Bosnian Journal of Basic Medical Sciences 2020-04-03T16:47:44+02:00 Faruk Skenderi Open Journal Systems <p>The BJBMS (Bosnian Journal of Basic Medical Sciences) is а premier venue for discoveries in basic and clinical biomedical science. The BJBMS was founded in 1998 and is published by the Association of Basic Medical Sciences, a nonprofit honor organization of physician-scientists.</p> <p>Broad readership and scope. The BJBMS reaches readers across a wide range of medical disciplines and sectors. The journal publishes basic and translational/clinical research submissions and reviews in all biomedical specialties, including Genetics and Molecular biology, Immunology, Microbiology, Pathology, Biochemistry, Pharmacology, Anatomy, Biomaterials, new and emerging research and diagnostic methods, new and emerging medical entities, and others.</p> Association of miRNA-145 with the occurrence and prognosis of hydrosalpinx-induced defective endometrial receptivity 2020-04-01T16:17:11+02:00 Qingli Wang Haiquan Ai Xia Li Haiqing Tian Bingxue Ning Meng Zhang Xiaolin La <p>MiR-145 is reported to facilitate inflammation and is also associated with unsuccessful embryonic implantation. Whether miR-145 mediates inflammatory response underlying hydrosalpinx-induced defective endometrial receptivity (ER) remains unclear, and this study attempted to clarify this point. Endometrium samples were collected from hydrosalpinx patients (case, n = 243) and patients with tubal patency/obstruction (control, n = 187). The peripheral blood samples of cases and controls were collected to determine the genotypes of miR-145 SNPs. The value of miR-145 expression in the diagnosis and prognostic estimation of hydrosalpinx was assessed using ROC curve and regression analysis, respectively. Lipopolysaccharide (LPS) cell model was established with endometrial cells, and cells were transfected with miR-145 mimic, inhibitor, or negative control. MiR-145 and cytokine levels were quantified by quantitative reverse transcription PCR or western blot. MiR-145 expression was significantly higher in hydrosalpinx compared to control group, and high miR-145 expression was significantly associated with moderate/severe tube lesion, high pulsatility index (&gt;1.06), and high resistance index (&gt;0.61) in hydrosalpinx patients. ROC curve analysis indicated that monitoring miR-145 expression may be useful for the diagnosis of hydrosalpinx (AUC = 0.704). A alleles of rs41291957 (G&gt;A) and rs353292 (G&gt;A) were significantly associated with an increased risk of hydrosalpinx compared to G allele (<em>p </em>&lt; 0.05), yet the mutant allele of rs353291 (A&gt;G) and rs4705343 (T&gt;C) significantly reduced susceptibility to hydrosalpinx compared to the wild type allele. Treatments with miR-145 mimic and LPS in endometrial cells significantly increased the levels of TGF-β1, TNF-α, IL-6, and IL-8 compared to negative control, while treatment with miR-145 inhibitor decreased the cytokine levels. In conclusion, abnormally expressed miR-145 may be involved in hydrosalpinx-induced ER defects by regulating inflammatory response.</p> 2020-04-01T12:46:29+02:00 Copyright (c) 2020 Qingli Wang, Haiquan Ai, Xia Li, Haiqing Tian, Bingxue Ning, Meng Zhang, Xiaolin La Immunohistochemical study of dental pulp cells with 3D collagen type I gel in demineralized dentin tubules in vivo 2020-03-31T16:38:10+02:00 Sultan Zeb Khan Sana Mirza Samina Karim Takashi Inoue Mohammed S. Bin-Shuwaish Laila Al Deeb Khold Al Ahdal Rana S. Al-Hamdan Ahmed M. Maawadh Fahim Vohra Tariq Abduljabbar <p>Dental pulp cells (DPCs) represent good candidates for the regeneration of dental tissue. This study aimed to evaluate the growth and differentiation potential of DPCs cultured inside demineralized dentin tubules <em>in vivo</em>. Six green fluorescent protein (GFP)-transgenic rats (body weight 100 g each) and thirty-two Sprague-Dawley (SD) male rats (body weight 250 g each) were used for DPC collection and dentin tubules preparation and transplantation, respectively. Third-passage DPCs with or without collagen gels were loaded into demineralized dentin tubules. Both types of grafts were transplanted into the rectus abdominis muscles of SD rats and were harvested after 21 days. The expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), osteopontin (OPN), nestin, and dentin sialoprotein (DSP) were analyzed by immunohistochemistry. Histological analysis showed that DPCs in the collagen gel formed an osteodentin-like hard tissue matrix after 21 days. Increased positive immunoreactivity for ALP, BSP, OPN, nestin, and DSP was observed in experimental groups compared with control. Our results demonstrate that DPCs in collagen gel inside demineralized dentin tubules show increased growth and differentiation.