فهرست مطالب
Reports of Biochemistry and Molecular Biology
Volume:11 Issue: 2, Jul 2022
- تاریخ انتشار: 1401/06/23
- تعداد عناوین: 20
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Pages 190-199Background
Oxidative stress is defined as the condition in which balance between the synthesis and detoxification of reactive oxygen species in cells is disrupted. This research explored the effects of intermittent and prolonged fasting on malondialdehyde (MDA), carbonyl, reduced glutathione (GSH), and
specific activity of catalase as biomarkers for oxidative stress in hearts, brains, and kidneys of New Zealand White (NZW) rabbits.MethodsFifteen NZW rabbits were divided into control, intermittent fasting (IF), and prolonged fasting (PF) groups. The controls were fed ad lib. IF and PF groups were fasted for 16 and 40 hours, respectively, followed by eight hours of non-fasting, for six days and were sacrificed on the 7th day. One hundred mg of heart, brain, and kidney tissues were homogenized in 1 ml of phosphate-buffered saline. MDA, carbonyl, GSH, and catalase were analyzed by spectrophotometry. Data were analyzed using One-way ANOVA and post hoc test.
ResultsIn heart, MDA was significantly greater in the control than in the IF and PF groups. In brain, GSH was greater in the IF than in the PF and control groups. Also, in brain, catalase specific activity was significantly greater in the control than in the IF and PF groups. In kidney, catalase specific activity was
significantly less in the PF than in the control group.ConclusionsThe effect of fasting on oxidative stress in various organs showed various responses, however fasting reduced oxidative stress based on MDA and GSH levels in the heart and brain, respectively.
Keywords: Fasting, Oxidative stress, Vital organs, Rabbit -
Pages 200-208Background
The oral squamous cell carcinoma (OSCC) composes about 90% of all head and neck cancers. The toll-like receptor (TLR)+ immune cells have potential of invasion and malignancy transformation. The aim of this study was assessment of possible associations between clinicopathological indices and TLR2 and TLR9 gene expression in OSCC.
MethodsForty-two OSCC samples with related healthy margins including 25 early and 17 advanced stages were gathered. The samples were classified histologically from grade I to II. The expression of TLR2 and TLR2 was evaluated by Real-time PCR. The patient’s disease-free survival (DFS) and overall survival (OS) were analyzed using SPSS V.23 software.
ResultsThe expression of TLR2 and TLR9 genes in tumor tissues (especially in grade I and II) were higher than healthy surgical margin tissue (p< 0.001). TLR9 expression in grade II was statistically significant than grade I in tumor tissue (p< 0.001). TLR9 expression in advanced stage was statistically significant in compare to early stage (p= 0.012). In advanced stage both overall survival (p= 0.029) and disease-free survival (p= 0.012) were statistically lower than early stage. The follow-up time to recurrence in advanced stage was statistically lower than early stage (p= 0.007).
ConclusionsOverexpression of TLRs 2, 9 play role in the pathogenesis and tumor development of OSCC and can be applied as biomarker in prognostic approaches.
Keywords: Disease-free survival, Oral squamous cell carcinoma, Overall survival, TLR2, TLR9 -
Pages 209-215Background
Plasma protein profile test is a potential laboratory method to assess nutritional status especially albumin and globulins levels which reflects protein adequacy. The purpose of this study is to evaluate plasma protein profile of lactating women from two primary health centers in Jakarta.
MethodsA cross-sectional study was conducted involving lactating women attending routine maternal examinations in two public primary health centers in Jakarta, Indonesia. The mother’s plasma total protein, albumin, globulin, and immunoglobulin levels were measured.
ResultsSixty lactating women were recruited, mostly were 28 years old, slightly overweight, bearing two children, and their recent children were 2 months old. The mean total protein level was 8.13 g/dl, albumin 5.00 g/dl, globulin 3.18 g/dl, albumin: globulin ratio 1.558, mean total IgG level of 1255.98 and mean total IgM level of 135.819. All the measured plasma protein parameters were shown to be not correlated with maternal age, maternal BMI, or maternal parity.
DiscussionThe plasma total protein, albumin, globulin, as well as total IgG and IgM level of lactating women in Jakarta were within normal range. These biochemical parameters were shown to be not correlated with anthropometrical data such as maternal age and BMI. The small and relatively homogenous samples were supposed to be the cause of such findings.
ConclusionsThe plasma protein profile of lactating women in Jakarta was adequate. Further studies are required to evaluate the eligibility of plasma protein profile as biochemical parameter of nutritional status in lactating women.
Keywords: Blood protein, lactation, protein, albumin, globulin, immunoglobulin M, G -
Pages 216-223Background
A group of transcription factors involved in several cellular processes like cell growth, proliferation, cell cycle, differentiation and apoptosis which are critical to the cell biology of cancer is Forkhead Box O (FOXO) family. FOXOs are known as putative tumor suppressors. FOXO1 is a member
of FOXO family which its abnormal expression or function has been indicated to promote cell proliferation and tumorigenesis. The probable effects of FOXO1 rs17592236 polymorphism on Papillary thyroid carcinoma (PTC) and its clinical findings were evaluated.MethodsIn total, 156 PTC patients and 158 healthy subjects were participated in the study. Genotyping of FOXO1 rs17592236 polymorphism was carried out using RFLP-PCR method.
