MiR-339 and especially miR-766 reactivate the expression of tumor suppressor genes in colorectal cancer cell lines through DNA methyltransferase 3B gene inhibition

It is observed that upregulation of DNMT3B enzyme in some cancers, including colon cancer, could lead to silencing of tumor suppressor genes. MiR-339 and miR-766 have been predicted to target 3′UTR of DNMT3B gene. Luciferase reporter assay validated that individual and co-transfection of miR-766 and miR-339 into the HEK293T cell reduced luciferase activity to 26% ± 0.41%, 43% ± 0.42 and 64% ± 0.52%, respectively, compared to the control (P < 0.05). Furthermore, transduction of miR-339 and miR-766 expressing viruses into colon cancer cell lines (SW480 and HCT116) decreased DNMT3B expression (1.5, 3-fold) and (3, 4-fold), respectively. In addition, DNA methylation of some tumor suppressor genes decreased. Expression of these genes such as SFRP1 (2 and 1.6-fold), SFRP2 (0.07 and 4-fold), WIF1 (0.05 and 4-fold), and DKK2 (2 and 4-fold) increased in SW-339 and SW-766 cell lines; besides, expression increments for these genes in HCT-339 and HCT-766 cell lines were (2.8, 4-fold), (0.005, 1.5-fold), (1.7 and 3-fold) and (0.04, 1.7-fold), respectively. Also, while in SW-766, cell proliferation reduced to 2.8% and 21.7% after 24 and 48 hours, respectively, SW-339 showed no reduced proliferation. Meanwhile, HCT-766 and HCT-339 showed (3.5%, 12.8%) and (18.8%, 33.9%) reduced proliferation after 24 and 48 hours, respectively. Finally, targeting DNMT3B by these miRs, decreased methylation of tumor suppressor genes such as SFRP1, SFRP2, WIF1 and DKK2 in the mentioned cell lines, and returned the expression of these tumor suppressor genes which can contribute to lethal effect on colon cancer cells and reducing tumorigenicity of these cells.     

Gene expression comparison between primary estrogen receptor-positive and triple-negative breast cancer with paired axillary lymph node metastasis

The aim of this study is to characterize and compare changes in gene expression patterns of paired axillary lymph node (ALN) metastases from estrogen receptor (ER)-positive and triple-negative (TNBC) primary breast cancer (PBC). Patients with stage 2-3 PBC with macrometastasis to an ALN were selected. Gene expression of 2567 cancer-associated genes was analyzed with the HTG EdgeSeq system coupled with the Illumina Next Generation Sequencing (NGS) platform. Changes in gene expression between ER/PR-positive, HER2-negative PBC, and their paired ALN metastases were compared with TNBC and their paired ALN metastases. Fourteen pairs of ER-positive and paired ALN metastasis were analyzed. Compared with the PBC, ALN metastasis had 673 significant differentially expressed genes, including 348 upregulated genes and 325 downregulated genes. Seventeen pairs of TNBC and paired ALN metastasis were analyzed. ALN metastasis had 257 significant differentially expressed genes, including 123 upregulated genes and 134 downregulated genes. When gene expression of the ALN for ER-positive PBC was compared to that of TNBC, 97 genes were upregulated in both, and 115 genes were similarly downregulated. Common upregulated genes were associated with cell death, necrosis, and homeostasis. Common downregulated genes were those of migration, degradation of extracellular matrix, and invasion. Although ER-positive PBC and TNBC have a distinct gene expression profiles and distinct changes from PBC to ALN metastases, a significant number of genes are similarly up- or downregulated. Understanding the role of these common genomic changes may provide clues to understanding the metastatic process itself.     