</p> 2020-03-27T18:46:00+01:00 Copyright (c) 2020 Sultan Zeb Khan, Sana Mirza, Samina Karim, Takashi Inoue, Mohammed S. Bin-Shuwaish, Laila Al Deeb, Khold Al Ahdal, Rana S. Al-Hamdan, Ahmed M. Maawadh, Fahim Vohra, Tariq Abduljabbar Endothelial loss during the surgical procedure in saphenous veins harvested by open and endoscopic techniques in coronary artery bypass surgery 2020-03-26T15:46:04+01:00 Aleksandra Milutinović Ruda Zorc-Pleskovič <p>The patency of the vein graft in coronary artery bypass grafting could be dependent on conventional (vsO) or endoscopic (vsE) harvesting and on the hypoxic damage of endothelial cells. We aimed to evaluate both surgical techniques according to endothelial loss that occurs in the time between harvesting and implantation. Twenty-six saphenous veins were divided into vsO (n = 16) and vsE (n = 10) group. Three samples were taken from each vein. The first sample was taken after removal, the second before implantation of the distal part, and the third before the implantation of the proximal part, and they were stained with HE, Movat, and immunohistochemically with CD31. A significant loss of endothelial cells within both groups was found at the time of implantation of the distal and the proximal part of the vein graft compared to the endothelial cells at the time of harvesting. There were no significant differences in the endothelial loss between vsE and vsO groups at the time of harvesting and at the time before the implantation of the distal part. A higher number of endothelial cells was found in vsE group compared to vsO group at the time just before the implantation of the proximal part. The comparison of the implanted portions of vsE and vsO grafts to mammary arteries revealed a significant loss of endothelial cells only in vsO graft. We conclude that, at the time of implantation, the endothelial layer of the vein graft harvested endoscopically is more preserved than harvested openly.</p> 2020-03-19T10:05:58+01:00 Copyright (c) 2020 Aleksandra Milutinović, Ruda Zorc-Pleskovič Can telomere length predict bone health? A review of current evidence 2020-03-19T05:10:57+01:00 Sok Kuan Wong Soelaiman Ima-Nirwana Kok-Yong Chin <p>Telomeres are repetitive DNA sequences located at the end of chromosomes that serve as a protective barrier against chromosomal deterioration during cell division. Approximately 50–200 base pairs of nucleotides are lost per cell division, and new repetitive nucleotides are added by enzyme telomerase allowing telomere maintenance. Telomere shortening has been proposed as an indicator for biological ageing, but its relationship with age-related osteoporosis is ambiguous. We summarize the current evidence on the relationship between telomere length and bone health in experimental and epidemiological studies, which serve as a scientific reference for the development of novel diagnostic markers of osteoporosis or novel therapeutics targeting telomere and telomerase of bone cells to treat osteoporosis.</p> 2020-03-10T14:56:29+01:00 Copyright (c) 2020 Sok Kuan Wong, Soelaiman Ima-Nirwana, Kok-Yong Chin Inflammation-related cytokines and their roles in gastroenteropancreatic neuroendocrine neoplasms 2020-03-11T14:16:33+01:00 Davorka Herman Mahečić Maja Cigrovski Berković Vanja Zjačić-Rotkvić Tamara Čačev Sanja Kapitanović Monika Ulamec <p>Proinflammatory counterworks are important at different stages of tumor development, particularly during invasion and metastasis. Immune cells and their signal molecules can influence all stages of tumor progression, as well as therapeutic intervention. Proinflammatory cytokines are known triggers of growth in gastroenteropancreatic neuroendocrine neoplasms (GEP-NENs). In this study, we explored the immunohistochemical expression of tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1β), IL-2, and IL-6 in tissues from 43 GEP-NEN patients with tumors of gastric, duodenal, ileal, appendical, and colonic origin. The immunohistochemical expression of TNF-α was increased in tumor groups with high proliferation rates (Ki67; <em>p</em> = 0.034), as well as in those with higher tumor grades (<em>p</em> = 0.05). Moreover, the immunohistochemical expression of TNF-α positively correlated with death outcomes (<em>p</em> = 0.016). Expression of IL-6, IL-1β, and IL-2 displayed similar immunohistochemical expression patterns regardless of Ki67, although the expression between the ILs differed. Most GEP-NENs had high levels of IL-6 and lower levels of IL-1β and IL-2. Although further comprehensive studies are required for a complete understanding of activated mechanisms in proinflammatory protumoral microenvironment of GEP-NENs, TNF-α is a potential marker in the prognosis of those tumors.