DNA Repair

Characterization of rare NEIL1 variants found in East Asian populations Publication date: July 2019 Source: DNA Repair, Volume 79 Author(s): Irina G. Minko, Vladimir L. Vartanian, Naoto N. Tozaki, Oskar K. Linde, Pawel Jaruga, Sanem Hosbas Coskun, Erdem Coskun, Chunfeng Qu, Huan He, Chungui Xu, Taoyang Chen, Qianqian Song, Yuchen Jiao, Michael P. Stone, Martin Egli, Miral Dizdaroglu, Amanda K. McCullough, R. Stephen Lloyd Abstract The combination of chronic dietary exposure to the fungal toxin, aflatoxin B1 (AFB1), and hepatitis B viral (HBV) infection is associated with an increased risk for early onset hepatocellular carcinomas (HCCs). An in-depth knowledge of the mechanisms driving carcinogenesis is critical for the identification of genetic risk factors affecting the susceptibility of individuals who are HBV infected and AFB1 exposed. AFB1-induced mutagenesis is characterized by G to T transversions. Hence, the DNA repair pathways that function on AFB1-induced DNA adducts or base damage from HBV-induced inflammation are anticipated to have a strong role in limiting carcinogenesis. These pathways define the mutagenic burden in the target tissues and ultimately limit cellular progression to cancer. Murine data have demonstrated that NEIL1 in the DNA base excision repair pathway was significantly more important than nucleotide excision repair relative to elevated risk for induction of HCCs. These data suggest that deficiencies in NEIL1 could contribute to the initiation of HCCs in humans. To investigate this hypothesis, publicly-available data on variant alleles of NEIL1 were analyzed and compared with genome sequencing data from HCC tissues derived from individuals residing in Qidong County (China). Three variant alleles were identified and the corresponding A51V, P68H, and G245R enzymes were characterized for glycosylase activity on genomic DNA containing a spectrum of oxidatively-induced base damage and an oligodeoxynucleotide containing a site-specific AFB1-formamidopyrimidine guanine adduct. Although the efficiency of the P68H variant was modestly decreased, the A51V and G245R variants showed nearly wild-type activities. Consistent with biochemical findings, molecular modeling of these variants demonstrated only slight local structural alterations. However, A51V was highly temperature sensitive suggesting that its biological activity would be greatly reduced. Overall, these studies have direct human health relevance pertaining to genetic risk factors and biochemical pathways previously not recognized as germane to induction of HCCs.

Regulation of GLI1 by cis DNA elements and epigenetic marks Publication date: July 2019 Source: DNA Repair, Volume 79 Author(s): Robert Taylor, Jun Long, Joon Won Yoon, Ronnie Childs, Kathrine B. Sylvestersen, Michael L. Nielsen, King-Fu Leong, Stephen Iannaccone, David O. Walterhouse, David J. Robbins, Philip Iannaccone Abstract GLI1 is one of three transcription factors (GLI1, GLI2 and GLI3) that mediate the Hedgehog signal transduction pathway and play important roles in normal development. GLI1 and GLI2 form a positive-feedback loop and function as human oncogenes. The mouse and human GLI1 genes have untranslated 5′ exons and large introns 5′ of the translational start. Here we show that Sonic Hedgehog (SHH) stimulates occupancy in the introns by H3K27ac, H3K4me3 and the histone reader protein BRD4. H3K27ac and H3K4me3 occupancy is not significantly changed by removing BRD4 from the human intron and transcription start site (TSS) region. We identified six GLI binding sites (GBS) in the first intron of the human GLI1 gene that are in regions of high sequence conservation among mammals. GLI1 and GLI2 bind all of the GBS in vitro. Elimination of GBS1 and 4 attenuates transcriptional activation by GLI1. Elimination of GBS1, 2, and 4 attenuates transcriptional activation by GLI2. Eliminating all sites essentially eliminates reporter gene activation. Further, GLI1 binds the histone variant H2A.Z. These results suggest that GLI1 and GLI2 can regulate GLI1 expression through protein-protein interactions involving complexes of transcription factors, histone variants, and reader proteins in the regulatory intron of the GLI1 gene. GLI1 acting in trans on the GLI1 intron provides a mechanism for GLI1 positive feedback and auto-regulation. Understanding the combinatorial protein landscape in this locus will be important to interrupting the GLI positive feedback loop and providing new therapeutic approaches to cancers associated with GLI1 overexpression. Graphical abstract

DNA-PKc deficiency drives pre-malignant transformation by reducing DNA repair capacity in concert with reprogramming the epigenome in human bronchial epithelial cells Publication date: July 2019 Source: DNA Repair, Volume 79 Author(s): Ivo Teneng, Maria A. Picchi, Shuguang Leng, Christopher P. Dagucon, Suresh Ramalingam, Carmen S. Tellez, Steven A. Belinsky Abstract The expression of DNA-dependent protein kinase catalytic subunit (DNA-PKc) is highly variable in smokers and reduced enzyme activity has been associated with risk for lung cancer. An in vitro model of lung pre-malignancy was used to evaluate the role of double-strand break DNA repair capacity in transformation of hTERT/CDK4 immortalized human bronchial epithelial cells (HBECs) and reprograming of the epigenome. Here we show that knockdown of DNA-PKc to levels simulating haploinsufficiency dramatically reduced DNA repair capacity following challenge with bleomycin and significantly increased transformation efficiency of HBEC lines exposed weekly for 12 weeks to this radiomimetic. Transformed HBEC lines with wild type or knockdown of DNA-PKc showed altered expression of more than 1,000 genes linked to major cell regulatory pathways involved in lung cancer. While lung cancer driver mutations were not detected in transformed clones, more than 300 genes that showed reduced expression associated with promoter methylation in transformed clones or predictive for methylation in malignant tumors were identified. These studies support reduced DNA repair capacity as a key factor in the initiation and clonal expansion of pre-neoplastic cells and double-strand break DNA damage as causal for epigenetic mediated silencing of many lung cancer-associated genes. The fact that DNA damage, repair, and epigenetic silencing of genes are causal for many other cancers that include colon and prostate extends the generalizability and impact of these findings.

Transcriptional responses to DNA damage Publication date: July 2019 Source: DNA Repair, Volume 79 Author(s): Erica Silva, Trey Ideker Abstract In response to the threat of DNA damage, cells exhibit a dramatic and multi-factorial response spanning from transcriptional changes to protein modifications, collectively known as the DNA damage response (DDR). Here, we review the literature surrounding the transcriptional response to DNA damage. We review differences in observed transcriptional responses as a function of cell cycle stage and emphasize the importance of experimental design in these transcriptional response studies. We additionally consider topics including structural challenges in the transcriptional response to DNA damage as well as the connection between transcription and protein abundance.

Acetylation of Werner protein at K1127 and K1117 is important for nuclear trafficking and DNA repair Publication date: July 2019 Source: DNA Repair, Volume 79 Author(s): Deblina Ghosh, Vilhelm A. Bohr, Parimal Karmakar Abstract Werner syndrome is a rare autosomal recessive disorder where Werner (WRN) gene is mutated. Being a nucleolar protein, during DNA damage, WRN translocates at the damage site where its catalytic function is required in DNA repair. Several studies have indicated that WRN acetylation may modulate WRN trafficking and catalytic function (Blander et al., 2002; Lozada et al., 2014). Among the six acetylation sites in WRN protein identified by mass-spectrometry analysis (Li et al., 2010) we here explore the role of acetylation sites in C-terminal of WRN (K1127, K1117, K1389, K1413) because the C- terminal domain is the hub for protein- protein interaction and DNA binding activity (Brosh et al. [4]; Muftuoglu et al., 2008; Huang et al., 2006). To explore their functional activity, we created mutations in these sites by changing the acetylation residue lysine (K) to a non-acetylation residue arginine (R) and expressed them in WRN mutant cell lines. We observed that K1127R and K1117R mutants are sensitive to the DNA damaging agents etoposide and mitomycin C and display deficient DNA repair. Importantly, deacetylation of WRN by SIRT1 (Mammalian Sir2) is necessary for restoration of WRN localization at nucleoli after completion of DNA repair. Among all putative acetylation sites, K1127R, K1117R and the double mutant K1127R/K1117R showed significantly delayed re-entry to the nucleolus after damage recovery, even when SIRT1 is overexpressed. These mutants showed partial interaction with SIRT1 compared to WT WRN. Thus, our results suggest that K1127 and K1117 are the major sites of acetylation, necessary for DNA repair. These results elucidate the mechanism by which SIRT1 regulates WRN trafficking via these acetylation sites during DNA damage. Graphical abstract

