فهرست مطالب

Reports of Biochemistry and Molecular Biology - Volume:11 Issue: 4, Jan 2023

Reports of Biochemistry and Molecular Biology
Volume:11 Issue: 4, Jan 2023

  • تاریخ انتشار: 1402/03/03
  • تعداد عناوین: 20
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  • Sanaa Hagag, Ahmad Kodous*, Hebatallah Shaaban Pages 532-546
    Background

    Breast cancer (BC) plays a major public health in Egyptian woman. In Upper Egypt, there is an increase in incidence of BC compared to other Egyptian areas. Triple-negative BC, estrogen receptor (ER)-negative, progesterone receptor (PR)-negative, and HER2-neu-negative, is a high-risk BC that lacks the benefit of specific therapy that targets these proteins. Accurate determination of Caveolin-1(Cav-1), Caveolin-2 (Cav-2) and HER-2/neu status have become of major clinical significance in BC by focusing about its role as a tumor marker for response to different therapies.

    Methods

    The present study was performed on 73 female BC patients in the South Egypt Cancer Institute. Blood samples were used for Cav-1, Cav-2, and HER-2/neu genes amplification and expression. In addition, immunohistological analysis of mammaglobin, GATA3, ER, PR, and HER-2/neu was done.

    Results

    There was a statistically significant association between Cav-1, 2 and HER-2/neu genes expression and the age of patients (P< 0.001). There are increase in the level of Cav-1, 2 and increase in HER-2/neu mRNA expression in groups treated with chemotherapy and group treated with both chemotherapy and radiotherapy compared to each group baseline level of genes mRNA expression before treatment. On the contrary, the group treated with chemotherapy, radiotherapy and hormonal therapy revealed increase on the level of Cav-1, 2 and HER-2/neu mRNA expression when compared with their baseline for the same patients before treatment.

    Conclusions

    Noninvasive molecular biomarkers such as Cav-1 and Cav-2 have been proposed for use in the diagnosis and prognosis for women with BC.

    Keywords: Breast Cancer, Caveolin-1, ER, GATA-3, HER-2, neu, Mammaglobin, PR, 2
  • Reza Zare, Hamidreza Abdolsamadi, Sara Soleimani Asl, Shahrbanoo Radi, Hossein Bahrami, Shokoofeh Jamshidi* Pages 547-552
    Background

    The role of the basic fibroblast growth factor (bFGF) has well known in the angiogenesis and ulcer healing. In this study, we aimed to evaluate the effects of bFGF on tissue repair in a rat oral mucosal wound.

    Methods

    Musosal wound induced on the lip mucosa of rats and bFGF was injected along the edge of the mucosal defect immediately after surgery. The tissues were collected on days 3, 7 and 14 after the wound induction. The micro vessel density (MVD) and CD34 expression were done by histochemical studies.

    Results

    The bFGF significantly accelerated granulation tissue formation and MVD was increased three days after ulcer induction but decreased 14 days after surgery. The MVD was significantly higher in the bFGF-treated group. The wound area was decreased in all groups time-dependently and a statistically significant difference (p value?) was observed between the bFGF-treated group and untreated group. The wound area was smaller in the bFGF-treated group compared to the untreated group.

    Conclusions

    Our data demonstrated that bFGF can accelerated and facilitated wound healing.

    Keywords: bFGF, Healing, Angiogenesis, Wound, Micro Vessel Density
  • Abbas Khosravi, MohammadReza Deyhim*, Fatemeh Yari, Mahin Nikougoftar Zarif Pages 553-564
    Background

    In the current study we have aimed to find the effects of Resveratrol treatment on platelet concentrates (PCs) at the dose dependent manner. We have also attempted to find the molecular mechanism of the effects.

    Methods

    The PCs, have received from Iranian blood transfusion organization (IBTO). Totally 10 PCs were studied. The PCs divided into 4 groups including untreated (control) and treated by different dose of Resveratrol; 10, 30 and 50 µM. Platelet aggregation and total reactive oxygen species (ROS) levels were evaluated at day 3 of PCs storage. In silico analysis was carried out to find out the potential involved mechanisms.

