Retraction Note: Blockage of miR-92a-3p with locked nucleic acid induces apoptosis and prevents cell proliferation in human acute megakaryoblastic leukemia.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Serogroups, and drug resistance of nontyphoidal Salmonella in symptomatic patients with community-acquired diarrhea and chicken meat samples in Tehran.

BACKGROUND: Salmonella is considered as a main cause of community-acquired diarrhea in humans, however, sources of the multi-drug resistant (MDR) strains and their link with the disease are not well known. AIMS: This study aimed to investigate the frequency, serogroup diversity, and antimicrobial susceptibility patterns of Salmonella strains in poultry meat and stool samples of patients with community acquired diarrhea in Tehran. METHODS: We compared the frequency of non-typhoidal Salmonella serogroups, the similarities of their resistance patterns to 10 antimicrobial compounds, the prevalence of extended spectrum ß-lactamase (ESBL) and ampicillinase C (AmpC) genetic determinants, and class 1 and 2 integrons in 100 chicken meat and 400 stool samples of symptomatic patients in Tehran during June 2018 to March 2019. RESULTS: Salmonella was isolated from 75% and 5.5% of the chicken meats and human stool samples, respectively. The chicken meat isolates mainly belonged to serogroup C (88%, 66/75), while the human stool isolates were mainly related to serogroup D (59.1%, 13/22). The MDR phenotype and the most common rates of resistance to antibiotics, including tetracycline, trimethoprim/sulfamethoxazole (TS) and azithromycin, were detected in 4.5% and 45.3%, 59% and 13.6%, 43% and 9.1%, 42% and 9.1% of the human stool and chicken meat samples, respectively. Carriage of bla CTX, bla SHV, and bla PER genes in the meat isolate with ESBL resistance phenotype and bla ACC, bla FOX, and bla CMY-2 among the 7 meat strains with AmpC resistance phenotype was not confirmed using polymerase chain reaction (PCR). High prevalence of class 1 and 2 integrons was characterized and showed a correlation with resistance to TS and chloramphenicol. CONCLUSION: These findings showed a lack of association between chicken meats and human isolates due to discrepancy between the characterized serogroups and resistance phenotypes.

The correlation between Pax5 deletion and patients survival in Iranian children with precursor B-cell acute lymphocytic leukemia.

Despite advances in treatment, children with acute lymphoblastic leukemia (ALL) still experience drug resistance and relapse. Several gene mutations are involved in the onset of this disease and resistance to therapy. The present study examines the incidence of IKZF1, CDKN2A/B, PAX5, EBF1, ETV6, BTG1, RB1, JAK2, and Xp22.33 gene deletions/duplications associated with pediatric ALL in Iran and investigates the possible effect of these mutations on drug resistance. Three-year disease-free survival (3DFS) was evaluated for children diagnosed with Philadelphia negative precursor-B-cell ALL hospitalized at Sayed-al-Shohada Hospital, Isfahan-Iran, from January 2009 until December 2012. DNA was extracted from bone marrow slides, and ALL correlated gene deletions and duplications were measured using Multiplex Ligation-dependent Probe Amplification (MLPA) method. The correlation between gene mutations and 3DFS was then assessed. Among the nine aforementioned investigated genes, 63% of samples showed at least one gene mutation. At least two concomitant genomic mutations were observed in 42% of samples. Pax5 deletion was the most prevalent gene mutation observed in 45% of cases, and showed significant negative impact on response to treatment. CDKN2A/B (9p21.3) gene deletion, and ETV6 (12p13.2) gene duplication also demonstrated negative effect on patient survival and contributed to a worse prognosis if concomitant with Pax5 gene deletion. ALL patients with one of the gene deletions including Pax5  and CDKN2A/B (9p21.3) or ETV6 (12p13.2) gene duplication are classified as high-risk patients and need more intensified protocols of treatment to improve their chance of survival.

Circulating exosomes and exosomal microRNAs as biomarkers in gastrointestinal cancer.