DNA repair in personalized brain cancer therapy with temozolomide and nitrosoureas Publication date: June 2019 Source: DNA Repair, Volume 78 Author(s): Bernd Kaina, Markus Christmann Abstract Alkylating agents have been used since the 60ties in brain cancer chemotherapy. Their target is the DNA and, although the DNA of normal and cancer cells is damaged unselectively, they exert tumor-specific killing effects because of downregulation of some DNA repair activities in cancer cells. Agents exhibiting methylating properties (temozolomide, procarbazine, dacarbazine, streptozotocine) induce at least 12 different DNA lesions. These are repaired by damage reversal mechanisms involving the alkyltransferase MGMT and the alkB homologous protein ALKBH2, and through base excision repair (BER). There is a strong correlation between the MGMT expression level and therapeutic response in high-grade malignant glioma, supporting the notion that O6-methylguanine and, for nitrosoureas, O6-chloroethylguanine are the most relevant toxic damages at therapeutically relevant doses. Since MGMT has a significant impact on the outcome of anti-cancer therapy, it is a predictive marker of the effectiveness of methylating anticancer drugs, and clinical trials are underway aimed at assessing the influence of MGMT inhibition on the therapeutic success. Other DNA repair factors involved in methylating drug resistance are mismatch repair, DNA double-strand break (DSB) repair by homologous recombination (HR) and DSB signaling. Base excision repair and ALKBH2 might also contribute to alkylating drug resistance and their downregulation may have an impact on drug sensitivity notably in cells expressing a high amount of MGMT and at high doses of temozolomide, but the importance in a therapeutic setting remains to be shown. MGMT is frequently downregulated in cancer cells (up to 40% in glioblastomas), which is due to CpG promoter methylation. Astrocytoma (grade III) are frequently mutated in isocitrate dehydrogenase (IDH1). These tumors show a surprisingly good therapeutic response. IDH1 mutation has an impact on ALKBH2 activity thus influencing DNA repair. A master switch between survival and death is p53, which often retains transactivation activity (wildtype) in malignant glioma. The role of p53 in regulating survival via DNA repair and the routes of death are discussed and conclusions as to cancer therapeutic options were drawn.

The mismatch repair-dependent DNA damage response: Mechanisms and implications Publication date: June 2019 Source: DNA Repair, Volume 78 Author(s): Dipika Gupta, Christopher D. Heinen Abstract An important role for the DNA mismatch repair (MMR) pathway in maintaining genomic stability is embodied in its conservation through evolution and the link between loss of MMR function and tumorigenesis. The latter is evident as inheritance of mutations within the major MMR genes give rise to the cancer predisposition condition, Lynch syndrome. Nonetheless, how MMR loss contributes to tumorigenesis is not completely understood. In addition to preventing the accumulation of mutations, MMR also directs cellular responses, such as cell cycle checkpoint or apoptosis activation, to different forms of DNA damage. Understanding this MMR-dependent DNA damage response may provide insight into the full tumor suppressing capabilities of the MMR pathway. Here, we delve into the proposed mechanisms for the MMR-dependent response to DNA damaging agents. We discuss how these pre-clinical findings extend to the clinical treatment of cancers, emphasizing MMR status as a crucial variable in selection of chemotherapeutic regimens. Also, we discuss how loss of the MMR-dependent damage response could promote tumorigenesis via the establishment of a survival advantage to endogenous levels of stress in MMR-deficient cells.

Role of Y-family translesion DNA polymerases in replication stress: Implications for new cancer therapeutic targets Publication date: June 2019 Source: DNA Repair, Volume 78 Author(s): Peter Tonzi, Tony T. Huang Abstract DNA replication stress, defined as the slowing or stalling of replication forks, is considered an emerging hallmark of cancer and a major contributor to genomic instability associated with tumorigenesis (Macheret and Halazonetis, 2015). Recent advances have been made in attempting to target DNA repair factors involved in alleviating replication stress to potentiate genotoxic treatments. Various inhibitors of ATR and Chk1, the two major kinases involved in the intra-S-phase checkpoint, are currently in Phase I and II clinical trials [2]. In addition, currently approved inhibitors of Poly-ADP Ribose Polymerase (PARP) show synthetic lethality in cells that lack double-strand break repair such as in BRCA1/2 deficient tumors [3]. These drugs have also been shown to exacerbate replication stress by creating a DNA-protein crosslink, termed PARP 'trapping', and this is now thought to contribute to the therapeutic efficacy. Translesion synthesis (TLS) is a mechanism whereby special repair DNA polymerases accommodate and tolerate various DNA lesions to allow for damage bypass and continuation of DNA replication (Yang and Gao, 2018). This class of proteins is best characterized by the Y-family, encompassing DNA polymerases (Pols) Kappa, Eta, Iota, and Rev1. While best studied for their ability to bypass physical lesions on the DNA, there is accumulating evidence for these proteins in coping with various natural replication fork barriers and alleviating replication stress. In this mini-review, we will highlight some of these recent advances, and discuss why targeting the TLS pathway may be a mechanism of enhancing cancer-associated replication stress. Exacerbation of replication stress can lead to increased genome instability, which can be toxic to cancer cells and represent a therapeutic vulnerability.

Deciphering the interstrand crosslink DNA repair network expressed by Trypanosoma brucei Publication date: June 2019 Source: DNA Repair, Volume 78 Author(s): Ambika Dattani, Shane R. Wilkinson Abstract Interstrand crosslinks (ICLs) represent a highly toxic form of DNA damage that can block essential biological processes including DNA replication and transcription. To combat their deleterious effects all eukaryotes have developed cell cycle-dependent repair strategies that co-opt various factors from 'classical' DNA repair pathways to resolve such lesions. Here, we report the first systematic dissection of how ICL repair might operate in the Trypanosoma brucei, the causative agent of African trypanosomiasis, and demonstrated that this diverged eukaryote expresses systems that show some intriguing differences to those mechanisms present in other organisms. Following the identification of trypanosomal homologues encoding for CSB, EXO1, SNM1, MRE11, RAD51 and BRCA2, gene deletion coupled with phenotypic studies demonstrated that all the above factors contribute to this pathogen's ICL REPAIRtoire with their activities split across two epistatic groups. We postulate that one network, which encompasses TbCSB, TbEXO1 and TbSNM1, may operate throughout the cell cycle to repair ICLs encountered by transcriptional detection mechanisms while the other relies on homologous recombination enzymes (MRE11, RAD51 and BRCA2) that together help resolve lesions responsible for the stalling of DNA replication forks. This study not only sheds light on the conservation and divergence of ICL repair in one of only a handful of protists that can be studied genetically, but offers the promise of developing or exploiting ICL-causing agents as new anti-parasite therapies.

Transcriptional landscape of DNA repair genes underpins a pan-cancer prognostic signature associated with cell cycle dysregulation and tumor hypoxia Publication date: June 2019 Source: DNA Repair, Volume 78 Author(s): Wai Hoong Chang, Alvina G. Lai Abstract Overactive DNA repair contributes to therapeutic resistance in cancer. However, pan-cancer comparative studies investigating the contribution of all DNA repair genes in cancer progression employing an integrated approach have remained limited. We performed a multi-cohort retrospective analysis to determine the prognostic significance of 138 DNA repair genes in 16 cancer types (n = 16,225). Cox proportional hazards analyses revealed a significant variation in the number of prognostic genes between cancers; 81 genes were prognostic in clear cell renal cell carcinoma while only two genes were prognostic in glioblastoma. We reasoned that genes that were commonly prognostic in highly correlated cancers revealed by Spearman's correlation analysis could be harnessed as a molecular signature for risk assessment. A 10-gene signature, uniting prognostic genes that were common in highly correlated cancers, was significantly associated with overall survival in patients with clear cell renal cell (P < 0.0001), papillary renal cell (P = 0.0007), liver (P = 0.002), lung (P = 0.028), pancreas (P = 0.00013) or endometrial (P = 0.00063) cancers. Receiver operating characteristic analyses revealed that a combined model of the 10-gene signature and tumor staging outperformed either classifier when considered alone. Multivariate Cox regression models incorporating additional clinicopathological features showed that the signature was an independent predictor of overall survival. Tumor hypoxia is associated with adverse outcomes. Consistent across all six cancers, patients with high 10-gene and high hypoxia scores had significantly higher mortality rates compared to those with low 10-gene and low hypoxia scores. Functional enrichment analyses revealed that high mortality rates in patients with high 10-gene scores were attributable to an overproliferation phenotype. Death risk in these patients was further exacerbated by concurrent mutations of a cell cycle checkpoint protein, TP53. The 10-gene signature identified tumors with heightened DNA repair ability. This information has the potential to radically change prognosis through the use of adjuvant DNA repair inhibitors with chemotherapeutic drugs.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Apoptosis

Correction to: Autophagy inhibition with chloroquine reverts paclitaxel resistance and attenuates metastatic potential in human nonsmall lung adenocarcinoma A549 cells via ROS mediated modulation of β-catenin pathway The original version of this article unfortunately contained an error in acknowledgment text. The authors would like to include a statement: "Moumita Dasgupta is supported by Junior Research Fellowship from University Grant Commission, India." in acknowledgment section.