    Results

    The aggregation against collagen has fallen dramatically in all studied groups but at the same time, aggregation was significantly higher in the control versus treated groups (p<0.05). The inhibitory effect was dose dependent. The aggregation against Ristocetin did not significantly affect by Resveratrol treatment. The mean of total ROS significantly increased in all studied groups except those PCs treated with 10 µM of Resveratrol (P=0.9). The ROS level significantly increased with increasing Resveratrol concentration even more than control group (slope=11.6, P=0.0034). Resveratrol could potently interact with more than 15 different genes which, 10 of them enrolled in cellular regulation of the oxidative stress.

    Conclusions

    Our findings indicated that the Resveratrol affect the platelet aggregation at the dose dependent manner. Moreover, we have also found that the Resveratrol play as double-edged sword in the controlling oxidative state of the cells. Therefore, Using the optimal dose of Resveratrol is the great of importance.

    Keywords: Aggregation, Oxidative stress, Platelet storage lesion, Resveratrol
  • Constantine Anetakis*, Stella Mitka, Maria Chatzidimitriou, Konstantinos Anagnostopoulos, Phaedra Eleftheriou, Theodoros Lialiaris Pages 565-576
    Background

    Vitamin D deficiency is recognised as a pandemic in the developed world. However, the importance of prudent sun exposure tends to be overlooked, which is responsible for this pandemic.

    Methods

    We investigated the vitamin D status in 326 adults, 165 females and 161 males: 99 Osteoporosis patients, 53 Type 1 Diabetes patients, 51 Type 2 Diabetes patients, and 123 Athletic Healthy individuals, from Northern Greece, through the measurement of total calcidiol in winter and summer by immunoenzymatic assay.

    Results

    In the Whole Sample 23.31% had severe deficiency, 13.50% mild deficiency, 17.48% insufficiency, and 45.71% adequacy at the end of winter. Mean concentrations differed significantly (p <0.001) between males and females. The prevalence of deficiency in the young was significantly lower than in the middle-aged (p = 0.004) and in the elderly (p <0.001), while it was significantly lower (p = 0.014) in the middle-aged than in the elderly. The best vitamin D status was found in the Athletic Healthy individuals, followed by the Type 1 and Type 2 Diabetic patients, while Osteoporotic patients had the poorest status. The difference in mean concentrations between winter and summer was significant (p <0.001).

    Conclusions

    Vitamin D status deteriorated with increasing age and it was better in males than in females. Our findings suggest that outdoor physical activity in a Mediterranean country can cover the vitamin D needs of the young and the middle-aged, but not of the elderly, without the need for dietary supplements.

    Keywords: Total calcidiol, osteoporosis, Type 1 diabetes, Type 2 diabetes, Vitamin D status
  • Anastasia Proskurina, Valeriy Nikolin, Nelly Popova, Nikolay Varaksin, Tatiana Ryabicheva, Elizaveta Ershova, Svetlana Kostyuk, Olga Leplina, Alexandr Ostanin, Elena Chernykh, Sergey Bogachev* Pages 577-589
    Background

    Double-stranded fragmented extracellular DNA is a participant, inducer, and indicator of various processes occurring in the organism. When investigating the properties of extracellular DNA, the question regarding the specificity of exposure to DNA from different sources has always been raised. The aim of this study was to perform comparative assessment of biological properties of double-stranded DNA obtained from the human placenta, porcine placenta and salmon sperm.

    Methods

    The intensity of leukocyte-stimulating effect of different dsDNA was assessed in mice after cyclophosphamide-induced cytoreduction. The stimulatory effect of different dsDNA on maturation and functions of human dendritic cells and the intensity of cytokine production by human whole blood cells was analyzed ex vivo. The oxidation level of the dsDNA was also compared.

    Results

    Human placental DNA exhibited the strongest leukocyte-stimulating effect. DNA extracted from human and porcine placenta exhibited similar stimulatory action on maturation of dendritic cells, allostimulatory capacity, and ability of dendritic cells to induce generation of cytotoxic CD8+CD107a+ T cells in the mixed leukocyte reaction. DNA extracted from salmon sperm stimulated the maturation of dendritic cells, while having no effect on their allostimulatory capacity. DNA extracted from human and porcine placenta was shown to exhibit a stimulatory effect on cytokine secretion by human whole blood cells. The observed differences between the DNA preparations can be caused by the total methylation level and are not related to differences in oxidation level of DNA molecules.

    Conclusions

    Human placental DNA exhibited the maximum combination of all biological effects.

