Mycobacterium tuberculosis (M. tuberculosis) is the causative agent of tuberculosis in human. One of the major M. tuberculosis virulence factors is early secretory antigenic target of 6-kDa (ESAT-6), and EccB5 protein encoded by eccB5 is one of its components. EccB5 protein is a transmembrane protein in ESX-5 system. The aim of this study is to explore the characteristics of wild-type EccB5 and its mutant form N426I. We expressed the EccB5 protein by cloning the mutant and wild-type eccB5 gene in Escherichia coli (E. coli). We compared the protein structure of wild type and mutant form of EccB5 and found changes in structure around Asn426 (loop structure) in wild type and around Ile426 (β-strand) in the mutant. The truncated recombinant protein of EccB5 was successfully cloned and expressed using plasmid pCold I in E. coli DH5α and E. coli strain Rosetta-gami B (DE3) and purified as a 38.6 kDa protein by using the affinity column. There was no detectable adenosine triphosphatase activity in truncated forms of EccB5 and its mutant. In conclusion, our study reveals successful cloning and protein expression of truncated form of eccB5 gene of M. tuberculosis. EccB5 protein in ESX-5 system may be an important membrane component involved in the transport machinery of type VII secretion system, which is essential for growth and virulence.
Ovarian carcinogenesis can be induced by a large number of somatic gene mutations. Circulating tumor DNA (ctDNA) released into peripheral blood can provide insights into the genomic landscape of cancer cells and monitor their dynamics. Our aim was to detect and compare the genetic profiles in tumor tissue and plasma before and after tumor resection in ovarian cancer patients. All three samples were collected from each patient. In this study, we used a commercial cancer panel to identify somatic mutations in 26 genes in seven selected patients through next-generation sequencing on the Illumina platform. Overall, 16 variants with pathogenic effect were identified in the TP53, PIK3CA, PTEN, APC, NRAS, KRAS, GNAS, and MET genes involved in important signaling pathways. The genetic alterations found in the presurgical plasma in six of seven ovarian cancer patients were no longer present in the plasma after tumor surgical removal. Identical variants in formalin-fixed paraffin embedded (FFPE) tissues and preoperative plasma specimens were observed in only two cases. These findings suggest that the detected presurgical pathogenic variants absent in postsurgery plasma are associated with the primary ovarian tumor. Finally, the low-identified concordance between FFPE and plasma can be due to various factors, but most likely to high tumor heterogeneity and low ctDNA level.
Osteoarthritis (OA) of the knee causes disability, pain, and progressive destruction of cartilage in adult women. The objective of the study was to evaluate the concentrations of the urinary biomarker C-terminal telopeptide of type II collagen (CTX-II) and pain by radiographic grade in women with knee OA in northeastern Mexico: Cross-sectional study of 155 women with knee OA. Concentrations of biochemical parameters were evaluated and urine samples were collected to measure biomarker levels (uCTX-II) ng/mmol by competitive enzyme-linked immunoabsorbent assay (ELISA) technique and the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scale was used for pain classification; median age of 49 years and 29.1 kg/m2 of body mass index (BMI). uCTX-II biomarker levels were grade 2 (210.7 ng/mmol), grade 3 (314.8 ng/mmol), and grade 4 (478.8 ng/mmol) relative to Kellgren and Lawrence, uCTX-II levels were compared with WOMAC scale and presented significant statistical difference (p = 0.0001). An association of the biomarker CTX-II and an increase in BMI was found in female patients with knee OA (odds ratio = 1.01; 95% confidence interval 1.001-1.005; p = 0.047).This study demonstrates an increase in the levels of the biomarker uCTX-II, the degree of pain, and radiographic grade in women with knee OA in northeastern Mexico.
Breast cancer is the major mortality cause of women worldwide. In the course of management of breast cancer, the identification of a biomarker is important in enhancing our knowledge on cancer pathology, predicting the response to treatment, and selecting the patients who are more favorable to receive certain treatments. These biomarkers have a prognostic value. In addition to traditional breast cancer prognosis factors such as the tumor size and grade, the axillary lymph node micrometastasis, and biomarkers such as HER2/neu, newly discovered biomarkers have been discovered. Some of these factors are genetic signature in tissue or in peripheral blood. Lipid profil, a simple and accessible biological examination, has been a novel path on the prediction of breast cancer risk of occurrence and recurrence in many studies. The main goal of our review is to evaluate lipid profile and breast cancer risk with an emphasis on the prognosis value of lipid profiles in breast cancer patient management.
Eight percent of women suffer from vulvodynia, a chronic pain condition with unknown etiology. Inflammation and dysregulation of estrogen signaling have been suggested to play a role in the pathogenesis of localized provoked vulvodynia (LPV). Therefore, the aim of the study was to analyze protein expression levels of estrogen-related receptors ERRα, ERRß, ERRγ, estrogen receptor (ERα), and progesterone receptor (PRα) and CD3-positive T cells in the vulvar vestibulum obtained from women suffering from LPV in comparison to healthy, unaffected controls. Vulvar vestibulum tissue specimens were obtained from LPV patients (n = 12) who had undergone modified posterior vestibulectomy and from 15 healthy controls. Protein expression of ERRα, ERRß, ERRγ, ERα, and PRα and CD3-positive T cells was analyzed by immunohistochemistry (IHC). Expression of ERRß was significantly more pronounced in samples from LPV compared to healthy controls (p = 0.006). No significant difference in the expression patterns of ERRα, ERRγ, ERα, PRα, or CD3 cells was detected. To our knowledge, this is the first study reporting ERR expression in normal vestibulum and in vestibulectomy samples from LPV patients. The higher level of ERRß expression detected by IHC may reflect dysregulation of estrogen signaling in LPV.