The most important biological function of exosomes is their possible use as biomarkers in clinical diagnosis. Compared with biomarkers identified in conventional specimens such as serum or urine, exosomal biomarkers provide the highest amount of sensitivity and specificity, which can be attributed to their excellent stability. Exosomes, which harbor different types of proteins, nucleic acids and lipids, are present in almost all bodily fluids. The molecular constituents of exosomes, especially exosomal proteins and microRNAs (miRNAs), are promising as biomarkers in clinical diagnosis. This discovery that exosomes also contain messenger RNAs and miRNAs shows that they could be carriers of genetic information. Although the majority of RNAs found in exosomes are degraded RNA fragments with a length of <200 nucleotides, some full-length RNAs might be present that may affect protein production in the recipient cell. In addition, exosomal miRNAs have been found to be associated with certain diseases. Several studies have pointed out miRNA contents of circulating exosomes that are similar to those of originating cancer cells. In this review, the recent advances in circulating exosomal miRNAs as biomarkers in gastrointestinal cancers are discussed. These studies indicated that miRNAs can be detected in exosomes isolated from body fluids such as saliva, which suggests potential advantages of using exosomal miRNAs as noninvasive novel biomarkers.

Locked nucleic acid anti-miR-21 inhibits cell growth and invasive behaviors of a colorectal adenocarcinoma cell line: LNA-anti-miR as a novel approach.

Colorectal cancer (CRC) is the third leading cause of cancer-related death and has an extremely poor prognosis. Dysregulation of microRNAs (miRNAs) has been shown to be involved in the pathogenesis and progression of many malignancies. Recent data suggest that microRNA-21 (miR-21) is significantly elevated in different types of cancer, especially colon adenocarcinoma. Against this background, locked nucleic acid (LNA)-modified oligonucleotides have recently been suggested as a novel approach for targeting miRNAs as antisense-based gene silencing. The aim of the current study was to explore the functional role of LNA-anti-miR-21 in a colon adenocarcinoma LS174T cell line. LS174T cells were transfected with LNA-anti-miR-21 for 24, 48 and 72 h. Quantitative real-time reverse transcriptase-PCR (qRT-PCR) was performed to assess miR-21 expression by LNA-anti-miR-21. The viability of the cells was evaluated by MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide) assay and Annexin V/propidium iodide staining assay was used to detect apoptosis. Moreover, invasive behavior of the cells was evaluated before and after therapy by transwell assay. LNA-anti-miR-21 was successfully transfected in human LS174T cells and suppressed the endogenous miR-21. LNA-anti-miR-21 inhibited the cells' growth followed by induction of apoptosis. LNA-anti-miR-21 (50 pmol/µl) reduced the invasive behaviors of LS174T cells after 24 h, compared with untreated cells and scrambled LNA-transfected cells. However, this effect was more pronounced after 72 h. Our findings suggest the therapeutic potential of LNA-anti-miR-21 in a colon adenocarcinoma for targeting miR-21 expression. Further studies are warranted to investigate the molecular mechanisms underlying this novel inhibitor in colorectal cancer to establish its potential value for treatment of CRC patients with high miR-21 expression.

Categorization of endometritis and its association with ovarian follicular growth and ovulation, reproductive performance, dry matter intake, and milk yield in dairy cattle.

The objectives were to evaluate the effect of different categories of endometritis on follicular growth and ovulation, reproductive performance, dry matter intake (DMI), and milk yield (MY) in dairy cows. Lactating Holstein cows (n = 126) were examined for endometritis on 25 ± 1 day postpartum (DPP) using vaginoscopy, transrectal ultrasonography, and endometrial cytology to determine the presence and type of vaginal discharge, uterine fluid, and proportion of polymorphonuclear (PMN) cells, respectively. Cows that had mucopurulent vaginal discharge and/or presence of uterine fluid, no mucopurulent vaginal discharge or uterine fluid but 8% or more PMN, and mucopurulent vaginal discharge and/or uterine fluid and 8% or more of PMN were defined as having clinical (CLIN; n = 45), cytological (CYTO; n = 15), and clinical and cytological (CLINCYTO; n = 30) endometritis, respectively. Cows that had none of the above pathological conditions were classified as unaffected (UNAF; n = 36). The diameter of the largest follicle at first examination, intervals from calving to first dominant (diameter = 10 mm) follicle, preovulatory size (diameter = 16 mm) follicle, ovulation, presence of follicular cyst, and proportion of ovular cows at 35 and 65 DPP were recorded as the measures of follicular growth and ovulation. None of the ovarian follicular parameters analyzed was affected by categories of endometritis. The first service conception rate tended (P = 0.06) to differ among categories of endometritis; cows that had CLIN and CLINCYTO endometritis were four times less likely to conceive to the first insemination compared to UNAF cows. Cows that had CLIN (hazard ratio: 0.52) and CLINCYTO (hazard ratio: 0.40) endometritis had decreased likelihood of pregnancy at 150 DPP compared to UNAF cows. Similarly, cows diagnosed as having CLINCYTO endometritis had decreased likelihood (hazard ratio: 0.48) of pregnancy at 250 DPP compared to UNAF cows. The DMI and MY up to 5 weeks postpartum were not affected by categories of endometritis. In summary, categories of endometritis as determined at 25 DPP did not affect follicular growth and ovulation, DMI, or MY. However, the combined (CLINCYTO endometritis) category had a negative impact on first service conception rate and subsequent services.