    Keywords: Cytokines, Cytotoxic T cells, Dendritic cells, Double-stranded DNA
  • Omid Salemi, Zahra Noormohammadi, Fariborz Bahrami, Seyed Davar Siadat, Soheila Ajdary* Pages 590-598
    Background

    Two newly identified proteins, EspB and EspC are involved in the pathogenesis of Mycobacterium tuberculosis. The objective of the present study was to evaluate the immunogenicity of recombinant EspC, EspB, and EspC/EspB fusion proteins in mice.

    Methods

    BALB/c mice were immunized subcutaneously with recombinant EspC, EspB, and fusion EspC/EspB proteins, three times with along with Quil-A as an adjuvant. The cellular and humoral immune responses were evaluated by quantifying IFN-g, IL-4, IgG, IgG1, and IgG2a antibodies against the antigens.

    Results

    The results showed that the mice immunized with recombinant EspC, EspB, and EspC/EspB proteins did not produce IL-4, whereas IFN-g was secreted in response to all three proteins. EspC/EspB group produced significant amounts of IFN-g in response to stimulation with all the three recombinant proteins (P<0.001). In mice immunized with EspC, high levels of IFN-g were detected in response to EspC/EspB, and EspC (P<0.0001); while mice immunized with EspB produced lower levels of IFN- g in response to EspC/EspB, and EspB (P<0.05).

    Conclusions

    All the three recombinant proteins induced Th1-type immune responses in mice against EspB and EspC; however, EspC/EspB protein is more desirable due to the presence of epitopes from both EspC and EspB proteins and the production of immune responses against both.

    Keywords: EspB, EspC, ESX-1, Mycobacterium tuberculosis
  • Febriana Catur Iswanti*, Qarina Hasyala Putri, Ani Retno Prijanti, Samsuridjal Djauzi, Mohamad Sadikin, Arief Budi Witarto, Tomohiko Yamazaki Pages 599-613
    Background

    This study aims to prepare high stability chitosan nanoparticles (CNP) and examine the ability of CNP in CpG-ODN delivery when treating allergic mice model.

    Methods

    Preparation and characterization of CNP were performed by ionic gelation, dynamic light scattering, and zeta sizer. The CNP cytotoxicity and activation ability of CpG ODN delivered with CNP were tested using a cell counting kit-8 and Quanti blue method. Allergic mice were injected intraperitoneal with 10 ug ovalbumin on day 0 and 7, and then treated with intranasal CpG ODN/CpG ODN, delivered with CNP/CNP, on the third week three times per week for three weeks. The ELISA method measured cytokine and IgE profiles in the allergic mice’s plasma and spleen.

    Results

    CNP results have sizes 27.73 nm±3.67 dan 188.23 nm±53.47, spherical in shape and non-toxic, and did not alter the NF-κB activation of CpG ODN in RAW-blue cells. The application of CpG ODN delivered by chitosan nanoparticles shows no statistical difference between groups of IFN-γ, IL-10, and IL-13 in Balb/c mice’s plasma and spleen, in contrast with IgE level.

    Conclusions

    The results showed that using chitosan nanoparticles as a delivery system for CpG ODN has the potency to safely CpG ODN efficacy.

    Keywords: Allergy, Chitosan nanoparticle, CpG ODN, Immunotherapy, Mice spleen
  • Marwa Abdelgwad, Reem Zakaria*, Samar Marzouk, Dina Sabry, Rasha Ahmed, Hedy Ayman Badary, Mai Samir Pages 614-625
    Background

    Non-alcoholic fatty liver disease is a major problem worldwide that needs non-invasive biomarkers for early diagnosis and treatment response assessment. We aimed to assess the correlation between circRNA-HIPK3 and miRNA-29a expression and its role as miRNA-29a sponge, as well as the correlation between circRNA-0046367 and miRNA-34a expression and its role as miRNA-34a sponge and their effect on regulation of the Wnt/β catenin pathway, which may provide a new target for treatment of non-alcoholic steatohepatitis.

    Methods

    the research was performed on 110 participants: group (I): fifty-five healthy donors served as controls and group (II): fifty-five patients with fatty liver pattern on abdominal ultrasound. Lipid profile and liver functions were assessed. RT-PCR was performed to assess the RNAs: circRNA-HIPK3, circRNA-0046367, miRNA-29a, miRNA-34a and Wnt mRNA gene expression. ELISA was performed to determine β-catenin protein levels.

