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Leibniz Research Centre for Working Environment and Human Factors
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B. C. Behera, Pune, India
T. Chen, Stanford, CA/USA
E. Corsini, Milan, Italy
P. Diel, Cologne, Germany
C. Esser, Duesseldorf, Germany
P. B. Farmer, Leicester/UK
S. Hammad, South Valley/Egypt
P. Jennings, Innsbruck, Austria
M. Lotti, Bonn, Germany / Padova, Italy
P. Micke, Uppsala, Sweden
A. N. Misra, Ranchi, Jharkhand State, India
B. Ponugoti, Pennsylvania, PA/USA
C. Pope, Stillwater, OK/USA
K. Renganathan, Johnstown, PA/USA
S. D. Ray, Fort Wayne, IN/USA
M. Schwarz, Tuebingen, Germany
J. Timmer, Freiburg, Germany
H. van Steeg, Bilthoven, The Netherlands
A. Winterpacht, Erlangen, Germany
Y. Zhou, New Haven, CT/USA

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Detection of aflatoxin-producing fungi isolated from Nile Tilapia and fish feed
(2017-12-13) Mohamed, Hams M.A.; Emeish, Walaa F.A.; Braeuning, Albert; Hammad, Seddik
Contamination of fish by fungi and their mycotoxins poses major health concerns to human and animals. Therefore, our study was aimed to investigate Aspergillus flavus (A. flavus) infections and the levels of aflatoxins in Nile tilapia, Oreochromis niloticus (O. niloticus), and fish feed. Samples from O. niloticus and fish feed (n=25 for each) were randomly collected from private fish farms at Qena province, Egypt, during the winter season. Different Aspergillus spp. were detected in 60 % and 64 % of O. niloticus and fish feed, respectively. HPLC-based analysis revealed aflatoxin-producing activity in 75 % and 83 % of A. flavus isolates from fish and fish feed, respectively. While 96 % of O. niloticus muscles and fish feed samples were contaminated with aflatoxins, the detected levels were below the permissible limits, i.e. 20 μg/kg. Moreover, experimental infection with toxicogenic A. flavus isolates was conducted to evaluate their pathogenicity in O. niloticus. Expectedly, experimental infections of O. niloticus with A. flavus were associated with several clinical symptoms reported in naturally infected fish, e.g. yellow coloration with skin ulceration, hemorrhagic ulcerative patches on gills and skin, corneal opacity, fin rot and abdominal distention. Furthermore, aflatoxicogenic A. flavus isolates from fish were sensitive to herbal clove oil.Even though the measured levels of aflatoxin were below permissible limits, effort should be placed on furtherreduction of exposure to genotoxic and carcinogenic mycotoxins.
An in silico approach in predicting the possible mechanism involving restoration of wild-type p53 functions by small molecular weight compounds in tumor cells expressing R273H mutant p53
(2017-12-08) Malami, Ibrahim; Muhammad, Aliyu; Etti, Imaobong C.; Waziri, Peter M.; Alhassan, Alhassan M.
R273H mutant p53 is a DNA-contact mutant that renders p53 dysfunctional due to a single substitution of Arg273 for His273. Rescuing R273 mutant p53 implies that a competent molecule would have to bind to the site of DNA-contact hot spots to complement the loss of contact with the DNA-binding domain. Here, curcumin, flavokawain B, and alpinetin were docked against the crystal structure of R273H mutant p53 in silico. Consequently, all the compounds bind to the cavity of R273H mutant p53 with a dissociation constant estimated to have 36.57, 70.77, and 75.11 μM for curcumin, flavokawain B, and alpinetin, respectively. Subsequently, each molecule was able to bind to the R273H mutant p53 by interacting with the DNA-contact hot spot Arg248 and mutant R273H, thereby compensating for the loss of direct contact with the DNA-binding domain. Furthermore, all the molecules were able to induce a direct contact with the consensus site of the DNA binding domain, thus maintaining DNA-contact residues with the DNA. The present findings offer reliminary indirect supporting evidence that small molecular weight compounds may certainly rescue DNA-contact mutant p53, which may lay a foundation for designing a competent and effective molecule capable of rescuing mutant p53 in tumor cells expressing R273H mutant p53.