Locked nucleic acid inhibits miR-92a-3p in human colorectal cancer, induces apoptosis and inhibits cell proliferation.

MicroRNAs (miRNAs) are a type of small noncoding RNAs that have a vital role in basic biological processes such as cellular growth, division and apoptosis. A change in the expression of miRNAs can induce many diseases. Recently, the role of miRNA in some of the cancers as a tumor suppressor and oncogene has been recognized. Several studies have proved that miR-92a-3p acts as an oncogene in colorectal cancer (CRC). We studied CRC by inhibiting miR-92a-3p in SW48 cells (human colorectal cancer cell line) that were transfected with locked nucleic acid (LNA). At different times, the expression level of miR-92a-3p, cell vitality, apoptosis and necrosis were studied by qRT-PCR, MTT, Annexin-V and propidiumiodide. Our results showed that the expression of miR-92a-3p and proliferation of SW48 cells were decreased, and also a high percentage of SW48 cells were exposed to apoptosis and necrosis (P⩽0.005). Our study showed that the inhibition of miR-92a-3p with LNA inhibited cell proliferation and induced apoptosis and necrosis in CRC.

Association of TIM-1 5383-5397ins/del and TIM-3 -1541C>T polymorphisms with multiple sclerosis in Isfahan population.

Multiple sclerosis (MS) is an organ-specific autoimmune disease in central nervous system, affecting about 2.5 million people around the world. Probable involvement of two newly identified immunoregulator molecules, TIM-1 and TIM-3, has been reported in autoimmune diseases. In this study, for the first time, the association of TIM-1 5383-5397ins/del and TIM-3 -1541C>T polymorphisms with MS in an Iranian population was considered. The results of our study showed that there is no significant association between TIM-1 5383-5397ins/del and MS (P = 0.38); however, the frequency of CT genotype of TIM-3 -1541C>T in patient group was significantly higher than the control group, and there was a significant association between CT genotype and MS (P = 0.009, OR = 4.08).

Effects of prepartum diets supplemented with rolled oilseeds on calf birth weight, postpartum health, feed intake, milk yield, and reproductive performance of dairy cows.