Potential role of anastasis in cancer initiation and progression

Inhibition of TNF-α-induced neuronal apoptosis by antidepressants acting through the lysophosphatidic acid receptor LPA 1 Abstract Tumor necrosis factor-α (TNF-α), a pro-inflammatory cytokine considered to be implicated in the pathogenesis of major depressive disorder, is a critical regulator of neuronal cell fate. In the present study we found that TNF-α-induced apoptosis of HT22 hippocampal cells, a neuroblast-like cell line, was markedly attenuated by the antidepressants mianserin, mirtazapine and amitriptyline. The anti-apoptotic effect of the antidepressants was blocked by either pharmacological inhibition or gene silencing of the lysophosphatidic acid receptor LPA1. Mianserin failed to affect TNF-α-induced caspase 8 activation, but inhibited the loss of mitochondrial membrane potential, the release of cytochrome c from mitochondria, procaspase 9 cleavage and downstream activation of caspase 3 in response to the cytokine. By acting through LPA1, mianserin also attenuated the enhanced pro-apoptotic response induced by the combination of TNF-α with other pro-inflammatory cytokines. TNF-α appeared to counterbalance its own pro-apoptotic response by activating NF-kB, ERK1/2 and JNK. Antidepressants had no significant effects on NF-kB activation, but potentiated the TAK-1-dependent phosphorylation of ERK1/2 and JNK elicited by the cytokine. This synergistic interaction was associated with enhanced JNK-mediated phosphorylation of Bcl-2 at Ser70 and increased ERK1/2-dependent mitochondrial accumulation of Mcl-1, two anti-apoptotic proteins that promote mitochondrial outer membrane stability. These results indicate that certain antidepressants, by activating LPA1 signalling, protect HT22 hippocampal cells from TNF-α-induced apoptosis through a mechanism involving, at least in part, the potentiation of the pro-survival pathways activated by the cytokine.

BDNF-mediated mitophagy alleviates high-glucose-induced brain microvascular endothelial cell injury Abstract Endothelial cell dysfunction and diabetic vascular complications are intrinsically linked. Although BDNF plays a protective role in cerebral microvascular complications caused by diabetes, the mechanisms of this activity are not fully clear. In this study, we investigated the role of BDNF in the hyperglycemic injury of BMECs and its associated intracellular signal transduction pathways. BMECs were treated with 33 mM glucose to imitate the endothelium under hyperglycemic conditions. The high-glucose treatment caused cell dysfunction, as evaluated by oxidative stress and cell apoptosis, which could be alleviated by BDNF. In addition, BDNF preserved mitochondrial function as assessed by mPTP opening, mitochondrial membrane potential, calcium content, and mitochondrial biogenesis markers. Western blot analysis of LC3-II, p62, and TOMM20 and the detection of mRFP-GFP-LC3 adenovirus for autophagy flux revealed that BDNF enhanced autophagy flux. Furthermore, BDNF activated mitophagy, which was confirmed by the observed colocalization of LC3-II with BNIP3 and from transmission electron microscopy observations. The HIF-1α/BNIP3 signaling pathway was associated with BDNF/TrkB-induced mitophagy. In addition, BDNF-induced mitophagy played a protective role against BMEC damage under hyperglycemia. Thus, the results of this study suggest that BDNF/TrkB/HIF-1α/BNIP3-mediated mitophagy protects BMECs from hyperglycemia.

Modulation of CD95-mediated signaling by post-translational modifications: towards understanding CD95 signaling networks Abstract CD95 is a member of the death receptor family and is well-known to promote apoptosis. However, accumulating evidence indicates that in some context CD95 has not only the potential to induce apoptosis but also can trigger non-apoptotic signal leading to cell survival, proliferation, cancer growth and metastasis. Despite extensive investigations focused on alterations in the expression level of CD95 and associated signal molecules, very few studies, however, have investigated the effects of post-translational modifications such as glycosylation, phosphorylation, palmitoylation, nitrosylation and glutathionylation on CD95 function. Post-translational modifications of CD95 in mammalian systems are likely to play a more prominent role than anticipated in CD95 induced cell death. In this review we will focus on the alterations in CD95-mediated signaling caused by post-translational modifications of CD95.

Autophagy inhibition with chloroquine reverts paclitaxel resistance and attenuates metastatic potential in human nonsmall lung adenocarcinoma A549 cells via ROS mediated modulation of β-catenin pathway Abstract Paclitaxel is one of the most commonly used drugs for the treatment of nonsmall cell lung cancer (NSCLC). However acquired resistance to paclitaxel, epithelial to mesenchymal transition and cancer stem cell formation are the major obstacles for successful chemotherapy with this drug. Some of the major reasons behind chemoresistance development include increased ability of the cancer cells to survive under stress conditions by autophagy, increased expression of drug efflux pumps, tubulin mutations etc. In this study we found that inhibition of autophagy with chloroquine prevented development of paclitaxel resistance in A549 cells with time and potentiated the effect of paclitaxel by increased accumulation of superoxide-producing damaged mitochondria, with elevated ROS generation, it also increased the apoptotic rate and sub G0/ G1 phase arrest with time in A549 cells treated with paclitaxel and attenuated the metastatic potential and cancer stem cell population of the paclitaxel-resistant cells by ROS mediated modulation of the Wnt/β-catenin signaling pathway, thereby increasing paclitaxel sensitivity. ROS here played a crucial role in modulating Akt activity when autophagy process was hindered by chloroquine, excessive ROS accumulation in the cell inhibited Akt activity. In addition, chloroquine pre-treatment followed by taxol (10 nM) treatment did not show significant toxicity towards non-carcinomas WI38 cells (lung fibroblast cells). Thus autophagy inhibition by CQ pre-treatment can be used as a fruitful strategy to combat the phenomenon of paclitaxel resistance development as well as metastasis in lung cancer.

Improved in vivo targeting of BCL-2 phenotypic conversion through hollow gold nanoshell delivery Abstract Although new cancer therapeutics are discovered at a rapid pace, lack of effective means of delivery and cancer chemoresistance thwart many of the promising therapeutics. We demonstrate a method that confronts both of these issues with the light-activated delivery of a Bcl-2 functional converting peptide, NuBCP-9, using hollow gold nanoshells. This approach has shown not only to increase the efficacy of the peptide 30-fold in vitro but also has shown to reduce paclitaxel resistant H460 lung xenograft tumor growth by 56.4%.

Proteasome inhibitors trigger mutations via activation of caspases and CAD, but mutagenesis provoked by the HDAC inhibitors vorinostat and romidepsin is caspase/CAD-independent Abstract Genotoxic anti-cancer therapies such as chemotherapy and radiotherapy can contribute to an increase in second malignancies in cancer survivors due to their oncogenic effects on non-cancerous cells. Inhibition of histone deacetylase (HDAC) proteins or the proteasome differ from chemotherapy in that they eliminate cancer cells by regulating gene expression or cellular protein equilibrium, respectively. As members of these drug classes have been approved for clinical use in recent times, we investigated whether these two drug classes exhibit similar mutagenic capabilities as chemotherapy. The HDAC inhibitors vorinostat/SAHA and romidepsin/FK288 were found to induce DNA damage, and mis-repair of this damage manifested into mutations in clonogenically viable surviving cells. DNA damage and mutations were also detected in cells treated with the proteasome inhibitor bortezomib. Exposure to both drug classes stimulated caspase activation consistent with apoptotic cell death. Inhibition of caspases protected cells from bortezomib-induced acute (but not clonogenic) death and mutagenesis, implying caspases were required for the mutagenic action of bortezomib. This was also observed for second generation proteasome inhibitors. Cells deficient in caspase-activated DNase (CAD) also failed to acquire DNA damage or mutations following treatment with bortezomib. Surprisingly, vorinostat and romidepsin maintained an equivalent level of killing and mutagenic ability regardless of caspase or CAD activity. Our findings indicate that both drug classes harbour mutagenic potential in vitro. If recapitulated in vivo, the mutagenicity of these agents may influence the treatment of cancer patients who are more susceptible to oncogenic mutations due to dysfunctional DNA repair pathways.

Fibroblasts from patients with idiopathic pulmonary fibrosis are resistant to cisplatin-induced cell death via enhanced CK2-dependent XRCC1 activity Abstract Idiopathic pulmonary fibrosis (IPF) is a deadly and progressive fibrotic lung disease, but the precise etiology remains elusive. IPF is characterized by the presence of apoptosis-resistant (myo)fibroblasts that relentlessly produce a collagen-rich extracellular matrix (ECM). Recent studies showed that an anti-cancer chemotherapy drug cisplatin is implicated in the development of pulmonary fibrosis, suggesting that the treatment of cancer patients with cisplatin may alter fibroblast viability. To address this possibility, we investigated the cisplatin-induced cell death mechanism in lung fibroblasts derived from IPF and non-IPF patients in response to a collagen matrix. IPF fibroblasts showed enhanced resistance to cisplatin-induced cell death compared to non-IPF fibroblasts in a time- and dose-dependent manner. Molecular study showed that the expression of γH2AX, PUMA and caspase-3/7 activity was abnormally reduced in IPF fibroblasts, suggesting that DNA damage-induced apoptosis caused by cisplatin was suppressed in IPF fibroblasts. Our study further revealed that DNA repair protein XRCC1 activity was aberrantly increased as a result of CK2 hyper-activation in cisplatin-treated IPF fibroblasts, and this alteration protected IPF fibroblasts from cisplatin-induced cell death. Our results showed that IPF fibroblasts residing in a collagen rich matrix are resistance to cisplatin-induced cell death due to the aberrantly high CK2/XRCC1-dependent DNA repair activity. This finding suggests that pulmonary fibrosis may develop and worsen due to the presence of apoptosis-resistant lung fibroblasts in cisplatin-treated cancer patients.

