Effect of lactobacillus crispatus and lactobacillus rhamnosus culture supernatants on expression of autophagy genes and hpv e6 and e7 oncogenes in the hela cell line

Objective: The aim of this study was to clarify the mechanism by which lactobacilli exert their cytotoxic effects on cervical cancer cells. In addition, we aimed to evaluate the effect of lactobacilli on the expression of human papilloma virus [HPV] onco-genes

Materials and Methods: In this experimental study, using quantitative real-time polymerase chain reaction [PCR], we analyzed the expression of CASP3 and three autophagy genes [ATG14, BECN1 and alpha 2 catalytic subunit of AMPK [PRKAA2]] along with HPV18 E6 and E7 genes in HeLa cells before and after treatment with Lactobacillus crispatus and Lactobacillus rhamnosus culture supernatants

Results: The expression of CASP3 and autophagy genes in HeLa cells was decreased after treatment with lactobacilli culture supernatants. However, this de-crease was not significant for PRKAA2 when compared with controls. In addition, expression of HPV E6 was significantly decreased after treatment with lactobacilli culture supernatants

Conclusion: Lactobacilli culture supernatants can decrease expression of ATG14 and BECN1 as well as the HPV E6 oncogene. It has been demonstrated that the main changes occurring during cervical carcinogenesis in cell machinery can be reversed by suppression of HPV oncogenes. Therefore, downregulation of HPV E6 by lactobacilli may have therapeutic potential for cervical cancer. As the role of autophagy in cancer is complicated, further work is required to clarify the link between downregulation of autophagy genes and antiproliferative effects exerted by lactobacilli

Mitochondrial copy number and d-loop variants in Pompe patients

Objective: Pompe disease is a rare neuromuscular genetic disorder and is classified into two forms of early and late-onset. Over the past two decades, mitochondrial abnormalities have been recognized as an important contributor to an array of neuromuscular diseases. We therefore aimed to compare mitochondrial copy number and mitochondrial displacement-loop sequence variation in infantile and adult Pompe patients

Materials and Methods: in this retrospective study, the mitochondrial D-loop sequence was analyzed by polymerase chain reaction [PCR] and direct sequencing to detect possible variation in 28 Pompe patients [17 infants and 11 adults]. Results were compared with 100 healthy controls and sequences of all individuals were compared with the Cambridge reference sequence. Real-time PCR was used to quantify mitochondrial DNA copy number

Results: among 59 variants identified, 37[62.71%] were present in the infant group, 14[23.333%] in the adult group and 8[13.333%] in both groups. Mitochondrial copy number in infant patients was lower than adults [P<0.05]. A significant frequency difference was seen between the two groups for 12 single nucleotide polymorphism [SNP]. A novel insertion [317-318 ins CCC] was observed in patients and six SNPs were identified as neutral variants in controls. There was an inverse association between mitochondrial copy number and D-loop variant number [r=0.54]

Conclusion: the 317-318 ins CCC was detected as a new mitochondrial variant in Pompe patients

In vitro therapeutic effects of low level laser at mrna level on the release of skin growth factors from fibroblasts in diabetic mice

Numerous in vitro reports suggest that Low Level Laser Therapy [LLLT] affects cellular processes by biostimulation, however most of them emphasize on using visible light lasers which have low penetration. The aim of this study was to determine the effect of infrared laser light [which is more useful in clinic because of its higher penetration] on secretion of Fibroblast Growth Factor [FGF], Platelet Derived Growth Factor [PDGF] and Vascular Endothelial Growth Factor [VEGF], as important growth factors in wound healing. Fibroblasts were extracted from the skin of 7 diabetic and 7 nondiabetic mice and cultured. Cell cultures of experimental group were irradiated with single dose of LLLT [energy density of 1 J/ cm[2]] using an 810 nm continuous wave laser and the control group was not irradiated. Secretion of growth factors by skin fibroblasts were quantified through real time polymerase chain reaction. Diabetic irradiated group showed significant increase in FGF [p=0.017] expression, although PDGF increased and VEGF decreased in both diabetic and nondiabetic irradiated groups, but these variations were not statistically significant. These results suggest that LLLT may play an important role in wound healing by stimulating the fibroblasts