The objectives were to determine the effects of supplemental fat (no oilseed vs. oilseed) during late gestation and the source of fat (canola vs. sunflower seed), on dry matter intake (DMI), plasma metabolite concentrations, milk production and composition, calf birth weight, postpartum health disorders, ovarian function and reproductive performance in dairy cows. Pregnant Holstein cows, blocked by body condition and parity, were assigned to 1 of 3 diets containing rolled canola seed (high in oleic acid; n=43) or sunflower (high in linoleic acid; n=45) at 8% of dry matter, or no oilseed (control; n=43), for the last 35±2 d of pregnancy. After calving, all cows received a common lactation diet. Blood samples were collected at wk -3 (i.e., 2 wk after initiation of prepartum diets) and at wk +1, +2, +3, +4 and +5 postpartum to determine the concentration of fatty acids (mEq/dL), ß-hydroxybutyrate (mg/dL), and glucose (mg/dL). Ovarian ultrasonography was performed twice weekly to determine the first appearance of dominant (10mm) and preovulatory-size (≥16mm) follicles, and ovulation. Uterine inflammatory status based on the proportion of polymorphonuclear leukocytes (PMN; subclinical endometritis: >8% PMN) was assessed at d 25±1 postpartum. Significant parity by treatment interactions were observed for DMI and milk yield. Prepartum oilseed supplementation, more specifically sunflower seed supplementation, increased postpartum DMI in primiparous cows without affecting prepartum DMI or milk yield. Contrarily, in multiparous cows, prepartum oilseed supplementation decreased both prepartum and postpartum DMI and milk yield during the first 2 wk. Regardless of parity, prepartum feeding of canola reduced postpartum DMI compared with those fed sunflower. Mean fatty acids concentrations at wk -3 were greater in cows given supplemental oilseed than those fed no oilseeds. Gestation length and calf birth weight were increased in cows given supplemental oilseed prepartum compared with cows fed no oilseeds, and a disproportionate increase in the birth weight of female calves was evident in cows fed oilseed. Total reproductive disorders tended to be greater in cows fed supplemental oilseed than those fed no oilseed (42 vs. 23%). Furthermore, cows fed sunflower seed had greater incidences of dystocia (35 vs. 18%) and total health disorders (52 vs. 32%) than those fed canola seed. Added oilseed and type of oilseed did not affect uterine inflammation at 25±1 d postpartum. Oilseed supplementation did not alter the intervals from calving to establishment of the first dominant follicle, preovulatory-size follicle, and ovulation, nor did it affect fertility (conception rate to first artificial insemination and proportion of pregnant cows by 150 d after calving). In summary, prepartum oilseed supplementation (6.2 to 7.4% ether extract, % of dietary dry matter) decreased DMI during the entire experimental period (pre- and postpartum), decreased milk yield during early lactation in multiparous cows, and increased calf birth weight with no significant improvement in ovarian function and reproductive performance.

Short communication: Effects of prepartum diets supplemented with rolled oilseeds on Brix values and fatty acid profile of colostrum.

The objective of this study was to evaluate effects of oilseeds supplemented in prepartum diets on colostrum quality. Thirty-nine dry pregnant Holstein cows (14 primiparous and 25 multiparous cows) were blocked by body condition score and parity and assigned to 1 of 3 experimental diets containing rolled oilseeds at 8% of dietary dry matter (canola seed or sunflower seed) or no oilseed (control) at 35 d before the expected calving date. Canola seed is high in oleic acid and sunflower seed is high in linoleic acid content. Colostrum samples were collected at the first milking after calving, and concentrations of nutrient composition, fatty acid profile, and Brix value (an indicator IgG concentration) were determined. Cows fed sunflower seeds before calving produced colostrum with greater crude protein content (15.0 vs. 12.9%), colostral Brix values (24.3 vs. 20.3%), and conjugated linoleic acid concentration (18:2 cis-9,trans-11; 0.64 vs. 0.48%) compared with those fed canola seed. Positive effects of feeding sunflower seed might be mediated by ruminal metabolism of linoleic acid and subsequent enhanced production of conjugated linoleic acid. Oilseed supplementation in prepartum diets of dairy cows also altered fatty acid profile of colostrum in a way to reflect fatty acid profile of the supplemented oilseeds except for oleic acid. In conclusion, prepartum feeding of sunflower seed increased colostral Brix value, an indicator of colostral IgG concentration, compared with that of canola seed, but its mode of action and effects on health and productivity of calves need to be investigated.

PiggyBac as a novel vector in cancer gene therapy: current perspective.

Selection of suitable delivery system is one of the crucial aspects in gene therapy that determines the efficiency of gene therapy. The past two decades have witnessed extensive efforts for finding safe and efficient vectors to overcome the limitations of viral vectors. The utilization of DNA transposon-based vectors for gene therapy has emerged as a promising non-viral alternative. DNA 'cut-and-paste' is one of the main mechanisms of genome engineering by transposon elements. However, the lack of an efficient transposition system has limited the utilization of transposon vectors in mice and mammalian systems. PiggyBac (PB) is known as a highly efficient DNA transposon originally isolated from Trichoplusia ni as an alternative to Sleeping Beauty (SB). It has been shown that PB can be functional in various species including mammalian systems. This vector could overcome some limitations of other vectors in cancer gene therapy. Some advantages of PB include the capacity for integration into the genome and providing a stable expression, capacity to harbor 10 and 9.1 kb of foreign DNA into the host genome, without a significant reduction in their transposition activity and display non-overlapping targeting preferences. However, to advance PB to clinical applications, some obstacles still require to be overcome to improve its safety and efficiency. Hence, it seems that this vector could open new horizons in gene and cancer therapy.

