No abstract

This study investigated the influence of aeration and minimal medium conditions on antioxidant and antibacterial activities of 21 probiotic Lactobacillus strains isolated from dairy products. The probiotic potential of the isolates was evaluated by pH and bile tolerance. Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) was used to confirm the phenotypic identification of isolates. Antioxidant producer isolates were screened by resistance to reactive oxygen species (ROS). The antioxidant and antibacterial activities of extracellular materials after 48 h fermentation with antioxidative strains were determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and broth microdilution assays, respectively. The results indicate that the antioxidant capacity of supernatants was increased by using of both minimal medium and agitation. The antibacterial activity was increased in minimal medium, but there has nearly no change in the antibacterial properties by using both agitation and minimal medium. The maximum antibacterial activity was observed during mid-exponential phase until the beginning of the early-stationary phase, but the maximum antioxidant activity was detected at the stationary growth phase. There is a significant relationship between antioxidant and antibacterial activities of the cell-free probiotic extracts, and their production rates are closely related to the fermentation type. The bioactive materials from probiotics could be extracted in a large amount at an appropriate time under a suitable condition.

Background: It has been proven that probiotic Lactobacillus bacteria have inhibitory effects on human cancer cell lines. The aim of this study is to isolate and characterize the antioxidant probiotic Lactobacillus and determine the possible anticancer activities of the selected strain. Methods: One of the Lactobacillus strain isolated from camel doogh sample showed the high antioxidant activity by using of different methods such as resistance to hydrogen peroxide, hydroxyl radical and superoxide anions. The antioxidant strain was characterized by sequencing of 16S rRNA V2-V3 regions and the 16S-23S intergenic spacer (ITS). The methanol extract of this strain supernatant was fractionated using thin layer chromatography (TLC) and antioxidant activity of fractions was detected by 0.1% of DPPH through TLC-DPPH bioautography. In vitro anticancer activity of each fraction was investigated by using MTT and flow cytometry methods. Results: According to the phylogenetic results, the antioxidant Lactobacillus strain was closely related to Lactobacillus hilgardii strain E91 (Accession No. EF536365). After fractionation and anti-proliferation assessments of Lactobacillus hilgardii strain AG12a extracellular materials, one of the antioxidant fraction (F4) showed maximum DPPH radical scavenging activity (IC 50 of 535.27 μg/mL). MTT assay of the F4 fraction demonstrated cytotoxic activity against Caco-2 with the IC50 value of 299.05 μg/mL. The cell death activity of the fraction was confirmed by flow cytometry with 30.925. Conclusions: In this study, the anticancer and apoptotic properties of Lactobacillus hilgardii against Caco-2 cell line was reported for the first time. The isolated bioactive fraction from the extracellular methanol extract needs to be further investigated in human studies of cancer therapy.

Background: The presence of microbiome in the blood samples of healthy individuals has been addressed. However, no information can be found on the healthy human blood microbiome of Iranian subjects. The current study is thus aimed to investigate the existence of bacteria or bacterial DNA in healthy individuals. Methods: Blood samples of healthy subjects were incubated in BHI broth at 37 °C for 72 h. The 16S rRNA PCR and sequencing were performed to analyze bacterial isolates. The 16S rRNA PCR was directly carried out on DNA samples extracted from the blood of healthy individuals. NGS was conducted on blood samples with culture-positive results. Results: Fifty blood samples were tested, and six samples were positive by culture as confirmed by Gram staining and microscopy. The obtained 16S rRNA sequences of cultured bacterial isolates revealed the presence of Bacilli and Staphylococcus species by clustering in the GeneBank database (≥97% identity). The 16S rRNA gene sequencing results of one non-cultured blood specimen showed the presence of Burkholderia . NGS results illustrated the presence of Romboutsia , Lactobacillus , Streptococcus , Bacteroides, and Staphylococcus in the blood samples of positive cultures. Conclusion: The dormant blood microbiome of healthy individuals may give the idea that the steady transfer of bacteria into the blood does not necessarily lead to sepsis. However, the origins and identities of blood-associated bacterial rDNA sequences need more evaluation in the healthy population.

The delay in diagnosis and treatment of breast cancer results in low survival rates and high mortality. Thus, it is essential to characterize new therapeutic targets and prognostic breast cancer biomarkers. The rising evidence suggested that long non-coding RNAs (lncRNAs) expression levels are deregulated in human cancers and can use as biomarkers for the rapid diagnosis of breast cancer. In the present study, a Quantitative Real-time polymerase chain reaction (qRT-PCR) technique was used to measure twenty oncogenic and tumor suppressor lncRNAs expression levels in whole blood samples of breast cancer patients and normal controls. Blood samples from 30 healthy women and 30 female breast cancer patients were collected. Then cDNA was synthesized from the extracted RNA blood. The expression level of lncRNAs measured and analyzed by LinReg PCR and REST software and the correla tion between lncRNAs dysregulation and clinical characteristics and prognosis were also analyzed by SPSS software. The comparison between the expression levels of lncRNAs in the blood samples of breast cancer patients compared with healthy individuals revealed that some lncRNAs (MEG3, NBAT1, NKILA, GAS5, EPB41L4A-AS2, ZFAS1, MVIH, Z38, and BC040587) were down regulated. In contrast, other lncRNAs (H19, SPRY4-IT1, UCA1, AC026904.1, CCAT1) were up-regulated, signi cantly. It was shown that the expression levels of NKILA, NBAT1, and ZFAS1 lncRNAs were related to tumor size, and BC040587expression level related to age, node metastasis, tumor size, and grade (P<0.05). The association between H19 and SPRY4-IT1 lncRNAs with HER-2 was con rmed statistically (P<0.05). Our data highlighted the correlation of BC040587, H19, and SPRY4-IT1 lncRNAs with clinicopathological traits in breast cancer patients suggesting their future applications as novel biomarkers and therapeutic targets in breast cancer. In conclusion, circulating lncRNAs could consider as the prognostic and predictive markers in breast cancer.

