فهرست مطالب

Reports of Biochemistry and Molecular Biology - Volume:11 Issue: 1, Apr 2022
  • Volume:11 Issue: 1, Apr 2022
  • تاریخ انتشار: 1401/03/29
  • تعداد عناوین: 20
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  • Karim Parastouei, Mohammad Hossein Aarabi, Gholam Ali Hamidi, Zahra Nasehi, Shima Kabiri Arani, Faezeh Jozi, Mohamad Esmaeil Shahabodin* Pages 1-9
    Background

    Cannabinoids (CBs) have been found to regulate the immune system, affect innate and adaptive immune responses, and reduce inflammatory reactions. This study assessed the therapeutic effects of GW-405833 synthetic CB2 agonist on inflammatory factors as well as locomotor activity in experimental autoimmune encephalomyelitis (EAE).

    Methods

    In this experimental study, 48 adult male C57BL/6 mice were randomly and equally assigned to eight groups. By injecting 250 mg of MOG35-55 peptide, EAE was induced. Every other day for 17 days after EAE onset, EAE-afflicted mice in groups 1–3 received an intraperitoneal injection of GW-405833 at a dose of 3, 10, and 30 mg/kg, respectively. Clinical status and locomotor activity, measured using the beam walking assay, were assessed every other day during the first 17 days after EAE onset. Mice were euthanized in day 17th of treatment and the serum levels of the IL-1ß, IL-12, CRP, and TNF-α proinflammatory cytokines as well as IL-4 and TGF-ß anti-inflammatory cytokines were measured by ELISA method.

    Results

    Clinical manifestations of EAE in groups 2 and 3 were significantly milder than group 4 and locomotor activity in groups 1–3 was significantly better than group 4 in days 5–17 (p< 0.05). GW-405833 also significantly decreased the levels of IL-12, TNF-α, and CRP and significantly increased the levels of IL-4 and TGF-ß but had no significant effects on the level of IL-1ß. GW-405833 was not associated with significant side effects.

    Conclusions

    The CB2 receptor agonist GW-405833, improves clinical conditions and reduces inflammation in mice with EAE.

    Keywords: Clinical evaluation, Experimental autoimmune encephalomyelitis, GW-405833, Locomotor activity, Multiple sclerosis, Proinflammatory cytokines
  • Samaneh Beiki, Jalil Tavakkol-Afshari, Zhaleh Shariati-Sarabi, Javad Poursamimi, Arezoo Gowhari Shabgah, Mojgan Mohammadi* Pages 10-19
    Background

    inflammatory chemokines such as CCL2 and CCL5 are involved in the progress of osteoarthritis. Crocin with antioxidant and anti-inflammatory properties can reduce the symptoms of osteoarthritis (OA). This study was performed investigate the effect of Krocina™, on the gene expressions and plasma levels of CCL2 and CCL5 in OA patients.

    Methods

    The study included 35 patients that were randomized in the KrocinaTM and placebo groups. The intervention was Krocina™ 15 mg daily for four months. Clinical and paraclinical parameters were measured. CCL2 and CCL5 genes expression and plasma levels were determined using the SYBR Green Real-Time RT-PCR and Enzyme-linked Immunosorbent Assay (ELISA) techniques.

    Results

    The C-reactive protein (CRP) value in the KrocinaTM group and the visual analogue scale (VAS) value in the KrocinaTM and placebo groups decreased significantly after the intervention. The gene expression of CCL2 in the KrocinaTM and placebo groups decreased significantly. On the contrary, the gene expression of CCL5 in the KrocinaTM and placebo groups increased significantly. Moreover, the plasma levels of CCL2 in the KrocinaTM and placebo groups decreased meaningfully. There was no difference regarding the plasma levels of CCL5 within the Krocina™ and placebo groups before and after the intervention in either of the groups.

    Conclusions

    Administration of Krocina™ reduced the clinical signs of inflammation and CRP and VAS value. Also, Krocina™ significantly decreased the plasma levels and gene expression of CCL2 in osteoarthritis patients.