The predictive value of pre- and post-induction chemotherapy plasma EBV DNA level and tumor volume for the radiosensitivity of locally advanced nasopharyngeal carcinoma
(2017-11-28) Song, Yang; Xiao, He; Yang, Zhenzhou; Geng, Mingying; Ma, Jungang; Ren, Yujiang; Liu, Yun; Wang, Ge
This study was dedicated to investigate the predictive value of pre- and post-induction chemotherapy plasma EBV (Epstein-Barr Virus) DNA level and tumor volume for the radiosensitivity of locally advanced NPC. 129 previously untreated locally advanced NPC patients were enrolled. Plasma EBV-DNA copy number and tumor volume was detected before and after induction chemotherapy. The tumor volume was also measured after radiotherapy. Among 129 patients, 98 were positive for EBV DNA. The residual gross target volume of the primary tumor (GTVnx) and GTVnd after radiotherapy was positively correlated with post-induction chemotherapy EBV copy number (rho=0.357, P<0.001; rho=0.356, P<0.001, respectively). Univariate logistic regression analyses showed that the AUC of ROC curves of post-induction chemotherapy tumor volume, tumor regression rate before and after induction chemotherapy, post-induction EBV copy number, EBV decrease rate for predicting no residual nasopharyngeal tumor were 0.859, 0.782, 0.678 and 0.657, respectively. Multivariate logistic analyses showed that T stage, post-induction chemotherapy EBV copy number and tumor volume were independent predictors for no residual nasopharyngeal tumor after radiotherapy. The changes in plasma EBV DNA and tumor volume during treatment could be used to predict the sensitivity of locally advanced NPC patients in response to intensity-modulated radiation therapy (IMRT).
In vitro transfection of anti-tumor miR-101 induces BIM, a pro-apoptotic protein, expression in acute myeloid leukemia (AML)
(2017-11-27) Nikoonahad Lotfabadi, Narges; Mohseni Kouchesfahani, Homa; Sheikhha, Mohammad Hasan; Kalantar, Seyed Mehdi
Acute myeloid leukemia (AML) frequently relapses after initial treatment, though it is possible that drug resistance occurs. Hence, it seems necessary to develop novel therapies such as gene therapy specifically via miRNA trans- fection. MicroRNA-101 has been considered as a tumor suppressor in different types of cancer. It is demonstrated that exogenous miR-101 transfection is associated with decreased viability in AML in this paper. Besides, the increase of pro-apoptotic protein BIM expression in both mRNA and protein level has been illustrated. The recent findings provide an insight into the novel function of miR-101 in AML by activating BIM as an important mediator in intrinsic apoptosis pathways. Generally, miR-101 has been considered as a therapeutic target in our data and might have a valuable role in AML.
The relationship between serum levels of fibroblast growth factor 21 and diabetic retinopathy
(2017-11-22) Mousavi, Zohre; Bonakdaran, Shokoufeh; Sahebkar, Amirhossein; Yaghoubi, Gholamhossein; Yaghoubi, Mohammad Ali; Davoudian, Najmeh; Mohebbi, Masoud
Fibroblast growth factor 21 (FGF21) is a major metabolic regulator that has been shown to be elevated in a number of metabolic disturbances including type 2 diabetes mellitus (T2DM) and the metabolic syndrome, but few studies about the relationship between serum FGF21 and the complications of diabetes have been done. Since the association between FGF21 and diabetic retinopathy is not clear, this study was conducted to investi- gate this relationship. In this cross-sectional study, 61 subjects (14 healthy controls, 22 diabetic patients without retinopathy, and 25 patients with diabetic retinopathy) were evaluated. All patients in the study were examined for the presence of diabetic retinopathy. Various clinical and biochemical parameters including FGF21 were evaluated and analyzed and compared between the study groups. Serum levels of FGF21 showed a significant difference between the three groups (P=0.003) but the difference between diabetic patients with and without ret- inopathy was not significant (P=0.122). Regression model was used to evaluate the role of FGF21 in predicting diabetic retinopathy. In the multivariate logistic regression model after adjustment of systolic blood pressure and fasting blood glucose, the level of FGF21 was not associated with diabetic retinopathy. In the multivariate mod- el, only fasting blood glucose was associated with diabetic retinopathy (P=0.009). According to the results of this study, serum levels of FGF21 in diabetic patients was higher than the control group but these raised levels could not predict the presence of diabetic retinopathy.
