Dual Suppressive Effect of miR-34a on the FOXM1/eEF2-Kinase Axis Regulates Triple-Negative Breast Cancer Growth and Invasion

Purpose: Recent studies indicated that dysregulation of noncoding KNAs (ncRNA) such as miRNAs is involved in pathogenesis of various human cancers. However, the molecular mechanisms underlying miR-34a are not fully understood in triple-negative breast cancer (TNBC). Experimental Design: We performed in vitro functional assays on TNBC cell lines to investigate the role of mi R-34a in FOLM1/eEF2K signaling axis. TNBC tumor xenograft models were used for in vivo therapeutic delivery of miR-34a. Results: In this study, we investigated the role of p53-driven ncRNA miR-34a and found that miR-34a is associated with significantly longer patient survival in TNBC and inversely correlated with levels of proto-oncogenic eEF2K, which was associated with significantly shorter overall patient survival, We showed that miR-34a directly binds to the 3'-untranslated region of eEF2K and FOXM1 mRNAs and suppresses their expression, leading to inhibition of TNBC cell proliferation, motility, and invasion. Notably, restoring miR-34a expression recapitulated the effects of inhibition of eEF2K and FOXM1, the transcription factor for eEF2K and the direct target of p53, in TNBC cell lines, whereas overexpression of eEF2K and FOXM1 rescued the effects and signaling pathways mediated by miR-34a. Moreover, in vivo therapeutic delivery of miR-34a nanopartides by systemic intravenous administration delayed tumor growth of two different orthotopic TNBC tumor xenograft models by inhibiting eEF2K and FOXM1, intratumoral proliferation and angiogenesis, and inducing apoptosis. Conclusions: Overall, our findings provide new insights into the tumor suppressor role of miR-34a by dual-targeting of FOXM1/eEF2K signaling axis and suggest that miR-34a-based gene therapy may be a potential therapeutic strategy in TNBC. (C)2018 AACR.NIH/NCIUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Cancer Institute (NCI) [R21CA199050, P30CA016672]; noncoding RNA center; NATIONAL CANCER INSTITUTEUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Cancer Institute (NCI) [P30CA016672] Funding Source: NIH RePORTERThis work was supported in part by grants from the NIH/NCI (R21CA199050 and P30CA016672) and the funding from noncoding RNA center and used the Functional Proteomics RPPA Core Facility

MicroRNA 603 acts as a tumor suppressor and inhibits triple-negative breast cancer tumorigenesis by targeting elongation factor 2 kinase

Triple negative breast cancer (TNBC) is an aggressive type of breast cancer characterized by the absence of defined molecular targets, including estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and is associated with high rates of relapse and distant metastasis despite surgery and adjuvant chemotherapy. The lack of effective targeted therapies for TNBC represents an unmet therapeutic challenge. Eukaryotic elongation factor 2 kinase (eEF2K) is an atypical calcium/calmodulin-dependent serine/threonine kinase that promotes TNBC tumorigenesis, progression, and drug resistance, representing a potential novel molecular target. However, the mechanisms regulating eEF2K expression are unknown. Here, we report that eEF2K protein expression is highly up-regulated in TNBC cells and patient tumors and it is associated with poor patient survival and clinical outcome. We found that loss/reduced expression of miR-603 leads to eEF2K overexpression in TNBC cell lines. Its expression results in inhibition of eEF2K by directly targeting the 3-UTR and the inhibition of tumor cell growth, migration and invasion in TNBC. In vivo therapeutic gene delivery of miR-603 into TNBC xenograft mouse models by systemic administration of miR-603-nanoparticles led to a significant inhibition of eEF2K expression and tumor growth, which was associated with decreased activity of the downstream targets of eEF2K, including Src, Akt, cyclin D1 and c-myc. Our findings suggest that miR-603 functions as a tumor suppressor and loss of miR-603 expression leads to increase in eEF2K expression and contributes to the growth, invasion, and progression of TNBC. Taken together, our data suggest that miR-603-based gene therapy is a potential strategy against TNBC

Gingival tissue human beta-defensin levels in relation to infection and inflammation.