</p> 2020-03-10T06:57:09+01:00 Copyright (c) 2020 Davorka Herman Mahečić, Maja Cigrovski Berković, Vanja Zjačić-Rotkvić, Tamara Čačev, Sanja Kapitanović, Monika Ulamec Asymptomatic neurotoxicity of amyloid β-peptides (Aβ1-42 and Aβ25-35) on mouse embryonic stem cell-derived neural cells 2020-03-10T12:25:22+01:00 Nur Izzati Mansor Carolindah Makena Ntimi Noraishah Mydin Abdul-Aziz King-Hwa Ling Aishah Adam Rozita Rosli Zurina Hassan Norshariza Nordin <p>One of the strategies in the establishment of <em>in vitro</em> oxidative stress models for neurodegenerative diseases, such as Alzheimer’s disease (AD), is to induce neurotoxicity by amyloid beta (Aβ) peptides in suitable neural cells. Presently, data on the neurotoxicity of Aβ in neural cells differentiated from stem cells are limited. In this study, we attempted to induce oxidative stress in transgenic 46C mouse embryonic stem cell-derived neurons via treatment with&nbsp;Ab peptides (Aβ<sub>1-42</sub> and Aβ<sub>25-35</sub>). 46C neural cells were generated by promoting the formation of multicellular aggregates, embryoid bodies (EBs) in the absence of leukemia inhibitory factor (LIF), followed by the addition of all-trans retinoic acid (ATRA) as the neural inducer.&nbsp;Mature neuronal cells were exposed to different concentrations of Aβ<sub>1-42</sub> and Aβ<sub>25-35</sub> for 24 h. Morphological changes, cell viability, and intracellular ROS production were assessed. We found that 100 µM Aβ<sub>1-42</sub> and 50 µM Aβ<sub>25-35</sub> only promoted 40% and 10%, respectively, of cell injury and death in the 46C-derived neuronal cells. Interestingly, treatment with each of the Aβ peptides resulted in a significant increase of intracellular ROS activity, as compared to untreated neurons. These findings indicate the potential of using neurons derived from stem cells and Aβ peptides in generating oxidative stress for the establishment of an <em>in vitro </em>AD model that could be useful for drug screening and natural product studies.</p> 2020-03-06T13:08:02+01:00 Copyright (c) 2020 Nur Izzati Mansor, Carolindah Makena Ntimi, Noraishah Mydin Abdul-Aziz, King-Hwa Ling, Aishah Adam, Rozita Rosli, Zurina Hassan, Norshariza Nordin Knockdown of TPT1-AS1 inhibits cell proliferation, cell cycle G1/S transition, and epithelial–mesenchymal transition in gastric cancer 2020-03-13T11:43:05+01:00 Jun Tang Fei Huang Hui Wang Feng Cheng Yaping Pi Juanjuan Zhao Zhihong Li <p>Long non-coding RNAs are considered to be critical regulators of tumor progression. Tumor protein translationally controlled 1 antisense RNA 1 (TPT1-AS1) was shown to have an oncogenic role in cervical and ovarian cancer. The clinical significance and biological function of TPT1-AS1 in gastric cancer (GC) are not clear. In this study, we analyzed the expression of TPT1-AS1 in GC tissues and cell lines and performed functional and mechanistic analysis of TPT1-AS1 effects on GC cell proliferation, migration, and invasion. TPT1-AS1 expression was determined in 76 pairs of GC tissues vs. matched adjacent normal tissues and in four GC cell lines (SGC-7901, AGS, BGC-823, and MGC-803) vs. GES-1 cell line by quantitative reverse transcription PCR. SGC-7901 and MGC-803 cells were transfected with small interfering RNA or scrambled negative control, and cell proliferation, colony formation, migration, invasion and cell cycle assays were performed. The expression of proteins involved in cell cycle progression and epithelial–mesenchymal transition was analyzed by Western blot. TPT1-AS1 expression was significantly higher in GC tissues and cell lines compared to controls. The overexpression of TPT1-AS1 was significantly correlated with TNM stage and lymph node metastasis, and it was associated with worse prognosis of GC patients according to the Kaplan–Meier survival analysis and Cox proportional hazard regression analysis. The knockdown of TPT1-AS1 significantly inhibited proliferation, cell cycle G1/S transition, migration, and invasion of SGC-7901 and MGC-803 cells. Moreover, TPT1-AS1 knockdown downregulated the expression of CDK4, cyclin D1, and vimentin and upregulated the expression of p21 and E-cadherin. Our findings suggest that TPT1-AS1 may be a promising therapeutic target in GC.</p> 2020-02-28T12:22:29+01:00 Copyright (c) 2020 Jun Tang, Fei Huang, Hui Wang, Feng Cheng, Yaping Pi, Juanjuan Zhao, Zhihong Li Long non-coding RNA PVT1 regulates the migration of hepatocellular carcinoma HepG2 cells via miR-3619-5p/MKL1 axis 2020-04-03T16:47:44+02:00 Hua Liu Yan Yin Ting Liu Yanying Gao Qing Ye Junqing Yan Fushuang Ha <p>Hepatocellular carcinoma (HCC) is the third most common malignant tumor of the digestive system. Plasma cell tumor heterotopic gene 1 (PVT1) is an intergenic long non-coding RNA that is aberrantly expressed in different cancers. Myocardin related transcription factor A (MKL1) is a transcriptional coactivator of serum response factor that has been shown to promote cancer cell migration and invasion. In this study, we investigated the relationship between PVT1 and MKL1 as a novel regulatory mechanism underlying HCC progression. We used HepG2 and Cos‑7 cell lines. Transfection experiments with miR-3619-5p mimics/inhibitor, PVT1, siRNA-PVT1, MKL1, or