TB chemotherapy. Arabinosyltransferase EmbB, which catalyzes the transfer of arabinose from the donor decaprenyl-phosphate-arabinose (DPA) to its arabinosyl acceptor is an essential enzyme for Mtb cell wall synthesis. Analysis of drug resistance mutations suggests that EmbB is the main target of the front-line anti-TB drug, ethambutol. Herein, we report the cryo-EM structures of Mycobacterium smegmatis EmbB in its "resting state" and DPA-bound "active state". EmbB is a fifteen-transmembrane-spanning protein, assembled as a dimer. Each protomer has an associated acyl-carrier-protein (AcpM) on their cytoplasmic surface. Conformational changes upon DPA binding indicate an asymmetric movement within the EmbB dimer during catalysis. Functional studies have identified critical residues in substrate recognition and catalysis, and demonstrated that ethambutol inhibits transferase activity of EmbB by competing with DPA. The structures represent the first step directed towards a rational approach for anti-TB drug discovery.

Resistance of bioenergy sorghum to Diatraea saccharalis (Lepidoptera: Crambidae) / Resistência de sorgo bioenergia à diatraea saccharalis (lepidoptera: crambidae)

This study evaluated the effects of the sugarcane borer Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) on cultivars of sweet and biomass sorghum for the selection of resistant cultivars. The present work consisted of two trials, with natural pest infestation. In the first one, 10 sweet sorghum cultivars were analyzed for the following variables: plant height, number of healthy and damaged internodes, gallery position and size, stem infestation level and soluble solids content (°Brix). In the second trial, it was analyzed 16 genotypes of high biomass sorghum, with the same variables above mentioned, in addition to the lignin, cellulose and hemicellulose contents. Among sweet sorghum genotypes evaluated, the genotype CMSXS647 stood out due to the traits: plant height, infestation level, gallery size and soluble solids content. Among the sorghum genotypes evaluated, CMSXS7030, CMSXS7012 and CMSXS7028 presented ideal characteristics for infestation level, plant height and number of lignocellulosic compounds. Such information, in addition to supporting the bioenergy sorghum breeding program, will assist in integrated pest management for sorghum cultivation.

Foram estudados os efeitos causados pela broca-do-colmo Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae), em cultivares de sorgo sacarino e biomassa visando seleção de cultivares resistentes à praga. O presente trabalho foi constituído de dois ensaios, com infestação natural da praga. No primeiro, 10 cultivares de sorgo sacarino foram analisadas quanto às seguintes variáveis: altura das plantas, quantidade de internódios sadios e com injúrias, posição e tamanho da galeria, intensidade de infestação de colmos e teor de sólidos solúveis (°Brix). No segundo ensaio, foram analisados 16 genótipos de sorgo biomassa, com as mesmas variáveis supracitadas, além dos teores de lignina, celulose e hemicelulose. Entre os genótipos de sorgo sacarino avaliados, o genótipo CMSXS647 foi o que se destacou em função das características: altura de plantas, intensidade de infestação, tamanho de galerias e teor de sólidos solúveis. Entre os genótipos de sorgo biomassa avaliados: CMSXS7030, CMSXS7012 e CMSXS7028 apresentaram características ideais para intensidade de infestação, altura de plantas e quantidade de compostos lignocelulósico. Tais informações, além de prover o programa de melhoramento de sorgo energia podem ajudar o programa de MIP para a cultura do sorgo, uma vez que o produtor conhece a suscetibilidade dos materiais escolhidos.

Microalgae protoplasts isolation and fusion for biotechnology research / Aislamiento y fusión de protoplastos de microalgas para aplicaciones biotecnológicas

ABSTRACT Protoplasts are microbial or vegetable cells lacking a cell wall. These can be obtained from microalgae by an enzymatic hydrolysis process in the presence of an osmotic stabilizer. In general, protoplasts are experimentally useful in physiological, genetic and biochemical studies, so their acquisition and fusion will continue to be an active research area in modern biotechnology. The fusion of protoplasts in microalgae constitutes a tool for strain improvement because it allows both intra and interspecific genetic recombination, resulting in organisms with new or improved characteristics of industrial interest. In this review we briefly describe the methodology for obtaining protoplasts, as well as fusion methods and the main applications of microalgal platforms.

RESUMEN Los protoplastos son células microbianas o vegetales que carecen de pared celular. Estos pueden obtenerse a partir de microalgas por un proceso de hidrólisis enzimática en presencia de un estabilizador osmótico. En general, los protoplastos son experimentalmente útiles en estudios fisiológicos, genéticos y bioquímicos, por lo que su obtención y fusión continuarán siendo un área de investigación activa en la biotecnología moderna. La fusión de protoplastos en microalgas constituye una herramienta para el mejoramiento de cepas pues permite la recombinación genética intra e interespecífica, logrando así organismos con nuevas características de interés industrial. En esta revisión, describimos brevemente la metodología para obtener protoplastos, métodos de fusión y las principales aplicaciones de las plataformas basadas en microalgas.