The neuroprotective action of 3,3′-diindolylmethane against ischemia involves an inhibition of apoptosis and autophagy that depends on HDAC and AhR/CYP1A1 but not ERα/CYP19A1 signaling Abstract There are no studies examining the effects of 3,3′-diindolylmethane (DIM) in neuronal cells subjected to ischemia. Little is also known about the roles of apoptosis and autophagy as well as AhR and ERα signaling and HDACs in DIM action. We demonstrated for the first time the strong neuroprotective capacity of DIM in mouse primary hippocampal cell cultures exposed to ischemia at early and later stages of neuronal development. The protective effects of DIM were mediated via inhibition of ischemia-induced apoptosis and autophagy that was accompanied by a decrease in AhR/CYP1A1 signaling and an increase in HDAC activity. DIM decreased the levels of pro-apoptotic factors, i.e., Fas, Caspase-3, and p38 mitogen-activated protein kinase (MAPK). DIM also reduced the protein levels of autophagy-related Beclin-1 (BECN1) and microtubule-associated proteins 1A/1B light chain (LC3), partially reversed the ischemia-induced decrease in Nucleoporin 62 (NUP62) and inhibited autophagosome formation. In addition, DIM completely reversed the ischemia-induced decrease in histone deacetylase (HDAC) activity in hippocampal neurons. Although DIM inhibited AhR/CYP1A1 signaling, it did not influence the protein expression levels of ERα and ERα-regulated CYP19A1 which are known to be controlled by AhR. This study demonstrated for the first time, that the neuroprotective action of 3,3′-diindolylmethane against ischemia involves an inhibition of apoptosis and autophagy and depends on AhR/CYP1A1 signaling and HDAC activity, thus creating the possibility of developing new therapeutic strategies that target neuronal degeneration at specific molecular levels.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Psychiatric Quarterly

Emotion Regulation and Desire Thinking as Predictors of Problematic Facebook Use Abstract Research evidence suggests that problematic Facebook use (PFU) affects large numbers of people worldwide. A variety of studies have investigated the relationship between PFU and psychological dysfunction, revealing that negative mood and cognitive self-regulation are common among problematic Facebook users. The aim of the present study was to examine the role of craving, emotion regulation and desire thinking in predicting PFU. An online survey was administered to 750 participants (Mage = 23.38 years; SDage = 5.72). Correlation analyses indicated that all predictor variables were positively associated with PFU. Path analysis showed that imaginal prefiguration was associated with craving, which, in turn, was associated with verbal prefiguration that was directly linked to PFU, beyond the direct effect of emotion regulation. These findings provide further support for the importance of desire thinking in predicting problematic behaviours. Desire thinking and emotion regulation should be considered in the modification of problematic Facebook use.

Intervention Effects of Motivation Interviewing Chinese Modified on the Mental Health of College Students with Exercise Dependence Abstract Exercise dependence is a psychological problem that cannot be ignored and is positively related to anxiety and depression of college students. However, only a few effective intervention methods are available to deal with exercise dependence. This study aims to investigate the intervention effects of motivation interviewing Chinese modified on the mental health of college students with exercise dependence. Thirty college students with exercise dependence were selected from Hunan University of Science and Technology in Hunan province of China to participate in the experiment. The participants were divided equally into the intervention and control groups. A three-week motivation interviewing Chinese modified session was conducted in the intervention group, whereas no intervention was carried out in the control group. This strategy allowed for the vertical and the horizontal comparison of the intervention objects' situation before and after the experiment. The State-Trait Anxiety Inventory and other Scales were used to evaluate the effects of the intervention and explore the intervention effects of motivation interviewing Chinese modified on the mental health of college students with exercise dependence. After three weeks of motivation interviewing Chinese modified, differences in state anxiety, depression, self-satisfaction, negative emotion, energy, and positive emotion in the intervention and control groups before and after the intervention appear to be statistically significant (P < 0.05). Motivation interviewing Chinese modified can improve the mental health level of college students with exercise dependence. Hence, motivation interviewing Chinese modified is good for the treatment of addiction behaviors and provides a reliable intervention method for exercise dependence.

Media Use Is Linked to Lower Psychological Well-Being: Evidence from Three Datasets Abstract Adolescents spend a substantial and increasing amount of time using digital media (smartphones, computers, social media, gaming, Internet), but existing studies do not agree on whether time spent on digital media is associated with lower psychological well-being (including happiness, general well-being, and indicators of low well-being such as depression, suicidal ideation, and suicide attempts). Across three large surveys of adolescents in two countries (n = 221,096), light users (<1 h a day) of digital media reported substantially higher psychological well-being than heavy users (5+ hours a day). Datasets initially presented as supporting opposite conclusions produced similar effect sizes when analyzed using the same strategy. Heavy users (vs. light) of digital media were 48% to 171% more likely to be unhappy, to be in low in well-being, or to have suicide risk factors such as depression, suicidal ideation, or past suicide attempts. Heavy users (vs. light) were twice as likely to report having attempted suicide. Light users (rather than non- or moderate users) were highest in well-being, and for most digital media use the largest drop in well-being occurred between moderate use and heavy use. The limitations of using percent variance explained as a gauge of practical impact are discussed.

PG-13 Rated Movie Violence and Societal Violence: is there a Link? Abstract Recent scholarship has suggested that the frequency of violence in PG-13 rated movies has increased in recent years. Although some scholars have expressed concern that such an increase may have public health implications, this has remained untested. In the current article, trends in PG-13 movie violence are tested against trends in violence in society, including both homicides and youth violence. Raw correlations suggest that PG-13 rated movie violence is inversely related to actual violence in society. However, controlling for autocorrelations suggests that the best interpretation is that PG-13 rated movie violence is unrelated to violence in society. Caution is advised for scholars to avoid implying that PG-13 rated movie violence may have a causal effect on crime in society.

Hospitalization Patterns over 30 Years Across a Statewide System of Public Mental Health Hospitals: Readmission Predictors, Optimal Follow-Up Period, Readmission Clusters and Individuals with Statistically Significant High Healthcare Utilization Abstract Four related hospital utilization questions (optimal follow-up period, predictors of readmission, definition of individuals with statistically significant high healthcare utilization, and patterns of readmissions) were examined using data for 491,094 hospital discharges for 250,091 patients across a statewide public mental health hospital system for 30 years (1987 to 2016). Using survival analysis, the first quartile of the survival time, the time when 25% of the entire population of discharges had a readmission was 229 days. Using observed readmissions, rather than the population as in survival analysis, revealed that 50% of all observed readmissions occurred by 222 days. Both suggest that using a one year observation period for determining high utilization may be reasonable. Major predictors of readmission were diagnoses of schizophrenia (OR = 2.11) or bipolar disorder (OR = 1.57) as well as total number of previous discharges (OR = 1.23). Statistically significant z scores (p < .01) were used to determine annual (3 or more discharges) and lifetime (7 or more discharges) criteria for individuals with statistically significant high healthcare utilization that were somewhat lower than in previous research. Cluster analysis of all readmissions revealed four relatively distinct clusters of patients: short stay-quick readmission, extremely long stay, long time in community between readmissions and frequent readmissions. While no cluster corresponded exactly with the annual statistically significant high healthcare utilization criteria, the frequent readmission cluster was somewhat similar to the lifetime statistically significant high healthcare utilization criteria with 46% of this cluster's patients having 7 or more discharges.

Working Nursing Students Willing to Seek Psychological Services Abstract Work-related psychological stress may result in reduced coping abilities. Working nursing students can develop work-related psychological stress. This study's purpose was to describe first-year working rural nursing students with work-related psychological stress and their perception of stigma to psychological services and perceived willingness and openness to seek such services. Results showed over 55% of students reported work-related stress and a willingness to seek psychological services. Students in a licensed practical nursing program showed less stigma to seeking psychological services (F (2, 23) = 10.09, p = 0.001) as compared to higher degree seeking students in associate and bachelor nursing programs. Stigma appeared not to be a factor in rural nursing students seeking psychological services. In conclusion, working nursing students are willing to seek psychological services regardless of stigma.