Comparative proteomics analysis of Neisseria gonorrhoeae strains in response to extended-spectrum cephalosporins
(2017-11-08) Nabu, Sunanta; Lawung, Ratana; Isarankura-Na-Ayudhya, Patcharee; Roytrakul, Sittiruk; Dolprasit, Supamas; Sengyee, Sineenart; Isarankura-Na-Ayudhya, Chartchalerm; Prachayasittikul, Virapong
Neisseria gonorrhoeae strains displaying reduced susceptibility and resistance to extended-spectrum cephalo- sporins (ESCs) are major public health concerns. Although resistance mechanisms of ESCs have extensively been studied, the proteome-wide investigation on the biological response to the antibiotic stress is still limited. Herein, a proteomics approach based on two-dimensional gel electrophoresis and MALDI-TOF/TOF-MS analy- sis was applied to investigate the global protein expression under ESC stresses of ESC-susceptible and ESC- reduced susceptible N. gonorrhoeae strains. Upon exposure to ceftriaxone, 14 and 21 proteins of ESC- susceptible and ESC-reduced susceptible strains, respectively, were shown to be differentially expressed. In the meanwhile, differential expressions of 13 and 17 proteins were detected under cefixime stress for ESC- susceptible and ESC-reduced susceptible strains, respectively. ESC antibiotics have been proven to trigger the expression of several proteins implicated in a variety of biological functions including transport system, energy metabolism, stress response and pathogenic virulence factors. Interestingly, macrophage infectivity potentiators (Ng-MIP) showed increased expression for ESC-reduced susceptible strain under ESC stress. The altered ex- pression of Ng-MIP was found to be a unique response to ESC stresses. Our finding proposes a broad view on proteomic changes in N. gonorrhoeae in response to ESC antibiotics that provides further insights into the gono- coccal antimicrobial resistance and physiological adaptation mechanism.
Higher risk of progressing breast cancer in Kurdish population associated to CDH1 -160 C/A polymorphism
(2017-11-06) Zarei, Farzaneh; Menbari, Mohammad Nazir; Ghaderi, Bayazid; Abdi, Mohammad; Vahabzadeh, Zakaria
There is an increasing interest about studying possible effects of genetic polymorphisms and risk of cancer pro- gression. E-cadherin (CDH1) involves in many important cellular processes including cell-cell interactions, cell development and genetic changes of this molecule has been associated with greater tumor metastasis. The present study was aimed to evaluate the possible role of CDH1 -160 C/A polymorphism as a potential risk factor for breast cancer in Kurdish population. This case-control study consisted of 100 breast cancer patients and 200 healthy controls. Clinicopathological findings of all individuals were reported and immunohistochemistry staining was carried out on tissue samples. The CDH1 -160 C/A genotype was determined by polymerase chain reaction- re- striction fragment length polymorphism method (PCR-RFLP). CDH1 -160 C/A polymorphism was differently distributed between patient and control groups. The A allele of CDH1 -160 C/A polymorphism significantly in- creased in patients compared to controls. In addition we found that the A allele of this polymorphism might be a potential risk factor for progression of breast cancer in our studied population. Patients with A allele of CDH1 -160 C/A was in higher risk to progress invasive ductal carcinoma. The A allele was also correlated with high grade and stage IV and also with metastatic tumors in studied subjects. The CDH1 -160 C/A polymorphism is correlated with clinicopathologial findings of breast cancer patients. The A allele of CDH1 -160 C/A may be a risk factor for progression of breast cancer in Kurdish patients.