siRNA-MKL1 were performed. RNA and protein levels were analyzed by quantitative reverse transcription PCR and Western blot, respectively. Cell migration was assessed by Transwell assay. Luciferase assays, RNA-FISH, RNA immunoprecipitation, and chromatin immunoprecipitation assays were performed to confirm the interaction between PVT1, miR-3619-5p, and MKL1 in HCC cells. Overexpression of PVT1 was positively correlated with MKL1 upregulation, which promoted HepG2 cell migration. miR-3619-5p inhibited MKL1 expression in HCC cells by acting on its 3' UTR. Furthermore, PVT1 promoted MKL1 expression and migration in HCC cells by directly binding to miR-3619-5p. In a positive feedback loop, MKL1 could activate PVT1 transcription by binding to the CArG box in the promoter region. Our findings may provide a basis for the development of novel targeted therapies in HCC.</p> 2020-02-26T14:14:04+01:00 Copyright (c) 2020 Hua Liu, Yan Yin, Ting Liu, Yanying Gao, Qing Ye, Junqing Yan, Fushuang Ha Neural stem cell-conditioned medium ameliorates Aβ25–35-induced damage in SH-SY5Y cells by protecting mitochondrial function 2020-03-06T04:53:24+01:00 Guoyong Jia Zengyan Diao Ying Liu Congcong Sun Cuilan Wang <p>Inhibition of amyloid β (Aβ)-induced mitochondrial damage is considered crucial for reducing the pathological damage in Alzheimer’s disease (AD). We evaluated the effect of neural stem cell-conditioned medium (NSC-CDM) on Aβ<sub>25–35</sub>-induced damage in SH-SY5Y cells. An <em>in</em><em> vitro</em> model of AD was established by treating SH-SY5Y cells with 40 μM Aβ<sub>25–35</sub> for 24 h. SH-SY5Y cells were divided into control, Aβ<sub>25–35</sub> (40 μM), Aβ<sub>25–35</sub> (40 μM) + NSC-CDM, and Aβ<sub>25–35</sub> (40 μM) + neural stem cell-complete medium (NSC-CPM) groups. Cell viability was detected by CCK-8 assay. Apoptosis, reactive oxygen species (ROS) production, and mitochondrial membrane potential (MMP) were detected by flow cytometry. Malondialdehyde (MDA) content was detected by ELISA assay. Western blot analysis was used to detect cytochrome c release and apoptosis-related proteins. Transmission electron microscopy (TEM) was used to observe mitochondrial morphology. Cell viability significantly decreased and apoptosis significantly increased in SH-SY5Y cells treated with Aβ<sub>25–35</sub>, and both effects were rescued by NSC-CDM. In addition, NSC-CDM reduced ROS production and significantly inhibited the reduction of MMP caused by Aβ<sub>25–35</sub>. Furthermore, NSC-CDM ameliorated Aβ<sub>25–35</sub>-induced reduction in Bcl-2 expression levels and increased the expression levels of cytochrome c, caspase-9, caspase-3, and Bax. Moreover, Aβ<sub>25–35</sub> induced the destruction of mitochondrial ultrastructure and this effect was reversed by NSC-CDM. Collectively, our findings demonstrated the protective effect of NCS-CDM against Aβ<sub>25–35</sub>-induced SH-SY5Y cell damage and clarified the mechanism of action of Aβ<sub>25–35</sub> in terms of mitochondrial maintenance and mitochondria-associated apoptosis signaling pathways, thus providing a theoretical basis for the development of novel anti-AD treatments.</p> 2020-02-24T09:18:49+01:00 Copyright (c) 2020 Guoyong Jia, Zengyan Diao, Ying Liu, Congcong Sun, Cuilan Wang Neurological outcome in patients after successful resuscitation in out-of-hospital settings 2020-02-26T11:39:19+01:00 Martin Marinšek Andreja Sinkovič David Šuran <p>Neurological outcome is an important determinant of death in admitted survivors after out-of-hospital cardiac arrest (OHCA). Studies demonstrated several significant pre-hospital predictors of ischemic brain injury (time to resuscitation, time of resuscitation, and cause of OHCA). Our aim was to evaluate the relationship between post-resuscitation clinical parameters and neurological outcome in OHCA patients, when all recommended therapeutic strategies, including hypothermia, were on board. We retrospectively included consecutive 110 patients, admitted to medical ICU after successful resuscitation due to OHCA. Neurological outcome was defined by cerebral performance category (CPC) scale I-V. CPC categories I-II defined good neurological outcome and CPC categories III-V severe ischemic brain injury. Therapeutic mesures were aimed to achieve optimal circulation and oxygenation, early percutaneous coronary interventions (PCI) in acute coronary syndromes (ACS), and therapeutic hypothermia to improve survival and neurological outcome of OHCA patients. We observed good neurological outcome in 37.2% and severe ischemic brain injury in 62.7% of patients. Severe ischemic brain injury was associated significantly with known pre-hospital data (older age, cause of OHCA, and longer resuscitations), but also with increased admission lactate, in-hospital complications (involuntary muscular contractions/seizures, heart failure, cardiogenic shock, acute kidney injury, and mortality), inotropic and vasopressor support. Good neurological outcome was associated with early PCI, dual antiplatelet therapy, and better survival. We conclude that in OHCA patients, post-resuscitation early PCI and dual antiplatelet therapy in ACS were significantly associated with good neurological outcome, but severe ischemic brain injury was associated with several in-hospital complications and the need of vasopressor and inotropic support.