Neutrophils play a crucial role in the pathogenesis of joint inflammatory diseases such as rheumatoid arthritis (RA). Therefore their elimination and/or a functional inhibition might have beneficial or even therapeutic effects in these diseases. In the present study we exploited the cytotoxic action of etoposide to deplete neutrophils. We administrated the drug twice (at day -3 and day -1) to SCID mice having intact innate immunity and a fail in T- and B-cell maturation. Ankle inflammation was induced by the injection of zymosan (ZY). Joint damage was evaluated by histology grading system for cell infiltration and proteoglycan loss and degree of cartilage erosion. The frequencies of mature Ly6G+CD11b+ cells in bone marrow (BM) were monitored at days -4, -2 and 0 by flow cytometry. At day 7 of ankle inflammation the amounts of pro-inflammatory cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-17 were measured by enzyme-linked immunosorbent assay (ELISA). Histological analysis of the joint sections showed decreased scores for cell infiltration and cartilage proteoglycan loss and reduced cartilage erosion in drug-treated zymosan injected mice in comparison to untreated group with ankle inflammation. Etoposide diminished cell numbers in BM, inhibits granulopoiesis triggered by zymosan and decreased the frequencies of mature Ly6G+CD11b+ cells in BM and eliminated Ly6G+ cells from blood and synovial fluid. We observed reduced TNF-α and impaired IL-17 production in etoposide-treated ZY group. Our data provide a proof-of principle that the elimination of neutrophils might be exploited in a design of new therapeutic approaches for joint inflammatory diseases.

The combination of Raman and Infrared spectroscopic signatures were used to find the different vibrational modes of individual carotenoid as their spectral fingerprint. Both have been previously demonstrated to be highly useful methodology for the identification and/or typing of microorganisms. In this study, we set out to evaluate whether these technologies could be applied to detect the presence of carotenoids in psychrotrophic bacterial isolates. FTIR and Raman spectra of four psychrotrophic bacteria viz. Kocuria rosea, K. turfanensis, Sanguibacter suarezii and Planococcus maritimus were examined during the investigation. FTIR spectra bands at 1653-1661cm-1 in different samples were assigned as part of chlorophyll, 1424-1426 cm-1 as -C-H- (CH2) bending vibration from methylene of carotenoids or lycopene, 1366-1367 cm-1 band as the b-ionone ring of β-carotene due to the C-H, (–CH3) symmetrical bending. Interestingly, Raman spectra revealed intense Raman bands in the range of 1511-1530, 1153-1159 and 1003-1010 cm-1 representing bacterial carotenoids. We hypothesize the biosynthesis of carotenoid as adaptive strategy to cope up inhospitable cold environments of Leh and Ladakh. The strong, scattering bands by different isolates attributable to ν(C=C) phase stretching, ν(C-C) and δ(C-CH3) methyl components systems, which could be probably membrane-associated C50 carotenoids. Their high intensities are due to resonance enhancement. It can be concluded that Raman spectroscopy is a sensitive and convenient detection tool for typing of the bacterial biomarkers with less time consumption.

Freshwater macroalgae, Cladophora glomerata Kützing and Microspora floccosa (Vaucher) Thuret, harvested from Nan River in northern Thailand, were extracted with hexane, ethyl acetate, methanol and hot water. The extracts were screened for antibacterial activities. Hexane and ethyl acetate extracts of both algae showed the activities against Bacillus cereus and Vibrio parahaemolyticus. The extracts were further separated using column chromatography and chemically characterized by GC–MS in order to be tentative identify the compounds responsible for such activities. The main compositions were fatty acid and other organic compounds, in which have not been reported in these algae. These results indicate that extracts of C. glomerata and M. floccosa exhibited appreciable antimicrobial activity and could be a source of valuable bioactive materials for health products.