    Results

    miRNA-34a and circRNA-HIPK3 expression were significantly greater, while miRNA-29a and circRNA-0046367 expression were significantly less, in patients than in controls. Wnt/β-catenin regulated by miRNA-29a and miRNA-34a showed a significant decrease that leads to its abnormal effect on lipid metabolism.

    Conclusions

     our results imply that miRNA-29a can be investigated as a target for circRNA-HIPK3, while miRNA-34a can be investigated as a target for circRNA-0046367, and that circRNA-HIPK3 and circRNA-0046367 may have emerging roles that can affect the pathogenesis of nonalcoholic steatohepatitis through the Wnt/β-catenin pathway and thus be used as therapeutic targets for the disease.

    Keywords: Circrna-HIPK3, circRNA-0046367, miRNA-29a, miRNA-34a, Wnt, β-catenin NASH
  • Sajad Dehnavi, Ali Khodadadi, Ali Asadirad, Ata Ghadiri* Pages 626-634
    Background

    Exosomes are nanoscale vesicles widely used as drug delivery systems. Mesenchymal stem cell (MSC)-derived exosomes have shown immunomodulatory potential. This study optimized loading OVA into the mice adipose tissue-derived MSC-isolated exosomes to prepare the OVA-MSC-exosome complex for allergen-specific immunotherapy.

    Methods

    MSCs were harvested from mice adipose tissue and characterized by flow cytometry and evaluating differentiation potential. The exosomes were isolated and characterized via Dynamic Light Scattering, Scanning Electron Microscopy, and flow cytometry. Different concentrations of ovalbumin were incubated with MSC-exosome in various durations to optimize a more suitable protocol. BCA and HPLC analysis were used to quantify, and DLS was applied to qualify the prepared formulation of the OVA-exosome complex.

    Results

    The harvested MSCs and isolated exosomes were characterized. Analysis of the OVA-exosome complex revealed that OVA in primary 500 μg/ml concentration and incubation for 6 h results in higher efficacy.

    Conclusions

    Loading OVA into MSC-derived exosomes was successfully optimized and could be administrated for allergen-specific immunotherapy in the animal model.

    Keywords: Delivery system, Exosome, Mesenchymal stem cell (MSC), Ovalbumin
  • Soha Mohamed Hamdy, Marwa Saad Ali, Rehab Galal Abd El-Hmid, Noha Khalifa Abdelghaffar, Omayma Owees Abdelaleem* Pages 635-643
    Background

    Pediatric immune thrombocytopenic purpura (ITP) is an autoimmune disease; whose etiology is unknown. lncRNAs are regulators of numerous actions, which participate in the development of autoimmune diseases. We evaluated the expression ofNEAT1 and Lnc-RNA in dendritic cell (Lnc-DC) in pediatric ITP.

    Methods

    Sixty ITP patients and 60 healthy subjects were enrolled in the present study; Real-time PCR was performed to assess the expression levels of NEAT1 and Lnc-DC in sera of children with ITP as well as healthy children.

    Results

    Both lncRNAs, NEAT1 and Lnc-DC were significantly upregulated in ITP patients in comparison to controls (p <0.0001 and P= 0.001 respectively). Furthermore, significant upregulation of the expression levels of NEAT1 and Lnc-DC were observed in the non-chronic compared with chronic ITP patients. Also, there was significant negative correlation between each of NEAT1 and Lnc-DC and platelet counts before treatment (r= -0.38; P= 0.003 and r= -0.461; P< 0.0001, respectively).

    Conclusions

    serum lncRNAs, NEAT1 and Lnc-DC could be used as potential biomarkers in differentiating childhood ITP patients and healthy controls in addition to differentiating non-chronic from chronic ITP which may provide a theoretical basis for the mechanism and treatment of immune thrombocytopenia.

    Keywords: Lnc-DC, NEAT1, Pediatric, ITP
  • Maryam Jafar Sameri*, Rafie Belali, Niloofar Neisi, Reza Noei Razliqi *, Seyed Ali Mard, Feryal Savari, Seyyed Saeed Azandeh Pages 644-655
    Background

    Liver diseases and injuries are important medical problems worldwide. Acute liver failure (ALF) is a clinical syndrome characterized by severe functional impairment and widespread death of hepatocytes. Liver transplantation is the only treatment available so far. Exosomes are nanovesicles originating from intracellular organelles. They regulate the cellular and molecular mechanisms of their recipient cells and have promising potential for clinical application in acute and chronic liver injuries. This study compares the effect of Sodium hydrosulfide (NaHS) modified exosomes with non-modified exosomes in CCL4-induced acute liver injury to ascertain their role in ameliorating hepatic injury.