</p> 2020-02-24T07:33:22+01:00 Copyright (c) 2020 Martin Marinšek, Andreja Sinkovič, David Šuran Laryngeal tuberculosis in renal transplant recipients: A case report and review of the literature 2020-02-22T13:47:08+01:00 Fabrizio Cialente Michele Grasso Massimo Ralli Marco de Vincentiis Antonio Minni Griselda Agolli Michele Dello Spedale Venti Mara Riminucci Alessandro Corsi Antonio Greco <p>Renal allograft recipients are at greater risk of developing tuberculosis than the general population. A woman with a kidney transplant was admitted to the emergency department with high temperature, dysphonia, odynophagia, and asthenia. The final diagnosis was laryngeal tuberculosis. Multidisciplinary collaboration enabled accurate diagnosis and successful treatment. Laryngeal tuberculosis should be considered in renal allograft recipients with hoarseness. A more rapid diagnosis of tuberculosis in renal transplant recipients is desirable when the site involved, such as the larynx, exhibits specific manifestations and the patient exhibits specific symptoms. In these cases, prognosis is excellent, and with adequate treatment, a complete recovery is often achieved.</p> 2020-02-21T08:26:51+01:00 Copyright (c) 2020 Fabrizio Cialente, Michele Grasso, Massimo Ralli, Marco de Vincentiis, Antonio Minni, Griselda Agolli, Michele Dello Spedale Venti, Mara Riminucci, Alessandro Corsi, Antonio Greco P2X7R as an independent prognostic indicator in gastric cancer 2020-04-02T08:11:15+02:00 Ilknur Calik Muhammet Calik Burcu Sarikaya Ibrahim Hanifi Ozercan Ramazan Arslan Gokhan Artas Adile Ferda Dagli <p>Gastric cancer (GC) is one of the foremost causes of cancer-related death around the world. The P2X7 receptor (P2X7R), a member of the P2X7R subfamily of P2 receptors, is a unique molecule that has been shown to affect tumor growth and progression as well as various inflammatory processes, including proliferation of T lymphocytes, release of cytokines, and production of free oxygen radicals. P2X7R has been established as a prognostic parameter in some cancers, and recently, it has been investigated in the development of new targeted therapies. In the present study, we aimed to investigate the prognostic value of P2X7R expression in GC. The expression profile of P2X7R was evaluated immunohistochemically in 156 paraffin-embedded human GC specimens. P2X7R expression was higher in patients with lymph node metastasis than in those without (<em>p </em>&lt; 0.001). P2X7R overexpression was closely related with tumor-infiltrating lymphocytes (TILs) (<em>p </em>= 0.001), vascular invasion (<em>p </em>= 0.006), depth of invasion (<em>p </em>&lt; 0.001), distant metastasis (<em>p </em>&lt; 0.001), and advanced tumor, node, metastasis stage (<em>p </em>&lt; 0.001). Moreover, univariate (hazard ratio [HR] 3.98; 95% confidence interval (CI) 1.89-11.82; <em>p </em>&lt; 0.001) and multivariate (HR 2.24; 95% CI 3.53-12.50; <em>p </em>&lt; 0.001) Cox regression analysis showed that upregulated P2X7R expression clearly correlated with worsened overall survival. In summary, our data revealed that P2X7R may serve as a reliable prognostic parameter and promising therapeutic target for GC.</p> 2020-02-19T06:10:55+01:00 Copyright (c) 2020 Ilknur Calik, Muhammet Calik, Burcu Sarikaya, Ibrahim Hanifi Ozercan, Ramazan Arslan, Gokhan Artas, Adile Ferda Dagli Post-operative functional neurological symptom disorder after anesthesia 2020-02-18T07:24:04+01:00 Ryan S D'Souza Matthew NP Vogt Edwin Rho <p>A rare manifestation during the post-anesthetic period may include the occurrence of functional neurological symptom disorder (FNSD). FNSD is described as neurological symptoms that are not consistently explained by neurological or medical conditions. We report a case series consisting of six patients who underwent a general anesthetic at a tertiary referral hospital and experienced FNSD in the immediate post-anesthetic period. Life-threatening causes were excluded based on benign physical exam findings and knowledge of past history. Five of six cases manifested with FNSD only in the immediate post-operative setting after exposure to anesthesia, and never otherwise experienced these symptoms during their normal daily lives. MEDLINE and Google Scholar were searched through October 2019 using a highly-sensitive search strategy and identified 38 published cases of post-anesthetic FNSD. Meta-analysis of pooled clinical data revealed that a