Enhancing Completion of Cognitive Processing Therapy for Posttraumatic Stress Disorder with Quetiapine in Veterans with Mild Traumatic Brain Injury: a Case Series Abstract To evaluate the outcomes of the antiarousal medications valproate, risperidone, and quetiapine on completion of treatment of cognitive processing therapy (CPT) for PTSD. A case series of fifty treatment-seeking adult (≥18 years) veterans with mild traumatic brain injury and combat-related PTSD who had unsuccessful trials of 2 or more first-line agents and previously declined treatment with trauma-focused therapy, seen at the psychiatric outpatient services of the local Polytrauma Rehabilitation Center from January 1, 2014, through December 31, 2017. Patients were prescribed valproate (n = 8), risperidone (n = 17), or quetiapine (n = 25) and were referred for individual weekly treatment with CPT. Outcome measurements of interest were measures of engagement and completion rate of CPT, PTSD Checklist total score (range, 0–80; higher scores indicate greater PTSD severity) and arousal subscale score (range, 0–24; higher scores indicate greater arousal severity), and clinical observations of sleep variables. Of the 50 patients included in the study, 48 (96%) were men; mean (SD) age was 36 (8) years. Eighteen (86%) patients initially receiving quetiapine and none taking valproate or risperidone became adequately engaged in and completed CPT. Among patients who completed CPT, the mean decrease in the PTSD Checklist score was 25 [95% CI, 30 to 20] and 9 (50%) patients no longer met criteria for PTSD. These preliminary findings support quetiapine as an adjunctive medication to facilitate CPT. A pragmatic trial is needed to evaluate the efficacy, safety, and feasibility of quetiapine to improve engagement in and completion rate of CPT.

Gender Differentiation of Indirect Self-Destructiveness in Drug Addicted Individuals (Indirect Self-Destructiveness in Addicted Women and Men) Abstract The use of psychoactive substances is considered to be a typical self-destructive behaviour with addiction itself regarded as one of the self-destructiveness forms. The aim of this work was to explore the gender differentiation of the indirect self-destructiveness syndrome (and its particular categories) in drug addicted individuals treated in drug addiction treatment centres. 172 drug addicted individuals (116 men and 56 women, M age = 23,5), ranged from 19 to 28 years, was recruited. In order to examine indirect self-destructiveness and its manifestations, the Polish version of the "Chronic Self-Destructiveness Scale" by Kelley (CS-DS) was administered. The statistical processing of scores used the Mann-Whitney U significance test. Women treated for drug addiction achieved significantly higher scores on indirect self-destructiveness: general score (p = 0.001), subscales of Transgression and Risk (p = 0.001), Personal and Social Neglects (p = 0.02), and Lack of Planfulness (p < 0.001). They scored lower on Poor Health Maintenance (p < 0.002) and Helplessness (p < 0.001). There is a need for specific, gender-adjusted manners of intervention and treatment in addicted women. Optimistically, after an addiction treatment, women cope and feel better psychologically and socially. They also care more about their health.

Folie du système? Preventing Violence Against Nurses in In-patient Psychiatry Abstract Violence against psychiatric nurses is a difficult reality of work on in-patient psychiatry units. Health care providers and managers, nursing unions, and workplace protection agencies are looking for solutions to improve safety and quality of care. We are suggesting that simultaneous to this solution-seeking, there is also a need to critically reflect on the nature of violence itself within in-patient psychiatric settings. In this article we consider the gendered dynamics of power and violence within the in-patient psychiatric setting. The nursing profession is over 90% female. Given that violence in society often has a 'gendered' nature, and in light of a report from the Ontario Council of Hospital Unions which likened violence against nurses to domestic violence, we have put forth a view of the acute in-patient psychiatric milieu that considers gender and power in its analysis of violence against nurses. Intended to encourage enquiry into our pre-suppositions as health care providers, we use Foucauldian and feminist theories to up-end our notions of "anti-violence technologies", and to consider the unique and risky position that psychiatric nurses occupy as carers, care providers, and "anti-violence officers". We conclude by posing ethical questions which may be of interest for professional development, care planning, team building, and clinical ethics and education.

Insight and Symptom Severity in an Inpatient Psychiatric Sample Abstract Individuals with a severe mental illness, particularly a psychotic disorder, often lack insight into having a mental illness. This study sought to examine the differences in insight and symptom severity between individuals with psychotic, bipolar, and depressive disorders in an inpatient psychiatric sample. 199 participants were interviewed and medical records were consulted. Results show that participants with a psychotic disorder had significantly less insight into their illness, more debilitating symptoms, and reported less depression symptoms after controlling for education, race, marital status, homelessness, age, gender, and history of incarceration. Insight was shown to be a mediator between having a psychotic disorder and symptom severity. Subjective quality of life did not differ by diagnosis. Substance use was not associated with insight or overall symptom severity, while homelessness was associated with having a psychotic disorder and more severe symptoms. Fostering insight during an inpatient stay may be an important part of reducing symptom severity and preventing patient relapse. However, greater insight may increase depression and suicidality, indicating a need for mood management and safety planning along with psychoeducation of symptoms.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Research in Pharmaceutical Sciences

Siamese neem flower extract suppresses cholesterol absorption by interfering NPC1L1 and micellar property in vitro and in intestinal Caco-2 cells Acharaporn Duangjai, Atcharaporn Ontawong, Chutima Srimaroeng Research in Pharmaceutical Sciences 2019 14(3):190-200 Siamese neem (Azadirachta indica A. Juss var. siamensis Valeton) (A. indica) leaf extract, a traditional ayurvedic medicine, has been reported to exhibit antipyretic, antibacterial, antidyslipidemic, and antihyperglycemia effects. This study investigated the mechanism of hypocholesterolemic effect of methanolic extract of Siamese neem flowers in in vitro studies and in Caco-2 cells. Pancreatic cholesterol esterase and 3-hydroxy 3-methylglutaryl-CoA (HMG-CoA) reductase activities were assessed. Cholesterol micelle formation was prepared for in vitro cholesterol physicochemical property analyses, micelle size and solubility, and transport of cholesterol into the Caco-2 cells. The expression of niemann-pick C1 like 1 (NPC1L1), and its major regulator, peroxisome proliferator-activated receptor δ (PPARδ), were determined by western blot and real time polymerase chain reaction, respectively. A. indica flower extract inhibited pancreatic cholesterol esterase activity and increased cholesterol micelles size. Uptake of cholesterol into Caco-2 cells was inhibited by A. indica flower extract in a dose-dependent manner. In addition, A. indica extract inhibited HMG-CoA reductase activity, resulting in low level of intracellular cholesterol accumulation, together with increased cytosolic NPC1L1 protein expression and decreased PPARδ gene expression. In conclusion, A. indica flower extract has cholesterol-lowering effects by inhibiting intestinal cholesterol absorption, interfering micellar cholesterol formation, and attenuating cholesterol synthesis. As such, A. indica flower extract has potential for developing into nutraceutical product for prevention of hypocholesterolemia.

Protective effects of melatonin solid lipid nanoparticles on testis histology after testicular trauma in rats Mehri Mirhoseini, Zahra Rezanejad Gatabi, Majid Saeedi, Katayoun Morteza-Semnani, Fereshteh Talebpour Amiri, Hamid Reza Kelidari, Abbas Ali Karimpour Malekshah Research in Pharmaceutical Sciences 2019 14(3):201-208 Testicular traumatic injuries occur frequently, which can result in an alteration in spermatogenesis. These injuries can also cause oxidative stress and male infertility. Antioxidant efficiency of melatonin (MLT), known as a potent antioxidant, will be improved if used in a form of solid lipid nanoparticles (MLT-SLN). The aim of the current study is to evaluate the effect of MLT-loaded SLN on traumatic testis in rats. In this study 32 adult male Wistar rats were divided into 4 groups. Group 1 (sham group), right testicle was drawn out from the scrotum and returned without manipulation. Group 2, right testicle was dropped by 25 g sinker for 4 times. Group 3, animals were received a single dose (25 mg/kg) of MLT intraperitoneally after trauma. Group 4, animals were received a single dose of MLT-SLN intraperitoneally after trauma. Under anaesthesia, rats were sacrificed, and their testicles were removed three days after the surgery. After tissue processing, the sample sections were H&E stained. MLT and MLT-SLN could partially repair spermatogenesis by Johnson’s criteria but the repairs were significant only in MLT-SLN group (P = 0.02). Trauma decreased seminiferous tubule diameter and its epithelium height. MLT could restore epithelium height (P ≤ 0.05) but its NPs improved both epithelium diameter (P ≤ 0.05) and thickness (P ≤ 0.001). The Malondialdehyde increased significantly in trauma group (P = 0.002), but decreased in MLT and NPs groups compared to trauma group (P = 0.098 and P = 0.002 respectively). This decrease was significant only in NPs group. Testicular trauma disturbed spermatogenesis, morphometric, and oxidative parameters. MLT and specially MLT-SLN improved traumatic damages.

