Bosnian Journal of Basic Medical Sciences <p>The Bosnian Journal of Basic Medical Sciences (BJBMS) is an international, English-language, peer-reviewed journal, publishing articles in different disciplines of basic medical sciences. BJBMS welcomes original research and comprehensive reviews as well as short research communications in the field of molecular biology, biochemistry, genetics, immunology, microbiology, pathology, pharmacology, pharmaceutical sciences, physiology and translational research.</p> en-US <p>© Association of Basic Medical Sciences of FBIH.</p> (Faruk Skenderi) (Faruk Skenderi) Tue, 12 Feb 2019 23:38:59 +0100 OJS 60 Expression patterns and prognostic value of miR-210, miR-494, and miR-205 in middle-aged and old patients with sepsis-induced acute kidney injury <p>Septic patients suffer a ‘cytokine storm’ from proinflammatory cytokines, chemokines and other inflammatory mediators, resulting in acute kidney injury (AKI) and death. The purpose of the present study was to determine the expression patterns of microRNA-210 (miR-210), miR-494, and miR-205 in middle-aged and old patients with sepsis-induced AKI and to evaluate their association with patient prognosis. Serum blood urea nitrogen (BUN), creatinine (Cr) and cystatin C levels were determined in peripheral venous blood collected from 110 patients with sepsis-induced AKI and 110 healthy controls. The expression profile of miRNAs was analyzed by TaqMan low-density array (TLDA) in plasma samples from patients and controls. Association of miRNAs with prognosis and survival of patients was analyzed by Spearman’s rank correlation coefficient, Cox multivariate analysis, and ROC curve analysis. TILDA analysis showed 11 upregulated and 11 downregulated miRNAs in patients with sepsis-induced AKI. MiR-210 and miR-494 were the most upregulated and miR-205 was the most downregulated miRNA. High expression of miR-210 and miR-494 was positively correlated with BUN, Cr and cystatin C levels of patients, while low expression of miR-205 was negatively correlated. MiR-210 and miR-494 expression was significantly decreased and miR-205 expression was increased in survivors with sepsis-induced AKI (28-day survival, n = 68) vs. non-survivors (n = 42). BUN, Cr and miR-205 were independent risk factors for prognosis in sepsis-induced AKI. Our study showed the predictive value of miR-210, miR-494, and miR-205 in prognosis and survival of patients with sepsis-induced AKI. MiR-205 is an independent risk factor for sepsis-induced AKI and its decreased expression is associated with shorter patient survival.</p> Yongjun Lin, Ying Ding, Shuping Song, Man Li, Tao Wang, Feng Guo Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Thu, 18 Apr 2019 10:56:20 +0200 An exceptional group of non-small cell lung cancer difficult to diagnose: Evaluation of lipid-poor adrenal lesions <p>In some non-small cell lung cancer (NSCLC) patients, lipid-poor adrenal adenomas cannot be adequately differentiated from metastases using imaging methods. Invasive diagnostic procedures also have a low negative predictive value (NPV) in such cases. The current study aims to establish a specific and clinically practical metabolic parameter for lipid-poor adrenal lesions (ALs) in NSCLC patients. This diagnostic approach may prevent unnecessary abdominal enhanced computed tomography (CT), magnetic resonance imaging, or invasive diagnostic procedures. Sixty-four NSCLC patients with 69 lipid-poor ALs and 28 control patients with 30 benign lipid-poor ALs, who underwent FDG-PET/CT, were retrospectively reviewed. Two morphological and four metabolic parameters were analyzed in FDG-PET/CT images of NSCLC and control patients. Baseline and post-chemotherapy images of 64 NSCLC patients were re-evaluated according to the PERCIST 1.0. In cases where ALs could not be differentiated, follow-up FDG-PET/CT images were re-examined. The receiver operating characteristic (ROC) curve method was used for the evaluation of diagnostic parameters. Out of 69 ALs, 39 were determined as metastatic lesions (adrenal metastasis), while 30 lesions were considered non-metastatic (adrenal adenomas). The mean attenuation value, SUVmax AL/SUVmax primary tumor, SUVmax, SUVmax AL/liver, and SUVmax AL/SUVmean liver were significantly different between metastatic and benign ALs from NSCLC patients. The SUVmax AL/SUVmean liver ≥1.81 had the best positive (PPV, 94.3%) and negative (NPV, 82.4%) predictive values, and the highest specificity (93.3%), sensitivity (84.6%) and accuracy (86.9%). Lipid-poor ALs with SUVmax AL/SUVmean liver ≥1.81 can be accepted as malignant in NSCLC. However, if SUVmax AL/SUVmean liver is &lt;1.81, a pathologic examination is required. Utilizing this cut-off value to decide on adrenal core biopsy may prevent its unnecessary use. Moreover, this diagnostic approach can save time and reduce the healthcare costs.