Evaluation of glutathione S-transferase P1 (GSTP1) Ile105Val polymorphism and susceptibility to type 2 diabetes mellitus
(2017-11-06) Saadat, Mostafa
It is well established that type 2 diabetes mellitus (T2DM) is associated with oxidative stress and glutathione S- transferases (GSTs) protect cells against oxidative stress. The missense substitution Ile105Val (rs1695) of the glutathione S-transferase P1 (GSTP1, OMIM: 134660) results from an A/G base substitution at nucleotide 313. Many studies have evaluated the correlation between the rs1695 polymorphism and T2DM, but the results remain inconclusive. The aim of the present meta-analysis was to investigate the association between GSTP1 Ile105Val polymorphism and the susceptibility risk of T2DM. Eligible studies (published before August 2017) were identi- fied in several databases. The heterogeneity between studies was evaluated with the chi-square based Q test and the I2 test. The strengths of the association were assessed by pooled odds ratios (ORs) and the corresponding 95 % confidence interval (95 % CI) using either a fixed or random-effects models. Eighteen studies documenting a total of 2595 T2DM cases and 2888 controls were included in this meta-analysis. In the overall analysis there was no significant association between the rs1695 polymorphism and the risk of T2DM. The subgroup analyses stratified by ethnicity, publication year and sample size did not reveal significant association between the study polymor- phism and the risk of T2DM and any sources contributing to the substantial heterogeneity between studies. The present meta-analysis suggested that there was significant heterogeneity between studies. Considering some limi tations of our meta-analysis, further large-scale studies should be done to reach a more comprehensive understanding.
microRNA-503 contribute to pancreatic ß cell dysfunction by targeting the mTOR pathway in gestational diabetes mellitus
(2017-10-27) Xu, Ke; Bian, Dezhi; Hao, Lanxiang; Huang, Fei; Xu, Min; Qin, Jie; Liu, Yanmei
Loss of pancreatic β cells is involved in pathogenesis of gestational diabetes mellitus (GDM). Recently, several studies have elucidated the connection between microRNAs (miRNAs) and diabetes mellitus (DM), but the role of miRNAs in GDM remains unclear. The aim of this study was to evaluate the potential functions of miRNAs in GDM and to investigate the underlying mechanisms of action. First, we explored the expression profile of miRNAs in placenta tissue from GDM patients using microarray. Validation analysis was performed in peripheral blood specimens using quantitative reverse transcription PCR (qRT-PCR). Then the role and regulating mechanism of miR-503 in weaken the function of pancreatic β cell was investigated. We found that miR-503 was markedly upregulated in placenta tissue from GDM patients, as elevated in peripheral blood specimens, and the high level was positively correlated to blood glucose concentration. Knockdown of miR-503 enhanced insulin secretion of pancreatic β-cells, promoted cell proliferation and protected cells from apoptosis, whereas overexpression of miR- 503 showed the opposite effects. Furthermore, mammalian target of rapamycin (mTOR) was identified as a direct target of miR-503 and mTOR silencing could reverse the improving effects of miR-503 knockdown on insulin secretion and pancreatic β-cells proliferation. High expression of miR-503 in peripheral blood may be acted as a unctions of pancreatic β-cells by targeting the mTOR pathway, suggesting that targeting miR-503/mTOR axis could serve as a novel therapeutic target for GDM.
Knockdown of SLC39A7 suppresses cell proliferation, migration and invasion in cervical cancer
(2017-10-24) Wei, Yongqing; Dong, Jie; Li, Fuli; Wei, Zhuqing; Tian, Yuling
Cervical cancer is the fourth leading cause of malignancy related mortality in women worldwide. SLC39A7 (ZIP7) is a zinc transporter that plays a key role in intestinal epithelial self-renewal. However, whether or not SLC39A7 is involved in human cervical cancer remains unclear. In this study, we investigated the effects of SLC39A7 in cervical cancer in vitro and elucidate related underlying mechanisms. Using Oncomine data analysis, we first found SLC39A7 is commonly upregulated in cervical cancer tissues in comparison with corresponding normal controls. The in vitro experiments indicated that silencing of SLC39A7 expression resulted in decreased cell pro- liferation, increased cell apoptosis, and attenuated migratory and invasive ability using CCK-8, colony formation, flow cytometry, transwell assays, respectively in cervical cancer cell lines, HeLa and ME-180 cells. In molecular levels, Western blot further demonstrated that silencing of SLC39A7 significantly upregulated the expression of Bax and E-cadherin, downregulated the expression of Bcl-2 and MMP-2 in both HeLa and ME-180 cells. These findings provide evidence that SLC39A7 plays a positive role in the progression of cervical cancer and its knock- down might be as a potential therapeutic target for cervical cancer treatment.