    Keywords: CCL2, CCL5, Krocina™, Osteoarthritis
  • Sara Ahmed Aglan, Ahmad Mohamad Zaki, Amel Sobhy El Sedfy, Heba Gaber El-Sheredy*, Ola Hussein Elgaddar Pages 20-29
    Background

    ATP-binding cassette membrane transporter G2 (ABCG2) gene is one of transporter family and well characterized for their association with chemoresistance. Promoter methylation is a mechanism for regulation of gene expression. O6-Methyl guanine DNA methyl transferase (MGMT) gene plays a fundamental role in DNA repair. MGMT has the ability to remove alkyl adducts from DNA at the O6 position of guanine. Alkylating agents exert their function through adding these alkyls adducts to DNA leading to cell death unless it is repaired by MGMT. MGMT promoter was found to be methylated in several malignancies. The aim of the present work is to study the relation of MGMT and ABCG2 promoter methylation status in advanced breast cancer patients to response to cyclophosphamide–doxorubicin (AC) based therapeutic regime.

    Methods

    This retrospective study included Forty-two female patients with advanced breast cancer assessed before receiving chemotherapy and after the completion of regimens. They were grouped into responders and non-responders according to RECIST criteria. Methylation analysis of MGMT and ABCG2 genes were performed on breast cancer tissues.

    Results

    MGMT promoter was methylated in 40.5% of the cases. ABCG2 promoter was methylated in 14.3% of cases. There was no statistically significant association between MGMT and ABCG2 promoter methylation status and clinicopathological parameters. There was statistically significant association between methylation status of both promoters and response to AC when followed by Taxane.

    Conclusions

    Methylation of MGMT and ABCG2 promoters combined could be a potential predictive factor for response to cyclophosphamide-doxorubicin based therapeutic regime.

    Keywords: ABCG2, Breast cancer, Chemoresistance, DNA methylation, MGMT
  • Saad Shakir Mahdi Al-Amara* Pages 30-35
    Background

    Antibiotics called macrolide, lincosamide and streptogramin B (MLSB) are being used to treat staphylococci infections. Multiple pathways that impart resistance to MLSB antibiotics have been confirmed to cause clinical failure. The present work aimed to determine the frequency of constitutive and inducible clindamycin resistant among coagulase-negative staphylococci (CoNS) isolates of different clinical samples in Al-Basrah governorate, Iraq.

    Methods

    The 28 CoNS, traditional techniques and the Vitek®2 system were used to identify the isolates. The disk diffusion technique was used to detect methicillin resistance and antibiotic sensitivity patterns via cefoxitin, gentamicin, ciprofloxacin, amikacin, teicoplanin, linezolid, doxycycline and vancomycin disks. Erythromycin and clindamycin antibiotic disks was used to detect the inducible and constitutive clindamycin resistance as well as a D-test according to CLSI guidelines.

    Results

    Among 28 CoNS isolated, the Staphylococcus aureus 11(39.29%), Staphylococcus epidermidis 7(25 %), Staphylococcus haemolyticus 4(14.29%) and Staphylococcus saprophyticus 3 (10.71%) were predominant isolated species. Out of 28 CoNS isolates, 15(53.57%) were methicillin resistant coagulasenegative staphylococci (MRCoNS) isolates and 13(46.43%) were methicillin sensitive coagulase-negative staphylococci (MSCoNS) isolates. The 15(53.57%) isolates out of 28 CoNS, showed erythromycin resistance while 6(40%) isolates out of 15 CoNS, showed inducible macrolide-lincosamide-streptogramin B (iMLSB) and 2(13.3%) of CONS isolated showed constitutive macrolide-lincosamide-streptogramin B (cMLSB).

    Conclusions

    In order to achive the best result in choosing the suitable treatment and avoiding the loses the money and time, it is better to use the D-test for inducible clindamycin resistance in the daily routine work of antibiotic susceptibility testing in hospital and private clinical laboratories.

    Keywords: Anti-Bacterial Agents, Clindamycin, Staphylococcus
  • Thanaa Abdulmahdi Mokif, Zena Abdul-Ameer Mahdi, Raheem Tuama Obayes Al- Mammori, Hussein Oleiwi Muttaleb Al-Dahmoshi*, Noor Salman Kadhim Al-Khafaji Pages 36-43
    Background

    Menopause is a unique event in women's life it usually occurs naturally, most often after age 50 when woman has not menstruated in 12 consecutive months. This study was planned to assess the relationship between Vitamin D3 level, PAI-1 and HCG in Babylon women at age <50 years as pre-menopausal and> 50 years as post-menopausal.

    Methods

    The sample were selected from a group of pre- and post-menopausal women, 30 and 50 respectively. All the tests were evaluated to measure Vitamin D3 level, PAI-1 and HCG level. The sample was collected between July 2019 and January 2020 at Merjan medical city GIT and Liver Center, Babylon province, Iraq.