</p> Fikri Selcuk Simsek, Muhammet Arslan, Yusuf Dag Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Thu, 18 Apr 2019 10:51:00 +0200 Optogenetics: Therapeutic spark in neuropathic pain <p>Optogenetics is an emerging field, which uses light and molecular genetics to manipulate the activity of live cells by expressing light-sensitive proteins. With the discovery of bacteriorhodopsin, a light-sensitive bacterial protein, in 1971 Oesterhelt and Stoeckenius laid the pavement of optogenetics. However, the cross-integration of different disciplines is a little more than a decade old. The toolbox contains fluorescent sensors and optogenetic actuators which enable visualization of signaling events and manipulation of cellular activities, respectively. Neuropathic pain is a pain caused either by damage or disease that affects the somatosensory system. The exact mechanism for the neuropathic pain is not known, however proposed mechanisms include immune reactions, ion channel expressions, and inflammation. Current regimen for the disease provides about 50% relief for only 40-60% of patients. Recent <em>in vivo</em> and <em>in vitro</em> studies demonstrate the potential therapeutic applications of optogenetics by manipulating the activity of neurons. This review summarizes the basic concept, therapeutic applications for neuropathy and potential of optogenetics to reach from bench to bedside in the near future.</p> Kang Liu, Long Wang Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Thu, 18 Apr 2019 09:36:56 +0200 EP1 receptor is involved in prostaglandin E2-induced osteosarcoma growth <p>Recent studies showed that the activation of prostaglandin (PG) receptor EP1 promotes cell migration and invasion in different cancers. The aim of this study was to investigate the role of EP1 in the proliferation of osteosarcoma (OS) cells <em>in vitro</em> and <em>in vivo</em>. EP1 mRNA and protein levels were analyzed by real-time RT-PCR and Western blot, respectively in human OS cell lines MG63, OS732, U-2OS, HOS and SAOS-2 compared to human fetal osteoblastic hFOB 1.19 cells. MG63 cells were treated with PGE<sub>2</sub>, EP1 specific agonist 17-PT-PGE<sub>2</sub>, 17-PT-PGE<sub>2</sub> + EP1 specific antagonist SC51089, or DMSO (control). EP1R-siRNA or a non-silencing irrelevant RNA duplex (negative control) were used for the transfection of MG63 cells, followed by PGE<sub>2</sub> treatment. Nude mice carrying MG63 xenografts were treated with SC51089 (2 mg/kg/day). MG63 cells/xenografts were analyzed by MTT assay, TUNEL assay, PKC enzyme activity assay, and Western blot (EP1 and apoptotic proteins), and tumor growth/volume was evaluated in mice. EP1 levels were significantly higher in OS cells compared to osteoblasts. PGE<sub>2</sub> or 17-PT-PGE<sub>2</sub> treatment increased the proliferation and decreased the apoptosis of MG63 cells. Inhibition of EP1 by SC51089 or siRNA markedly decreased the viability of MG63 cells. Similarly, SC51089 treatment significantly inhibited MG63 cell proliferation and promoted apoptosis <em>in</em> <em>vivo</em>. The silencing of EP1 receptor by siRNA or blockade of EP1 signaling by SC51089 activated extrinsic and intrinsic apoptotic pathways both <em>in</em> <em>vivo</em> and <em>in</em> <em>vitro</em>, as evidenced by increased levels of Bax, cyt-c, cleaved caspase-3, caspase-8 and caspase-9. EP1 appears to be involved in PGE<sub>2</sub>-induced proliferative activity of MG63 cells. Antagonizing EP1 may provide a novel therapeutic approach to the treatment of OS.</p> Jing-cai Niu, Nan Ma, Wei Liu, Pei-ji Wang Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Thu, 18 Apr 2019 09:26:35 +0200 Age- and gender-independent association of glutathione S-transferase null polymorphisms with chronic myeloid leukemia <p>The glutathione S-transferase (GST) genes encode enzymes that mediate the detoxification of xenobiotics by catalyzing the conjugation of glutathione (GSH) to xenobiotic substrates. The aim of the current study is to investigate the association between <em>GSTT1</em> and <em>GSTM1</em> polymorphisms and chronic myeloid leukemia (CML) among Sudanese patients. Patients with CML (n = 115) were recruited to the study from the Radiation and Isotope Centre Khartoum (RICK)-Sudan. Healthy individuals (n = 104) were included as controls. Genotyping of <em>GSTT1</em> and <em>GSTM1</em> polymorphisms was performed using multiplex PCR. Null deletions in the <em>GSTT1</em> and <em>GSTM1</em> genes are common in the Sudanese population (control group), with frequencies of 33.9% and 38.2%, respectively. The frequencies of <em>GSTT1</em> (OR: 3.25, 95% CI: 1.87–5.65, <em>p</em> &lt; 0.001) and <em>GSTM1</em> (OR: 2.14, 95% CI: 1.25–3.67, <em>p</em> &lt; 0.005) null genotypes were significantly higher in CML patients vs. controls. The distribution of <em>GSTT1</em> and <em>GSTM1</em> null polymorphisms was not different between male and female (<em>p</em> &gt; 0.01) and young and old CML patients (<em>p</em> &gt; 0.05). Hematological parameters were not affected by null polymorphisms in the patient group (<em>p</em> &gt; 0.05). In addition, the frequency of <em>GSTM1</em> null polymorphism was lower in advanced-phase CML patients compared to chronic-phase patients (<em>p</em> &lt; 0.05). The <em>GSTT1</em> and <em>GSTM1</em> null polymorphisms are associated with CML among Sudanese patients, independently of their age and gender.