    Methods

    Human Mesenchymal stem cells (MSCs) were treated with or without NaHS (1 μmol) and exosomes were isolated using an exosome isolation kit. Male mice (8-12 weeks old) were randomly divided into four groups (n=6): 1-control, 2-PBS, 3- MSC-Exo, and 4- H2S-Exo. Animals received 2.8 ml/kg body weight of CCL4 solution intraperitoneally, and 24 h later MSC-Exo (non-modified), H2S-Exo (NaHS-modified), or PBS, was injected in the tail vein. Moreover, 24 h after Exo administration, mice were sacrificed for tissue and blood collection.

    Results

    Administration of both MSC-Exo and H2S-Exo reduced inflammatory cytokines (IL-6, TNF-α), total oxidant levels, liver aminotransferases, and cellular apoptosis.

    Conclusions

    MSC-Exo and H2S-Exo had hepato-protective effects against CCL4-induced liver injury in mice. Modification of cell culture medium with NaHS as an H2S donor enhances the therapeutic effects of MSC exosomes.

    Keywords: Acute liver failure (ALF), CCL4-induced liver injury, Exosomes, Mesenchymal stem cells (MSCs), Sodium hydrosulfide (NaHS)
  • Pouyan Razavi, Seyed Abdolrahim Rezaee, Salehe Akhondian, Nastaran Asgari, Kazem Fatemi*, Farnaz Mohajertehran* Pages 656-662
    Background

    Periodontal disease is an inflammatory condition affecting the tooth's supporting tissues, resulting in gradual loss of periodontal ligament (PDL), alveolar bone, and gum resorption. Neutrophil and monocyte/macrophage, destructive proteases like matrix metalloproteinase (MMP)-3 and MMP-9 play pivotal roles in such lesions in periodontitis. Therefore, this study aims to compare the level of MMP-3 and MMP-9 gene expression in patients with or without periodontitis in an Iranian population.

    Methods

    This cross-sectional study was carried out on 22 chronic periodontitis patients and 17 healthy control subjects referred to the department of periodontology, Mashhad Dental School. In both groups, the gingival tissue was removed during surgery and transferred to the Molecular Biology Laboratory for MMP-3 and MMP-9 gene expression evaluation. The qRT-PCR, TaqMan method was used for gene expression assessments.

    Results

    The average age of periodontitis patients was 33± 5 years, and in controls, 34.7± 6 with no significant differences. The mean MMP-3 expression in periodontitis patients was 146.67±38.7, and in controls, 63.4±9.1. The difference was statistically significant (P=0.04). The mean expression of MMP-9 in periodontitis patients and controls were 103.8± 21.66 and 87.57± 16.05, respectively. Although the target gene expression in patients was higher, the difference was insignificant. Furthermore, there was not any significant correlation between age or gender with the expression of MMP3 or MMP9.

    Conclusions

    The study demonstrated that the MMP3 seems to have a destructive impact on the gingival tissue in chronic periodontitis, but not MMP9.

    Keywords: Chronic periodontitis, Matrix metalloproteinase-3, Matrix metalloproteinase-9
  • Yones Guntoro Aji, Stella Melita, Rendy Dijaya, Dionysius Subali*, Felicia Kartawidjajaputra, Antonius Suwanto Pages 663-671
    Background

    Caffeine is generally suggested to increase VO2max in endurance performance. Nevertheless, the response to caffeine ingestion does not seem to be uniform across individuals. Therefore, caffeine ingested timing on endurance performance based on the type of CYP1A2 single nucleotide polymorphism rs762551, that were classified as fast and slow metabolizers, need to be evaluated.

    Methods

    Thirty participants participated in this study. DNA was obtained from saliva samples and genotyped using polymerase chain reaction-restriction fragment length polymorphism. Each respondent completed beep tests under three treatments blindly: placebo, 4 mg/kg body mass of caffeine one hour, and two hours before test.  

    Results

    Caffeine increased estimated VO2max in fast metabolizers (caffeine=29.39±4.79, placebo=27.33±4.02, p<0.05) and slow metabolizers (caffeine=31.25±6.19, placebo=29.17±5.32, p<0.05) in one hour before test. Caffeine also increased estimated VO2max in fast metabolizers (caffeine=28.91±4.65, placebo=27.33±4.02, p<0.05) and slow metabolizers (caffeine=32.53±6.68, placebo=29.17±5.32, p<0.05) in two hour before test. However, for slow metabolizers, the increasing was greater when caffeine was administered two hours before test (slow=3.37±2.07, fast=1.57±1.62, p<0.05).