    Results

    The result of current study revealed that there are significant differences in vitamin D3 level in various age categories within postmenopausal women (p= 0.02) also there is no significant differences in PAI-1 and HCG with in these two groups, p= 0.08 and 0.07, respectively. Also, there is significant negative correlation between vitamin D3 and PAI-1 in postmenopausal women (p. value is 0.01).

    Conclusions

    Indeed, postmenopausal women regarded as elderly, but they have sufficient vitamin D3 and normal PAI-I levels as markers for normal non fibrosis status.

    Keywords: _ PAI-1, HCG, Menopause, Vitamin D3
  • Sheida Ebrahimi, Fariba Rezaei Fakhrnezhad, Sanaz Jahangiri, Mahdis Borjkhani, Rosa Behboodi, Amir Monfaredan* Pages 44-53
    Background

    Prostate cancer is considered as the second leading cause of cancer related death in men worldwide and the third frequent cancer among Iranian men. Despite the use of PSA as the only biomarker for early diagnosis of prostate cancer, its application in clinical settings is under debate. Therefore, the introduction of new molecular markers for early detection of prostate cancer is needed.

    Methods

    In the present study we intended to evaluate the expression of IGSF1, Wnt5a, FGF14, and ITPR1 in prostate cancer specimens by real time PCR. Biopsy samples of 40 prostate cancer cases and 41 healthy Iranian men were compared to determine the relative gene expression of IGSF1, Wnt5a, FGF14, and ITPR1 by real time PCR.

    Results

    Our results showed that Wnt5a, FGF14, and IGSF1 were significantly overexpressed in the prostate cancer patients while the mean relative expression of ITPR1 showed a significant decrease in PCa samples compared to healthy controls.

    Conclusions

    According to results of the present study, the combination panel of IGSF1, Wnt5a, FGF14, and ITPR1 genes could be considered as potential genetic markers for prostate cancer diagnosis. However further studies on larger populations and investigating the clinicopathological relevance of these genes is needed.

    Keywords: FGF14, IGSF1, ITPR1, Prostate Cancer, Wnt5a
  • Charalambos Papadopoulos, Konstantinos Mimidis, Dimitris Papazoglou, George Kolios, Ioannis Tentes, Konstantinos Anagnostopoulos* Pages 54-62
    Background

    Non-alcoholic fatty liver disease (NAFLD) constitutes a global pandemic. An intricate network among cytokines and lipids possesses a central role in NAFLD pathogenesis. Red blood cells comprise an important source of both cytokines and signaling lipids and have an important role in molecular crosstalk during immunometabolic deregulation. However, their role in NAFLD has not been thoroughly investigated.

    Methods

    Conditioned media from erythrocytes derived from 10 NAFLD patients (4 men, 6 women, aged 57.875±15.16) and 10 healthy controls (4 men, 6 women, aged 39.3±15.55) was analyzed for the cytokines IFN-γ, TNF-α, CCL2, CCL5, IL-8, IL-1β, IL-12p40, IL-17, MIP-1β, the signaling lipids sphingosine-1-phosphate (S1P) and lysophosphatidic acid (LPA), and cholesterol. Their effect on the cytokine profile released by RAW 264.7 macrophages was also studied.

    Results

    MCP1 levels were greater in conditioned growth medium from NAFLD patient erythrocytes than in that from healthy controls (37±40 vs 6.51±5.63 pg/ml). No statistically significant differences were found between patients and healthy controls with regard to S1P, LPA, cholesterol, or eight other cytokines. TNFa release by RAW 264.7 cells was greater after incubation with patient-derived erythrocyte-conditioned medium than in medium without RAW 264.7 cells from either healthy or NAFLD subjects.

    Conclusions

    Erythrocytes may contribute to liver infiltration by monocytes, and macrophage activation, partially due to CCL2 release, in the context of NAFLD.

    Keywords: Cytokines, Erythrocytes, Lipids, Non-alcoholic fatty liver disease, Signaling
  • Navishaa Govindasamy, Kian Chung Chok, Pei Ying Ng, Rhun Yian Koh, Soi Moi Chye* Pages 63-73
    Background

    Peripheral nerve injury (PNI) is a common condition that compromises motor and sensory functions. Peripheral nerves are known to have regenerative capability and the pineal hormone, melatonin, is known to aid nerve regeneration. However, the role of Schwann cells and the pathways involved remain unclear. Thus, the aim of this study is to identify the effects of melatonin on Schwann cell proliferation, dedifferentiation, and the involvement of nuclear factor kappa light chain enhancer of activated B cells (NFĸB), focal adhesion kinase (FAK) and proto-oncogene tyrosine-protein kinase, Src pathways in this process.

