فهرست مطالب

Basic Medical Sciences - Volume:21 Issue: 7, Jul 2018

Iranian Journal of Basic Medical Sciences
Volume:21 Issue: 7, Jul 2018

  • تاریخ انتشار: 1397/03/30
  • تعداد عناوین: 16
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  • Sayyad Khanizadeh *, Banafshe Hasanvand, Hamed Esmaeil Lashgarian, Mohammad Almasian, Gholamreza Goudarzi Pages 651-659
    It is estimated that up to 20% of all types of human cancers worldwide are attributed to viruses. The genome of oncogenic viruses carries genes that have protein products that act as oncoproteins in cell proliferation and transformation. The modulation of cell cycle control mechanisms, cellular regulatory and signaling pathways by oncogenic viruses, plays an important role in viral carcinogenesis. Different signaling pathways play a part in the carcinogenesis that occurs in a cell. Among these pathways, the Wnt signaling pathway plays a predominant role in carcinogenesis and is known as a central cellular pathway in the development of tumors. There are three Wnt signaling pathways that are well identified, including the canonical or Wnt/β-catenin dependent pathway, the noncanonical or β-catenin-independent planar cell polarity (PCP) pathway, and the noncanonical Wnt/Ca2 pathway. Most of the oncogenic viruses modulate the canonical Wnt signaling pathway. This review discusses the interaction between proteins of several human oncogenic viruses with the Wnt signaling pathway.
    Keywords: ?-catenin, Canonical pathway, Carcinogenesis, Oncogenic viruses, Wnt signaling
  • Negar Karimaghai, Amin Tamadon, Farhad Rahmanifar, Davood Mehrabani, Alireza Raayat Jahromi, Shahrokh Zare, Zahra Khodabandeh, Iman Razeghian Jahromi, Omid Koohi-Hosseinabadi, Mehdi Dianatpour* Pages 660-667
    Objective(s)
    Adipose tissue-derived mesenchymal stem cells (AT-MSCs) with more potent immunomodulatory effects, greater proliferative potential and secretion of growth factors and cytokines in comparison with bone marrow derived MSCs are more appropriate for cell therapy. The aims of the present study were to evaluate the histomorphometric effect of AT-MSCs allotransplantation on regeneration of germinal layer cells of seminiferous tubules in busulfan-induced azoospermic hamsters.
    Materials And Methods
    In the present experimental case-control study, AT-MSCs were isolated from adipose tissue of two female and six male donor albino hamsters, and testes of the males were simultaneously used as negative control group. Six mature male recipient hamsters received two doses of busulfan with three weeks interval to stop endogenous spermatogenesis. Right testis of hamsters was intratubular injected with AT-MSCs via efferent duct 35 days after induction of azoospermia and was used as cell therapy group. The left testis without cell therapy was served as azoospermia group.
    Results
    After 35 days, testes and epididymis in all groups were removed for histological evaluation. Histomorphometric analyses of AT-MSCs-treated testes and epididymis showed that the epithelial tissue of seminiferous tubules was normally repaired in most cell-treated seminiferous tubules, and spermatozoa were present in epididymis tubes in comparison with intact testes. The untreated seminiferous tubules and epididymis tubes of azoospermia group were empty.
    Conclusion
    Allotransplanted AT-MSCs could successfully induce spermatogenesis in azoospermic seminiferous tubules of hamster. Therefore, AT-MSCs can be suggested as an attractive candidate in cell transplantation of azoospermia.
    Keywords: Adipose tissue, Azoospermia, Cell therapy, Hamster, Mesenchymal stem cells
  • Hojat Anbara, Rasoul Shahrooz *, Mazdak Razi, Hassan Malekinejad, Gholamreza Najafi Pages 668-677
    Objective(s)
    The aim of the present study was to assess the effects of vitamin C (Vit C) on hemolytic anemia induced by phenylhydrazine (PHZ).