Evaluation of in vitro and in vivo anticancer potential of two 5-acetamido chalcones against breast cancer
(2017-10-19) Wankhede, Sonal; Kumar, Nitesh; Simon, Lalitha; Biswas, Subhankar; Gourishetti, Karthik; Ramalingayya, Grandhi Venkata; Joshi, Mit; Rao, C. Mallikarjuna
Two 5’acetamido chalcones, C1 and C2 were synthesized by Claisen-Schmidt condensation method and characterized by IR, LC-MS, 1H NMR and 13C NMR. These compounds were evaluated for anticancer activity in vitro in breast cancer cell lines (MCF-7 and MDA-MB-231) using MTT assay, anti-metastatic assay, apoptotic screening by AO/EB staining and in vivo in N-Methyl-N-nitrosourea (MNU) induced breast carcinoma model. Sprague-Dawley rats with developed tumors (50 mg/kg MNU i.p.) were grouped in four, namely MNU control (0.25 % of CMC p.o.), standard group (doxorubicin 2 mg/kg once in 4 days, i.p.), C1 and C2 groups (50 mg/kg p.o. each). After 21 days of treatments, tumor volume and weight were assessed. Excised tumors were subjected to DNA fragmentation study. MTT assay showed IC50 values of 62.56 and 37.8 µM by for C1 and C2. Both compounds increased apoptotic bodies more than 3 fold compared to normal control in AO/EB staining. Antimeta- static (scratch wound) assay showed a significant (p<0.05) reduction in cell migration after 24 h and 48 h treat- ments compared to normal control. In in vivo studies, tumor weight and tumor volume were significantly (p<0.05) reduced in the doxorubicin group as well as in test groups compared to MNU control. DNA fragmentation assay showed an increase in the number of bands formed in C1 and C2 compared to normal control. Results obtained from in vitro and in vivo studies demonstrated the significant anticancer potentials of C1 and C2.
Microwave-assisted synthesis and antitumor evaluation of a new series of thiazolylcoumarin derivatives
(2017-08-30) Gabr, Moustafa T.; El-Gohary, Nadia S.; El-Bendary, Eman R.; El-Kerdawy, Mohamed M.; Ni, Nanting
A new series of thiazolylcoumarin derivatives was synthesized. The designed strategy embraced a molecular hybridization approach which involves the combination of the thiazole and coumarin pharmacophores together. The new hybrid compounds were tested for in vitro antitumor efficacy over cervical (Hela) and kidney fibroblast (COS-7) cancer cells. Compounds 5f, 5h, 5m and 5r displayed promising efficacy toward Hela cell line. In addition, 5h and 5r were found to be the most active candidates toward COS-7 cell line. The four active analogs, 5f, 5h, 5m and 5r were screened for in vivo antitumor activity over EAC cells in mice, as well as in vitro cytotoxicity toward W138 normal cells. Results illustrated that 5r has the highest in vivo activity, and that the four analogs are less cytotoxic than 5-FU toward W138 normal cells. In this study, 3D pharmacophore analysis was performed to investigate the matching pharmacophoric features of the synthesized compounds with trichostatin A. In silico studies showed that the investigated compounds meet the optimal needs for good oral absorption with no expected toxicity hazards.