Aim To profile gingival tissue levels of human beta-defensin (hBD)-2 and hBD-3 in relation to gingival inflammation, Th17-related cytokine concentrations, Porphyromonas gingivalis counts, and gingipain and total protease activities. Materials and Methods Gingival tissue and subgingival plaque samples were collected from 21 periodontitis patients including 48 periodontal pocket sites with marginal, mild, or moderate to severe inflammation. hBD levels were determined by immunodetection, P. gingivalis counts with real-time polymerase chain reaction, protease activities with fluorogenic substrates, and cytokine concentrations with Luminex technique. Data were statistically analysed using Kruskal-Wallis and Mann-Whitney U tests and Spearman correlation coefficients. Results Subgingival plaque counts of P. gingivalis (p = .001) and gingipain activity (p <.001), as well as interleukin (IL)-1 beta (p = .012), IL-10 (p = .024), IL-17A (p = .002), IL-17F (p = .006), and IL-23 (p = .036) concentrations were elevated in severely inflamed sites, whereas no change was observed in hBD-2 and hBD-3 levels. Negative correlations were found between protease activity and hBD-2 (p = .033) and hBD-3(p = .003) levels. Conclusions Shift in gingival inflammation from marginal to mild stage is related to elevations in subgingival plaque P. gingivalis counts and gingipain activity, but not to tissue hBD levels. Negative correlations between hBDs and total protease activity suggest the degradation of these antimicrobial peptides in progressed inflammation.Peer reviewe

Gingival Tissue Human Beta-Defensin Levels in Relation to Infection and Inflammation

AimTo profile gingival tissue levels of human beta‐defensin (hBD)‐2 and hBD‐3 in relation to gingival inflammation, Th17‐related cytokine concentrations, Porphyromonas gingivalis counts, and gingipain and total protease activities.Materials and MethodsGingival tissue and subgingival plaque samples were collected from 21 periodontitis patients including 48 periodontal pocket sites with marginal, mild, or moderate to severe inflammation. hBD levels were determined by immunodetection, P. gingivalis counts with real‐time polymerase chain reaction, protease activities with fluorogenic substrates, and cytokine concentrations with Luminex technique. Data were statistically analysed using Kruskal–Wallis and Mann–Whitney U tests and Spearman correlation coefficients.ResultsSubgingival plaque counts of P. gingivalis (p = .001) and gingipain activity (p p = .012), IL‐10 (p = .024), IL‐17A (p = .002), IL‐17F (p = .006), and IL‐23 (p = .036) concentrations were elevated in severely inflamed sites, whereas no change was observed in hBD‐2 and hBD‐3 levels. Negative correlations were found between protease activity and hBD‐2 (p = .033) and hBD‐3(p = .003) levels.ConclusionsShift in gingival inflammation from marginal to mild stage is related to elevations in subgingival plaque P. gingivalis counts and gingipain activity, but not to tissue hBD levels. Negative correlations between hBDs and total protease activity suggest the degradation of these antimicrobial peptides in progressed inflammation.</p

Assessment the effect of diabetes education on self-care behaviors and glycemic control in the Turkey Nursing Diabetes Education Evaluating Project (TURNUDEP): a multi-center study

Background: Diabetes education in Turkey is provided by diabetes nurse educators in almost all healthcare organizations. However, the education is not standardized in terms of learning content, duration, and methods. This multi-center study was performed to assess the self-care behaviors and glycemic control following education provided to the patients with type 2 diabetes mellitus by diabetes nurse educators. Methods: This was a descriptive and cross-sectional study and included 1535 patients admitted to 28 public hospitals for the treatment of type 2 diabetes mellitus. The education was assessed by using a Patient Identification Form and Self-care Scale. Results: The proportion of individuals who received diabetes education within the last year was 78.5%, with 46.7% of them having received it once. Of the patients, 84.8% reported that they received diabetes education individually. It was found that the proportion of individuals who received education about oral antidiabetics (78.5%) and glucose testing at home (78.5%) was higher than the proportion of individuals who received education about exercise (58.8%) and foot care (61.6%). The status of diabetes education, education intervals, and the correlation of the education method with self-care and glycemic control was evaluated. Self-care and glycemic control levels were better among the patients who received diabetes education thrice or more and in patients who received education both individually and in a group (p < 0.05). Conclusions: Approximately three-quarters of individuals with type 2 diabetes mellitus received education by diabetes nurse educators in Turkey. Diabetes education is positively correlated with self-care and glycemic control levels among patients with type 2 diabetes mellitus. Efforts for generalization and standardized education for all diabetes patients are necessary. © 2022, The Author(s)