</p> Abdel Rahim Mahmoud Muddathir, Elharam I. Abdallah, Omar Falah Khabour, Ream Elzain Abdelgader, Mahmoud Mohamed Elgari Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Mon, 15 Apr 2019 11:15:36 +0200 Valspodar-modulated chemotherapy in human ovarian cancer cells SK-OV-3 and MDAH-2774 <p>Overcoming drug resistance in ovarian cancer is the overarching goal in gynecologic oncology. One way to increase drug cytotoxicity without increasing the drug dose is to simultaneously apply multidrug resistance modulator. Valspodar is the second generation P-glycoprotein 1 modulator capable of reversing multidrug resistance in different cancers. In this study, we evaluated the effect of valspodar and cisplatin co-treatment on cell viability, cell death and oxidative status in ovarian cancer cells. The two cisplatin-resistant human ovarian cancer cell lines SK-OV-3 and MDAH-2774 were treated with cisplatin, valspodar, or cisplatin+valspodar for 24 and 48 hours. Untreated cells were used as control group. Cell viability was evaluated by MTT assay. Cell death was assessed by TUNEL and comet assay. Lipid peroxidation (malondialdehyde) and protein thiol groups were analyzed as oxidative stress markers. The expression of mitochondrial superoxide dismutase (MnSOD) was assessed by immunocytochemistry. Valspodar effectively reduced the resistance of SK-OV-3 cells to cisplatin, as demonstrated by increased oxidative stress, decreased cell viability and increased apoptosis in SK-OV-3 cells co-treated with valspodar and cisplatin compared to other groups. However, valspodar did not affect the resistance of MDAH-2774 cells to cisplatin. Stronger staining for MnSOD in MDAH-2774 vs. SK-OV-3 cells after co-treatment with cisplatin and valspodar may determine the resistance of MDAH-2774 cell line to cisplatin.</p> Maciej Zalewski, Julita Kulbacka, Jolanta Saczko, Małgorzata Drag-Zalesinska, Anna Choromanska Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Mon, 08 Apr 2019 10:21:21 +0200 Internodal HER2 heterogeneity of axillary lymph node metastases in breast cancer patients <p>Determination of human epidermal growth factor receptor 2 (HER2) status is important for adequate treatment of breast cancer (BC) patients. The novel HER2 gene protein assay (GPA) is particularly convenient, as it allows the simultaneous assessment of HER2 protein expression and gene amplification at individual cell level. Here we investigated the frequency of internodal HER2 heterogeneity in axillary lymph node macrometastases of BC patients and compared HER2 status between primary breast tumor and its metastases. We included a total of 41 female patients operated between 2014 and 2015 for primary BC with axillary lymph node macrometastases. Representative paraffin blocks of metastatic lymph nodes were sectioned and the slides were stained using the GPA in 38 BC cases. GPA results were assessed according to the ASCO/CAP 2013 criteria. We analyzed 12586 individual tumor cells, 120 cells per section of each metastatic lymph node. HER2 status differed between the primary tumor and its metastases in 5/38 cases (13.2%). In patients with at least two metastatic nodes, the HER2 status of lymph node metastases was only slightly different in 4/23 cases (17.4%). Our results indicate rare but substantial differences in HER2 status between primary breast tumor and its axillary lymph node metastases that may direct the choice and outcomes of targeted therapy in BC patients. The impact of the rare and subtle internodal HER2 heterogeneity evidenced in this study remains uncertain. Determining the HER2 status of lymph node metastases in BC seems to be rational, but assessing a limited number of metastatic nodes may be sufficient.</p> Ilija Vladimir Baroš, Nataša Tanasković, Ulrika Pellas, Živka Eri, Ljiljana Tadić Latinović, Tibor Tot Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 02 Apr 2019 14:54:06 +0200 Clear cell urothelial carcinoma of the urinary bladder - a rare pathological entity. A case report and a systematic review of the literature <p>The most common histological type of urinary bladder cancer is urothelial carcinoma (UC). In contrast, the clear cell variant of urothelial carcinoma (CCUC) is quite a rare neoplasm. In this study, we report a case of an 81-year-old male, presenting with gross hematuria and acute urinary retention, which was subsequently diagnosed with CCUC at our pathology department. Furthermore, we provide a short systematic review of the literature (PubMed, Scopus, Science Citation Index) for this rare histopathological entity and a brief discussion about its morphological and immunohistochemical (IHC) characteristics.