    Methods

    Schwann cells was treated with melatonin and its proliferation and dedifferentiation were identified using MTT assay and immunofluorescence staining for SRY (sex determining region Y)-box 2 (SOX2). Next, the protein expressions of NF-ĸB, FAK and Src pathways were identified by Western blot.

    Results

    MTT results confirmed increased proliferation of Schwann cells with melatonin treatment, and it was highest at 10 μM melatonin. Immunofluorescent staining revealed an increase in the green fluorescence staining for SOX2 in melatonin-treated cells, showing enhanced dedifferentiation. Western blot assay revealed melatonin increased phospho-NF-ĸB (PNF-ĸB), IKK-α, FAK (D2R2E), phospho-FAK (Tyr 576/577 and Tyr 397) protein expressions as compared with control. However, Src (32G6), Lyn (C13F9), Fyn, Csk (C74C1) protein expressions were not increased as compared with control.

    Conclusions

    Melatonin promotes Schwann cell proliferation and dedifferentiation via NF-ĸB, FAKdependent but Src-independent pathways.

    Keywords: Dedifferentiation, Melatonin, Peripheral nerve injury, Proliferation, Schwann cells. Introduction Peripheral
  • Augusta Chinyere Nsonwu-Anyanwu*, Augusta Ndudi Idenyi, Sunday Jeremiah Offor, Caroline Chinenyenwa Thomas, Friday Okputu, Clement Emmanuel Edet, Chinyere Adanna Opara Usoro Pages 74-82
    Background

    Multiple organ dysfunctions have been linked to exposure to polycyclic aromatic hydrocarbons (PAH) and oxidative stress (OS), oxidative DNA damage, and inflammatory response to PAH have been implicated. The biomarkers of OS (malondialdehyde (MDA), total plasma peroxide (TPP), total antioxidant capacity (TAC), glutathione (GSH), nitric oxide (NO), oxidative stress index (OSI)); 8-hydroxy-2-deoxyguanosine (8-OHdG)); tumor necrosis factor-alpha (TNF-α)); 1-hydroxy pyrene (1-HOP)), serum and urine creatinine, uric acid (UA), estimated glomerular filtration rate (eGFR) and peak expiratory flow rate (PEFR) were assessed in barbecue makers.

    Methods

    One hundred barbecue makers and 50 controls were enrolled into the study. Serum and urine creatinine, UA, TAC, MDA, GSH, NO and TPP were estimated by colorimetry, 8-OHdG and TNF-α by ELISA, PEFR using peak flow meter, 1-HOP by HPLC, eGFR and OSI by calculation.

    Results

    Barbecue makers had lower TAC, PEFR, and higher TNF-α and OS compared to controls (P<0.05). Higher TNF-α, lipid peroxidation, and lower antioxidants were observed in barbecue makers who had worked for >5years compared to <5years (P<0.05). Increasing number of working hours was associated with higher NO, lipid peroxidation, OS and lower antioxidants in barbecue makers (P<0.05). Positive associations were observed between 1-HOP and TPP (r=0.570, P=0.000), OSI (r=0.299, P=0.035) and negative association between TAC and TNF-α (r=-0.209, P=0.037), MDA (r=-0.265, P=0.008) in barbecue makers.

    Conclusion

    Increased lipid peroxidation, OS, inflammation and depressed antioxidants and lung function observed in barbecue makers suggest increased risk of chronic lung conditions which may be associated with exposure to PAH in barbecue fumes.

    Keywords: Inflammation, Kidney, Lipid Peroxidation, Lung, Oxidative Stress, Polycyclic Aromatic Hydrocarbon
  • Thekra Abdulaali Abed, Anwar Ali Abdulla* Pages 83-88
    Background

    The aim of present study is to asset the IL-2 promoter gene (SNP -475) as a candidate gene for multiple sclerosis (MS) susceptibility.

    Methods

    This study included 70 patients with relapsing – remitting multiple sclerosis (RRMS) and 50 healthy controls. Following the extraction of genomic DNA from peripheral blood, frequency of genotypes and alleles of SNP -475 was calculated using Restriction fragment length polymorphism-polymer chain reaction (RFLP-PCR) and then the results were analyzed statistically.

    Results

    The results revealed the unusual ratio for the heterozygous (AT) was 1.6972 indicating that heterozygous patients were at higher risk of multiple sclerosis than wild homozygous (AA), and homomutant (TT). The results show protective role for - 475 IL-2 promoter among individuals with multiple sclerosis, (O.R: 0.4872; C.I. 95%: 0.1617- 1.4680) and (O.R: 0.9275; C.I. 95%: 0.2476 - 3.4745) for both AA and TT genotypes, respectively.