Anti-inflammatory effect and mechanism of action of Lindera erythrocarpa essential oil in lipopolysaccharide-stimulated RAW264.7 cells
(2017-08-29) Ko, Yeong-Jong; Ahn, Ginnae; Ham, Young-Min; Song, Sang-Mock; Ko, Eun-Yi; Cho, Su-Hyeon; Yoon, Weon-Jong; Kim, Kil-Nam
The aim of this study was to investigate the chemical constituents of Lindera erythrocarpa essential oil (LEO) by gas chromatography-mass spectrometry and evaluate their inhibitory effect on the expression of pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Fifteen compounds, accounting for 63.7 % of the composition of LEO, were identified. The main compounds were nerolidol (18.73 %), caryophyllene (14.41 %), α-humulene (7.73 %), germacrene-D (4.82 %), and α-pinene (4.47 %). LEO significantly inhibited the expression of inducible nitric oxide (NO) synthase and cyclooxygenase-2, and subsequent production of NO and prostaglandin E2. In addition, it reduced the release of pro-inflammatory cytokines in LPS-activated RAW264.7 cells. The molecular mechanism underlying the effect of LEO was associated with inhibition of the phosphorylation of mitogen-activated protein kinase (MAPK). Furthermore, LEO inhibited LPS-induced phosphorylation and degradation of inhibitor of kappa B-α, which is required for the activation of the p50 and p65 nuclear factor (NF)-κB subunits in RAW264.7 cells. Taken together, these data suggest that LEO exerted its anti-inflammatory effect by downregulating LPS-induced production of pro-inflammatory mediators through the inhibition of NF-κB and MAPK signaling in RAW264.7 cells.
A novel medium size lactam ring analoges as antibacterial agents
(2017-08-14) Putcha A. N. V., Harita; Putcha, Seshi Kumar; Guduru, Shiva Krishna Reddy; Ravula, Parameshwar; Sharath Chandra, J N Narendra
A novel series of medium size (S)-3-alkyl-3,4,6,7-tetrahydro-1H-benzo[e][1,4]diazonine-2,5-dione (6a-f) analogues were synthesized from (E)-3-(2-nitrophenyl)acrylicacid (2) reacting with various amino acid esters using Di-isopropyl Carbodiimide as a coupling agent. The final cyclization is carried out by using reagent 1-Ethyl-3(3- dimethylaminopropyl) Carbodiimide Hydrochloride. The synthesized compounds have been supported by Mass, 1H-NMR and 13C-NMR. Further antibacterial studies were conducted, where some molecules are noticed with potent activity, especially molecule 6d shown highest activity which was also supported by molecular docking studies. All final molecules were docked with enzyme peptide deformylase to determine the probable binding conformation.
Troxerutin protects hippocampal neurons against amyloid beta-induced oxidative stress and apoptosis
(2017-08-09) Farajdokht, Fereshteh; Amani, Mohammad; Mirzaei Bavil, Fariba; Alihemmati, Alireza; Mohaddes, Gisou; Babri, Shirin
Alzheimer’s disease (AD) is an age-related neurodegenerative disease linked with increased production and/or deposition of amyloid-beta (Aβ) in the brain. The aim of the present study was to investigate the possible neuroprotective effect of troxerutin on an animal model of Alzheimer's disease. Alzheimer model was induced by a single dose intracerebroventricular (ICV) injection of Aβ 1–42 (5 nmol/5 μl). Thereafter, troxerutin (300 mg/kg) was gavaged for 14 days. The hippocampal malondialdehyde (MDA) levels and enzymatic activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and acetylcholinesterase (AChE) were measured using enzymelinked immunosorbent assay (ELISA) method. In addition, the number of apoptotic cells in the dentate gyrus (DG) was assessed by TUNEL kit. The results showed that ICV microinjection of Aβ 1-42 increased MDA levels, reduced SOD and GPx, and increased AChE activities in the hippocampus. Chronic administration of troxerutin significantly attenuated MDA levels and AChE activity and increased SOD and GPx activities in the hippocampus. Moreover, the number of apoptotic cells was decreased by troxerutin treatment. Taken together, our study demonstrated that troxerutin could increase the resistance of hippocampal neurons against apoptosis, at least in part, by diminishing the activity of AChE and oxidative stress. Therefore, troxerutin may have beneficial effects in the management of Alzheimer's disease.