    Materials And Methods
    Twenty-four healthy male mice were divided into four groups, randomly: Control group (0.1 ml/day, normal slaine, IP), PHZ group that received only PHZ 8 mg/100 g/48 hr, IP, PHZ嘫 C group that received PHZ 8 mg/100 g/48 hr, IP and Vit C 100 mg/kg BW-1/day by gavage and Vit C group that received 100 mg/kg BW-1/day Vit C by gavage. After 35 days, germinal cells, RNA damage, sperm parameters, testis malondialdehyde (MDA) content, serum total antioxidant capacity (TAC), pre-implantation embryo development and mRNA levels of cyclin D1 and c-myc in two-cell, and morula and blastocyst stages were assessed.
    Results
    Vit C reduced the RNA damage, enhanced sperm quality, promoted pre-implantation embryo development and improved testicular antioxidant and endocrine status (P
    Conclusion
    Vit C enhanced the fertilizing potential by ameliorating the endocrine status, antioxidant capacity, and sperm quality. Finally, the cyclin D1 and c-myc gene expressions were regulated in PHZ嘫 C treated group that promoted the embryo development.
    Keywords: Hemolytic anemia, Histology, Mice, Testis, Vitamin C
  • Tahereh Barati, Mahnaz Haddadi, Fatemeh Sadeghi, Samad Muhammadnejad, Ahad Muhammadnejad, Ronak Heidarian, Motahareh Arjomandnejad, Saeid Amanpour * Pages 678-681
    Objective(s)
    Gastric cancer is the third leading cause of cancer-related death worldwide. The overall survival rate of patients is poor because gastric cancers are usually diagnosed at the late stages. Therefore, further research is needed and appropriate research tools are required to develop novel therapeutic approaches.
    Materials And Methods
    Eight female athymic nude mice with a C57BL/6 background were used in this study. AGS cells were inoculated into the flank. The tumor volumes were calculated and growth curves were drawn. When the volume of the tumors reached 1000 mm3, the animals were humanely euthanized with CO2 gas. After harvesting, tumors were analyzed with Hematoxylin and Eosin (H&E) and immunohistochemistry (IHC). A pathologist confirmed tumor entity through H&E staining. Tumors were evaluated for expression of HER-2, P53, Ki-67, CD34, cytokeratin 8 (CK8), vimentin, estrogen receptor (ER), and progesterone receptor (PR) utilizing immunohistochemistry.
    Results
    The tumor take rate was 62.5%, mean doubling time was 40.984 d, and the latency period was 30.62 days. The H&E staining results showed highly malignant hyperchromatin epithelial cells. IHC assessment showed the mutation status of P53 gene. The expression score of the CK8 protein in the tumor cells was . Vimentin protein was not expressed and changes in mesenchymal phenotype were not observed. Ki-67 IHC indicated that the proliferation rate was >43% and angiogenesis was defined as high MVD.
    Conclusion
    The respective AGS xenograft model provides an opportunity to understand the pattern of tumor growth as well as to evaluate new gastric cancer therapies in in vivo studies.
    Keywords: AGS, Athymic nude mice, Immunohistochemistry, Stomach neoplasm, Xenograft model
  • Rogaye Diba, Gisou Mohaddes, Fariba Mirzaie Bavil, Fereshteh Farajdokht, Parvin Bayandor, Maryam Hosseindoost, Keyvan Mehri, Zohreh Zavvari Oskuye, Shirin Babri * Pages 682-687
    Objective(s)

    Maternal high-fat diet (HFD) is linked with metabolic and cognitive deficits in offspring. Neuroprotective effects of troxerutin, a natural bioflavonoid, have been reported recently. This study aimed to investigate the effects of troxerutin on spatial memory and serum and hippocampal apelin levels in the male offspring of HFD fed mothers.

    Materials and Methods

    Three-week-old female Wistar rats (n= 40) received HFD or control diet (CD) for 8 weeks. After mating, pregnant animals were divided into two subgroups according to the troxerutin (TRO) supplementation: CD, CD+TRO, HFD, and HFD+TRO. HFD continued to the end of lactation in HFD and HFD+TRO groups. TRO was gavaged (150 mg/kg/day) during pregnancy. After weaning, the male offspring were fed a normal diet until 12 weeks of age. Spatial memory was evaluated in the Morris water maze (MWM) on postnatal day (PND) 90. Total apelin concentration was measured in the serum of maternal rats before mating and after lactation and also in the serum and hippocampus of their male offspring.