The inheritance and gene effects structure were investigated for ten quantitative traits in field cultivars by a full diallel model of four parents – the cultivars Kerpo, Pleven 10, Mir and EFB33. Overdominance was prevalent for most of the traits in both hybrid generations (F1 and F2) except for 1000-seed weigth. A non-additive gene action appeared to be more predominant for the inheritance of most traits studied. The fraction of h2/H2 indicated that at least one group of genes controlled dominance for all the traits, namely for seed weight per plant (6), seeds per pod (3). The Н2/4Н1 ratio was close to the expected value of 0.25 for seeds per plant and seed weight per plant, indicating a symmetrical distribution of dominant and recessive genes. Dominant alleles were more frequent in parental forms for the inheritance of most traits except seeds per pod in F1. In the case of seeds per pod (F1) and 1000-seed weight (F1 and F2) a comparatively low difference was established between the coefficient of heritability in narrow and broad sense, indicating that the selection in early generations may be effective. The genotypes with exhibited dominant alleles were EFB33 for plant height, first pod height, internode distance and fertile nodes per plant, Mir for pods per plant, nodes per plant and 1000-seed weight and Pleven 10 for seeds per plant and seed weight per plant. Kerpo had recessive alleles for all the tested traits except seeds per pod.

The expression of bacterial gusA gene encoding for E. coli enzyme β-glucuronidase in haploid and diploid cells of methylotrophic yeast Hansenula (Ogataea) polymorpha was studied. Haploid strains were transformed with plasmid carrying gusA gene under the control of strong alcohol oxidase (AOX) promoter and several Gus+ transformants were isolated. They were crossed with wild type strains lacking Gus activity. The resulting diploids were sporulated and several meiotic segregants with increased Gus activity were isolated. Some of these segregants were crossed and the corresponding diploid strains were obtained. It was found that Gus activity of diploid strains was twice higher compared to haploids, whereas Gus activity of diploids was increased about six fold compared to those of haploids. Our results can be used as an approach for development of yeast strains with elevated expression of heterologous proteins.

This study elucidated the individual and mixed mycorrhizal effects of two arbuscular mycorrhizal (AM) isolates on growth and biochemical status of Acacia arabica under salinity stress gradients. Salt treatment provided in soil hampered legume growth and its biochemical status. But, mycorrhizal colonizations in plant root system reduced the extent of deleterious salt effect and also helped in plant growth enhancement. Additionally, mixed mycorrhizal association (Glomus mosseae + Glomus fasciculatum) responded better towards osmolyte accumulation and in salt stress alleviation. Due to individual and mixed mycorrhizal colonizations in A. arabica; protein, carbohydrate and reducing sugar acquisitions were found maximum at soil salinity of 5.94 dS/m over corresponding non-mycorrhizal plant. However, mixed AM inoculation accumulated proline content and improved dry biomass to a higher magnitude at the highest soil salinity level. Mixed AM (G. mosseae + G. fasciculatum) colonization improved maximum amount of total chlorophyll (20.94%), protein (19.72%), carbohydrate (23.83%), reducing sugar (17.60%) at soil salinity of 5.94 dS/m and dry biomass (20.35%), proline content (10.99%) at salinity level of 8.26 dS/m when compared with non-mycorrhizal counterpart. Greater magnitude of AM root colonization was found in mixed AM treated plant and may be responsible for more improvement in growth and biochemical status and consequently mitigated adverse salt effect better.

Olive oil mill wastewaters (OMWW) are the main liquid effluents generated by the olive oil production industry. This liquid, considered pollutant and toxic, is characterised by its high content of organic matter including mainly sugars and fats, and phenols compounds, which can be used in ruminants feeding. The purpose of this study is to valorise this agricultural by-product in ruminant feeding by estimation its in vitro degradability in presence of ovine ruminale microbiota comparatively to vetch-oat hay, using in vitro gas production technique coupled with NH3-N and protozoa measurements. Cumulative gas production was recorded at 3, 6, 9, 24, 48, 72 and 96 hours of incubation. The determination of gazes produced (carbon dioxide and methane) was recorded at 6, 9, 24, 48 and 96 hours. However, Ammonia and protozoa number were recorded after 24 hours of incubation. Fermentation profile was fitted to the exponential model y = a + b (1 – e-kt). The OMWW are characterized by their high sugars content (39.91%) and their low content in ash (1.99%) and crude protein (2.70%). This by-product is also characterized by its high concentration in total phenols (7.2%) and tannins (4.5%). However, they contain a very small amount of condensed tannins (0.89%). Comparatively to vetch-oat hay, OMWW produced low amount of gas (-23.6 units). Furthermore, its in vitro fermentation generates low volume of methane (9.83%, V/V), suggesting that the OMWW nature enhanced the efficiency of ruminale microbiota towards microbial biomass production and inhibition of ruminale methanogenesis pathway. This result is reinforced by the reduction of ammonia production (-0.35 units) and protozoa proliferation (-1 unit) comparatively to vetch-oat hay. The anaerobic biodegradation of OMWW reveal their significant use by the rumen microbiota, allowing us to strongly recommend its use as a supplement in feed ruminant. In addition, it allows considering using this residue as a feed additive in diets of ruminants for the reduction of ruminal methanogenesis.