    Conclusions

    Genetic variance may affect the optimal caffeine ingestion timing, sedentary individuals who want to enhance their endurance performance may ingest caffeine 1 hour before exercise for fast metabolizers and 2 hours before exercise for slow metabolizers.

    Keywords: Caffeine, CYP1A2, Performance Enhancer, Sedentary, VO2max
  • Seyed MohammadAli Hashemi, Abdolvahab Moradi, Seyed Younes Hosseini, Hadi Razavi Nikoo, Taravat Bamdad, Mahboobeh Razmkhah, Jamal Sarvari*, Alijan Tabarraei* Pages 672-683
    Background

    Suppression of p53 is an important mechanism in Epstein-Barr virus associate-tumors and described as EBNA1-USP7 which is a key axis in p53 suppression. Thus, in this study, we aimed to evaluate the function of EBNA1 on the expression of p53-inhibiting genes including HDAC-1, MDM2, MDM4, Sirt-3, and PSMD10 and the influence of USP7 inhibition using GNE-6776 on p53 at protein/mRNA level.

    Methods

      The electroporation method was used to transfect the BL28 cell line with EBNA1. Cells with stable EBNA1 expression were selected by Hygromycin B treatment. The expression of seven genes, including PSMD10, HDAC-1, USP7, MDM2, P53, Sirt-3, and MDM4, was evaluated using a real-time PCR assay. For evaluating the effects of USP7 inhibition, the cells were treated with GNE-6776; after 24 hours and 4 days, the cells were collected and again expression of interest genes was evaluated.

    Results

    MDM2 (P=0.028), MDM4 (P=0.028), USP7 (P=0.028), and HDAC1 (P=0.015) all showed significantly higher expression in EBNA1-harboring cells compared to control plasmid transfected cells, while p53 mRNA expression was only marginally downregulated in EBNA1 harboring cells (P=0.685). Four-day after treatment, none of the studied genes was significantly changed. Also, in the first 24-hour after treatment, mRNA expression of p53 was downregulated (P=0.685), but after 4 days it was upregulated (P=0.7) insignificantly.

    Conclusions

    It seems that EBNA1 could strongly upregulate p53-inhibiting genes including HDAC1, MDM2, MDM4, and USP7. Moreover, it appears that the effects of USP7 suppression on p53 at protein/mRNA level depend on the cell nature; however, further research is needed.

    Keywords: Epstein-Barr virus, EBNA1, P53, USP7, P53-inhibiting Genes
  • Octavio Galindo-Hernández, Lizbeth Angelina García-Salazar, Victor Guadalupe García-González, Raúl Díaz-Molina, José Luis Vique-Sánchez* Pages 684-693
    Background

    Cancer continues worldwide. It has been reported that OTUB1, a cysteine protease, plays a critical role in a variety of tumors and is strongly related to tumor proliferation, migration, and clinical prognosis by its functions on deubiquitination. Drug advances continue against new therapeutic targets. In this study we used OTUB1 to develop a specific pharmacological treatment to regulate deubiquitination by OTUB1. The aim of this research is to regulate OTUB1 functions.

    Methods

    By molecular docking in a specific potential OTUB1 interaction site between Asp88, Cys91, and His26 amino acids, using a chemical library of over 500,000 compounds, we selected potential inhibitors of the OTUB1 catalytic site.

    Results

    Ten compounds (OT1 - OT10) were selected by molecular docking to develop a new anti-cancer drug to decrease OTUB1 functions in cancer processes.

    Conclusions

    OT1 – OT10 compounds could be interacting in the potential site between Asp88, Cys91, and His265 amino acids in OTUB1. This site is necessary for the deubiquitinating function of OTUB1. Therefore, this study shows another way to attack cancer.

    Keywords: Anti-cancer, Dub inhibitor, Molecular docking, OTUB1
  • Malika Nurtleu, Zhansaya Adish, Kasym Mukanov, Kanat Tursunov*, Yerlan Ramankulov, Kanatbek Mukantayev Pages 694-701
    Background

    Macrophages are essential cellular components in various body tissues and tumor microenvironments. The high infiltration of macrophages into the tumor microenvironment determines the importance of ex vivo treatment of personalized macrophages with recombinant cytotoxic T-lymphocyte-associated protein 4 (rCTLA-4), programmed death-ligand 1 (rPD-L1), and programmed cell death protein 1 (rPD-1) proteins to block immune checkpoints.