    Results

    Both traveled distance (P<0.05) and time spent (P<0.05) in the target quadrant were significantly decreased in the offspring of HFD-fed dams, which were reversed by TRO treatment. Moreover, TRO significantly (P<0.05) decreased serum apelin levels in dams. Furthermore, TRO treatment in dams significantly (P<0.05) increased serum and hippocampal levels of apelin in their offspring.

    Conclusion

    These results indicated that TRO treatment during pregnancy improved maternal HFDinduced spatial memory impairments of the offspring possibly through modulation of serum and hippocampal apelin levels.

    Keywords: Apelin, Maternal high fat diet, Memory, Morris Water Maze, Troxerutin
  • Mohsen Abedi, Seyed Alireza Mesbah-Namin *, Ali Noori-Zadeh, Taki Tiraihi, Taher Taheri Pages 688-694
    Objective(s)
    Human superoxide dismutase 1 (SOD1) is the cytosolic form of this enzyme it detoxifies superoxide anions and attenuates their toxicities and concomitant detrimental effects on the cells. It is believed that the amount of these enzymes present in the oxidative stress-induced diseases is crucial for preventing disease progression. Transfection of rat bone marrow stromal cells (BMSCs) by a constructed vector carrying the human wild-type SOD1 gene, a non-viral gene transfer method, was the main aim of this study.
    Materials And Methods
    For this purpose, the rat BMSCs were transfected with the vector using Turbofect reagent and then stabilized. Western-blot and real-time PCR were also used for evaluation of SOD1 expression.
    Results
    Data analysis from RT-PCR and Western-blot techniques revealed that the stable transfected cells could secrete human wild-type SOD1 in the supernatant. Also, the total activity of SOD1 was about 0.5±0.09 U/ml and 0.005±0.002 U/ml in the supernatants of the transfected and not-transfected of rat BMSCs, respectively.
    Conclusion
    This study showed that expansion of the stable transfected rat BMSCs by a constructed vector carrying the human wild-type SOD1 gene is capable of secreting the active SOD1 enzyme under ex-vivo conditions. The recommendation of this study is that the same experiment would be applicable for expression of the other form of this enzyme, SOD3, as well. More valuable information could probably be provided about the variety of the diseases caused by superoxide anions toxicities by intervention and application of the non-viral method for expressions of SOD1 and SOD3 enzymes.
    Keywords: Vector, Expression, Ex-vivo, Gene delivery, Human SOD1, Rat BMSCs
  • Ayse Aydogan, Seyit Ali Bingol* Pages 695-700
    Objective(s)
    The aim of this study was to determine Oxytocin receptor (OTR) gene expression and localization in diabetic and non-diabetic mouse testes by RT-PCR and immunohistochemistry, respectively.
    Materials And Methods
    In this study, 18 male BALB/c mice (8–12 weeks old) were used and divided into three groups: diabetic, sham, and control. Streptozotocin (STZ) was applied to the diabetic group and sodium citrate was administered to the sham group in the same way, however, the control group was left untouched. The testicular tissues were removed on the thirtieth day of testing; the right testis tissues were passed through a routine histologic process and sections were stained with H&E and PAS staining techniques. The avidin-biotin-peroxidase method was applied to determine OTR immunoreactivity, while the left testis tissues were used for RT-PCR.
    Results
    It was found that the body weight had decreased in the diabetic group and the diameter of the seminiferous tubules in the said group was shorter than those of the other groups. There were no obvious differences with regard to the histologic appearance between the groups. The immunohistochemical examination showed that the OTR immunoreactivity was strong in the control and sham groups but weak in the diabetic group, and the immunoreactivity was only seen in the Leydig cells. In addition, the OTR gene expression was lower in the diabetic group than in the other groups.
    Conclusion
    We concluded that diabetes reduces the OTR expression in the testis. It is suggested that OTR protection should be researched in diabetes for healthy reproduction and sexuality.
    Keywords: Diabetes, Immunohistochemistry, Oxytocin receptor, RT-PCR, Testis
  • Majid Nejati, Mohammad Ali Atlasi, Mohammad Karimian*, Hossein Nikzad, Abolfazl Azami Pages 701-708
    Objective(s)
    Stroke is the most common neurological disorder and genetic susceptibility has an important role in its etiology. Polymorphism in several genes such as lipoprotein lipase (LPL) is propounded as a risk for stroke. This meta-analysis investigated the association of rs285 and rs320 LPL polymorphism with stroke risk.