In Silico Characterization of Meloidogyne Genus Nematode Cellulose Binding Proteins

Abstract Root-knot nematodes are a group of endoparasites species that induce the formation of giant cells in the hosts, by which they guarantee their feeding and development. Meloidogyne species infect over 2000 plant species, and are highly destructive, causing damage to many crops around the world. M. enterolobii is considered the most aggressive species in tropical regions, such as Africa and South America. Phytonematodes are able to penetrate and migrate within plant tissues, establishing a sophisticated interaction with their hosts through parasitism factors, which include a series of cell wall degradation enzymes and plant cell modification. Among the parasitism factors documented in the M. enterolobii species, cellulose binding protein (CBP), a nematode excretion protein that appears to be associated with the breakdown of cellulose present in the plant cell wall. In silico analysis can be of great importance for the identification, structural and functional characterization of genomic sequences, besides making possible the prediction of structures and functions of proteins. The present work characterized 12 sequences of the CBP protein of nematodes of the genus Meloidogyne present in genomic databases. The results showed that all CBP sequences had signal peptide and that, after their removal, they had an isoelectric point that characterized them as unstable in an acid medium. The values of the average hydrophilicity demonstrated the hydrophilic character of the analyzed sequences. Phylogenetic analyzes were also consistent with the taxonomic classification of the nematode species of this study. Five motifs were identified, which are present in all sequences analyzed. These results may provide theoretical grounds for future studies of plant resistance to nematode infection.

Β-Glucan of candida albicans cell wall extract inhibits salmonella typhimurium colonization by potentiating cellular immunity (cd8 + and cd4 + t cells)

Abstract INTRODUCTION: Antimicrobial resistance has been reported in the drugs used for the treatment of typhoid fever. The immunomodulatory substance β-glucan can be used as an alternative therapy as it potentiates host immunity. The aims of this study are to observe the effect of Candida albicans cell wall (CCW) extract towards host immunity (TCD8+ and TCD4+ cells in spleen, intestinal sIgA) and its capacity to kill Salmonella in the intestine and liver of typhoid fever mice models. METHODS: Typhoid fever mice models were created by infecting mice with S. Typhimurium orally. Mice were divided into four groups: the Non-Infected, Infected, CCW (infected mice treated with 300 µg CCW extract/mouse once a day), and Ciprofloxacin groups (infected mice treated with 15 mg/kg BW ciprofloxacin twice a day). RESULTS: Secretory IgA (sIgA) concentrations of mice in the CCW group remained unchanged. However, their TCD4+ and TCD8+ cells increased substantially compared to those in the Non-Infected group. In the Ciprofloxacin group, sIgA concentrations increased markedly compared to those in the Non-Infected and CCW groups; TCD4+ and TCD8+ cells also increased significantly compared to those in the Infected Group, but not significant compared to those in the CCW group. Colonization of S. Typhimurium in the intestine and liver decreased significantly in the CCW and Ciprofloxacin groups compared to that in the Infected group, with the lowest reduction being found in the Ciprofloxacin group. CONCLUSIONS The inhibition of S. Typhimurium colonization by CCW is associated with the increase in TCD4+ and TCD8+ cells.

Inhibitory effect of Corynebacterium parvum cell wall on bladder cancer cells / 中国医师进修杂志

Objective To test the inhibitory effect of Corynebacterium cell wall extract on bladder cancer cells. Methods The bladder RNA was extracted from bladder cancer rats, and concentration and purity of RNA was detected. The extracted RNA was reversely transcribed into cDNA, then the primers were designed and the Grim19 gene, β-actin gene, Stat3 gene was amplified. Finaly the PCR product was subjected to agarose gel electrophoresis. Results OD260 was 0.07, and OD260/OD280 was 2.03 in the group of Corynebacterium parvm extract(3-5# ); OD260 was 0.12, and OD260/OD280 was 2.07 in the group of MNU( 6-5#);OD260 was 0.08, and OD260/OD280 was 2.07 in the group of physiological saline(7-5#) . The results of agarose gelelectrophoresis showed that Grim19 gene and Stat3 gene was expressed highly in the bladder of rats from the group of CP cell wall extract (3-5# ). Grim19 gene was expressed lowly, while Stat3 gene was expressed highly in the bladder of rats from the group of MNU (6-5#). Grim19 gene and Stat3 gene was expressed normally in the bladder of rats from physiological saline(7-5). Conclusions The expression situation of antitumor gene Grim19 and tumor gene Stat3 in the bladder of rats was inhibited by the cell wall extract of Corynebacterium parvm, which indicates that the cell wall extract of Corynebacterium parvm has inhibitory effect on bladder cancer cells.