    Conclusions

    Our results showed that in this population of Iraqi patients, the AT genotype / A allele of -475 IL-2 promoter gene SNP may include attributed factors for MS predisposition.

    Keywords: IL-2, Multiple sclerosis, PCR-RFLP SNP
  • Moataza Moataza Hassan Omran*, Basma El-Sayed Fotouh, Wafaa Ghoneim Shousha, Abeer Ismail, Shimaa Shawki Ramadan Pages 89-101
    Background

    Breast Cancer (BC), the second leading cause of cancer mortality after lung cancer and varied across the world due to genetic and environmental factors. In this study, we evaluated the interaction between the polymorphisms in genes encoding enzymes of folate metabolism: methylenetetrahydrofolate reductase (MTHFR), methionine synthesis reductase (MTR) with the BC prognostic factors.

    Methods

    This study was conducted on 160 Egyptian subjects, 60 controls and 100 cases. Sequencing, RFLP analysis in addition to statistical analysis including Chi‐squared test, haplotype analysis was used to evaluate associations with BC risk and its clinicopathological parameters. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using unconditional logistic regression.

    Results

    Strong significant association with breast cancer risk was observed for the haplotype (T-C-G) of MTHFR C677T/ MTHFR A1289C and MTRA2576G and hormonal receptor expression (ER-/PR- /HER2+), bigger and advanced tumor and metastatic lymph nodes. However, no significant difference was
    observed for age.

    Conclusions

    The combination of SNPs from MTHFR and MTR genes has a more synergistically genetic effect on BC disease progression. These SNPs could be used as tumor aggressiveness markers among Egyptian females with BC and could help in saving money and time.

    Keywords: Breast cancer, Methionine synthesis reductase, MTHFR, PCR-RFLP, SNPs
  • Maytham Hassan Jasim Al-Thabhawee, Hussein Oleiwi Muttaleb Al-Dahmoshi* Pages 102-110
    Background

    Multidrug resistance Pseudomonas aeruginosa (MDRPA) is most important issue in healthcare setting. It can secrete many virulence effector proteins via its secretion system type (T1SS-T6SS). They are using them as conductor for delivering the effector proteins outside to begins harmful effect on host cell increasing pathogenicity, competition against other microorganism and nutrient acquisition. 

    Methods

    The study include investigation of 50 isolates of MDRPA for transport secretion system and resistance for antibiotics. Molecular diagnosis using P. aeruginosa specific primer pairs, investigation of AprF, HasF, XcpQ, HxcQ, PscC, CdrB, CupB3, and Hcp using specific primer pairs by PCR were also
    performed.

    Results

    The results revealed high resistance to beta lactam antibiotics (78% for ceftazidime, 78% for cefepime and 46% for piperacillin) can indicate possessing of isolates for beta lactamases and this confirmed by dropping resistance to piperacillin to 16% when combined with tazobactam. Also, the results shown the ability of MDRPA for pyocyanin biosynthesis using the system of genes.

    Conclusions

    The current study conclude that all isolates of P. aeruginosa were highly virulent due to their possessing of all transport secretion system to deliver different effector proteins with possible harmful effects of these proteins.

    Keywords: Drug resistance, MDR, Efflux pump, Pseudomonas aeruginosa
  • Abbas Ghotaslou, Amir Samii, Hassan Boustani, Omid Kiani Ghalesardi, Minoo Shahidi* Pages 111-124
    Background

    Doxorubicin (DOX)-induced cardiotoxicity appears to be a growing concern for extensive use in acute lymphoblastic leukemia (ALL). The new combination treatment strategies, therefore might be an effective way of decreasing its side effects as well as improving efficacy. AMG232 (KRT-232) is a potential MDM-2 inhibitor, increasing available p53 through disturbing p53-MDM-2 interaction. In this study, we examined the effects of AMG232 on DOX-induced apoptosis of NALM-6 cells.

    Methods

    The anti-leukemic effects of Doxorubicin on NALM-6 cells, either alone or in combination with AMG232, were confirmed by MTT assay, Annexin/PI apoptosis assay, and cell cycle analysis. Expression of apoptosis and autophagy-related genes were further evaluated by Real time-PCR method. To investigate the effect of AMG232 on NALM-6 cells, the activation of p53, p21, MDM-2, cleaved Caspase-3 proteins was evaluated using western blot analysis.