    Materials And Methods
    We searched PubMed, Clarivate Analytics Web of Science, Google Scholar, and Science Direct databases for appropriate studies. The odds ratios (ORs) with 95% confidence intervals (CIs) were calculated to estimate the strength of this association. Also, the effects of four common polymorphisms (rs268, rs285, rs320, and rs328) on the molecular aspects of LPL were evaluated by in silico tools. Five studies were included in meta-analysis after screening.
    Results
    Our data indicated that rs320 significantly decreased the risk of stroke (G vs. T: OR= 0.64, 95%CI=0.54-0.76; GG vs. TT: OR=0.47, 95%CI=0.29-0.75; TG vs. TT: OR=0.65, 95%CI=0.53-0.80; TG vs. TT: OR=0.62, 95%CI=0.51-0.75; GG vs. TT: OR=0.51, 95%CI=0.32-0.82). Moreover, a significant association between rs285 and diminution of stroke risk was seen (P- vs. P: OR=0.72, 95%CI=0.58-0.91; P-P- vs. P: OR=0.50, 95%CI=0.31-0.82; PP- vs. P: OR=0.72, 95%CI=0.53-0.96; P-P- vs. P㿯迸: OR=0.581, 95%CI=0.369-0.916). Also, the same results were observed after stratifying, without any publication bias (PEgger>0.05). Furthermore, computational analysis revealed that rs268 and rs328 may affect the protein structure (prediction: non-neutral; score=19; expected accuracy=59%) while rs320 could affect the RNA structure (distance=0.2264, P-value=0.0534; P
    Conclusion
    This meta-analysis indicated that risk of stroke was decreased in rs320 and rs285 polymorphisms in the LPL gene.
    Keywords: Computational biology, Genetic polymorphism, Lipoprotein lipase, Meta-analysis, Stroke
  • Fatemeh Mohammadali, Saeid Abroun *, Amir Atashi Pages 709-716
    Objective(s)
    Cord blood (CB) is known as a valuable source of hematopoietic stem cells (HSC). Identifying strategies that enhance expansion and maintain engraftment and homing capacity of HSCs can improve transplant efficiency. In this study, we examined different culture conditions on ex vivo expansion and homing capacity of CB-HSCs.
    Materials And Methods
    In this study, 4-5 different units of human CB in each of 3 independent experiments were collected.CD34 HSC was isolated, cultured in the serum-free medium(Stem line II) and supplemented with cytokines: FMS-like tyrosine kinase 3 ligand (FLt3L), Thrombopoietin (TPO), stem cell factor (SCF) with/without bone marrow mesenchymal stem cell (BM-MSC) feeder layer in normoxia (20% O2) and mild hypoxia (5% O2) for 7 days. Before and after this period, total nucleated cell count (TNC), CD34 cells count, Colony-forming cell (CFC) assay, migration assay and CXCR4 expression were evaluated by real time PCR. Data analysis was performed with t- test and ANOVA. P-value less than 0.05 was considered as statistically significant differences.
    Results
    At the end of 7 days of culture, the highest count of TNC, CD34 cells, CFUs, migration percentage and CXCR4 mRNA level were observed in feeder猫梒⧞ group at 5% O2 tension. Our findings demonstrated statistically significant (1.7-3.2 fold) increase of CXCR4 gene expression in hypoxia versus normoxia.
    Conclusion
    Combination of BM-MSC and mild hypoxia (5% O2) not only improves HSC expansion but also enhances homing capacity of HSC and better mimickes the niche microenvironment conditions.
    Keywords: CD34+ cells, Cord blood, CXCR4, Hematopoietic stem cell, Hypoxia, Mesenchymal stem cell
  • Ma Jianjun, Kangmao Huang, Yan Ma, Shuai Chen, Chao Liu, Zhi Shan, Xiangqian Fang * Pages 717-723
    Objective(s)
    In traditional Chinese medicine, gamboge can detoxify bodies, kill parasites, and act as a hemostatic agent. Recent studies have demonstrated that gambogic acid (GBA) suppressed inflammation in arthritis, and also presented antitumor effect. Thus, this study investigated the new biological properties of GBA on macrophages.