Differentiation of adult human mesenchymal stem cells into dopaminergic neurons Marjan Khademizadeh, Manoochehr Messripour, Nazem Ghasemi, Fariborz Momen beik, Ahmad Movahedian Attar Research in Pharmaceutical Sciences 2019 14(3):209-215 The striatal dopamine (DA) deficiency is known as the main cause of the clinical picture of Parkinson’s disease (PD). The disease is a progressive degeneration of dopaminergic neurons in the striatum. The treatment of PD is based on compensation for the brain’s supply of DA lost by drug therapy, deep brain stimulation, surgery, gene and cell therapies. Clinical studies have focused on the utility of stem cell-based therapies in PD. Embryonic and mesenchymal stem cells (MSCs) are widely used. Recently, human adipose derived stem cells (hADSCs) have been considered as a suitable source of tissue for this purpose. In this project, hADSCs differentiated into dopaminergic neurons and the specificity of the cell preparations was examined. Human adipose tissues were collected from healthy volunteers undergoing liposuction and hADSCs were isolated by collagenase-based enzymatic method. Flow cytometry was performed using the surface cluster of differentiation (CD) markers to confirm the cell typical properties. Then hADSCs were differentiated to dopaminergic neurons in neurobasal medium in the presence of differentiation factors and confirmed by immunocytochemistry via neuronal and dopaminergic markers. The isolated hADSCs were cultured and identified by the expression of MSCs surface markers including CD90, and CD44. These cells did not express hematopoietic surface markers such as CD45 and CD14. Differentiated cells express neuronal marker NeuN and dopaminergic marker tyrosine hydroxylase (TH). It is concluded that hADSCs can be easily taken from the patient’s own body and differentiated into dopaminergic cells having a lower risk of transplant rejection.

β-lactoglobulin-irinotecan inclusion complex as a new targeted nanocarrier for colorectal cancer cells Nooshin Bijari, Sirous Ghobadi, Katayoun Derakhshandeh Research in Pharmaceutical Sciences 2019 14(3):216-227 Beta-lactoglobulin (β-LG) is a lipocalin family member whose general function appears to be solubilizing and transport of hydrophobic molecules. Some properties such as avalability, ease of purification, and peculiar resistance to acidic environments can make β-LG as a carrier for hydrophobic and acid labile drugs for oral administration. In this protein vehicle, drug could be protected in acidic environment of stomach and then released within the basic small intestine. In this study, the potential of β-LG as a nanocarrier for oral delivery of a potent agent in colorectal cancer treatment, irinotecan, was evaluated. The nanoparticle was prepared by the physical inclusion complex method. Size, drug loading, encapsulation efficiency, and in vitro drug release at various pH values were investigated. The optimum formulation showed a narrow size distribution with an average diameter of 139.86 ± 13.75 nm and drug loading about 84.33 ± 5.03%. Based on the results obtained from docking simulation of irinotecan-complex, there are two distinct binding sites in this nanocarrier. Cytotoxicity of this nanocarrier on the HT-29 cancer cell line and AGS was measured by MTT assay. The cytotoxicity experiment showed that the drug-loaded nanocarrier was more effective than free drug. The higher release percent of drug from the β-LG complex at pH 7.4 compared to pH 1.2 indicated that the proposed nanocarrier could be introduced as a suitable nanovehicle for labile drugs in acidic medium targeted for colorectal segment.

Anti-inflammatory effects of alosetron mediated through 5-HT3 receptors on experimental colitis Azadeh Motavallian, Mohsen Minaiyan, Mohammad Rabbani, Parvin Mahzouni, Sasan Andalib Research in Pharmaceutical Sciences 2019 14(3):228-236 Development of new medicine with fewer deleterious effects and more efficacies for treatment of inflammatory bowel disease is needed. 5-Hydroxytryptamine 3 receptor (5-HT3R) antagonists have exhibited analgesic and anti-inflammatory features in vitro and in vivo. The present study was designed to evaluate the anti-inflammatory effect of alosetron, a 5-HT3R antagonist, on trinitrobenzenesulfonic acid (TNBS)-induced ulcerative colitis in rats. Two h subsequent to induce colitis (intracolonic instillation of TNBS, 50 mg/kg) in male Wistar rats, alosetron (1 mg/kg), dexamethasone (1 mg/kg), meta-chlorophenylbiguanide (mCPBG, a 5-HT3R agonist, 5 mg/kg), or alosetron + mCPBG were administrated intraperitoneally for 6 days. Animals were thereafter sacrificed and the efficacy of drugs was evaluated macroscopically, histologically, and biochemically (myeloperoxidase, tumor necrosis factor-alpha, interleukin-6, and interleukin-1 beta) on distal colon samples. Treatment with alosetron and dexamethasone improved macroscopic and microscopic colonic damages significantly and decreased myeloperoxidase activity and colonic levels of inflammatory cytokines. The profitable effects of alosetron were antagonized by concurrent administration of mCPBG. Our data provided evidence that the protective effects of alosetron on TNBS-induced colitis can be mediated by 5- HT3R.

Effects of thyroxine on adhesion molecules and proinflammatory cytokines secretion on human umbilical vein endothelial cells Attabak Milani, Mohammad Hassan Khadem-Ansari, Yousef Rasmi Research in Pharmaceutical Sciences 2019 14(3):237-246 Thyroid dysfunction is associated with elevated cardiovascular risk factors and atherosclerosis. It could be suggested that, hyperthyroidism is related to a higher prevalence of arterial abnormalities. Therefore, evaluating the endothelial dysfunction (ED) related biomarkers seem to be an important issue. It is not clear whether endothelial cells are biologically responsive to thyroid hormones (THs) or how THs induces the production of endothelial cells (EC)-derived proinflammatory mediators. Hence, in this study the effects of thyroxine (T4) on ED and inflammatory related mediators were evaluated. Human umbilical vein endothelial cells was used as endothelial cell model which was treated with concentrations of 50, 100, 200 nmol/L of T4 in various exposure times. In the following, gene and protein expression levels of EC-related markers including intercellular adhesion molecule-1 (ICAM-1), vascular endothelial growth factor (VEGF), and E-selectin were determined using real time polymerase chain reaction (RT-PCR) and western blotting methods. Also, interleukin-6 (IL-6) and tumor necrosis factor (TNF-α) protein levels as proinflammatory cytokines were determined by enzyme linked immunosorbent assay (ELISA) method. Gene and protein expression analysis revealed that T4 treatments up regulated the levels of VEGF, ICAM-1, and E-selectin as ED markers. In addition, T4-treated cells had higher significant levels of IL-6 and TNF-α versus untreated cells in different incubation times. This study proposed the atherosclerotic effects of thyroid hormone. Based on our findings, T4 had strong effects on the gene and protein expression levels of pro-inflammatory, angiogenesis, and ED major mediators associated with atherosclerosis development.

Synthesis and cytotoxic evaluation of novel quinozalinone derivatives with substituted benzimidazole in position 3 Elham Taherian, Ghadamali Khodarahmi, Marzieh Khajouei, Farshid Hassanzadeh, Nasim Dana Research in Pharmaceutical Sciences 2019 14(3):247-254 Quinazolinone and benzimidazole are both fused heterocyclic compounds which have shown valuable biological properties including cytotoxic, antibacterial, and antifungal activities. In this study, a series of novel quinazolinone derivatives substituted with benzimidazole were synthesized in two parts. In the first part 2 - phenyl - 1H - benzimidazol - 6 - amine (4) was synthesized from the reaction of 4-nitro-o-phenylenediamine and benzoic acid. In the second part, new 3-(2-phenyl-1H benzoimidazol-5-yl)- 3H-quinazolin-4-one derivatives (8a-8f) were also prepared. Finally compound 4 was reacted with the different benzoxazinone derivatives (8a-8f) to give the target compounds. The structures of the synthesized compounds were confirmed by IR and 1HNMR. Cytotoxic activities of the final compounds were assessed at 100, 200, 300, 400, and 500 μM against MCF-7 and HeLa cell lines using the MTT colorimetric assay. Almost all compounds exhibited good cytotoxic activity against both cell lines. Compound 9d demonstrated the highest cytotoxic activity against MCF7 and Hela cell lines with IC50 70 μM and 50 μM, respectively.

The regulatory effect of saffron stigma on the gene expression of the glucose metabolism key enzymes and stress proteins in streptozotocin-induced diabetic rats Maryam Motamedrad, Alireza Shokouhifar, Mina Hemmati, Maryam Moossavi Research in Pharmaceutical Sciences 2019 14(3):255-262 Oxidative stress plays a crucial role in the pathogenesis of hyperglycemia mediated complications. Since a great number of researches have reported antioxidant features of saffron, this study investigated the antioxidant effect of saffron stigma extract (SSE) in streptozotocin-induced diabetic rats. Twenty eight diabetic male Wistar rats were divided in four groups containing: two diabetic groups receiving 25 and 100 mg/kg SSE respectively, one diabetic group receiving glibenclamide (0.6 mg/kg) and one diabetic control group receiving normal saline. Seven healthy adult male Wistar rats were also used as normal control group. After treatment (21 days), fasting blood glucose, insulin, oxidative stress markers, and pancreatic regeneration were assessed. The gene expression level of heat shock factor1, heat shock protein 27, and heat shock protein 70, also glucokinase (GK), and glucose 6-phosphatase (G6Pase) were determined using real-time polymerase chain reaction (RT-PCR). SSE in high dose (100 mg/kg) reduced fasting blood glucose (8.3 ± 0.4 mmol/L) compared with diabetic control (24.6 ± 1.2 mmol/L) (P < 0.05). Furthermore, SSE in high dose increased insulin level compared with diabetic control group (12.7 ± 0.6 vs 7.1 ± 0.3 μϋ/mL). RT-PCR analysis revealed decline in mRNA levels of stress proteins and G6Pase and increase in mRNA level of GK in treatment diabetic groups compared with diabetic control group. Data showed antioxidant and antidiabetic effects of SSE through altering insulin release and glucose metabolism pathways. Hypoglycemic potential of SSE may be due to change in GK and G6Pase enzymes expression. These findings provide a basis for the therapeutic potential of saffron in treatment of diabetes.