    Results

    The results showed that AMG232 inhibition of MDM-2 enhances Doxorubicin-induced apoptosis in NALM-6 cells through caspase-3 activation in a time and dose-dependent manner. Furthermore, cotreatment of AMG232 with Doxorubicin hampered the transition of NALM-6 cells from G1 phase throughincreasing p21 protein. In addition, this combination treatment led to enhanced expression of apoptosis and autophagy-related genes in ALL cell lines.

    Conclusions

    The results declared that AMG232 as an MDM-2 inhibitor could be an effective approach to enhance antitumor effects of Doxorubicin on NALM-6 cells as well as an effective future treatment for ALL patients.

    Keywords: Acute Lymphoblastic Leukemia, AMG 232, Autophagy, Doxorubicin, p53
  • Chee Hoe Cheah, Anna Pick Kiong Ling*, Ying Pei Wong, Rhun Yian Koh, Sobri Hussein Pages 125-137
    Background

    It is believed that activation of microglia in the central nervous system upon detection of stimulus like lipopolysaccharides provokes neuroinflammation via the production of pro-inflammatory mediators and cytokines. The cytoprotective and anti-inflammatory properties of various folk medicine has been gaining attention as a strategy to combat various disease. This study aimed to assess the antineuroinflammatory properties of chloroform extract of in vitro Panax ginseng root culture based on nitric oxide and cytokines production.

    Methods

    The study was initiated with the determination of maximum non-toxic dose (MNTD) of P.ginseng root culture chloroform extract using the MTT assay. The lipopolysaccharides-stimulated BV2 microglia cells were treated with MNTD and ½MNTD of the extract and its anti-neuroinflammatory properties were assessed by measuring the production of nitric oxide (NO) via Griess assay, as well as TNF-α, IL-6 and IL-10 using Quantikine ELISA.

    Results

    It was found that the MNTD and ½MNTD of the extract did not play a significant role in the production of pro-inflammatory cytokines such as NO, TNF-α and IL-6. However, the MNTD and ½MNTD of chloroform extract significantly increased the anti-inflammatory IL-10 compared to the untreated cells.

    Conclusions

    With this, the chloroform extract of P. ginseng root culture potentially exerts antineuroinflammatory properties.

    Keywords: Anti-inflammatory cytokines, Ginsenosides, Microglia cells, Neuroinflammation, Pro-inflammatory cytokines
  • Hawraa Saad Al-Kawaz*, Oda Mizil Yasser, Mazin Jaafar Mousa Pages 138-145
    Background

    Pre-eclampsia is an idiopathic pregnancy disorder characterized by appearance proteinuria and hypertension, with poorly understood etiology. It has been linked to a variety of system abnormalities, including ion transport disorders in neonatal, maternal, and placental cell lines. A new method was described to evaluate the inhibition percentage of endogenous digitalis in plasma of pre-eclampsia patients compared with normal pregnancies, with the estimation of sensitivity and specificity of the proposed test.

    Methods

    This was a case-control study consisting of 130 cases that were divided into three groups, 55 normal pregnancies (positive control), 30 non-pregnant women (negative control), and 45 preeclampsia (patients). The new method included the estimation of the percentage inhibition of endogenous digitalis by measuring specific enzyme activity of Na-K ATPase for the patient and positive control. The results were analyzed using the statistical package for social sciences (SPSS®) software version 26.0. A p-value of≤ 0.05 was considered significant.

    Results

    In the pre-eclampsia patient, the specific activity of Na-K ATPase was significantly lower with mean= 0.239 mg/g±0.043 compared to the control group which was 0.397 mg/g±0.021, p<0.001. While the result of inhibition percentage of endogenous digitalis showed significantly higher in the pre-eclampsia patient (mean= 35.852 mg/g %±2.692%) compared to the control group (mean=17.964%±1.784), with a p< 0.001.

    Conclusions

    Pre-eclampsia is linked with lower erythrocyte sodium pump activity significantly in pre-eclampsia patients than in normal pregnancies. Also, results show the inhibited percentage of endogenous digitalis elevation in patients with pre-eclampsia compared with normal pregnancy.

    Keywords: Endogenous Digitalis, Hypertension, Inhibition Percentage, Pre-eclampsia
  • Hakam Hasan Alkhateeb*, Nasser Mohammed Kaplan, Mohammed Al-Duais Pages 146-156
    Background

    Oleuropein, the main constituent of olive fruit and leaves, has been reported to protect against insulin resistance and diabetes. While many experimental investigations have examined the mechanisms by which oleuropein improves insulin resistance and diabetes, much of these investigations have been carried out in either muscle cell lines or in vivo models two scenarios with many drawbacks. Accordingly, to simplify identification of  mechanisms by which oleuropein regulates specific cellular processes, we resort, in the present study, to isolated muscle preparation which enables better metabolic milieu control and permit more detailed analyses.