    Materials And Methods
    RAW 264.7 cells were pretreated with GBA at different concentrations (10, 20, 40, 80, 160, 320 nM) for 24 hrs, and then cell viability was measured using Cell Counting Kit (CCK)-8 assays. Pro-inflammatory cytokines such as TNF-α, IL-6 and IL-1β were determined using ELISA kits and qPCR. Then nitrite concentration was calculated according to a standard curve. At last, the effect of GBA on MAPK and NF-κB signaling pathways was assessed by western blot and luciferase reporter gene assay.
    Results
    GBA (IC50: 260 nM) suppressed the TNF-α, IL-6 and IL-1β expression induced by lipopolysaccharide (LPS) in RAW 264.7 cells. The expression of TNF-α, IL-6 and IL-1β decreased to 30-50% and 70-75% in the high-dose (160 nM) and low-dose (40 and 80 nM) GBA groups, respectively. Furthermore, the nitric oxide (NO) production and the activation of NF-κB, ERK, and JNK pathways were significantly reduced in high-dose (160 nM) GBA only, while p38 pathway was inhibited at both low (40 and 80 nM) and high (160 nM) concentration of GBA.
    Conclusion
    These data suggested that GBA inhibited LPS-induced production of pro-inflammatory cytokines including TNF-α, IL-6 and IL-1β mainly through the suppression of the p38 pathway.
    Keywords: Anti-inflammatory agents, Gambogic acid, MAPK, NF-?B, p38, RAW 264.7 cells
  • Nazanin Mozafari, Ali Shamsizadeh, Iman Fatemi, Mohammad Allahtavakoli, Amir Moghadam-Ahmadi, Elham Kaviani, Ayat Kaeidi * Pages 724-730
    Objective(s)
    Growing evidence suggests that dysfunction of the glutamatergic system and α-amino-3-hydroxy-5-methyl-4-isoazolepropionic acid (AMPA) receptors are involved in pathology of Alzheimer’s disease (AD). Because AMPA receptors play a key role in plasticity synaptic regulation, positive modulation of these receptors may rescue the cognitive deficits in the AD. The aim of this study was to explore the effect of CX691, a specific positive allosteric modulator of the AMPA-type glutamate receptors (Ampakine), on spatial learning and memory in a rat model of AD.
    Materials And Methods
    For induction of AD, amyloid-beta 1-42 (Aβ1-42) was microinjected into the hippocampus of male Wistar rats (250-300 g). The Morris water maze (MWM) test was used to evaluate the effect of CX691 (0.03 and 0.3 mg/kg, twice a day for 10 days, orally) on spatial learning and memory of rats. In order to evaluate the protein expression of brain-derived neurotrophic factor (BDNF) in hippocampus tissue, ELISA test was used.
    Results
    The obtained data showed that treatment with CX691 (0.3 mg/kg) improves the impairment of spatial learning and memory in AD rats. Also, treatment with CX691 (0.3 mg/kg), increased the BDNF protein level in hippocampus tissue of AD rats compared to non-treated animals.
    Conclusion
    The CX691 can improve the BDNF protein expression as well as spatial performance of learning and memory in AD rats.
    Keywords: Alzheimer's disease, AMPA receptors, BDNF, CX691, Memory, Rat
  • Hong-Liang Kong *, Ai-Jie Hou, Ning-Ning Liu, Bo-Han Chen, Hua-Ting Huang, Sheng-Nan Dai Pages 731-737
    Objective(s)
    This study intended to investigate the effects of Ginsenoside-Rbl (Gs-Rbl) on fatty acid β-oxidation (FAO) in rat failing heart and to identify potential mechanisms of Gs-Rbl improving heart failure (HF) by FAO pathway dependent on AMP-activated protein kinase (AMPK).
    Materials And Methods
    Rats with chronic HF, induced by adriamycin (Adr), were randomly grouped into 7 groups. Gs-Rb1, adenine 9-β-D-arabinofuranoside (Ara A, specific AMPK inhibitor), and 5'-aminoimidazole-4-carboxamide riboside (Aicar, specific AMPK activator) were administered to rats with HF, singly and/or combinedly. Myocardial high-energy phosphate (such as phosphocreatine, ADP, and ATP), free L-Carnitine, malonyl-CoA, and the activity of FAO-related enzymes in left ventricle from different groups were measured by using the corresponding molecular biological techniques.