Elevated antibiotic resistance of Sudanese urinary tract infection bacteria
(2017-08-07) Saeed, Amir; Hamid, Shadia A.; Bayoumi, Magdi; Shanan, Salah; Alouffi, Sultan; Alharbi, Samir A.; Alshammari, Fawaz D.; Abd, Hadi
This study determined the prevalence of urinary tract infections in the Sudanese state of Khartoum and antimicrobial susceptibility pattern of isolated bacterial species. 200 adult patient urine specimens were collected and cultivated to identify the growing bacteria and their susceptibility to antibiotics. 35 % of specimens had significant bacterial growth. The most frequent isolates in this study were E. coli, E. faecalis and S. aureus. Most of the isolates were resistant to many antibiotics; Gram-negative and Gram-positive isolates were resistant to 67 % and 44 % of the examined antibiotics, respectively. E. coli was the most frequent bacterium in the studied samples and it was highly resistant to first-line antibiotics. The most resistant bacteria isolated were Pseudomonas species and the lowest was for S. saprophyticus. The results highlighted the need for knowledge about antibiotic susceptibility profile of the bacteria causing UTI prior to antibiotic prescription in order to ensure optimal treatment.
Cancer chemoprevention by oleaster (Elaeagnus angustifoli L.) fruit extract in a model of hepatocellular carcinoma induced by diethylnitrosamine in rats
(2017-07-18) Amereh, Zahra; Hatami, Nasrin; Shirazi, Farshad H.; Gholami, Saman; Hosseini, Seyed Hojjat; Noubarani, Maryam; Kamalinejad, Mohammad; Andalib, Sina; Keyhanfar, Fariborz; Eskandari, Mohammad Reza
Hepatocellular carcinoma (HCC) is a frequent and fatal human cancer with poor diagnosis that accounts for over half a million deaths each year worldwide. Elaeagnus angustifolia L. known as oleaster has a wide range of pharmacological activities. This study aimed to investigate the chemopreventive effect of aqueous extract of E. angustifolia fruit (AEA) against diethylnitrosamine (DEN)-induced HCC in rats. HCC was induced in rats by a single injection of DEN (200 mg/kg) as an initiator. After two weeks, rats were orally administered 2-acetylaminofluorene or 2-AAF (30 mg/kg) as a promoter for two weeks. Oleaster-treated rats were orally pretreated with the increasing doses of AEA two weeks prior to DEN injection that continued until the end of the experiment. In the current study, a significant decrease in serum biomarkers of liver damage and cancer, including alfa-fetoprotein (AFP), gamma glutamyl transpeptidase (GGT), alanine transaminase (ALT), and aspartate transaminase (AST) was observed in AEA-treated rats when compared to HCC rats. Furthermore, the oleaster extract exhibited in vivo antioxidant activity by elevating reduced glutathione (GSH) contents as well as preventing lipid peroxidation in the liver tissues of DEN-treated rats. The relative weight of liver, a prognostic marker of HCC, was also reduced in oleaster-treated rats. To conclude, our results clearly demonstrated that oleaster fruit possesses a significant chemopreventive effect against primary liver cancer induced by DEN in rats. It can be suggested that the preventive activity of oleaster against hepatocarcinogenesis may be mediated through the antioxidant, anti-inflammation, and antimutagenic effects of the fruit.
Effects of platelet rich plasma and chondrocyte co-culture on MSC chondrogenesis, hypertrophy and pathological responses
(2017-07-16) Ramezanifard, Rouhallah; Kabiri, Mahboubeh; Hanaee Ahvaz, Hana
Regarding the inadequate healing capability of cartilage tissue, cell-based therapy is making the future of cartilage repair and regeneration. Mesenchymal stem cells (MSC) have shown great promise in cartilage regeneration. However, a yet-unresolved issue is the emergence of hypertrophic and pathologic markers during in vitro MSC chondrogenesis. Articular chondrocytes (AC) can suppress the undesired hypertrophy when co-cultured with MSC. On the other hand, platelet rich plasma (PRP), is considered potentially effective for cartilage repair and in-vitro chondrogenesis. We thus aimed to harness chondro-promotive effects of PRP and hypertrophic-suppressive effects of AC:MSC co-culture to achieve a more functional cartilage neo-tissue. We used PRP or conventional-differentiation chondrogenic media (ConvDiff) in MSC mono-cultures and AC:MSC co-cultures. We assessed gene expression of chondrogenic and hypertrophic markers using real-time RT-PCR and immunostaining. Alkaline-phosphatase activity (ALP) and calcium content of the pellets were quantified. We also measured VEGF and TNF-α secretion via ELISA. We showed PRP had higher chondrogenic potential (in mRNA and protein level) and hypertrophic-suppressive effects than Conv-Diff (mRNA level). Co-culturing reduced ALP while PRP increased calcium deposition. In all four groups, TNF-α was down-regulated compared to MSC controls, with co-cultures receiving ConvDiff media secreting the least. Meanwhile, the only group with increased VEGF secretion was PRP-mono-cultures. We observed synergistic effects for PRP and AC:MSC co-culture in enhancing chondrogenesis. Inclusion of AC reduced hypertrophic markers and angiogenic potential in PRP groups. We thus propose that combination of PRP and co-culture would favor chondrogenesis while alleviate but not totally eradicate undesired hypertrophic and pathologic responses.