    Methods

    For this purpose, soleus muscles were incubated for 12 h without or with palmitate (1.5 mM) in the presence or absence of oleuropein (1.5 mM), and compound C. Insulin-stimulated glucose transport, glucose transporter type 4 (GLUT4) translocation, Akt substrate of 160 kDa (AS160) hosphorylation and adenosine monophosphate-activated protein kinase (AMPK) phosphorylation were examined.

    Results

    Palmitate treatment reduced insulin-stimulated glucose transport, GLUT4 translocation and AS160 phosphorylation, but AMPK phosphorylation was not changed. Oleuropein administration (12 h) fully rescued insulin-stimulated glucose transport, but partially restored GLUT4 translocation. However, it fully restored AS160 phosphorylation, raising the possibility that oleuropein may also have contributed to the restoration of glucose transport by increased GLUT4 intrinsic activity. Inhibition of AMPK phosphorylation with compound C (50 μM) prevented oleuropein -induced improvements in insulin-stimulated glucose transport, GLUT4 translocation, and AS160 phosphorylation.

    Conclusions

    Our results clearly indicate that oleuropein alleviates palmitate-induced insulin resistance appears to occur via an AMPK-dependent mechanism involving improvements in the functionality of the AS160-GLUT4 signaling system.

    Keywords: AMPK, GLUT4, Muscle, Insulin resistance, Oleuropein
  • Mohamadreza Savaee, Ali Bakhshi, Fatemeh Yaghoubi, Fatemeh Pourrajab*, Koorosh Goodarzvand Chegini Pages 157-165
    Background

    Prostate cancer is known as one of the most prevalent health disorders in the male population globally. The aim of the current study was to evaluate the effects of separate and concomitant use of MK-2206 and salinomycin on prostate cancer cell line.

    Methods

    The antitumor potential of separate and concomitant use of MK-2206 and salinomycin was evaluated in a panel of prostate cancer cell line (PC-3). To get insights into the underlying mechanism of action, different assays including the rate of apoptosis, cell viability, and gene expression were performed in treated prostate cancer cells.

    Results

    A significant reduction was detected in the viability percentage of prostate cancer cells (p< 0.001) and the rate of Akt expression (p< 0.001) in all salinomycin, MK-2206, and salinomycin+MK-2206 groups compared to the negative control group. Furthermore, in comparison with the negative control group, there was a notable increase in both the rate of Bad expression (p< 0.001) and prostate cancer cells apoptosis after salinomycin, MK-2206, and salinomycin+MK-2206 treatments. Moreover, the concomitant use of salinomycin+MK-2206 revealed synergistic improvements regarding the viability of prostate cancer cells and the rate of the Akt and Bad expressions compared to the separate administration of salinomycin and MK-2206 (all p< 0.05).

    Conclusions

    The findings of the present study may contribute to improving the efficacy of the therapies regarding the management of prostate cancer and providing a beneficial strategy in clinical trials.

    Keywords: Apoptosis, Gene Expression, MK 2206, Prostatic Neoplasms, Salinomycin
  • Ruaa Ali Muhammed Ali, Jamal Mohammed Ridha Alshara, Nabil Salim Saaid Tuwaij*, Huda Jameel Baker Al-Khilkhali Pages 166-175
    Background

    Escherichia coli (E. coli) remains one of the leading agents of urinary tract infection (UTIs), it has become resistant to many drugs. Current work aimed to evaluate some chemical substances as antibacterial agents and molecular study of virulence factors associated with UTIs.

    Methods

    This work involved 133 urine specimens obtained from females’ patients suffering from UTIs, Methods of well diffusion and disk diffusion were achieved to assay the effect of some chemical substances and antibiogram profiles toward Sulfamethoxazole-trimethoprim (SXT)-resistant E. coli respectively. Virulence genes were done based on the technique of Polymerase Chain Reaction (PCR).