Saving space: Comparing mini - logMAR with standard logMAR visual acuity

Purpose:Assessing visual acuity (VA) is the cornerstone of an ophthalmic workup and needs VA charts in a four or six meters space. The objective of this study was to compare the performance of distant VA (DVA) on one meter mini-logMAR (MLM) with a standard six meter logMAR (SLM) chart.Methods:We developed a MLM chart to be used at 1 m with +1.0 D spectacles, by reducing the SLM chart designed for 6 meters, to 1/6^th its size, using AutoCAD version 2014. On an initial cohort, we obtained DVA on the two charts by optometrist trainees, masked to the outcomes on the different tests. We performed regression and checked agreement between the two measurements. Subsequently, on a new cohort, we validated the performance of the MLM.Results:Of the 56 initial subjects, mean DVA with SLM was 0.44 ± 0.13 and with MLM was 0.45 ± 0.13; mean difference of -0.01 ± 0.02, 95%CI: 0.007 to 0.018; P < 0.0001 on paired t-test. There was a significant correlation: r = 0.99; r² = 0.98, P < 0.0001. On an average, DVA with MLM was less than a letter worse than with SLM. The regression formula obtained: SLM DVA = -0.1312 + 1.0014 x (MLM DVA). The validation study revealed no significant difference (P = 0.29) between the predicted standard DVA calculated by the regression formula and the actual standard.Conclusion:We suggest that we can deduce distance logMAR VA from a mini-logMAR chart as devised and used by us. This will take less space, be portable and allow congenial interaction with patients.     

Vulvar squamous cell carcinoma associated with Fanconi’s anemia

Fanconi’s anemia is a rare autosomal recessive disorder which is rarely associated with squamous cell carcinoma (SCC) of the vulva. We report a 23-year-old virgin female with Fanconi’s anemia and diabetes mellitus who presented with a history of 6-month ulcerative lesions of the vulva. Gynecologic examination disclosed a 1 × 2 cm ulcerated tumor lesion at the right labia minor near to the urethra. The biopsy showed a high-grade vulvar intraepithelial neoplasia (VIN III). She underwent wide local excision for this lesion. Pathologic examination of the surgically removed specimen revealed SCC of the vulva. Therefore, radical vulvectomy and bilateral inguino-femoral lymphadenectomy were performed. Due to the involvement of right inguinal lymph node, radiotherapy with a total dose of 45 Gy was delivered to mid-pelvis through antero-posterior/postero-anterior fields with 18 mV photon energies. Until her last follow-up about 1 year after the treatment, the patient was free of disease or any recurrence at the site of operation. Patients with Fanconi’s anemia have the risk of developing SCC of the genital tract. Radical vulvectomy and lymphadenectomy along with radiotherapy were associated with a satisfactory outcome in 1-year follow-up period in the presented patient.     

Mathematical beta function formulation for maxillary arch form prediction in normal occlusion population

The aim of this study was to assess the dental arch curvature in subjects with normal occlusion in an Iranian population and propose a beta function formula to predict maxillary arch form using the mandibular intermolar widths (IMW) and intermolar depths (IMD). The materials used were study casts of 54 adolescents with normal occlusion and mean age of 14.1 years (25 males, 29 females, age range 12–16 years). Curve-fitting analyses were carried out and the curves passing through the facial-axis point of the canines, premolars, first molars, and the incisal edges of the anterior teeth were studied using a 3D laser scanner. Using the measured IMW and IMD of the dental arches at the maxillary and mandibular first molar region, a beta function formula proposed for predicting maxillary arch form. The accuracy of the proposed formula was assessed on 10 randomly selected dental casts. The mean (SD) of the maxillary and mandibular IMW and IMD were 57.92 (4.75), 54.19 (5.31), and 31.59 (2.90) and 28.10 (2.59) mm, respectively. There was no gender dimorphism (P > 0.05) for both variables (IMW, IMD). There was a strong positive association (n = 10, Pearson r = 0.98, P < 0.05) between the measured (actual) maxillary arch length and proposed arch length derived from generated formula. The goodness of fit (whole arch) for the proposed beta function formula, using adjusted r square measure and root mean square in 10 patients averaged 0.97 and 1.49 mm, respectively. The corresponding figures for the maxillary anterior arch (canine to canine) were 0.90 and 0.92 mm, respectively. The proposed beta function formula used for predicting maxillary arch form based on two mandibular measures (IMW, IMD) was found to have a high accuracy for maxillary arch prediction in the Iranian population and may be used as a guide to fabricate customized arch wires or as an aid in maxillary reconstructive surgery.