High occurrence of potentially-pathogenic free-living amoebae in tap water and recreational water sources in South-West Iran.

The present study was conducted to evaluate the presence of potentially pathogenic free living amoebae in drinking and recreational water sources in south-western Iran. From 75 collected water samples, 40 samples (53.3%) were positive for free living amoebae identified using morphological tools. Interestingly, all recreational waters in Ilam city included in the present study were positive for Acanthamoeba, Vahlkampfidae and Vermamoeba. Thirty percent of tap water samples in Ahvaz city were also positive for potentially pathogenic Free Living Amoebae. Moreover, the three genera identified in the present study have been previously reported as keratitis causative agents in Iran. The present research highlights the need to improve filtration methods for tap waters and to establish awareness in recreational water sources in Iran, in order to prevent Free Living Amoebae related infections. To the best of our knowledge, this is the first report of the isolation of potentially pathogenic free living amoebae including Acanthamoebae, Vermamoebae and Vahlkampfiids in the South-West of Iran.

Blockage of miR-92a-3p with locked nucleic acid induces apoptosis and prevents cell proliferation in human acute megakaryoblastic leukemia.

MicroRNAs (miRNAs) are non-coding RNAs involved in post-transcriptional regulation of gene expression. In many cancers, up- or downregulation of different miRNAs is reported. In acute myeloid leukemia, upregulation of miR-92a-3p was reported in human in vitro studies. We performed blockage of miR-29a-3p in human acute megakaryoblastic leukemia cell line (M-07e) by using locked nucleic acid (LNA) and cell proliferation; apoptosis and necrosis were assessed. At different time points after LNA-anti-miR92a-3p transfection, miR-92a-3p quantitation was assessed by qRT-real-time PCR, MTT assay and annexin/propidium iodide staining were performed. The data were processed using the ANOVA test. At all three time points, the expression of miR-92a-3p was lower in the LNA-anti-miR group compared with the control groups. Cell viability between LNA-Anti-miR and the control group was statistically significant. Blockage of miR-92a-3p was associated with increment of the ratio of apoptotic cells in the LNA-anti-miR group was higher than the other group. The ratio of necrotic cells in the LNA-antimiR group was higher than the other groups. These assessments indicate that miR-92a-3p blockage can decrease the viability of M-07e cells, which is mainly due to induction of apoptosis and necrosis. Our findings could open up a path to a miRNA based therapeutic approach for treatment of acute megakaryoblastic leukemia.

High occurrence of potentially-pathogenic free-living amoebae in tap water and recreational water sources in South-West Iran

The present study was conducted to evaluate the presence of potentially pathogenic free living amoebae in drinking and recreational water sources in south-western Iran. From 75 collected water samples, 40 samples (53.3%) were positive for free living amoebae identified using morphological tools. Interestingly, all recreational waters in Ilam city included in the present study were positive for Acanthamoeba, Vahlkampfidae and Vermamoeba. Thirty percent of tap water samples in Ahvaz city were also positive for potentially pathogenic Free Living Amoebae. Moreover, the three genera identified in the present study have been previously reported as keratitis causative agents in Iran. The present research highlights the need to improve filtration methods for tap waters and to establish awareness in recreational water sources in Iran, in order to prevent Free Living Amoebae related infections. To the best of our knowledge, this is the first report of the isolation of potentially pathogenic free living amoebae including Acanthamoebae, Vermamoebae and Vahlkampfiids in the South-West of Iran.

[The HHEX rs1111875A/G gene polymorphism is associated with susceptibility to type 2 diabetes in the Iranian population].