    Results

    The results recorded 49/133 (36.84%) E. coli among women suffering UTIs, 28/49 (57.14%) were resistant to SXT drug. imipenem, meropenem, and nitrofurantoin were recorded more effectively. Chemicals substances at the concentration 0.3 (g/ml) recorded percentages of inhibition, reaching 9.143±1.442, 15.36±0.5914, and 21.82±0.8699 for NaHCO3, Ch4c, and Viroxide Super™ respectively. PCR demonstrated that 28/28 (100%) of SXT-resistant E. coli isolates were harbored Sul-2, FeoB and PapC genes, while 14/28 (50%), 15/28 (53.57%), 19/28 (67.85%) and 26/28 (92.85%) in U250 (pet), FumC, Sul-1 and IutA genes, respectively. Sul-3 gene was not observed.

    Conclusions

    Observed a high percentage of E. coli that were resistant to SXT drug, and having several virulence genes, poses a real threat, it requires a real pause to create substitutions to limit the spreading of this threat.

    Keywords: Chemical substance, PCR, SXT-resistant E. coli, UTIs
  • Masar Riyadh Rashid Al-Mousawi, May Mohammed Ali, Noor Salman Kadhim Al-Khafaji, Hussein Oleiwi Muttaleb Al-Dahmoshi* Pages 176-181
    Background

    Human Adenovirus species D (HAdV-D) was common human viral pathogen especially in eye infection, consists of several types of which HAdV-D8, -D19 and –D37 were common in eye infection. This study includes detection of HAdV-D types implicated in conjunctivitis based on L2 (Penton protein) gene similarity.

    Methods

    Conjunctival swabs were collected from Keratoconjunctivitis patients as eye infection related to adenovirus. Viral nucleic acids were extracted and specific primer pairs for HAdV-D L2 gene (encoding for penton base protein) was used to amplify the target gene and only positive samples were sent to sequencing.

    Results

    The results revealed that only 6 samples give positive results for L2 gene amplification and then sent for sequencing for L2 (penton protein) gene-based typing. The results show that 4 local isolates (S1, S2, S3, S6) were similar to HAdV-D8 and 2 local isolates (S4, S5) were similar to HAdVD20. Also the results display that the HAdV-37, prominent HAdV-D type of human eye infection, may be variant of HAdV-D20 due to that six variation were seen in S4and S5 local isolates nucleotide sequence in relation to HAdV-D37: T>C at position 14364, A>C at position 14411, T>C at position 14427, C>A at position 14448, G>A at position 14540 and T>C at position 14617, leading to only 2 amino acid change in resulted penton protein: T (Threonine) instead of K (Lysine) at position 204 and N (Asparagine) instead of D (Aspartic acid) at position 247.

    Conclusions

    The current study concludes the possibility of implication of HAdV-D20 in eye infections especially conjunctivitis.

    Keywords: HAdV-D8, HAdV-D20, HAdV-D37, conjunctivitis, Iraq
  • Shaymaa Obaid Abdullah, Ghazi Mohamad Ramadan, Hayder Abdul-Amir Makki Al-Hindy*, Mazin Jaafar Mousa, Amir Al-Mumin, Safa Jihad, Suhad Hafidh, Yaqeen Kadhum Pages 182-189
    Background

    The contribution of neutrophils is still indistinct in the inflammatory response of bronchial asthma (BAs). Myeloperoxidase (MPO) is an  enzyme released from the primary azurophilic granules of the neutrophils. The study aimed to evaluate the levels of serum MPO as a biomarker for the assessment of the level of asthma control. 

    Methods

    The study participants included 94 asthmatic patients and 86 healthy controls. The identification of asthma severity had assessed using the ''Global Initiative for Asthma guidelines''. Asthmatic adults had divided into three groups: Good (n= 22), partial (n= 30), and poor control (n=44). Also, patients have been divided again into two groups (treated and untreated) for BAs.

    Results

    The predicted FEV1% and the peak expiratory flow (PEF/L) of all participants had verified by spirometry. The mean patients’ age was 31.9±15.1 year, with a predominance of females. The mean asthma duration was 10.5±8.6 years. Mean spirometric parameters (FEV1 and PEF) were significantly lower among the patients (0.00). Significant higher MPO levels had observed among BAs patients (p-0.00). The MPO levels have not differed significantly with asthma levels and had significant differences with the history of treatment. There was a nonsignificant negative correlation between the mean MPO levels and the spirometry variables among the patients. ROC curves revealed a sensitivity, specificity, accuracy for MPO (80.9%, 72.1%, and 84.3%), respectively to predict asthmatic severity.

    Conclusions

    There were significantly higher MPO levels compared to healthy controls. Levels of serum MPO had a non-significant positive correlation with levels of asthma control, but a negative non-significant correlation with spirometric results.

    Keywords: Asthma, And Neutrophils, FEV1, MPO, PEF