miRNA-21 expression analysis in 35 colorectal cancer

Colorectal cancer is the third most common cancer in the world. Non-coding RNA especially miRNAs have important regulatory roles in cancer. miRNAs are small non coding RNA 21-23 nucleotides long which have different levels of expression between tumors and normal tissues. This study was designed to compare expression level of miRNA-21 between Iranian population colorectal cancer tissues and normal tissue. This case-control study has performed in medical genetics department of Tehran University of Medical Sciences from January to November 2013. We used 35 samples. The samples were isolated from tumor and adjacent normal tissues of colon. Thirty-five samples were divided into different groups according to cliniopathologic features including tumor size [>4 and <4 cm], metastasis [+ and -] and stage. After small RNA extraction from tissues by small RNA purification kit the quality and quantity of extracted RNA was determined using spectrophotometry. cDNAs were synthesized and real-time polymerase chain reaction carried out. Finally expression levels were statistically analyzed by LinRegPCR and REST software. miRNA-21 expression ratio in stages I, II and III were 1/804 and 4/574, respectively, the increase from stage III was statistically significant [P= 0.037]. The expression were also studied according to different clinicopathologic status of colon cancer, tumor size [>4 and <4 cm] and metastatic [+ and -], miRNA-21 over expressed in both groups, however the increase was not statistically significant. In this study, we found miR-21 over-expression in advanced stage in tumoral tissue comparing with normal adjacent tissue. This means perhaps in the future it would be possible to use miRNA-21 as an informative prognostic biomarker to guide for better treatment strategies for colorectal cancer patients. Our findings also indicate that miRNA-21 is a promising new molecular target for designing novel therapeutic strategies to control colorectal cancer

Effect of heavy metals on silencing of engineered long interspersed element-1 retrotransposon in nondividing neuroblastoma cell line

L1 retrotransposons are the most active mobile DNA elements in human genome. Unregulated L1 retrotransposition may have deleterious effect by disrupting vital genes and inducing genomic instabilities. Therefore, human cells control L1 elements by silencing their activities through epigenetic mechanisms. It has been shown that cell division and heavy metals stimulate the frequency of L1 activities. Removal of silencing by L1 motivators may restart L1 element functions. Here, we have proposed that weather neurotoxic environmental heavy metals [as L1 stimulating factors] have a role in removing L1 silencing and restating its activities in nondividing neuronal cells. L1-RP green fluorescent protein [GFP]-tagged knock-in human neuroblastoma clones were prepared. Single-cell clone was treated with mitomycin-c combined with nontoxic and toxic concentrations of iron [Fe], copper [Cu], and mercury [Hg]. Silencing status of engineered L1 elements in dividing and nondividing cells was determined through measuring the amount of GFP expressing cells with flow cytometry. The cytotoxic effect of mitomycin-c combined with metals was measured by MTT assay. Hg in nondividing cells and Fe, Cu, and Hg in dividing neuroblastoma cells could significantly remove L1 silencing. Also, mitomycin-c treatment did not have any effect on metal toxicity status in neuroblastoma cells.Totally, our findings have shown that cell division has a role in removing L1 silencing as well as L1 retrotransposition induced by environmental heavy metals. It has been also indicated that Hg at all concentrations could remove silencing of engineered L1 element regardless of cell cycle state

[Evaluating the pre-, peri- and post-natal conditions of neonates affected by multiple congenital anomalies: a brief report]

Neonatal deaths stand for almost two-thirds of all deaths occurring in infants under one year of age. Congenital anomalies are responsible for 24.5% of these cases forming a highly important issue for health policy-makers. We studied the pre-, peri- and post-natal conditions of 77 patients with multiple congenital anomalies [MCA] through genetic counseling at Several university Hospitals, in Tehran, Iran. The collected data were subsequently analyzed using SPSS software. The patients did not have a good prognosis, demonstrating the need for the diagnosis of such diseases early in pregnancy to be of utmost importance. We screened for trisomy and nuchal translucency, which the first showed a low risk and the second showed normal results in most cases. Establishment of standards for prenatal diagnosis of congenital anomalies and monitoring their implementation seem to be necessary for the reduction of deaths due to congenital anomalies and infant mortality rate [IMR]