    Results
    Gs-Rb1 improved HF significantly, accompanied by a significant increase in phosphocreatine (PCr), ADP, ATP, PCr/ATP ratio, free carnitine, malonyl-CoA, mRNA, activity of carnitine palmitoyltransferase (Cpt), medium-chain Acyl-CoA Dehydrogenase (MCAD) and long-chain acyl-CoA Synthetase (ACSL) and a significant decrease of the ADP/ATP ratio in the left ventricular myocardium. However, all those effects were almost abolished by Ara A and were not further improved by Aicar.
    Conclusion
    Taken together, it suggests that Gs-Rb1 may modulate cardiac metabolic remodeling by improving myocardial fatty acid β-oxidation in failing heart. In addition, the effects of Gs-Rb1 may be mediated via activating AMPK.
    Keywords: AMP-activated protein kinase, Carnitine palmitoyl-transferase 1, Ginsenosides-Rbl, Heart failure, L-carnitine, Long-chain acyl-CoA synthetase, Malonyl-CoA, Medium-chain acyl-CoA dehydrogenase
  • Hamid Sadeghian *, Seyed Mohammad Seyedi, Zeinab Jafari Pages 738-744
    Objective(s)
    15-Lipoxygenases are one of the iron-containing proteins capable of performing peroxidation of unsaturated fatty acids in animals and plants. The critical role of enzymes in the formation of inflammations, sensitivities, and some cancers has been demonstrated in mammals. The importance of enzymes has led to the development of mechanistic studies, product analysis, and synthesis of inhibitors.
    Materials And Methods
    The inhibitory activity of all synthetic compounds against SLO (soybean 15-lipoxygenase: L1; EC 1,13,11,12) was determined using the peroxide formation method. In this method, the basis of evaluation of lipoxygenase activity is measuring the concentration of fatty acid peroxide. All measurements were compared with 4-​methyl-​2-​(4-​methylpiperazinyl)pyrimido[4,​5-​b]benzothiazine (4-MMPB) as one of the known lipoxygenase inhibitors. The radical scavenging ability of all synthetic compounds using stable free radicals (DPPH: 2,2-diphenyl-1-picrylhydrazyl) was measured for further investigation.
    Results
    In this study, a series of esters from phenolic acids with terpenoid alcohols was synthesized and their inhibitory potency against soybean 15-lipoxygenase and their free radical scavenging properties were determined. Among the synthetic compounds, adamantyl protocatetuate 2j and bornyl protocatetuate 2o showed the most potent inhibitory activity with IC50 values of 0.95 and 0.78 μm, respectively.
    Conclusion
    By changing the alcohol and acyl portions of stylosin, it was found that electronic properties play main role in lipoxygenase inhibition potency in contrast with steric features. Insertion of more reductive phenolic moiety such as catechuate and gallate lead to more lipoxygenase inhibition potency of the esters as observed in their radical scavenging activity.
    Keywords: Inhibitors, Phenolic acid, Radical scavenging, Terpenoids, 15-lipoxygenase
  • Wahyu Widowati *, Ervi Afifah, Tjandrawati Mozef, Ferry Sandra, Rizal Rizal, Annisa Amalia, Yukko Arinta, Indra Bachtiar, Harry Murti Pages 745-752
    Objective(s)
    This study aimed to determine the collagen type II (COL2) and SOX9 expression in interleukin growth factor (IGF-1)-induced Wharton’s Jelly mesenchymal stem cells (WJMSCs) and the level of chondrogenic markers in co-culture IGF1-WJMSCs and IL1β-CHON002 as osteoarthritis (OA) cells model.
    Materials And Methods
    WJMSCs were induced with IGF1 (75, 150, and 300 ng/ml) to enhance their chondrogenesis capability. The gene expression of SOX9 and COL2 was evaluated with quantitative RT-PCR. Furthermore, IGF1-WJMSCs were co-cultured with IL1β-CHON002 cells in varied ratios (1:2, 1:1, 2:1). Chondrogenic markers ADAMTS1, ADAMTS5, MMP3, MMP1, and RANKL were measured with ELISA.