</p> Ioana Mihai, Sorina Taban, Alin Cumpanas, Emilian Gh. Olteanu, Alis Dema Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Thu, 28 Mar 2019 09:55:41 +0100 Targeted immunotherapy with a checkpoint inhibitor in combination with chemotherapy: A new clinical paradigm in the treatment of triple-negative breast cancer <p>The treatment of several solid and hematologic malignancies with immune checkpoint inhibitors (against programmed death receptor-1/ligand-1 [PD-1/PD-L1]) has dramatically changed the cancer treatment paradigm. However, no checkpoint inhibitors were previously approved for the treatment of triple-negative breast cancer (TNBC), a difficult-to-treat disease with a high unmet therapeutic need. Based on IMpassion130 clinical trial (NCT02425891), the Food and Drug Administration (FDA) has recently granted an accelerated approval for atezolizumab (TECENTRIQ®), a monoclonal antibody drug targeting PD-L1, plus chemotherapy (Abraxane; nab®-Paclitaxel) for the treatment of adults with PD-L1-positive, unresectable, locally advanced or metastatic TNBC. The FDA has also approved the Ventana diagnostic antibody SP142 as a companion test for selecting TNBC patients for treatment with atezolizumab. In the present review, we briefly discuss the importance of this breakthrough as the first cancer immunotherapy regimen to be approved for the management of breast cancer.</p> Farhan S. Cyprian, Saghir Akhtar, Zoran Gatalica, Semir Vranic Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Wed, 27 Mar 2019 08:10:02 +0100 Histological and biochemical effects of Cinnamomum cassia nanoparticles in kidneys of diabetic Sprague-Dawley rats <p>This study investigated the antidiabetic activity of<em> Cinnamomum cassia</em> (<em>C. cassia</em>, Cc) silver nanoparticles (CcAgNPS) and effects of <em>C. cassia</em> on the kidneys of rats with induced type 2 diabetes. Twenty-four Sprague-Dawley rats weighing 250 ± 20 g were induced with diabetes by intraperitoneal injection of streptozotocin (STZ, 60 mg/kg). Animals were randomly assigned to one of four groups (n = 6) and treated for eight weeks with normal saline (control, group A), 5 mg/kg of CcAgNPs (group B), 10 mg/kg of CcAgNPs (group C), or 200 mg/kg of Cc (group D). Body weight and fasting blood glucose (FBG) was measured weekly and fortnightly, respectively. At the end of experiments animals were euthanized, blood and kidney tissue samples were collected for biochemistry (oxidative stress markers and renal function parameters) and kidneys were harvested for histology (PAS and H&amp;E staining). Body weight was significantly higher in group B and C vs. control (<em>p</em> &lt; 0.05), while no significant differences were observed in the kidney-to-body weight ratio between groups. FBG, glutathione, malondialdehyde, alanine aminotransferase, aspartate aminotransferase, serum urea and creatinine were significantly lower in group B, C and/or D vs. control (all <em>p</em> &lt; 0.05). In group A, severe distortion of the glomerular network was observed, marked by the loss of capsular integrity, thickened basement membrane, tubular cells with pyknotic nuclei, vacuolization, and interstitial space with infiltrations. These adverse effects were mitigated by 5 mg/kg and 10 mg/kg of CcAgNPs. Our study confirms structural and functional damage to kidneys caused by diabetes. CcAgNPs have a regenerative potential in diabetes-induced kidney damage and may be used as an antidiabetic agent.</p> Koffi Kouame, Aniekan Imo Peter, Edidiong Nnamso Akang, Roshila Moodley, Edwin Coleridge Naidu, Onyemaechi Okpara Azu Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Fri, 22 Mar 2019 10:55:49 +0100 Recombinant deoxyribonucleoside kinase from Drosophila melanogaster can improve gemcitabine based combined gene/chemotherapy for targeting cancer cells <p>A recombinant deoxyribonucleoside kinase from <em>Drosophila melanogaster</em> with a deletion of the last 20 amino acid residues (named <em>Dm</em>dNKΔC20) was hypothesized as a potential therapeutic tool for gene therapy due to its broad substrate specificity and better catalytic efficiency towards nucleosides and nucleoside analogs. This study was designed to evaluate the effect of <em>Dm</em>dNKΔC20 for sensitizing human cancer cell lines towards gemcitabine and to further investigate its role in reversal of acquired drug resistance in gemcitabine-resistant cancer cell line. The <em>DmdNKΔC20</em> gene was delivered to three different cancer cell lines, including breast, colon and liver cancer