    Methods

    We investigated the development of humoral immunity against CTLA-4, PD-L1, and PD-1 receptors by introducing macrophages treated ex vivo with the corresponding proteins into mice. Peritoneal macrophages from BALB/c mice were cultured in medium containing recombinant human CTLA-4, PD-L1, and PD-1 proteins. Macrophages processing recombinant proteins were analyzed via immunofluorescence staining using antibodies against CTLA-4, PD-L1, and PD-1. The treated macrophages were administered intraperitoneally to mice to induce anti-CTLA-4, anti-PD-L1, and anti-PD-1 antibodies. The antibody titer in vaccinated mice was determined via enzyme-linked immunosorbent assays, followed by statistical analysis of the results.
    The specificity of the antibodies was determined via immunofluorescence staining in MCF7 cells.

    Results

    The ex vivo treatment of macrophages with rCTLA-4, rPD-L1, and rPD-1 induced the formation of specific antibodies in vaccinated mice. The various rPD-L1 and rPD-1 concentrations used to treat macrophages had no significant effect on the specific antibody titers, while the anti-rCTLA-4 titer was dependent on the protein concentration in the culture medium. Immunofluorescence analysis revealed that anti-CTLA-4 and PD-L1 antibodies reacted with MCF7 cells.

    Conclusions

    The ex vivo treatment of macrophages with rCTLA-4, rPD-L1, and rPD-1 can help induce humoral immunity and develop new approaches for cancer immunotherapy.

    Keywords: Cytotoxic T Lymphocyte-Associated Protein 4, Immunotherapy, Macrophages, Programmed Death-Ligand 1, Programmed Cell Death Protein 1
  • Assem Abdeen Radwan*, Nour El-Dean Abd-Elazeem Hefney, Emad Farah Mohammed Kholef, Abdallah Elebidi, Hala Mahmoud Pages 702-709
    Background

    The Transforming Growth Factor-beta (TGF-β) is one of the main growth factors associated with fibrosis or cirrhosis progression in the liver, but its role in hepatocarcinogenesis is controversial. To highlight the role of Transforming Growth Factor β as a marker of Hepatocellular carcinoma (HCC) in patients with chronic hepatitis C virus (HCV) infection.

    Methods

    Ninety subjects were enrolled in this study, classified into three groups: Group I (chronic HCV group) included 30 patients with chronic HCV infection; Group II (HCC group) include 30 patients having HCC and chronic HCV infection and Group III consisted of 30 age and sex-matched healthy controls. TGF-β was evaluated in all the enrollees and its levels were correlated to liver function and other clinical parameters.

    Results

    TGF-β was found significantly higher in HCC group than in control and chronic HCV (P <0.001). In addition, it was correlated with biochemical and clinical parameters of cancer.

    Conclusions

    Patients with HCC showed increased level of TGF-β compared to chronic HCV infection patients and controls.

    Keywords: Chronic HCV, Liver cirrhosis, Transforming Growth Factor β
  • Azam Rasti, Masoud Khalili, Ali Mohammad Fakhr Yasseri, Neda Nasirian, Reza Shirkoohi, Mohammad Reza Nowroozi, MohammadHossein Modarressi* Pages 710-719
    Background

    Many researchers have tried to identify bladder cancer biomarkers to reduce the need for cystoscopy. The aim of this study was to identify and measure appropriate transcripts in patient urine to develop a non-invasive screening test.

    Methods

     From February 2020 to May 2022, 49 samples were obtained from Velayat Hospital, Qazvin University of Medical Sciences, Qazvin, Iran. Twenty-two samples were obtained from bladder cancer patients and 27 from bladder cancer-free subjects. RNA was extracted from participant samples, quantitative RT-PCR was performed, and TNP plots were used to assess IGF2 (NCBI Gene ID: 3481), KRT14 (NCBI Gene ID: 3861) and KRT20 (NCBI Gene ID: 54474) expression. For UCSC Xena analysis, Dataset ID: TCGA-BLCA was used to compare transitional cell carcinoma (TCC) and normal samples for survival rates.