    Results
    The IGF1-WJMSCs had an increased expression of COL2 and SOX9. ADAMTS1, ADAMTS5, MMP1, MMP3, and RANKL levels were decreased in the co-culture IGF1-WJMSCs and IL1β-CHON002.
    Conclusion
    The IGF1-induced WJMSCs were capable to enhance chondrogenesis, indicated by increased expression of SOX9 and COL2 and decreased expression of ADAMTS1, ADAMTS5, MMP3, MMP1, and RANKL. These findings can be further used in the osteoarthritis treatment.
    Keywords: Chondrogenesis, IGF-1, Mesenchymal stem cell MMPs, Osteoarthritis
  • Ali Torkashvand, Fariborz Bahrami, Minoo Adib, Soheila Ajdary * Pages 753-759
    Objective(s)
    After decades of containment, pertussis disease, caused by Bordetella pertussis seems to be re-emerging and still remains a major cause of reported vaccine-preventable deaths worldwide. The current licensed whole-cell vaccines display reactogenicity while acellular vaccines are expensive and do not induce Th1-type immune responses that are required for optimum protection against the disease. Thus, there is an urgent need to develop new vaccines and the recombinant technology seems to be the method of choice for this purpose. The present study was an attempt to develop a new, simplified, cost-effective and well-defined vaccine against Bordetella pertussis, with capacity to induce a Th1 response.
    Materials And Methods
    A fusion DNA fragment encoding the N-terminal region of pertussis toxin S1 subunit and filamentous hemagglutinin type 1 immunodominant domain was constructed and the corresponding fusion protein (F1S1) was produced in Escherichia coli. F1S1 in conjunction with imiquimod was administered by subcutaneous (SC) and intranasal (IN) routes to BALB/c mice.
    Results
    This vaccine formulation could elicit high levels of IFN-γ, serum IgG (with higher IgG2a/IgG1 ratio) and lung IgA after the SC and, to a lesser extent, following the IN administration.
    Conclusion
    Our results indicate that the above-mentioned important proteins of B. pertussis could be successfully produced in E. coli as a single fusion protein. Furthermore, this protein could induce proper systemic and mucosal immune responses after administration via SC or IN routes.
    Keywords: Bordetella, Filamentous hemagglutinin, Imiquimod, Pertussis, Pertussis toxin, Recombinant
  • Zulfiqar Ali Mirani *, Aiman Fatima, Shaista Urooj, Mubashir Aziz, Muhammad Khan, Tanveer Abbas Pages 760-769
    Objective(s)
    This study was designed to determine the relationship of Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli isolates in multispecies biofilms and their individual phenotypic characters in biofilm consortia.
    Materials And Methods
    The subject isolates were recovered from different food samples and identified on the basis of growth on differential and selective media. Tube methods, Congo-red agar method, and scanning electron microscopy (SEM) were used to study biofilms phenotypes. The hydrophobicity of the strains was evaluated by the adhesion to apolar solvent.
    Results
    The results showed that E. coli dominated the pre-biofilm stage. It has been observed that E. coli adopted biofilm life much before S. aureus and P. aeruginosa. However, after adopting biofilm lifestyle, slowly and gradually, P. aeruginosa dominated the consortia and dispersed other stakeholders. The subject isolates of P. aeruginosa produce cis-2-decanoic acid to disperse or inhibit S. aureus and E. coli biofilms. Gas-chromatography and mass spectrometry results showed that cis-2-decanoic was higher in the co-culture condition and increased at late log-phase or at stationary phase. Although majority of S. aureus were unable to compete with P. aeruginosa, however, a minor population competed, survived, and persisted in biofilm consortia as small colony variants. The survivors showed higher expression of sigB and sarA genes. P. aeruginosa showed comparatively higher hydrophobic surface properties.
    Conclusion
    Comparative analysis showed that cell surface hydrophobicity, growth rate, and small colony variants (SCVs) are correlated in biofilm consortia of the P. aeruginosa, S. aureus, and E. coli.
    Keywords: Biofilms, Escherichia coli, Hydrophobicity, Pseudomonas aeruginosa, Small colony variants, Staphylococcus aureus