The illuminating picture of genetic mechanisms underlying the development of type 2 diabetes (T2DM) includes differently accumulated genetic polymorphisms that increase the risk along with environmental factors. A number of single nucleotide polymorphisms (SNPs) are indicated to be linked with T2DM, but also conflicting results have been found. To examine the contribution of these polymorphisms in conferring susceptibility to T2DM, the association of HHEX rs1111875A/G and CDKN2A/B rs10811661C/T common gene polymorphisms with the risk of T2DM in an Iranian population was evaluated. In this study participated 140 patients and 140 controls. Genomic DNA was extracted from samples and genotyping of the polymorphisms was performed by the Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) technique. A significant association was found with the G allele (OR = 1.729, CI = 1.184-2.523, P = 0.004) and GG genotype (OR = 2.921, 95% CI = 1.789-4.771, P < 0.001) of the rs1111875A/G SNP for susceptibility to T2DM in the recessive model. Furthermore, compared with the GG genotype, individuals with the GA genotype had a lower risk to develop T2DM (OR = 0.237, 95% CI = 0.137-0.408, P< 0.001) in the additive model. In addition, an association between the polymorphism and BMI in regard to the risk of T2DM was identified. The genotype and allele frequencies of the rs10811661C/T polymorphism did not show a statistically significant association with T2DM in any genetic model. Our results show that the rs1111875A/G polymorphism is an important susceptibility polymorphism for the development of T2DM in the Iranian population. Also, these findings support that this polymorphism is a key genetic risk factor for the development of T2DM in multiple ethnic populations.

Association analysis of -416 G>C polymorphism of T-cell immunoglobulin and mucin domain-1 gene with asthma in Iran.

TIM (T-cell immunoglobulin (Ig) and mucin domain)-1, one of the members of TIM family, expresses on Th2 cells and promotes the production of Th2 signature cytokines. This can increase a series of responses in these cells which could be one of the causes of asthma or asthma-related phenotypes. The aim of this study was to investigate whether a TIM-1 promoter single nucleotide polymorphism (SNP), -416 G>C, is associated with asthma in Iranian population. In this case-control study, existence of the -416 G>C polymorphism was assessed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) in 300 patients with asthma (97 atopic, 203 nonatopic) and 309 healthy volunteers. Additionally, the relationship between these polymorphism genotypes and total serum IgE levels in this Iranian population was evaluated. We discovered a significant association between the -416 G>C polymorphism and atopic asthma susceptibility in the population, but this SNP showed no connection with nonatopic asthma (P < 0.05). However, our results showed significant relation between this polymorphism and serum IgE level (P < 0.05). Our results suggest that -416 G>C polymorphism in TIM-1 gene could be a predisposing factor for atopic asthma in Iranian population, and CC genotype of this SNP can be associated with increased level of IgE in patients with asthma in the same population.

Synthesis, characterization and pH-controllable methotrexate release from biocompatible polymer/silica nanocomposite for anticancer drug delivery.

The objective of this study was to develop pH-responsive silica nanoparticles by imidazole-based ionic liquid for controlled release of methotrexate. In this article, we synthesized pH-responsive cationic silica nanoparticles by graft copolymerization of vinyl functionalized silica nanoparticles and methacrylic acid (MAA) monomer. Imidazole-based ionic liquid (Im-IL) was verified by (1)HNMR and Fourier-transform infrared (FTIR) spectroscopy. The synthesized functionalized silica particles were characterized and confirmed by various technologies including the scanning electron microscopy (SEM), the infrared spectroscopy (IR) and the thermogravimetric analysis (TGA). SEM results reveal the uniformity in size/shape of silica particles. This nanosystem is modified for targeted delivery of an anticancer agent methotrexate. The nanocomposite-MTX complex was formed at physiological pH (7.4) due to the electrostatic interactions between anionic carboxylic group of MTX molecules and cationic rings in carrier, while, the release of which can be achieved through the cleavage of the nanocomposite-MTX complex by protonation of carboxyl groups in the MTX segment that are sensitive to variations in external pH at weak acidic conditions. FT-IR spectroscopy showed the presence of light interactions between the silicate silanols and the drug. MCF7 cells were incubated with the MTX-free nanocomposite and MTX-loaded nanocomposite at various concentrations for 24, 48 and 72 h, and the data showed that the nanocomposites themselves did not affect the growth of MCF7 cells. Antitumor activity of the MTX-loaded nanocomposites against the cells was kept over the whole experiment process. The results showed that the MTX could be released from the fibers without losing cytotoxicity.