BAKIR – PİRİNÇ BORU İKİLİ MALZEME GRUPLARININ SÜRTÜNME KARIŞTIRMA KAYNAK YÖNTEMİ İLE BİRLEŞTİRİLMESİ

BAKIR &ndash; PİRİN&Ccedil; BORU İKİLİ MALZEME GRUPLARININ S&Uuml;RT&Uuml;NME KARIŞTIRMA KAYNAK Y&Ouml;NTEMİ İLE BİRLEŞTİRİLMESİDemir t&uuml;r&uuml; ve demir dışı metallerin birleştirilmesinde kullanılan olduk&ccedil;a farklı kaynak y&ouml;ntemleri mevcut olup, genel halde bunlar katı hal, ergitme y&ouml;ntemi ve optik kaynak teknikleridir. Son yıllarda &uuml;zerinde &ccedil;alışmalar yapılan katı hal tekniklerinden biriside s&uuml;rt&uuml;nme karıştırma kaynak&nbsp; (SKK) y&ouml;ntemidir. Yapmış olduğumuz &ccedil;alışmada değişken parametreler olarak karıştırıcı ucun devir sayısı ve iş par&ccedil;asının &ccedil;evresel ilerleme hız değerleri değişken parametreler olarak se&ccedil;ilmiştir. Elde edilen birleştirmelere ve sertlik testi uygulanmış ve mikro yapılar optik mikroskopla ve SEM (Taramalı Elektron Mikroskobu) mikroskobunda incelenmiştir.&nbsp;Yapılan &ccedil;alışmada, pirin&ccedil; boru malzemesi ile bakır boru malzemesi kullanılmıştır. Kullanılan borular 60 mm dış &ccedil;ap, 54 i&ccedil; &ccedil;ap ve 100 mm uzunluğunda hazırlanmıştır. Kaynak sonrasında ise toplamda 200 mm uzunlukta numune &uuml;retilmiştir. Kaynak işlemi i&ccedil;in &uuml;niversal freze, ilgili ilave ekipmanları ve boruların &uuml;zerine rijit halde bağlanması i&ccedil;in &ouml;zel bağlama aparatı yapılmıştır. Borular bu aparatın &uuml;zerine sabitlendikten sonra, iş tablası &uuml;zerine bağlanarak kaynak işlemi ger&ccedil;ekleştirilmiştir.&nbsp;Kaynak işleminde değişken parametreler olarak, is par&ccedil;ası &ccedil;evresel ilerleme hızları i&ccedil;in; 1.35 mm/sn, 1.70 mm/sn ve 2.10 mm/sn değerleri kullanılmıştır. Genel olarak uygun kaynak dikişleri elde edilmiştir. Ancak en uygun kaynak dikişleri i&ccedil;in optimum devir 710 d/d ve &ccedil;evresel ilerleme hızı olarak da 1.35 mm/s olduğu g&ouml;r&uuml;lm&uuml;şt&uuml;r. Artan kaynak hızı ve devir hızında kaynak metalinde spiral izlerin oluştuğu ve bu izlerin aralıklarının y&uuml;ksek kaynak hızlarında arttığı g&ouml;r&uuml;lm&uuml;şt&uuml;r.&nbsp;Deneysel &ccedil;alışmalar sonunda artan kaynak hızına bağlı olarak kaynaklı b&ouml;lgelerin sertliğinin artışında&nbsp; kısmen etki ettiği anlaşılmıştır.THE JOINING OF BRASS AND COPER PIPE DUAL MATERIAL GROUPS VIA THE FRICTION-STIR WELDING METHODThere are various different welding methods used to join iron and non-iron metals including solid state, melting and optical welding methods. One of them is friction stir welding that is studied on it, recently. The main focus of this study is friction stir welding method which is one of the solid state welding methods, and a great number of studies and research have been carried out on this method in recent years.&nbsp;During the welding the rotation number of the tip of the mixer and the speed of the welding are chosen as changeable parameters. The resulting combinations undergo the rigidity test and their micro-structures are examined with microscopes and SEM.&nbsp;In this study brass and copper pipe materials were used. The pipes used are 100 millimetres in length, 60 millimetres in outer diameter and 54 millimetres in internal diameter. After the welding a sample which is 200 millimetres in length was produced. For the welding, universal freeze, necessary additional equipment and special tieing apparatus to be rigidly tied on the pipes were done. After the pipes were fixed on this apparatus, they were tied to platform for welding. For the welding as changeable parameters 1.35 &nbsp;mm/sec.,1.70mm/sec. and 2.10 mm/sec. were used for running piece enviromental progression speed. Generally, suitable welding stitches were obtained. However, it was observed that the optimum rotation is 710 d/d and progression speed is 1.35 mm/sec for the most suitable welding stitch.&nbsp;It has been observed that as a result of the increased welding speed and rotation speed, spiral marks occur on the welded materials and the intervals of these marks have increased at high welding speed. &nbsp;&nbsp;</p