cells, using lipid-mediated transfection reagent. After transfection, gene expression of <em>DmdNKΔC20 </em>was confirmed by reverse transcription quantitative PCR (qRT-PCR) and the combined effect of <em>Dm</em>dNKΔC20 and gemcitabine based cytotoxicity was observed by cell viability assay. We further evolved a gemcitabine-resistant breast cancer cell line (named MCF7-R) through directed evolution in the laboratory, which showed 375-fold more resistance compared to parental MCF7 cells. Upon transfection with <em>DmdNKΔC20</em> gene, MCF7-R cells showed 83-fold higher sensitivity to gemcitabine compared to the control group of MCF7-R cells. Moreover, we observed 79% higher expression of p21 protein in transfected MCF7-R cells, which may indicate induction of apoptosis. Our findings highlight the importance and therapeutic potential of <em>DmdNKΔC20 </em>in combined gene/chemotherapy approach to target a wide range of cancers<em>,</em> particularly gemcitabine-resistant cancers.</p> Mahak Fatima, M. Mubasshar Iqbal Ahmed, Faiza Batool, Anjum Riaz, Moazzam Ali, Birgitte Munch-Petersen, Zeeshan Mutahir Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 19 Mar 2019 09:18:36 +0100 Epidemiology and mechanism of drug resistance of Mycoplasma pneumoniae in Beijing, China: A multi-center study <p><em>Mycoplasma pneumoniae</em> (<em>M. pneumoniae</em>) is one of the most common causes of community-acquired respiratory tract infections (RTIs). We aimed to investigate the prevalence of <em>M. pneumoniae </em>infection, antibiotic resistance and genetic diversity of <em>M. pneumoniae</em> isolates across multiple centers in Beijing, China. P1 protein was detected by Nested PCR to analyze the occurrence of <em>M. pneumoniae </em>in pediatric patients with RTI. <em>M. pneumoniae</em> isolates were cultured and analyzed by Nested-PCR to determine their genotypes. Broth microdilution method was used to determine the minimum inhibitory concentration (MIC) of antibiotics. Out of 822 children with RTI admitted to 11 hospitals in Beijing, 341 (41.48%) were positive for <em>M. pneumoniae</em> by Nested PCR and 236 (69.21%) samples had mutations in 23S rRNA domain V. The highest proportion of <em>M. pneumoniae </em>positive samples was observed in school-age children (118/190; 62.11%) and in pediatric patients with pneumonia (220/389; 56.56%). Out of 341 <em>M. pneumoniae</em> positive samples, 99 (12.04%) isolates were successfully cultured and the MIC values were determined for 65 <em>M. pneumoniae</em> strains. Out of these, 57 (87.69%) strains were resistant to macrolides, and all 65 strains were sensitive to tetracyclines or quinolones. <em>M. pneumoniae</em> P1 type I and P1 type II strains were found in 57/65 (87.69%) and 8/65 (12.31%) of cultured isolates, respectively. Overall, we demonstrated a high prevalence of <em>M. pneumoniae </em>infection and high macrolide resistance of<em> M. pneumoniae </em>strains in Beijing. School-age children were more susceptible to<em> M. pneumoniae</em>, particularly the children with pneumonia. Thus, establishment of a systematic surveillance program to fully understand the epidemiology of <em>M. pneumoniae</em> is critical for the standardized use of antibiotics in China.</p> Dong-Xing Guo, Wen-Juan Hu, Ran Wei, Hong Wang, Bao-Ping Xu, Wei Zhou, Shao-Jie Ma, Hui Huang, Xuan-Guang Qin, Yue Jiang, Xiao-Pei Dong, Xiao-Yan Fu, Da-Wei Shi, Liang-Yu Wang, A-Dong Shen, De-Li Xin Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Sun, 17 Mar 2019 21:06:48 +0100 Soft tissue grafts for dural reconstruction after meningioma surgery <p>The meninges are involved in various pathologies and are often directly or indirectly severed during surgical procedures, especially the dura mater. This can pose a real challenge for the surgeon, as a proper reconstruction of the meninges is important to prevent complications such as cerebrospinal fluid leak (CSF). A variety of techniques for dural reconstruction have been described, employing natural and artificial materials. A novel technique for dural reconstruction involves soft tissue grafts in the form of fibrous or fibromuscular flaps, which are placed on the dural defects to seal the gaps. These soft tissue grafts represent an appropriate scaffold for cell ingrowth and fibrosis, thus preventing CSF. In this pilot study, we described the application of soft tissue grafts for dural reconstruction in 10 patients who underwent convexity meningioma surgery.