Immunization enhances the natural antibody repertoire
(2017-07-10) Beinart, Dylan; Ren, Daniel; Pi, Cinthia; Poulton, Susan; Holzknecht, Zoie E.; Swanson, Chelsea; Parker, William
The role of immunization in the production of antibodies directed against immunogens is widely appreciated in laboratory animals and in humans. However, the role of immunization in the development of “natural antibodies” has not been investigated. Natural antibodies are those antibodies present without known history of infection or immunization, and react to a wide range of targets, including “cryptic” self-antigens that are exposed upon cell death. In this study, the ability of immunization to elicit the production of natural antibodies in laboratory rats was evaluated. Laboratory rats were immunized with a series of injections using peanut extracts (a common allergen), a high molecular weight protein conjugated to hapten (FITC-KLH), and a carbohydrate conjugated to hapten (DNPFicall). Significantly greater binding of antibodies from immunized animals compared to controls was observed to numerous autologous organ extracts (brain, kidney, liver, lung, prostate, and spleen) for both IgM and IgG, although the effect was more pronounced for IgM. These studies suggest that immunization may have at least one unforeseen benefit, enhancing networks of natural antibodies that may be important in such processes as wound repair and tumor surveillance. Such enhancement of natural antibody function may be particularly important in Western society, where decreased exposure to the environment may be associated with a weakened natural antibody repertoire.
The association of vimentin and fibronectin gene expression with epithelial-mesenchymal transition and tumor malignancy in colorectal carcinoma
(2017-07-10) Niknami, Zohreh; Eslamifar, Ali; Emamirazavi, Amirnader; Ebrahimi, Alireza; Shirkoohi, Reza
Colorectal cancer is the most common malignancy of the gastrointestinal tract with very high mortality. One of the most distinguishing features for the establishment of an epithelial-mesenchymal transition phenotype is the alteration of mesenchymal markers expression and structural adhesion proteins. We evaluated the significance of vimentin and fibronectin gene expression in relation to invasion and metastasis in CRC patients. Tissue specimens were collected consecutively from forty-five colorectal carcinoma patients during surgeries. Tissues were divided into two separate parts for pathological and molecular assays. In order to histological staging, tissue sections were prepared from formalin-fixed paraffin-embedded blocks and stained with Hematoxylin and Eosin. To quantify gene expression, specimens were dissected and homogenized. Moreover, SW480, SW48, SW948, Caco-2, HT-29 and LS174T as human colon cancer cell lines were obtained and cultured, then molecular analyzing was performed. As results the expression of VIM gene increased in SW480, SW48 and SW948 while it decreased in Caco-2, HT-29 and LS174T. Moreover, FN was up-regulated in Caco-2, HT-29 and SW948, while it was down-regulated in SW480, SW48 and LS174T. In tissues, vimentin and fibronectin expression significantly increased in stromal cells, whereas vimentin decreased in colonic epithelial cells and fibronectin had no significant change. Vimentin and fibronectin expression were changed in tumor tissues. It was found an association between vimentin expression with age and tumor size; over-expression in older age and decreasing in larger tumor size. Furthermore, fibronectin over-expression is correlated to older age and high tumor stages; up-regulation with increasing age and high tumor stages.

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