Regulations of RLM1 gene affect the anti-autolytic ability of lager yeast / 生物工程学报

The autolysis of brewer's yeast seriously affects the quality of beer and the quality of yeast is considered as one of the key factors in beer brewing. Previous studies on brewer's yeast autolysis showed that RLM1 gene, an important transcription factor in cell integrity pathway, is closely related to the autolysis of yeast. In this study, RLM1 was knocked out and overexpressed in a haploid brewer's yeast. RLM1 disruption resulted in poor anti-autolysis performance of yeast, whereas overexpression of RLM1 contributed to the anti-autolytic ability of yeast. In addition, RLM1 gene knockout affected the osmotic stress resistance, cell wall damage resistance, nitrogen starvation resistance and temperature tolerance of yeast strain. The transcriptional level of GAS1 involved in cell wall assembly and DNA damage response was regulated along with the expression of RLM1, whereas other genes in CWI pathway did not show apparent regularity. RLM1 might mainly affect the expression of GAS1 so as to improve the stress resistance of lager yeast in harsh environment. The result from this study help further understand the mechanism of yeast autolysis and lay a foundation for breeding brewer's yeast strain with better anti-autolytic ability.

Progress in brewer's yeast cell wall stress response / 生物工程学报

Yeast cell wall plays an important role in the establishment and maintenance of cell morphology upon the cell wall stress. The cell wall of yeast consists of β-glucans, mannoproteins and chitin. The composition and structure remodel due to cell wall stress. Brewer's yeast cell wall exhibits stress response during long-term acclimation in order to adapt to environmental changes. This paper reviews the composition and structure of yeast cell wall and the molecular mechanisms of cell wall remodeling and signal pathway regulation.

Bacillus Calmette-Guérin (BCG)-Cell Wall Skeleton as Immunotherapeutic Option for BCG-Refractory Superficial Bladder Cancer

Although intravesical instillation of Mycobacterium bovis bacillus Calmette-Guérin (BCG) is the most successful cancer immunotherapy for superficial bladder cancer, the serious side effects are frequently arisen by using live mycobacteria. To allow less toxic and more potent immunotherapeutic agents following intravesical BCG treatment for superficial bladder cancer, noninfectious immunotherapeutic drug instead of live BCG would be highly desirable. Recently, immune-enhancing adjuvants are considered an effective vaccine immunotherapy for cancer, providing enhanced antitumor effects and boosted immunity. The BCG-cell wall skeleton (BCG-CWS), the main immune active center of BCG, is a potent candidate as a noninfectious immunotherapeutic drug instead of live BCG against bladder cancer. However, the most limited application for anticancer therapy, it is difficult to formulate a water-soluble BCG-CWS due to the aggregation of BCG-CWS in both aqueous and nonaqueous solvents. To overcome the insolubility and improve the internalization of BCG-CWS into bladder cancer cells, it should be developed the lipid nanoparticulation of BCG-CWS, resulting in improved dispensability, stability, and small size. In addition, powerful technology of delivery systems should be applied to enhance the internalization of BCG-CWS, such as encapsulated into lipid nanoparticles using novel packaging methods. Here, we describe the progress in research on effects of BCG-CWS for cancer immunotherapy, development of lipid-based solvent, and packaging method using nanoparticles with drug delivery system.

In vivo investigation of antimicrobial effects of cinnamaldehyde using immunosuppressed ICR mice with invasive pulmonary aspergillosis / 中草药·英文版

Objective: Invasive pulmonary Aspergillus infection has the characteristics of high morbidity, difficult to be treated, poor prognosis and high mortality. This study aims to investigate the effects of cinnamaldehyde on 1,3-β-D-glucans in the pulmonary Aspergillus fumigatus cell wall to provide a basis for developing novel antifungal drugs. Methods: Immunosuppressed ICR mice were intranasally inoculated with 50 µL of A. fumigatus suspension (1 × 107 CFU/mL) and then separated into two groups, for the experimental group cinnamaldehyde was orally administered at 240 mg/kg/d consecutively for 14 d. While for the control group, voriconazole was used to treat the fungus infection. Pulmonary tissues were then extracted for 1,3-β-D-glucans assay and electron microscopy. Results: The concentration of 1,3-β-D-glucans was significantly different between the cinnamaldehyde and voriconazole groups, which was (1160.89 ± 364.96) pg/mL and (3885.94 ± 845.45) pg/mL, respectively (P < 0.01). Electron microscopy showed that 2−3 outer layers (1,3-β-D-glucan layer) of A. fumigatus cell wall were damaged and fell off, resulting in serious defect of the cell wall, but the cell membrane was clear and intact. Conclusion: Cinnamaldehyde has a significant influence on the integrity of 1,3-β-D-glucans in the pulmonary A. fumigatus cell wall, but the cell membrane is unaffected, suggesting that cinnamaldehyde has unique antifungal properties depending on its action against the 1,3-β-D-glucans on the pulmonary A. fumigatus cell wall.