</p> Tomaz Velnar, Lidija Gradisnik Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Mon, 11 Mar 2019 11:50:29 +0100 Outcomes and prognostic factors for patients with cervical esophageal cancer undergoing definitive radiotherapy or chemoradiotherapy <p>Cervical esophageal cancer (CEC) is uncommon, accounting for less than 5% of all esophageal cancers. The management of CEC is controversial. This study investigated treatment outcomes and prognostic factors of survival in CEC patients undergoing definitive radiotherapy or concurrent chemoradiotherapy (CCRT). Ninety-one CEC patients were treated by intensity-modulated radiation therapy (IMRT) and three-dimensional conformal radiation therapy (3DCRT) between July 2007 and September 2017. The mean prescription dose was 64 Gy (range 54-70 Gy) delivered as 1.8-2.2 Gy per fraction per day, 5 days a week. Out of 91 patients, 34 received concurrent cisplatin-based chemotherapy (CT) including 18 patients who also received neoadjuvant CT. Overall survival (OS), locoregional failure-free survival (LRFFS), and progression-free survival (PFS) were estimated by the Kaplan–Meier method. Prognostic factors of survival were determined in univariate (log-rank test) and multivariate (Cox proportional hazard model) analysis. Treatment-related toxicity was also assessed. Median follow-up time for all patients was 19 months. Two-year OS, LRFFS and PFS of all patients were 58.2%, 52.5% and 48.1%, respectively. Clinical stage was an independent prognostic factor for OS (HR = 2.35, 95% CI: 1.03-5.37, <em>p</em> = 0.042), LRFFS (HR = 3.84, 95% CI: 1.38-10.69, <em>p</em> = 0.011), and PFS (HR = 2.68, 95% CI: 1.11-6.45, <em>p</em> = 0.028). Hoarseness was an independent prognostic factor for OS (HR = 2.10, 95% CI: 1.05-4.19, <em>p</em> = 0.036). CCRT was independently associated with better LRFFS (HR = 0.33, 95% CI: 0.14-0.79, <em>p</em> = 0.012). 3DCRT and IMRT with concurrent CT is well-tolerated and may improve local tumor control in CEC patients. Advanced clinical stage and hoarseness are adverse prognostic factors for OS, LRFFS, and PFS in CEC.</p> Xin-xin Du, Rong Yu, Zhen-fei Wang, De-cheng Du, Qiao-yun Liu, Run-mei Wang, Shi-rong Kang, Hao Yang Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Mon, 11 Mar 2019 10:04:00 +0100 Thymic stromal lymphopoietin levels are increased in patients with celiac disease <p>Thymic stromal lymphopoietin (TSLP) is a cytokine produced by epithelial cells in the lungs, skin and intestinal mucosa, and is involved in several physiological and pathological processes. In this study, we evaluated serum TSLP levels in patients with celiac disease (CD). The prospective study was conducted at a gastroenterology outpatient clinic between March 2018 and August 2018. Eighty-nine participants aged between 18 and 75 years were classified into following groups: 22 patients with newly diagnosed CD; 20 patients with CD who were compliant with a gluten-free diet (GFD); 32 patients with CD who were not compliant with a GFD; and 15 healthy controls. Demographic characteristics, disease duration, and selected biochemical and hematologic parameters were recorded and compared between groups. Median serum TSLP levels were 1193.65 pg/mL (range: 480.1–1547.1) in newly diagnosed CD patients, 110.25 pg/mL (range: 60.3–216.7) in CD patients who were compliant with a GFD, 113.1 pg/mL (range: 76.3–303.4) in CD patients who were not compliant with a GFD, and 57 pg/mL (range: 49–67.8) in healthy controls. Overall, there was a significant difference in serum TSLP levels between groups (<em>p</em> = 0.001). Patients with newly diagnosed CD had the highest serum TSLP levels. There was no significant difference in serum TSLP levels between patients with CD who were or were not compliant with a GFD. TSLP appears to be involved in the pathogenesis of CD. Further studies are required to determine if the TSLP signaling pathway can be used in the treatment of CD.</p> Evrim Kahramanoğlu Aksoy, Muhammet Yener Akpınar, Ferdane Pirinççi Sapmaz, Özlem Doğan, Metin Uzman, Yaşar Nazlıgül Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Fri, 01 Mar 2019 11:43:19 +0100 Upregulated expression of MNX1-AS1 long noncoding RNA predicts poor prognosis in gastric cancer <p>As important regulators of gene expression long noncoding RNAs (lncRNAs) are implicated in various physiological and pathological processes, including cancer. An oncogenic role of MNX1 antisense RNA 1 (MNX1-AS1) lncRNA has been suggested in cervical cancer and glioblastoma. In this study, we investigated the clinicopathological significance and biological function of MNX1-AS1 in gastric cancer (GC). The expression of MNX1-AS1 was analyzed by qRT-PCR in 96 GC and adjacent non-tumor tissues in relation to clinicopathological features and overall survival (OS) of patients, and in five human GC cell lines compared to a normal gastric epithelial cell line. Loss-of-function experiments using small interfering RNA (siRNA) targeting MNX1-AS1 (si-MNX1-AS1) were carried out in AGS and MGC-803 GC cell lines. Cell proliferation (CCK-8 assay), migration (Transwell) and invasion (Transwell Matrigel), and protein expression of proliferating cell nuclear antigen (PCNA), E-cadherin, N-cadherin, vimentin and matrix metallopeptidase 9 (MMP-9) were analyzed in transfected GC cells. Expression of MNX1-AS1 was significantly higher in GC vs. adjacent non-tumor tissues. Higher MNX1-AS1 expression was significantly associated with tumor size, TNM stage and lymph node metastasis. Kaplan–Meier analysis showed that GC patients with higher MNX1-AS1 expression had worse OS compared to patients with lower MNX1-AS1 expression. Multivariate analysis showed that MNX1-AS1 is an independent poor prognostic factor in GC. Knockdown of MNX1-AS1 significantly inhibited proliferation, migration and invasion of AGS and MGC-803 cells, and resulted in increased E-cadherin and decreased PCNA, N-cadherin, vimentin and MMP-9 expression. Taken together, these results suggest that MNX1-AS1 has an oncogenic function in GC and potential as a molecular target in GC therapy.</p> Wei Zhang, Lunhua Huang, Xinyang Lu, Kecheng Wang, Xiaofei Ning, Zhiqiang Liu Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 26 Feb 2019 02:17:58 +0100 Isoflurane exposure in infant rats acutely increases aquaporin 4 and does not cause neurocognitive impairment <p>Isoflurane is commonly used in pediatric population, but its mechanism of action in cognition is unclear. Aquaporin 4 (AQP4) regulates water content in blood, brain, and cerebrospinal fluid. Various studies have provided evidence for the role of AQP4 in synaptic plasticity and neurocognition. In this study, we aimed to determine whether a prolonged exposure to isoflurane in infant rats is associated with cognition and what effect this exposure has on AQP4 expression. Ten-day-old [postnatal day (P) 10] Wistar albino rats were randomly allocated to isoflurane group (n = 32; 1.5% isoflurane in 50% oxygen for 6 hours) or control group (n = 32; only 50% oxygen for 6 hours). Acute (P11) and long-term (P33) effects of 6-hour anesthetic isoflurane exposure on AQP4 expression were analyzed in whole brains of P11 and P33 rats by RT-qPCR and Western blot. Spatial learning and memory were assessed on P28 to P33 days by Morris Water Maze (MWM) test. The analysis revealed that isoflurane increased acutely both mRNA (~4.5 fold) and protein (~90%) levels of AQP4 in P11 rats compared to control group. The increasing levels of AQP4 in P11 were not observed in P33. Also, no statistically significant change between isoflurane and control groups was observed in the latency to find the platform during MWM training and probe trial. Our results indicate that a single exposure to isoflurane anesthesia does not influence cognition in infant rats. In this case, acutely increased AQP4 after isoflurane anesthesia may have a protective role in neurocognition.</p> Serdar Demirgan, Onat Akyol, Zeynep Temel, Aslıhan Şengelen, Murat Pekmez, Recep Demirgan, Mehmet Salih Sevdi, Kerem Erkalp, Ayşin Selcan Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 26 Feb 2019 01:37:41 +0100 Enhancement of bone consolidation using high-frequency pulsed electromagnetic fields (HF-PEMFs): An experimental study on rats <p><em>In vitro</em> studies showed that high-frequency pulsed electromagnetic fields (HF-PEMFs) increase the activity/expression of early and late osteogenic markers and enhance bone mineralization. The main aim of this study was to investigate the <em>in vivo</em> effects of HF-PEMFs on fracture healing using a rat model. A femur fracture was established by surgery in 20 male Wistar rats. Titanium nails were implanted to reduce and stabilize the fracture. After surgery, 20 rats were equally divided into untreated control and treated group (from the first postoperative day HF-PEMFs at 400 pulses/sec [pps] were applied for 10 minutes/day, for two weeks). Quantitative and qualitative assessment of bone formation was made at two and eight weeks following surgery and included morphological and histological analysis, serological analysis by ELISA, micro-computed tomography (micro-CT), and three-point bending test. At two weeks in HF-PEMF group, soft callus was at a more advanced fibrocartilaginous stage and the bone volume/total tissue volume (BV/TV) ratio in the callus area was significantly higher compared to control group (<em>p</em> = 0.047). Serum concentration of alkaline phosphatase (ALP) and osteocalcin (OC) was significantly higher in HF-PEMF group (ALP <em>p</em> = 0.026, OC <em>p</em> = 0.006) as well as the mechanical strength of femurs (<em>p</em> = 0.03). At eight weeks, femurs from HF-PEMF group had a completely formed woven bone with dense trabeculae, active bone marrow, and had a significantly higher BV/TV ratio compared to control (<em>p</em> = 0.01). HF-PEMFs applied from the first postoperative day, 10 minutes/day for two weeks, enhance bone consolidation in rats, especially in the early phase of fracture healing.