FOXM1 regulates expression of eukaryotic elongation factor 2 kinase and promotes proliferation, invasion and tumorgenesis of human triple negative breast cancer cells

Eukaryotic elongation factor 2 kinase (eEF2K), an emerging molecular target for cancer therapy, contributes to cancer proliferation, cell survival, tumorigenesis, and invasion, disease progression and drug resistance. Although eEF2K is highly up-regulated in various cancers, the mechanism of gene regulation has not been elucidated. In this study, we examined the role of Forkhead Box M1 (FOXM1) proto-oncogenic transcription factor in triple negative breast cancer (TNBC) cells and the regulation of eEF2K. We found that FOXM1 is highly upregulated in TNBC and its knockdown by RNA interference (siRNA) significantly inhibited eEF2K expression and suppressed cell proliferation, colony formation, migration, invasion and induced apoptotic cell death, recapitulating the effects of eEF2K inhibition. Knockdown of FOXM1 inhibited regulators of cell cycle, migration/invasion and survival, including cyclin D1, Src and MAPK-ERK signaling pathways, respectively. We also demonstrated that FOXM1 (1B and 1C isoforms) directly binds to and transcriptionally regulates eEF2K gene expression by chromatin immunoprecipitation (ChIP) and luciferase gene reporter assays. Furthermore, in vivo inhibition of FOXM1 by liposomal siRNA-nanoparticles suppressed growth of MDA-MB-231 TNBC tumor xenografts in orthotopic models. In conclusion, our study provides the first evidence about the transcriptional regulation of eEF2K in TNBC and the role of FOXM1 in mediating breast cancer cell proliferation, survival, migration/invasion, progression and tumorgenesis and highlighting the potential of FOXM1/eEF2K axis as a molecular target in breast and other cancers

FOXM1 transcriptionally regulates expression of integrin beta 1 in triple-negative breast cancer

Triple-negative breast cancer (TNBC) is an aggressive type of breast cancer and associated with early metastasis, drug resistance, and poor patient survival. Fork head box M1 (FOXM1) is considered as an emerging molecular target due to its oncogenic role and high overexpression profile in 85% in TNBC. However, molecular mechanisms by which FOXM1 transcription factor mediate its oncogenic effects are not fully understood. Integrin beta 1 is often upregulated in invasive breast cancers and associated with poor clinical outcome and shorter overall patient survival in TNBC. However, the mechanisms regulating integrin beta 1 (ITGB1) gene expression have not been well elucidated