The aim of this study was profiling of primary metabolites of five Bulgarian Fumaria species (F. officinalis L., F. thuretii Boiss., F. kralikii Jord., F. rostellata Knaf. and F. schrammii Velen.) by GC-MS analyses. In polar fractions ten carbohydrates, one polyol, ten amino acids and six organic acids were identified. Apolar (lipid) fractions showed the presence of four free fatty acids, two esters of fatty acids with glycerol and two fatty alcohols. Sucrose and fructose were in the highest relative concentrations of identified carbohydrates. Citric acid was the dominant organic acid in polar fractions of five Fumaria species. Predominant compounds in lipid fractions were palmitic acid and 1-stearoyl-glycerol. Principal component analysis (PCA) of GC-MS data of polar and apolar fractions of five Bulgarian Fumaria species differentiates them in two groups (F. officinalis and F. thuretii; F. rostellata and F. schrammii, respectively), while F. kralikii had phythochemical similarity with plants of both distinguished groups. The obtained results of PCA of primary metabolites could be proposed as chemotaxonomic markers for plants of the genus Fumaria.

We cloned the cp23 gene coding P23 (glyco)protein from Cryptosporidium parvum isolated from Thua Thien Hue province, Vietnam. The coding region of cp23 gene from C. parvum is 99% similar with cp23 gene deposited in NCBI (accession number: U34390). SDS-PAGE and Western blot analysis showed that the cp23 gene in E. coli BL21 StarTM (DE3) produced polypeptides with molecular weights of approximately 37, 40 and 49 kDa. These molecules may be non-glycosylated or glycosylated P23 fusion polypeptides. Recombinant P23 protein purified by GST (glutathione S-transferase) affinity chromatography can be used as an antigen for C. parvum antibody production as well as to develop diagnostic kit for C. parvum.

Pectinase is an important enzyme that finds application in many food processing industries and solid state fermentation (SSF) is an attractive technology for enzyme production. In this work, design of experimental (DOE) methodology using Taguchi orthogonal array (OA) was applied to evaluate the influence of five factors (different levels of citrus pulp, initial pH of the medium, C/N ratio, type of solid substrate and citrus pulp) on the pectinase production by Aspergillus niger under solid-state fermentation. The results showed that citrus pulp concentration, type of solid substrate and citrus pulp were found to be the most effective factor for promoting enzyme production and the supplementation of the medium with citrus pulp caused a 23% increase in the pectinase production when compared with the basal medium. The study shows that the Taguchi's method is suitable to optimize the experiments for the production of pectinase (R2 = 0.946).

The tumor development in colorectal cancer is one of the most investigated tumorigenesis in the world. This study aims to analyze the impact of presence of IL-17, perforin and CD68 on colorectal cancer. During the period October 1st 2013 – March 1st 2014, samples from fifty patients with colon cancer (14 female and 36 male, 7-75 years old) from Al-Hussain Hospital City (Kerbala, Iraq), Hospital Medical City (Baghdad, Iraq) and Teaching Oncology Hospital, Baghdad Medical City (Baghdad, Iraq) were taken and analyzed. Immunohistochemical analyses of IL-17, perforin and CD68 of colorectal biopsies were performed by using Dako's Service (Denmark). The levels of IL-17 were significantly (p<0.01) higher in females (age group 41-75), and Duke C. Perforin was found around the crypts of the mucosa and submucosa. Their levels were significantly (p<0.01) higher in males (age group 41-75) with significance (p<0.01) in Duke B. CD68 was present around the crypts, but results showed no significant difference (p>0.05) between the genders, whereas there was a highly significant (p<0.01) increase in age group 41-75 when compared with the others groups. Also, the results revealed a highly significant raise (p<0.01) in Duke C when compared with the TNM groups and highly significant (p<0.01) correlation among study parameters. These results demonstrated the critical role of IL-17, perforin and CD68 highlighting tumorigenesis and inflammation as pro-inflammatory factors in colorectal cancer.