significant proportion of patients were females (86%), reported a history of psychiatric illness (49%), reported a prior history of FNSD (53%), and underwent general anesthesia as the primary anesthetic (93%). The majority of patients were exposed to diagnostic studies (66% received radiographic tests and 52% received electroencephalogram) as well as pharmacologic therapy (57%). While no deaths occurred, many patients had unanticipated admission to the hospital (53%) or to the intensive care unit (25%). These data may help inform the anesthesia literature on presentation, risk factors, and treatment outcomes of FNSD in the context of anesthetic administration. We contemplate whether anesthetic agents may predispose a vulnerable brain to manifest with involuntary motor and sensory control seen in FNSD.</p> 2020-02-12T16:27:07+01:00 Copyright (c) 2020 Ryan S D'Souza, Matthew NP Vogt, Edwin Rho Interventions of a clinical pharmacist in a medical intensive care unit – a retrospective analysis 2020-02-15T14:18:48+01:00 Maja Cvikl Andreja Sinkovič <p>Several studies demonstrated a significant decrease in prescription errors, adverse drug events, treatment costs and improved patient outcomes, when a clinical pharmacist (CP) was a full member of a multidisciplinary team in the intensive care unit (ICU). Our aim was to evaluate the activities of a CP, included in a 12-bed medical ICU team of a university hospital in the course of several months. We conducted a retrospective analysis of all the CP’s interventions from March to November 2017, carried out and documented after reviewing and discussing patients’ medical data with the treating ICU physicians. We identified four main categories of CP’s interventions: pharmacotherapy adjustments to kidney function (PAKF category), drug-drug interactions (DDIs category), therapeutic monitoring of drugs with narrow therapeutic index (TDM category), and drug administration by the nasogastric tube (NGT category). During the study period, 533 patients were admitted to the medical ICU. The CP reviewed the medical data of 321 patients and suggested 307 interventions in 95 patients. There were 147 interventions of the PAKF category, 57 interventions of the TDM category, 30 interventions of the NGT category, and 22 interventions of the DDIs category. Fifty-one interventions were unspecified. The majority of all interventions (203/307) were related to antimicrobial drugs. ICU physicians completely accepted 80.2% of the CP’s suggestions. We observed that regular participation of the CP in the medical ICU team contributed to more individualized and improved pharmacological treatment of patients. Therefore, ICU teams should be encouraged to include CPs as regular team members.</p> 2020-02-10T15:37:41+01:00 Copyright (c) 2020 Maja Cvikl, Andreja Sinkovič Significance of chromogranin A and synaptophysin in pancreatic neuroendocrine tumors 2020-02-11T15:59:22+01:00 Tatsuo Tomita <p>The two most commonly used immunohistochemical markers for neuroendocrine cells and their tumors are chromogranin A (CgA) and synaptophysin (SPY). CgA is a marker for neuroendocrine secretory granules of four pancreatic hormones and gastrin while SPY is a marker for synaptic vesicles in neuroendocrine cells, which release classic neurotransmitters such as acetylcholine and others. CgA is involved in synthesis and secretion of peptide hormones through exocytosis while function of SPY is elusive. Thirty-five pancreatic neuroendocrine tumors (Pan-NETs) were studied, consisting of 14 insulinomas, 8 gastrinomas, 2 glucagonomas, 6 pancreatic polypeptidomas and 5 non-functioning tumors, and were immunostained for four pancreatic hormones, gastrin, CgA and SPY. Majority of Pan-NETs were less immunostained for the endocrine hormones and CgA than the normal pancreatic endocrine cells. CgA immunostaining mostly correlates with each hormone staining in non-β-cell tumors while SPY immunostaining recognizes endocrine cells diffusely in the cytoplasm. CgA immunostaining is less in insulinomas than in non-β-cell tumors, and CgA immunostaining may distinguish CgA-weaker insulinomas from CgA-stronger non-β-cell tumors. CgA immunostaining may be used as an independent marker for biological aggressiveness in non-β-cell Pan-NETs. The serum CgA levels are higher in subjects harboring non-β-cell tumors than those harboring insulinomas, and the serum CgA elevate in parallel to the increasing metastatic tumor mass. Thus, CgA positive immunostaining in Pan-NETs correlate with the elevated serum levels of CgA for diagnosing CgA-positive non-β-cell Pan-NETs and the increasing serum CgA levels indicate increasing metastatic tumor mass.