    Results

    IGF and KRT14 were more greatly expressed in patient urine samples than in those of the normal group. However, KRT20 expression did not significantly differ between the two groups. IGF2 had 45.45 and 88.89% sensitivity and specificity, respectively, for detecting TCC in urine samples while KRT14 had 59 and 88.89% sensitivity and specificity, respectively. Also, these results infer that overexpression of IGF would be prognosticators of poor TCC outcomes.

    Conclusions

    Our study showed that IGF2 and KRT14 are overexpressed in bladder cancer patient urine, and IGF2 could be a potential biomarker for poor prognoses in TCC.

    Keywords: Biomarkers, Diagnosis, Genes, Liquid biopsy, Urinary bladder neoplasms
  • Albert Ananta, Rendy Dijaya, Dionysius Subali*, Felicia Kartawidjajaputra, Lina Antono Pages 720-729
    Background

    IgA is widely used as Upper Respiratory Tract Infection (URTI) risk marker, as a lower concentration in sIgA indicates a higher incidence of URTI. This study aimed to investigate the effect of different types of exercise; combined with Tempeh consumption in increasing sIgA concentration in saliva sample.

    Methods

    19 sedentary male subjects aged 20-23 were recruited and assigned into 2 groups based on the exercise type, endurance (n=9), and resistance (n=10). These subjects underwent 2 weeks of Tofu and Tempeh consumption, then were assigned to do exercises based on their groups.

    Results

    This study showed an increased mean value of sIgA concentrations in the endurance group; the baseline value, after food treatment, and after food and exercise treatment were 71.726 ng/mL, 73.266 ng/mL, and 73.921 ng/mL, respectively for Tofu treatment; and 71.726 ng/mL, 73.723 ng/mL, and 75.075 ng/mL, respectively for Tempeh treatment. While in the resistance group, there was also an increase in the mean value of sIgA concentrations; baseline, after food treatment, and after food and exercise treatments were 70.123 ng/mL, 71.801 ng/mL, and 74.430 ng/mL, respectively for Tofu treatment; and 70.123 ng/mL, 72.397 ng/mL, and 77.216 ng/mL, respectively for Tempeh treatment. These results indicated that combining both Tempeh consumption and moderate intensity resistance exercise was more effective to increase sIgA concentration.

    Conclusions

    This study showed that combining moderate intensity resistance exercise with consumption of 200 gr Tempeh for 2 weeks was more effective in increasing sIgA concentration; compared to endurance exercise and Tofu consumption.

    Keywords: : Exercise, IgA, Paraprobiotics, Tempeh, Upper Respiratory Tract Infection (URTI)
  • Latifa Mohamed Abdelgawad*, Ahmed Abdullatif Abdelaziz, Mahmoud Bawdy El-Begawey, Ali Mohamed Saafan Pages 730-738
    Background

    Oral squamous cell carcinoma (OSCC) is the sixth most common mouth cancer in the world. The aim of the present study is comparing the effects of using Nanocurcumin, and photodynamic therapy (PDT), alone or together in treatment of OSCC in rats.

    Methods

    Forty Wister male rats were divided into Control (group 1), 650 nm diode Laser only (group 2), Nanocurcumin alone (group 3), and PDT with a combination of laser with Nanocurcumin (group 4). Then, OSCC in the tongue induced by dimethylbenz anthracene (DMBA). The treatments were evaluated clinically, histopathologically, and immunohistochemically through BCL2 and Caspase-3 genes expression.

    Results

     Positive control with OSCC displayed significant weight loss, while PDT group gained more than nanocurcumin treated groups as well as laser groups comparing with control positive group.
    The histological examination of the tongue in PDT group showed improvement. In laser group, there were partial loss of surface epithelium with various ulcers and dysplasia and partial improvement by this type of treatment. The tongue in the positive control group showed ulcer in the dorsum surface with inflammatory cells, hyperplasia of the mucosa membrane around the ulcer (acanthosis) with increase of dentition, vacuolar degeneration of prickle cell layer and increase mitotic activity of basal cell layer together with dermal proliferation.

    Conclusions

    Under the condition of the present study, PDT using nanocurcumin photosensitizer was effective in the treatment of OSCC regarding clinical, histological and gene expression of BCL2 and Caspase-3.

    Keywords: BCL2, Caspase-3, Diode laser 650 nm, Histological analysis, Nanocurcumin, OSCC, PDT