Phytochemical analysis and antiproliferative activity of the aerial parts of Scrophularia subaphylla Abbas Delazar, Solmaz Asnaashari, Elhameh Nikkhah, Parina Asgharian Research in Pharmaceutical Sciences 2019 14(3):263-272 Scrophularia subaphylla (S. subaphylla) L., a medicinal plant from the Scrophulariaceae family, has been reported to possess potential profits in the treatment and prophylaxis of different diseases. Some phenolic compounds in this genus have been displayed decent effects on different types of cancer via multiple mechanisms. The current study aimed to bioassay guided isolation of cytotoxic constituents from the aerial parts of S. subaphylla against breast (MCF-7) and colon (HT-29) cancer cell lines as well as normal cells (L929). Different extracts of S. subaphylla were acquired by Soxhlet apparatus and then subjected to brine shrimp lethality test and MTT assay for assessing their cytotoxic characteristics. Cytotoxic extract subjected to further phytochemical fractionation using solid phase extraction, reversed-phase high pressure liquid chromatography (RP-HPLC), and one dimensional nuclear magnetic resonance (1D-NMR) spectroscopy. The biological activity of the isolated pure components, verbascoside and 3´ O rhamnosyl -4´ O para coumaryl 7- hydroxyl salidroside, was assessed using MTT assay against MCF-7 and HT-29 carcinoma cells. Two known phenylpropanoid compounds were isolated from this species. Their structures were elucidated by spectroscopic data (using 1H-NMR and 13C-NMR) and compared with the previous literature. Both pure compounds in comparison with control group demonstrated significant antiproliferative activity against cancerous cells (P < 0.001). In our study, verbascoside and its derivative could inhibit proliferation of cancerous cells without any side effects on normal cells.

Vitex rotundifolia fractions induce apoptosis in human breast cancer cell line, MCF-7, via extrinsic and intrinsic pathways Gul-e-Saba Chaudhry, Rehmat Jan, Habsah Mohamad, Tengku Sifzizul Tengku Muhammad Research in Pharmaceutical Sciences 2019 14(3):273-285 Breast cancer is amongst frequently diagnosed cancer type throughout the world. Due to reduced efficacy of current chemotherapeutics, several natural products have been screened for better alternatives. The cytotoxic activity of fractions prepared from leaves extract of Vitex rotundifolia (V. rotundifolia) on human breast cancer cell line, MCF-7 was studied. The fractions F1, F2, F3, and F5 of V. rotundifolia produced concentration-dependent cytotoxic effects on MCF-7 cell line. The relative potential of cytotoxicity of the fractions on MCF-7 cell line was found to be F3 > F2 > F5 > F1. The active fractions induce apoptosis in MCF-7 cell line determined by annexin V base assay. The phosphatidylserine externalization and the presence of DNA fragmentation in treated cells confirms the early and late apoptosis in treated cells. The V. rotundifolia fractions induced apoptosis by both pathways; extrinsic pathways via activation of caspase-8 and intrinsic pathways through enhanced bax/bcl-2 ratio and activation of caspase-3/7 and caspase-9 proapoptotic proteins. Furthermore, chemical profiling indicates various phenolic, flavonoids, and terpenoids compounds in the active fractions. Thus, V. rotundifolia might be a suitable candidate to investigate further and develop molecular targeted cancer therapeutics by understanding the fundamental mechanisms involved in the regulation of cell death in cancer cells.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Datta Meghe Institute of Medical Sciences University

Review of current global evidences for prevention of coronary heart disease Meenakshi Khapre, Vartika Saxena Journal of Datta Meghe Institute of Medical Sciences University 2019 14(1):1-5 India is experiencing epidemiologic transition with sharp increase in coronary heart disease (CHD). Fatality rate of CHD is alarmingly high leading to the most common cause of premature mortality in 2016. There is the growing burden of coronary risk factors owing to rapid urbanization and changes in lifestyle, including diabetes mellitus (DM), hypertension, dyslipidemia, smoking, alcohol consumption, dietary patterns, central obesity physical inactivity, and psychological factors. These risk factors attribute 90% of CHD in population. The global evidences of preventive strategies is being reviewed in this paper with the aim to get the glimpse of study on CHD and find gaps in the research.

To assess the utility of proliferative marker Ki-67 in surface epithelial ovarian tumor Sheronica Laishram, Vivek Gupta, Arvind Bhake, Akanksha Wankhede, Deepika Agrawal Journal of Datta Meghe Institute of Medical Sciences University 2019 14(1):6-10 Background: Surface epithelial ovarian tumor (SEOT) accounts for more than 90% of all the ovarian tumors and are the most lethal one. Cellular proliferation plays a significant role in the clinical behavior and aggressiveness of ovarian tumor. Proliferative activity of the tumor can be assessed using the proliferative marker Ki-67, which has both diagnostic and prognostic values. Aim and Objective: The aim and objective of this study was to assess the expression of proliferative marker Ki-67 in various SEOT and correlate its level of expression with clinicopathological parameters including the International Federation of Gynecology and Obstetrics (FIGO) staging and preoperative CA-125 level. Materials and Methods: The study was a cross-sectional and observational study carried out in the Department of Pathology, JNMC, Sawangi, Wardha, over 2 years. It included 74 patients with surgically resected specimen of SEOT. Ki-67 immunohistochemistry was performed in all the 74 cases, and the percentage of immunopositive cells was expressed as Ki-67 labeling index (Ki-67 LI). Results: Out of 74 cases, 54 were benign and 20 were malignant which comprised serous and mucinous histological subtypes. Ki-67 expression was found to be positive only in malignant tumors (Ki-67 LI >1%). High Ki-67 LI was associated with high-grade serous cystadenocarcinoma (48.5%), advanced FIGO staging (40.5%), and high CA-125 levels. However, there was no association between Ki-67 LI and histological subtype. Conclusion: Ki-67 is a cost-effective proliferative marker. Therefore, immunohistochemical assessment of Ki-67 expression can be included in routine histopathological report of SEOT for diagnosis and prognostication which will help in better understanding of the biologic behavior of the tumor and modifying treatment strategies.

To correlate histopathological changes and transvaginal sonography findings in the endometrium of patients with abnormal uterine bleeding Akanksha Wankhade, Sunita Vagha, Samarth Shukla, Arvind Bhake, Sheronica Laishram, Deepika Agrawal, Naincy Rastogi, Madhuri Wankhade Journal of Datta Meghe Institute of Medical Sciences University 2019 14(1):11-15 Background: Menstrual ailments are the common manifestation to call for medical visits among women of heavy menstrual bleeding of procreative age. Abnormal uterine bleeding (AUB) affects up to 30% of females in the society. Aim and Objective: The aim of this study was to establish the role of histopathological diagnosis of uterine lesions in patients of AUB and to correlate the transvaginal sonography (TVS) findings with histopathological examination. Materials and Methods: A observational and analytical study was carried out in 120 patients of AUB. All the patients underwent TVS, followed by a histopathological examination of specimens obtained from either D and C or hysterectomy. Results: Menorrhagia was the most common clinical finding. Mean endometrial thickness measured by TVS was 10.15 ± 3.86. Proliferative endometrium (28.33%) was the most frequent finding in histopathological examination, followed by endometrial hyperplasia comprised of 20.83%. Hyperplasia and carcinoma both had a low sensitivity of 36% and 50%, respectively, on TVS compared with histopathology. Conclusion: According to the histopathological diagnosis based on biopsy or D and C, the treatment plan is formulated and the surgery is planned. Hence, even the prognosis depends upon the histopathological examination, TVS is only the initial screening tool for AUB. We advised the clinicians not to rule out these entities only on the basis of TVS.