[Medical ethics in clinical diagnostic laboratory]

Medical Laboratory as a basis for disease diagnosis has significant role in the care, control and prevention of communicable and uncommunicable diseases. With recent laboratory science progress, the importance of this knowledge becomes clearer for both patients and physicians. Precision, speed, and ethics along with using updated standard scientific methods, are desirable characteristics of a clinical laboratory. Active employees in the medical laboratory are committed to the principles and ethical behavior. Due to the importance of ethics in the laboratory, the World Health Organization and International standard Organization have provided guidelines. Data collection, patient consent, confidentiality, testing, reporting results, archiving documents, access to documents, organized financial arrangement, some specific requests such as autopsy and testing of AIDS are some of the important issues in ethical behavior in medical laboratories discussed in this paper. Laboratory experts and technicians can improve their services using such information

role of Ile3434Thr XRCC7 gene polymorphism in differentiated thyroid cancer risk in an Iranian population

The aim of this study was to understand any association between differentiated thyroid carcinoma [DTC] and Ile3434Thr XRCC7 gene polymorphism [GenBank accession number: rs7830743]. DTC is the most prevalent thyroid neoplasm, which includes papillary and follicular cell carcinoma. XRCC7 gene encodes a protein that functions in non-homologous end joining DNA repair pathway. Non-synonymous polymorphisms in this gene may alter DNA repair capacity of the cell and change the risk of developing cancers. DTC patients [n = 173] and cancer free individuals [n = 204] were enrolled in a case-control study. The Ile3434Thr polymorphic alleles were discriminated by using amplification refractory mutation system-PCR method. The frequencies of this single nucleotide polymorphism in case and control groups were compared. Also, risk ratio for developing DTC in dichotomized genotypes was estimated by multivariate logistic regression analysis. Dichotomized genotypes into those with and without the 3434Thr allele showed that this allele was associated with DTC [OR [odd ratio]: 1.89, 95% confidence interval [CI] = 1.29-2.79, P<0.001]. Also, TC genotype was significantly associated with increased risk of DTC [OR: 2.42, 95% CI = 1.55-3.81, P = 0.0001] in individuals carrying this genotype. Allele 3434Thr in XRCC7 gene might be associated with differentiated thyroid cancer risk. Further studies with larger samples are needed to verify these initial findings

[Detection of RET proto-oncogene Cys634Arg mutation, the cause of medullary thyroid carcinoma, in an Iranian child]

Medullary Thyroid Carcinoma [MTC] is hereditary in 25% of cases, and occurs primarily in an autosomal dominant manner. Thus, siblings and offspring of a patient diagnosed with a germline mutation are at a 50% risk for carrying the same mutation. Germline mutations of RET proto-oncogene is the known cause of almost all hereditary MTCs. Mutation analysis of RET is necessary for all apparently sporadic MTC patients. Cases Presentation: In this study, three siblings diagnosed with MTC in a family were analyzed for RET mutations. The result of direct DNA sequencing showed Cys634Arg point mutation in all 3 of them. Screening of their offspring and 4th sibling resulted in identification of a 3-year-old boy as a mutation carrier. According to general consensus, he is at second risk level, which qualifies him for prophylactic total thyroidectomy. Molecular analysis of RET proto-oncogene should be done in a routine manner to differentiate hereditary MTC patients from sporadic ones. In case of finding a germline mutation, all siblings and offspring of the mutation carrier should be screened and asymptomatic carriers referred to total prophylactic thyroidectomy

Diabetes mellitus following pituitary adenomectomy in euglycemic patients with acromegaly

Insulin resistance, impaired glucose tolerance and diabetes mellitus [DM] secondary to acromegaly generally improve following treatment of the underlying disease. In rare cases, in spite of normoglycaemia in the presence of active acromegaly, patients develop diabetes mellitus following pituitary adenomectomy to the extent of requiring insulin therapy. We report 7 patients who developed diabetes mellitus after pituitary adenomectomy. Regular measuring of blood glucose in post-pituitary surgery patients is recommended as a necessary step even in the previously normoglycaemics