</p> Daniel Oltean-Dan, Gabriela Bombonica Dogaru, Dragos Apostu, Alexandru Mester, Horea Rares Ciprian Benea, Mihai Gheorghe Paiusan, Catalin Ovidiu Popa, Elena Mihaela Jianu, Gyorgy Istvan Bodizs, Cristian Berce, Alina Mihaela Toader, Gheorghe Tomoaia Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Wed, 20 Feb 2019 13:51:44 +0100 Vascular endothelial growth factor (VEGF)-related polymorphisms rs10738760 and rs6921438 are not risk factors for proliferative diabetic retinopathy (PDR) in patients with type 2 diabetes mellitus (T2DM) <p>Vascular endothelial growth factor (VEGF) is an important regulator of angiogenesis and has been investigated as a candidate gene in a number of conditions, including diabetes and its microvascular complications (e.g., retinopathy and nephropathy). Several <em>VEGF</em>-related polymorphisms have been shown to contribute to nearly half of the variability in circulating VEGF levels in healthy individuals. Our aim was to assess the association between <em>VEGF</em>-related rs10738760 and rs6921438 polymorphisms and proliferative diabetic retinopathy (PDR) in Slovenian patients with type 2 diabetes mellitus (T2DM). We also investigated the effect of these polymorphisms on VEGF receptor 2 (VEGFR-2) expression in fibrovascular membranes (FVMs) from patients with PDR. This case-control study enrolled 505 unrelated patients with T2DM: 143 diabetic patients with PDR as a study group, and 362 patients with T2DM of &gt;10 years duration and with no clinical signs of PDR as a control group. Patient clinical and laboratory data were obtained from their medical records. rs10738760 and rs6921438 polymorphisms were genotyped using TaqMan SNP Genotyping assay. VEGFR-2 expression was assessed by immunohistochemistry in 20 FVMs from patients with PDR, and numerical areal density of VEGFR-2-positive cells was calculated. The occurrence of PDR was 1.7 times higher in diabetic patients carrying GA genotype of rs6921438 compared to patients with GG genotype, with a borderline statistical significance (OR = 1.7, 95% CI = 1.00 – 2.86, <em>p</em> = 0.05). In addition, A allele of rs6921438 was associated with increased VEGFR-2 expression in FVMs from PDR patients. However, we observed no association between AA genotype of rs6921438 nor between rs10738760 variants and PDR, indicating that the two polymorphisms are not genetic risk factors for PDR.</p> Jana Sajovic, Ines Cilenšek, Sara Mankoč, Špela Tajnšek, Tanja Kunej, Daniel Petrovič, Mojca Globočnik Petrovič Copyright (c) 2018 Bosnian Journal of Basic Medical Sciences Tue, 12 Feb 2019 00:00:00 +0100 The association of early postoperative lactate levels with morbidity after elective major abdominal surgery <p>Lactate levels are widely used as an indicator of outcome in critically ill patients. We investigated the prognostic value of postoperative lactate levels for postoperative complications (POCs), mortality and length of hospital stay after elective major abdominal surgery. A total of 195 patients were prospectively evaluated. Lactate levels were assessed on admission to the intensive care unit (ICU) [L<sub>0</sub>], at 4 hours (L<sub>4</sub>), 12 hours (L<sub>12</sub>), and 24 hours (L<sub>24</sub>) after the operation. Demographic and perioperative clinical data were collected. Patients were monitored for complications until discharge or death. Receiver operating characteristic (ROC) curves were used to determine the predictive value of lactate levels for postoperative outcomes. The best cut-off lactate values were calculated to differentiate between patients with and without complications, and outcomes in patients with lactate levels above and below the cut-off thresholds were compared. Univariate and multivariate analyses were used to identify variables associated with POCs and mortality. Seventy-six patients developed 184 complications (18 deaths), while 119 had no complications. Serum lactate levels were higher in patients with complications at all time points compared to those without complications (<em>p</em> &lt; 0.001). L<sub>12</sub> had the highest predictive value for complications (AUROC<sub>12</sub> = 0.787; 95% CI: 0.719–0.854; <em>p</em> &lt; 0.001) and mortality (AUROC<sub>12</sub> = 0.872; 95% CI: 0.794–0.950; <em>p</em> &lt; 0.001). The best L<sub>12 </sub>cut-off value for complications and mortality was 1.35 mmol/l and 1.85 mmol/l, respectively. Multivariate analysis revealed that L<sub>12</sub> ≥ 1.35 mmol/l was an independent predictor of postoperative morbidity (OR 2.58; 95% CI 1.27–5.24, <em>p</em> = 0.001). L<sub>24 </sub>was predictive of POCs after major abdominal surgery. L<sub>12</sub> had the best power to discriminate between patients with and without POCs and was associated with a longer hospital stay.</p> Jelena Veličković, Ivan Palibrk, Biljana Miličić, Dejan Veličković, Bojan Jovanović, Goran Rakić, Milorad Petrović, Vesna Bumbaširević Copyright (c) 2018 Bosnian Journal of Basic Medical Sciences Tue, 12 Feb 2019 00:00:00 +0100