</p> 2020-02-05T13:42:14+01:00 Copyright (c) 2020 Tatsuo Tomita Cisplatin inhibits the proliferation of Saos-2 osteosarcoma cells via the miR-376c/TGFA pathway 2020-02-05T16:04:21+01:00 Yuan Wang Yichao Wu Awei Cai Chengxiao Ma Shang Cai Hao Wang Yukang Que Shenglin Xu Tangbing Xu Yong Hu <p>Transforming growth factor alpha (<em>TGFA</em>) gene is involved in the proliferation and metastasis of various tumors, but its role in cell sensitivity to cisplatin chemotherapy is unclear. In this study, we investigated the mechanism underlying inhibitory effects of cisplatin on growth and proliferation of osteosarcoma cells. Osteosarcoma and normal skeletal muscle tissues were collected from 26 patients by biopsy. <em>TGFA</em> was silenced or overexpressed in Saos-2 osteosarcoma cells by transfection with <em>TGFA</em>-shRNA or <em>TGFA</em> ORF clone, respectively. MiR-376c was inhibited or overexpressed by transfection of Saos-2 cells with miR-376c sponge or miR-376c mimics, respectively. Cell growth was analyzed by MTT assay and cell proliferation by BrdU assay. MiR-376c and <em>TGFA</em> mRNA expression was detected by quantitative reverse transcription PCR and <em>TGFA</em> protein expression by Western blot. The target relationship between miR-376c and <em>TGFA</em> was assessed by luciferase reporter assay. Both in osteosarcoma tissues and Saos-2 cells, miR-376c expression was significantly decreased and <em>TGFA</em> mRNA expression was significantly increased compared with control. Transfection of Saos-2 cells with <em>TGFA</em>-shRNA silenced <em>TGFA</em> expression and significantly inhibited cell growth and proliferation. <em>TGFA</em> mRNA and protein expression in Saos-2 cells significantly decreased with increasing cisplatin concentrations (2.5–10 mg/L). Transfection with <em>TGFA</em> ORF clone reversed the inhibitory effects of cisplatin on Saos-2 cell proliferation. Compared with cisplatin (10 mg/L) treatment alone, the combined treatment with cisplatin and miR-376c mimics inhibited the proliferation of Saos-2 cells more significantly. MiR-376c suppressed <em>TGFA</em> expression by directly interacting with its 3'-UTR region. Overall, cisplatin inhibited the proliferation of Saos-2 cells by upregulating miR-376c and downregulating <em>TGFA</em> expression.</p> 2020-02-05T09:11:04+01:00 Copyright (c) 2020 Yuan Wang, Yichao Wu, Awei Cai, Chengxiao Ma, Shang Cai, Hao Wang, Yukang Que, Shenglin Xu, Tangbing Xu, Yong Hu Expression of the anti-Mullerian hormone, kisspeptin 1, and kisspeptin 1 receptor in polycystic ovary syndrome and controlled ovarian stimulation rat models 2020-03-30T22:55:16+02:00 Ali Risvanli Halis Ocal Necati Timurkaan Pinar Ipek Ibrahim Seker Burak Karabulut <p>Polycystic ovary syndrome represents a significant cause of female infertility. The aim of this study was to investigate the expression of anti-Mul­lerian hormone (AMH), kisspeptin 1 (KISS-1), and kisspeptin 1 receptor (KISS1r) in rat models of polycystic ovary syndrome (PCOS) and controlled ovarian stimulation (COS). For this purpose, 28 rats were assigned into four groups. Estrus and Diestrus groups consisted of rats in estrus and diestrus phases, respectively, while COS and PCOS groups consisted of rats with induced COS and PCOS, respectively. The serum AMH, KISS-1, and estradiol levels, and ovarian KISS1r levels were analyzed by enzyme-linked immunosorbent assay. Furthermore, histopathological analysis of the ovary tissue was done and ovarian KISS-1 expression was determined by immunohistochemical assay. The results revealed that ovarian KISS1r levels were higher in the Estrus (1271.43±51.98 pg/mL) and COS (1191.43±85.67 pg/mL) groups, compared to Diestrus and PCOS groups. The highest level of AMH was found in the Estrus group (16.91±2.12 ng/mL). The results indicate that AMH had no effect on the development of COS and PCOS, while KISS-1 was found to affect the development of COS in rats.</p> 2020-02-05T00:00:00+01:00 Copyright (c) 2019 The Authors Effects of epinephrine on heart rate variability and cytokines in a rat sepsis model 2020-03-30T22:58:47+02:00 Yun-Te Chang Wei-Chun Huang Chin-Chang Cheng Meng-Wei Ke Jung-Shun Tsai Yao-Min Hung Neng-Chyan Huang Mu-Shun Huang Shue-Ren Wann <p>Catecholamines have both anti-inflammatory and vasoactive properties. A decreased cardiac response to catecholamines has been associated with a high risk of death in sepsis and septic shock. The aim of this study was to investigate the effects of epinephrine (EPI) on heart rate variability (HRV) and autonomic balance, as well as cytokine levels, in a rat sepsis model. Thirty-six male Sprague-Dawley rats were assigned to 4 experimental groups and 2 control groups of 6 rats each. The rats in the experimental groups were inoculated with a lipopolysaccharide (LPS, endotoxin) to establish a sepsis model. Group A received only LPS; group B received LPS, antecedent EPI and the nonselective β-blocker propranolol; group C received LPS and antecedent EPI; and group D received LPS, antecedent EPI and the selective β1-blocker esmolol. One control group received EPI and the other received saline placebo. Heart rate