The delay in diagnosis and treatment of breast cancer results in low survival rates and high mortality. Thus, it is essential to characterize new therapeutic targets and prognostic breast cancer biomarkers. The rising evidence suggested that long non-coding RNAs (lncRNAs) expression levels are deregulated in human cancers and can use as biomarkers for the rapid diagnosis of breast cancer. In the present study, a Quantitative Real-time polymerase chain reaction (qRT-PCR) technique was used to measure twenty oncogenic and tumor suppressor lncRNAs expression levels in whole blood samples of breast cancer patients and normal controls. Blood samples from 30 healthy women and 30 female breast cancer patients were collected. Then cDNA was synthesized from the extracted RNA blood. The expression level of lncRNAs measured and analyzed by LinReg PCR and REST software and the correla­tion between lncRNAs dysregulation and clinical characteristics and prognosis were also analyzed by SPSS software. The comparison between the expression levels of lncRNAs in the blood samples of breast cancer patients compared with healthy individuals revealed that some lncRNAs (MEG3, NBAT1, NKILA, GAS5, EPB41L4A-AS2, ZFAS1, MVIH, Z38, and BC040587) were down regulated. In contrast, other lncRNAs (H19, SPRY4-IT1, UCA1, AC026904.1, CCAT1) were up-regulated, significantly. It was shown that the expression levels of NKILA, NBAT1, and ZFAS1 lncRNAs were related to tumor size, and BC040587expression level related to age, node metastasis, tumor size, and grade (P<0.05). The association between H19 and SPRY4-IT1 lncRNAs with HER-2 was confirmed statistically (P<0.05). Our data highlighted the correlation of BC040587, H19, and SPRY4-IT1 lncRNAs with clinicopathological traits in breast cancer patients suggesting their future applications as novel biomarkers and therapeutic targets in breast cancer. In conclusion, circulating lncRNAs could consider as the prognostic and predictive markers in breast cancer.

Background: Nowadays, searching for natural bioactive compounds with potential use in food industries is a major issue. Because of simple purification, natural compounds from microbial sources attract more attention. These encompass antioxidant and antibacterial materials derived from probiotics. Methods: In this study, Lactobacillus strains were isolated from kefir specimens. The antioxidant and antibacterial activity of the methanol extract of the supernatants was determined using 2, 2-diphenyl-picyril hydrazil (DPPH) and minimum inhibitory concentration (MIC) methods, respectively. In order to increase the antioxidant properties, a minimum medium fermented aerobically was used. Results: Antibacterial activity of Lactobacillus supernatant increased against E. coli ATCC 11303 in case of minimum medium (25.32 mg/mL) compared to MRS broth (32 mg/mL); however, aerobic condition decreased antibacterial production (65.44 mg/mL). After fractionation by thin-layer chromatography (TLC), this value reached the highest level (500 µg/mL). Production analysis at different times showed that maximum antibacterial activity was obtained in the middle of the logarithmic growth phase until the beginning of the stationary growth phase. The antioxidant traits increased significantly in minimum culture media and anaerobic condition (492.1 ± 0.25 µg/mL) compared to the similar condition in MRS broth (880.96 ± 0.05 µg/mL). The highest antioxidant production was observed in the stationary growth phase of the aerobically fermented minimum medium (266.82 ± 0.17 µg/mL). Conclusions: The findings of this study showed that the best antibacterial and antioxidant-producing isolate, L. casei strain K1C (accession no.: KU954559), could be useful as a natural preservative in food industries.

Rising evidence proved the existence of dormant blood microbiota in healthy individuals, yet there is no information on the circulating microbiome of Iranian healthy women. The High-throughput Next-Generation Sequencing (NGS) provides a powerful method for the characterization of the blood microbiome. We investigated circulating bacterial composition in healthy individuals by 16S rDNA- based next-generation sequencing technique targeting the V3-V4 hypervariable region on the Illumina platform. We identified Operational Taxonomic Units (OTUs) similarity with various phyla, predominated by Proteobacteria (83%), Firmicutes (8%), Bacteroidetes (6%), and Actinobacteria (2%) phyla in healthy subjects. Based on the sequencing data, a diverse bacterial family was found in the whole blood of the healthy subjects, belongs to Burkholderiaceae (69%) and Enterobacteriaceae (13%) families on average. At the genus level, we illustrated the abundance of Ralstonia, Rhizobium, Ignatzchineria, Lactococcus, Stenotrophomonas, and Ideonella. Given the dominance of the putative blood microbiota within the different niches, it may be of considerable clinical significance and therapeutic strategies potential in the future. The blood microbiota can facilitate both the diagnosis and improved understanding of the onset of numerous human diseases as innovative biomarkers.