This study aimed to find candidate strains of Lactobacillus isolated from sheep dairy products (yogurt and ewe colostrum) with probiotic and anticancer activity. A total of 100 samples were randomly ...collected from yogurt and colostrum and 125 lactic acid bacteria were isolated. Of these, 17 Lactobacillus strains belonging to five species (L. delbrueckii, L. plantarum, L. rhamnosus, L. paracasei, and L. casei) were identified. L. plantarum 17C and 13C, which isolated from colostrums, demonstrated remarkable results such as resistant to low pH and high concentrations of bile salts, susceptible to some antibiotics and good antimicrobial activity that candidate them as potential probiotics. Seven strains (1C, 5C, 12C, 13C, 17C, 7M, and 40M), the most resistant to simulated digestion, were further investigated to evaluate their capability to adhere to human intestinal Caco‐2 cells. L. plantarum 17C was the most adherent strain. The bioactivity assessment of L. plantarum 17C showed anticancer effects via the induction of apoptosis on HT‐29 human cancer cells and negligible side effects on one human epithelial normal cell line (FHs 74). The metabolites produced by this strain can be used as alternative pharmaceutical compounds with promising therapeutic indices because they are not cytotoxic to normal mammalian cells.
Probiotics modify the gut microbial composition/homeostasis. Probiotics bind to mutagens and eliminate them from the body. Probiotics enhances the mucosal and system immune responses. Probiotics can control the cell proliferation and apoptosis. Probiotics protect the intestinal barrier from deleterious.
Sorghum (
Sorghum bicolor
L. Moench) is a rapidly emerging high biomass feedstock for bioethanol and lignocellulosic biomass production. The robust varietal germplasm of sorghum and its completed ...genome sequence provide the necessary genetic and molecular tools to study and engineer the biotic/abiotic stress tolerance. Traditional proteomics approaches for outlining the sorghum proteome have many limitations like, demand for high protein amounts, reproducibility and identification of only few differential proteins. In this study, we report a gel-free, quantitative proteomic method for in-depth coverage of the sorghum proteome. This novel method combining phenol extraction and methanol chloroform precipitation gives high total protein yields for both mature sorghum root and leaf tissues. We demonstrate successful application of this method in comparing proteomes of contrasting cultivars of sorghum, at two different phenological stages. Protein identification and relative quantification analyses were performed by a label-free liquid chromatography tandem mass spectrometry (LC/MS-MS) analyses. Several unique proteins were identified respectively from sorghum tissues, specifically 271 from leaf and 774 from root tissues, with 193 proteins common in both tissues. Using gene ontology analysis, the differential proteins identified were finely corroborated with their leaf/root tissue specific functions. This method of protein extraction and analysis would contribute substantially to generate in-depth differential protein data in sorghum as well as related species. It would also increase the repertoire of methods uniquely suited for gel-free plant proteomics that are increasingly being developed for studying abiotic and biotic stress responses.
Pulmonary emphysema is an important radiological finding in chronic obstructive pulmonary disease patients, but bacteriological differences in pneumonia patients according to the severity of ...emphysematous changes have not been reported. Therefore, we evaluated the bacteriological incidence in the bronchoalveolar lavage fluid (BALF) of pneumonia patients using cultivation and a culture-independent molecular method. Japanese patients with community-acquired pneumonia (83) and healthcare-associated pneumonia (94) between April 2010 and February 2014 were evaluated. The BALF obtained from pneumonia lesions was evaluated by both cultivation and a molecular method. In the molecular method, ~600 base pairs of bacterial 16S ribosomal RNA genes in the BALF were amplified by polymerase chain reaction, and clone libraries were constructed. The nucleotide sequences of 96 randomly selected colonies were determined, and a homology search was performed to identify the bacterial species. A qualitative radiological evaluation of pulmonary emphysema based on chest computed tomography (CT) images was performed using the Goddard classification. The severity of pulmonary emphysema based on the Goddard classification was none in 47.4% (84/177), mild in 36.2% (64/177), moderate in 10.2% (18/177), and severe in 6.2% (11/177). Using the culture-independent molecular method,
was significantly more frequently detected in moderate or severe emphysema patients than in patients with no or mild emphysematous changes. The detection rates of
and
were unrelated to the severity of pulmonary emphysematous changes, and
species - except for the
group and
- were detected more frequently using the molecular method we used for the BALF of patients with pneumonia than using culture methods. Our findings suggest that
is more frequently detected in pneumonia patients with moderate or severe emphysema than in those with no or mild emphysematous changes on chest CT.
may play a major role in patients with pneumonia complicating severe pulmonary emphysema.
Abstract Compared to culture-based method, the sensitivity, specificity, positive, and negative predictive values of the GenoType® HelicoDR for detecting Helicobacter pylori resistance were, ...respectively, 100, 86.2, 89.7%, and 100% to clarithromycin as well as 82.6, 95.1, 90.5%, and 90.7% to fluoroquinolones. This molecular assay detected a mixture of genotypes and could successfully analyze biopsies without transport/storage limitations.
Managing forests by selection cutting is a promising silvicultural technique for maintaining forest biodiversity. Despite the importance of fungi in decomposition and nutrient cycling in forest ...ecosystems, no study to date has investigated the effects of selection cutting on fungal communities, especially using a culture-independent molecular technique to assess more than just the species that are fruiting at the time of sampling. Based on operational taxonomic units (OTUs) found in coarse woody debris, we compared the richness and community composition of wood-inhabiting fungi from selection cutting, age-class, and unmanaged European beech-dominated forests. We found that fungal OTU richness in selection cutting and unmanaged forests was not significantly different (
P
> 0.05), but it was higher, in both cases, than that in the age-class forest (
P
= 0.0002). Fungal community composition was not significantly different among the three forest types (
P
> 0.05). Abundances of common, wood-inhabiting fungal OTUs in different forest types were significantly correlated: the highest and lowest correlations were found between unmanaged forests and selection cutting (
ρ
= 0.52,
P
< 0.0001,
n
= 94), and between unmanaged and age-class forests (
ρ
= 0.30,
P
= 0.0080,
n
= 79), respectively.