Methylation pattern of SFRP1 promoter in stool sample is a potential marker for early detection of colorectal cancer.

BACKGROUND: To setup a non-invasive genetic screening method for colorectal cancer, we evaluated the promoter methylation status of secreted frizzled-related protein1 (sfrp1) in stool samples of colorectal cancer with respect to a series of healthy individuals, using methylation-specific polymerase chain reaction. MATERIALS AND METHODS: In stool samples from 25 patients with colorectal cancer and 25 healthy control subjects, isolated DNA was treated with sodium bisulfite and analyzed by methylation-specific polymerase chain reaction with primers specific for methylated or unmethylated promoter sequences of the SFRP1 gene. RESULT: Methylation of the SFRP1 promoter was present in the stool DNA of patients with colorectal cancer. A sensitivity of 52% and specificity of 92% were achieved in the detection of colorectal neoplasia. The difference in methylation status of the SFRP1 promoter between the patients with colorectal neoplasia and the control group was statistically highly significant (P = 0.006). CONCLUSIONS: The results indicate that this DNA stool test of methylation of the SFRP1 promoter is a sensitive and specific method. It is assumed that the test is potentially useful for the early detection of colorectal cancer.

Effect of rs6983267 polymorphism in the 8q24 region and rs4444903 polymorphism in EGF gene on the risk of sporadic colorectal cancer in Iranian population.

Colorectal cancer (CRC) is among the major causes of cancer-related morbidity, mortality, and human health problem worldwide. Single-nucleotide polymorphisms (SNPs) in different genes are reported to be effective in increased risk of CRC in different ethnic population. We conducted a case-control study in patients diagnosed with sporadic colorectal cancer (n = 115) and healthy controls based on colonoscopy evidences (n = 120).In this replicative study, we aimed to investigate the association of two previously reported polymorphisms, rs6983267 and rs4444903, with sporadic colorectal cancer in a subset of Iranian patients. Genotyping was performed via polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. A significant relation was found between rs6983267 variant in the 8q24 region and colorectal cancer. The distribution of G/G genotypes among sporadic CRC patients was more frequent than that in the control group (P value = 0.001). The frequency of the G allele in the colorectal cancer patient group was also higher than that in the control group (65% vs. 48%; P value = 0.001). Compared with GG genotype, individuals with G/T and T/T genotypes had lower risk to develop sporadic CRC (OR = 0.357, 95% CI = 0.201-0.635). For the rs4444903 SNP, no significant association (P value = 0.149) was found with colorectal cancer risk. In conclusion, our findings suggest that the 8q24 rs6983267 SNP may play a pivotal role in the development of sporadic CRC in Iranian population. Therefore, it may be included as a potential genetic susceptibility marker for sporadic CRC.

The anti-biofouling effect of Lactobacillus fermentum-derived biosurfactant against Streptococcus mutans.

Biofouling in the oral cavity often causes serious problems. The ability of Streptococcus mutans to synthesize extracellular glucans from sucrose using glucosyltransferases (gtfs) is vital for the initiation and progression of dental caries. Recently, it was demonstrated that some biological compounds, such as secondary metabolites of probiotic bacteria, have an anti-biofouling effect. In this study, S. mutans was investigated for the anti-biofouling effect of Lactobacillus fermentum (L.f.)-derived biosurfactant. It was hypothesized that two enzymes produced by S. mutans, glucosyltransferases B and C, would be inhibited by the L.f.-biosurfactant. When these two enzymes were inhibited, fewer biofilms (or none) were formed. RNA was extracted from a 48-h biofilm of S. mutans formed in the presence or absence of L.f. biosurfactant, and the gene expression level of gtfB/C was quantified using the real-time polymerase chain reaction (RT-PCR). L.f. biosurfactant showed substantial anti-biofouling activity because it reduced the process of attachment and biofilm production and also showed a reduction in gtfB/C gene expression (P value < 0.05).