Impact of Living (Surface) Anatomy module as continuous professional development program for practicing physiotherapists Bincy M George, Satheesha B Nayak, Prem Venketesan, Sapna Marpalli, Mohandas K. G Rao Journal of Datta Meghe Institute of Medical Sciences University 2019 14(1):16-21 Background: It is often presumed that physiotherapists need to be thorough in their knowledge in gross anatomy prior to learning new skills or therapeutic techniques. We implemented a Living (Surface) Anatomy module for physiotherapists, prior to the teaching of therapeutic techniques, in one of the continuous professional development (CPD) programs as a supportive venture for their clinical practice. The present study intended to evaluate the impact of a CPD program on reinforcing practicing physiotherapists' knowledge, skills, and attitude for their clinical practice. Methods: A 1-day workshop was conducted on living (surface) anatomy for practicing physiotherapists (n = 27) through a CPD program. This training included manual muscle testing and body painting of selected muscles of trunk and limbs. Pre- and post-tests were conducted to analyze the impact of the module on improvement in participants' knowledge and skills. In addition, participants were requested to respond to a questionnaire (15 items) on a 5-point Likert scale. Results: Analysis of the pre- and post-test scores revealed a significant increase (34.6%) in surface anatomy knowledge. Majority of the participants opined that the workshop was organized effectively (100%) and the modules helped them to become aware of the lacunae in their knowledge (100%). They also echoed that they realized the need for continuous self-directed learning (100%) and responded that they would attempt to apply whatever they learned through the workshop in their clinical practice (96%). The overall satisfaction score reported by the participants was 9, on a rating scale ranging from 1 to 10 (1 = very poor; 10 = excellent). Conclusion: The CPD was well received by the participants, as evident from their feedback. The present study results demonstrated that the CPD had a positive impact on the participants' knowledge, skills, and attitude.

Profile of urinary tract infection in a rural tertiary care hospital: Two-year cross-sectional study Priyansh Bhayani, Rajendra Rawekar, Shilpa Bawankule, Sunil Kumar, Sourya Acharya, Abhay Gaidhane, Mahalaqua Nazli Khatib Journal of Datta Meghe Institute of Medical Sciences University 2019 14(1):22-26 Aim: The aim of this study was to study the profile of urinary tract infection (UTI) in patients admitted to the medicine department. Settings and Design: This was a cross-sectional study done in a rural tertiary care hospital conducted for a period of 2 years from September 2016 to September 2018. Materials and Methods: All the patients who were admitted (irrespective of the diagnosis at the time of admission) with onset of fever after 48 h of admission, and patients with catheter-associated UTI (CAUTI) and non-CAUTI were defined using the Centers for Disease Control and Prevention guidelines. Statistical Analysis: Statistical analysis was done using descriptive and inferential statistics using Chi-square test and Student's unpaired t-test, and software used in the analysis were SPSS 22.0 version (IBM, USA) and GraphPad Prism 6.0 version (Graphpad software, Inc. California, USA) and P < 0.05 is considered as level of significance. Results: Eighty-four patients developed UTI, the most common organism causing UTI was Escherichia coli. The risk factors associated with CAUTI were higher age, prolonged duration of catheterization, diabetes, and chronic kidney disease. The risk factors associated with non-CAUTI were higher age and benign prostate hyperplasia. The risk factors associated with mortality were prolonged duration of catheterization and diabetes. Twelve patients (14.28%) with CAUTI succumbed in the ICU to their primary illness. Conclusions: Diabetic and elderly patients are at high risk of developing UTI and patients with CAUTI had higher mortality and morbidity.

Dermatoglyphics: A prediction tool for malocclusion Smitha Sammith Shetty, Gloria Su Mi Li, Nurul Ashiqin Binti Babji, Liyana Syatrah Binti Mohd Yusof, Nicholas Ngieng Jiun Yang, Teong Dun Jun, Kuhan Magandran Journal of Datta Meghe Institute of Medical Sciences University 2019 14(1):27-30 Introduction: Dermatoglyphics has proved to have a potential role in predicting the anomalies related to orofacial regions. Aim: The aim of the present study was to explore an association if any, between the dermatoglyphic patterns and type of malocclusion among the Malaysian dental and medical students. Materials and Methods: A total of 104 Malaysian dental and medical students were included in the study. The fingerprints and palm prints were recorded to analyze the type of pattern. Occlusion status was clinically assessed using Angle's classification of malocclusion. Results: Statistically significant association was seen between the left thumb ridge pattern and type of malocclusion. Individuals with loop ridge pattern on their left thumb showed high frequency of Class I normal occlusion and Class III malocclusion, and those with whorl ridge pattern were witnessed to have Class I malocclusion. Conclusion: Dermatoglyphics serves to strengthen the diagnostic impression of malocclusion at an early age and hence can aid in predicting malocclusion and plan preventive and interceptive orthodontics in pediatric patients.

Role of sonoelastography in diagnosing endometrial lesions: Our initial experience Gulam Marfani, Suresh Vasant Phatak, Kaustubh Anil Madurwar, Samida Samad Journal of Datta Meghe Institute of Medical Sciences University 2019 14(1):31-35 Context: Real-time elastography is a novel and dynamic imaging technique that is based on the softness or hardness of tissues or organs of interest under the appropriate compression and can be used with conventional ultrasonography (USG) probes after performing grayscale imaging and Doppler USG. Aims: The aim was to analyze the role of sonoelastography in diagnosing endometrial lesions. Subjects and Methods: A prospective study was conducted from August 2016 to 2018. 30 females were included who underwent sonographic examination. Diagnoses were made and later elastograms were obtained. Strain ratios were calculated and the final diagnosis was compared to histopathological diagnoses to evaluate the role of sonoelastography in diagnosing endometrial pathologies. Statistical Analysis Used: Statistical analysis was done be using descriptive and inferential statistic's using chi square test, students unpaired test, sensitivity and specificity and software used in the analysis were SPSS 22.0 version and graft pad prism 6.0 version. Results: Of the total 30 patients, 5 cases were misdiagnosed on USG compared to 1 on elastography which was of atypical endometrial hyperplasia. Sensitivity, specificity, and diagnostic accuracy of ultrasound were found to be 90.28%, 80%, and 88.5%, respectively. Sensitivity, specificity, and diagnostic accuracy of sonography with elastography were 93.06%, 86.67%, and 91.95%, respectively, showing better results. Conclusions: Sonography coupled with elastography showed better results and can be used to avoid dependency on computed tomography and magnetic resonance imaging avoiding radiation exposures and high cost, especially in a developing country like India, or unnecessary surgical interventions can also be avoided.

Awareness of tuberculosis control program among health-care workers in a tertiary hospital, South India Satyavamsi Gadde, T Jaya Chandra Journal of Datta Meghe Institute of Medical Sciences University 2019 14(1):36-38 Objectives: The objective of the study is to access the awareness of tuberculosis control program (TCP) among health-care workers (HCWs) in a tertiary hospital and to compare the knowledge on TCP between HCWs and the public. Materials and Methods: The participants were provided with printed, multiple-choice questionnaire, containing 28 questions and options to mark the correct answer. All the questions were provided with “not aware” option also. The results were tabulated and analyzed by giving 1 for correct answer, −1 for wrong answer, and 0 for not aware option. The Mann–ANOVA test was used to find the statistical difference. Results: The mean average of scores was 18.52 in the test group and − 0.12 in the control group. Statistical mean significant difference was observed between the groups (P < 0.05). Conclusion: Awareness among the public and HCWs is important for the success of Revised National Tuberculosis Control Programme (RNTCP). To create the awareness in the public, it has to get advertised in cinema theaters, local channels, and newspapers. For awareness among HCWs, scientific updates should be advertised frequently in the journals and symposiums/seminars should be organized regularly by RNTCP.

A rare case of bilateral multiple ovarian dermoids with uterine fibroid and ectopic kidney Suvarna Satish Deshpande, Suresh Vasant Phatak Journal of Datta Meghe Institute of Medical Sciences University 2019 14(1):39-41 Ovarian dermoid cysts are made up of solid, cystic, and fat tissue arising from two of the three germ layers. The incidence of they being bilateral is rare and accounts for 10%–15%. A 42-year-old female came with chief complaints of increased bleeding during menses and pain in the abdomen. Ultrasonography revealed echogenic lesion with heterogeneous echotexture in both the ovaries with internal echogenic strands within which was s/o bilateral dermoid cyst. The patient had a left ectopic renal kidney on contrast-enhanced computed tomography of the abdomen; there was well–defined, rounded, mixed-density mass lesion seen in the pelvis showing fat density, areas of calcification (Rokitansky protuberance), soft tissue, and fluid components. After postoperative and histopathological correlation, it was found to be bilateral dermoid.

Plexiform unicystic ameloblastoma with adenoid differentiation: An unusual finding Monal Yuwanati, Ravi Dande, Amol Ramchandra Gadbail, Shailesh Gondivkar Journal of Datta Meghe Institute of Medical Sciences University 2019 14(1):42-44 Unicystic ameloblastoma (UA) shows clinical and radiologic characteristics of an odontogenic cyst, but histologically shows a typical ameloblastomatous epithelial lining part of the cyst cavity, with or without luminal and/or mural tumor proliferation. Very few cases were reported with variation in histological pattern but rarely with adenoid differentiation. Keeping this in mind, we here report a case report of plexiform UA with adenoid differentiation in maxilla.

Alexandros Sfakianakis
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