The purpose of this study was to use a novel method that was based on the application of chaperonin-60 sequencing to describe the vaginal microflora of 16 healthy women.
Asymptomatic women consented ...for vaginal swabs to be collected at the time of a clinical pelvic examination. Total genomic DNA was isolated from the vaginal swabs. Degenerate, universal polymerase chain reaction primers were used to amplify an approximately 555 base pair region of the universal chaperonin-60 gene, which is found in all eubacteria and eukaryotes, from the total genomic DNA and libraries of cloned polymerase chain reaction products were constructed. Library clones were sequenced, and the resulting sequences were assigned to taxonomic groups on the basis of similarity to reference sequence data. Presence of
Chlamydophila psittaci sequences in the samples was confirmed by species-specific polymerase chain reaction.
Sixteen of the 23 women who were enrolled had normal flora by Nugent's score of <4 and had adequate polymerase chain reaction product for assessment. Vaginal flora libraries were dominated by a variety of sequences with similarity to
Lactobacillus spp
L crispatus,
L iners,
L gasseri,
L jensenii, and
L buchneri. Other sequences that were identified included representatives of
Gardnerella spp, sequences with similarity to
Porphyromonas spp and
Megasphaera spp and sequences identical to
C psittaci.
Culture-independent, chaperonin-60 sequence-based molecular methods can lead to the identification of greater diversity within defined taxa compared with those that are identified by standard culture-based methods and to the identification of novel organisms that were not previously associated with vaginal flora.
Investigating the endophytic bacterial community in special moss species is fundamental to understanding the microbial-plant interactions and discovering the bacteria with stresses tolerance. Thus, ...the community structure of endophytic bacteria in the xerophilous moss Grimmia montana were estimated using a 16S rDNA library and traditional cultivation methods. In total, 212 sequences derived from the 16S rDNA library were used to assess the bacterial diversity. Sequence alignment showed that the endophytes were assigned to 54 genera in 4 phyla (Proteobacteria, Firmicutes, Actinobacteria and Cytophaga/Flexibacter/Bacteroids). Of them, the dominant phyla were Proteobacteria (45.9%) and Firmicutes (27.6%), the most abundant genera included Acinetobacter, Aeromonas, Enterobacter, Leclercia, Microvirga, Pseudomonas, Rhizobium, Planococcus, Paenisporosarcina and Planomicrobium. In addition, a total of 14 species belonging to 8 genera in 3 phyla (Proteobacteria, Firmicutes, Actinobacteria) were isolated, Curtobacterium, Massilia, Pseudomonas and Sphingomonas were the dominant genera. Although some of the genera isolated were inconsistent with those detected by molecular method, both of two methods proved that many different endophytic bacteria coexist in G. montana. According to the potential functional analyses of these bacteria, some species are known to have possible beneficial effects on hosts, but whether this is the case in G. montana needs to be confirmed.
Bromeliaceae is an important botany family that includes many species with economic value; demand for members of this family is increasing. However, illegal collection frequently occurs, drastically ...reducing the species populations; thus, it is necessary to collect and store Bromeliaceae genetic material. In this study, we identified and quantified genetic variability of the Bromeliad family using dominant markers to create the first Germplasm Bank in the northeast region of Brazil. Molecular tools were used to characterize the collected accessions. The combination of 11 inter-simple sequence repeats and 13 random amplified polymorphic DNA markers were used to detect the genetic variability of wild bromeliad accessions.
The composition of nitrifying bacteria in an aerated landfill bioreactor filled with municipal solid waste incineration bottom ashes and shredded incombustible wastes was monitored and analyzed as a ...function of time during 1 year operation using molecular techniques. Besides, the effects of differing operational conditions on the bioreactor performance and diversity of nitrifiers were evaluated thoroughly. The results confirmed the viability of rapid aerobic bio-stabilization in an aerated landfill bioreactor operated at various ORP levels (−400 to 150
mV). BOD
5 decreased faster than TOC and dropped below 10
mg/l after day 120. Subsequently, it remained quite constant until the end of the operational period. This rapid BOD
5 degradation in the aerated landfill bioreactor increased the possibility of nitrification by promoting nitrifying bacteria having high oxygen affinities. The nitrification activity in the bioreactor was approved by identification of intensive amounts of
Nitrosomonas-like ammonia oxidizers and
Nitrospira related nitrite oxidizers with 16S rDNA and
amoA based molecular microbiology techniques.
A total of 225 maize kernel samples were collected from Shandong Province in China from 2012 to 2014 and analysed for contamination with Fusarium spp. and fumonisins (FBs) using molecular methods and ...high-performance liquid chromatography with fluorescence detection. The results showed that the average incidences of Fusarium spp. in 2012, 2013 and 2014 were 23.3%, 37.1% and 36.5%, respectively, Fusarium verticillioides being the predominant species. In 2012, the average contamination level of FBs was 3071 ng g
−1
, which was higher than that in 2014 (2913 ng g
−1
) and 2013 (2072 ng g
−1
). Of all samples, 13% and 19% had FB contamination levels higher than 2000 and 4000 ng g
−1
, which are the maximum limits as set by the Food and Drug Administration of the United States and the European Commission, respectively. Therefore, efforts should be taken to minimise the potential risk of FBs to the health of humans and animals.
PDF