variability (HRV) was analyzed and tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) levels were measured. Measurements were carried out at baseline, at 0 hour after EPI infusion, and at 0.5, 2, and 4 hours after LPS inoculation. There were significant differences in HRV and cytokine levels between the groups, indicating that LPS infusion caused autonomic imbalance. Antecedent EPI significantly decreased the level of TNF-α in group C compared with group A in which TNF-α level peaked at 2 hours and then declined. Propranolol (group B) but not esmolol (group D) administration resulted in elevated TNF-α levels, comparable to those observed in group A. In conclusion, antecedent administration of EPI in a rat sepsis model inhibits the production of TNF-α possibly via the β<sub>2</sub>-adrenoceptor.</p> 2020-02-05T00:00:00+01:00 Copyright (c) 2018 The Authors Interleukins and inflammatory markers are useful in predicting the severity of acute pancreatitis 2020-03-30T23:03:58+02:00 Davorin Branislav Ćeranić Milan Zorman Pavel Skok <p>Acute pancreatitis (AP) is a disease with significant morbidity and mortality. The aim of this study was to evaluate the predictive role of inflammatory markers, particularly interleukins (ILs), in the course of AP and to determine the frequency of etiologic factors of AP. We included patients with AP who were treated at our institution from May 1, 2012 to January 31, 2015. Different laboratory parameters, including ILs, and the severity scoring systems Ranson’s criteria and Bedside Index of Severity in Acute Pancreatitis (BISAP) were analyzed. AP was classified into mild and severe, and independent parameters were compared between these groups. The predictive performance of each parameter was evaluated using receiver operating characteristic (ROC) curves and the area under the ROC curve (AUC). A binomial logistic regression was performed to evaluate Ranson’s criteria and IL6, IL8, and IL10 (at admission and after 48 hours) in the course of AP. Overall, 96 patients were treated, 59 (61.5%) males and 37 (38.5%) females, average age 62.5 ± 16.8 years (range 22–91 years). The best predictor for the severity of AP was IL6, measured 48 hours after admission (AUC = 0.84). Other useful predictors of the severity of AP were lactate dehydrogenase (<em>p</em> &lt; 0.001), serum glucose (<em>p</em> &lt; 0.006), and difference in the platelet count (<em>p</em> &lt; 0.001) between admission and after 48 hours (<em>p</em> &lt; 0.001), hemoglobin (<em>p</em> &lt; 0.027) and erythrocytes (<em>p</em> &lt; 0.029). The major causes of AP were gallstones and alcohol consumption. According to our results, IL6 and Ranson score are important predictors of the severity of AP.</p> 2020-02-05T00:00:00+01:00 Copyright (c) 2019 The Authors Integrin beta 4 (ITGB4) and its tyrosine-1510 phosphorylation promote pancreatic tumorigenesis and regulate the MEK1-ERK1/2 signaling pathway 2020-03-30T23:05:25+02:00 Xiangli Meng Peng Liu Yunhao Wu Xinlu Liu Yinpeng Huang Boqiang Yu Jiahong Han Haoyi Jin Xiaodong Tan <p>Pancreatic cancer is the fourth leading cause of cancer death, with a 5-year survival rate of only 1–4%. Integrin-mediated cell adhesion is critical for the initiation, progression, and metastasis of cancer. In this study we investigated the role of integrin b4 (ITGB4) and its phosphorylation at tyrosine Y1510 (p-ITGB4-Y1510) in the tumorigenesis of pancreatic cancer. We analyzed the expression of ITGB4 and p-ITGB4-Y1510 in pancreatic cancer tissue and cell lines using immunohistochemistry, Western blot, or semi-quantitative reverse transcription PCR. ITGB4 and p-ITGB4-Y1510 were highly expressed in pancreatic cancer (n = 176) compared with normal pancreatic tissue (n = 171). High p-ITGB4-Y1510 expression correlated with local invasion and distant metastasis of pancreatic cancer, and high ITGB4 was significantly associated with poor survival of patients. Inhibition of ITGB4 by siRNA significantly reduced migration and invasion of PC-1.0 and AsPC-1 cells. Overexpression of the mutant ITGB4-Y1510A (a mutation of tyrosine to alanine at 1510 position) in PC-1.0 and AsPC-1 cells not only blocked the ITGB4 phosphorylation at Y1510 but also suppressed the expression of ITGB4 (<em>p</em> &lt; 0.05 vs. wild-type ITGB4). The transfection of PC-1.0 and AsPC-1 cells with ITGB4-Y1510A significantly decreased the level of p-mitogen-activated protein kinase kinase (MEK)1 (T292) and p-extracellular signal-regulated kinase (ERK)1/2 but did not affect the level of p-MEK1 (T386) and p-MEK2 (T394). Overall, our study showed that ITGB4 and its phosphorylated form promote cell migration and invasion in pancreatic cancer and that p-ITGB4-Y1510 regulates the downstream MEK1-ERK1/2 signaling cascades. Targeting ITGB4 or its phosphorylation at Y1510 may be a novel therapeutic option for pancreatic cancer.</p> 2020-02-05T00:00:00+01:00 Copyright (c) 2019 The Authors