ResultsThere was no association between the FOXO1 rs17592236 polymorphism and PTC in codominant, recessive, dominant, overdominant, and log-additive models. The frequency of rs17592236A allele was 13% in PTC and 17% in control group and were not statistically significant (p= 0.15). The analysis of the relationship between FOXO1 rs17592236 polymorphism and clinical specifications of papillary thyroid carcinoma demonstrated no significant relationship between rs17592236 polymorphism and PTC in different ages (< 40 and≥ 40), gender (male/female), extrathyroidal expansion, N stage, vascular invasion and capsular invasion in PTC patients. There was a relationship between FOX1 rs17592236 polymorphism and a larger tumor size (≥ 1 cm) only in log-additive model (OR= 2.96, 95% CI= 0.88-9.98; p= 0.04).
ConclusionsFOXO1 rs17592236 polymorphism was not associated with PTC; however, this variant was associated with a larger tumor size (≥ 1 cm) only in log-additive model.
Keywords: FOXO1, Papillary thyroid carcinoma, Polymorphism, Tumor size -
Pages 224-237Background
The clinical effect of photodynamic therapy (PDT) may be correlated with the degree of dysplasia of cancer tissues. The aim of this study was to compare the effects of cisplatin, silver nanoparticles (AgNps), and photodynamic therapy (PDT) using methylene blue (MB) photosensitizer on Head and Neck squamous cell carcinoma - cell line (HNSCC), Hep-2, through genes expression.
MethodsHep-2 cells were divided into four groups: group I as control and without any treatment, group II and III were treated by cisplatin and AgNps, respectively, and group IV were incubated with MB for four minutes followed by PDT using laser irradiation at 650 nm for 8 minutes. The resulting toxicity was assessed in cell lines using MTT cytotoxicity assay. Further, apoptosis and the response to treatment was examined
via RT-qPCR.ResultsMB-PDT inhibited the proliferation of Hep-2 cells. Following PDT, compared with AgNps cells and via MTT assay, a highly significant decrease was observed in cell proliferation in Cancer cells treated with AgNps and MB- PDT groups compared to cancer group cells and cancer cells treated with Cisplatin
(p value< 0.001). Mechanistically, both the mRNA and protein expression levels of Bcl-2, Caspase-3, Cyclin-D, HIF-1, IL-8, MAPK-38, and ROS were found to be down regulated in Hep-2 cell line after MBPDT.ConclusionsMB-PDT effectively killed Hep-2 cells in vitro, however, under the same conditions, the susceptibilities of the cell line to cisplatin, AgNps, and MB-PDT were different. Further studies are necessary to confirm whether this difference is present in clinical oral cancer lesions.
Keywords: Head, Neck squamous cell carcinoma (HNSCC) silver nanoparticles (AgNps) photodynamic therapy (PDT), Methylene Blue (MB), Photosensitizer (PS) -
Pages 238-245Background
Tumor necrosis factor-alpha (TNF-α) may stimulate airway hyperresponsiveness in asthma, which is also affected by neutrophils activity. The latter can be determined indirectly by evaluating myeloperoxidase (MPO) activity. The insufficient studies that investigated the combined association of
serum TNF-α and MPO with asthma was objective of this study.MethodsA case-control study included 110-asthmatics besides 92-controls. All participants underwent venous sampling for TNF-α and MPO immunoassays. A percentage of predicted ''forced expiratory volume in one second (FEV1%)'', and the ''peak expiratory flow rate (PEF/L)'' of all participants were verified. The statistical analyses had done using SPSS V-25. The accuracy, specificity, sensitivity, and significance of both biomarkers to distinguish asthma examined ''under the ROC-curves''.
ResultsHigh TNF-α levels observed among the controls(p-0.006), opposing the higher MPO levels among the patients(p-0.00). There were nonsignificant variations of two biomarkers between the treatment groups and nonsignificant correlations of MPO with FEV1 and PEF. There was a significant correlation of MPO with the TNF-α levels of all participants. The TNF-α showed lower sensitivity, specificity, and accuracy to diagnose asthma. There were no MPO differences according to asthma levels. The TNF-α was higher among the severe asthmatics significantly.
ConclusionsTNF-α may be a contributory particle for neutrophilic inflammation of severe asthma. MPO levels were significantly higher among asthmatics, whereas TNF-α levels were lower. TNF-α levels were higher among those with severe compared to mild/moderate asthma. The MPO level has a significant
predictive capacity compared to TNF-α for distinguishing asthma from healthy subjects.Keywords: Asthma, Inflammation, MPO, Neutrophils, TNF-α -
Pages 246-251Background
During the gathering of demographic data for the biobank on Buerger’s Disease (BD), we found that, after the clinical manifestation of BD, the patients usually became infertile, and the age of their last child was compatible with the time of disease diagnosis. The aim of this study was to evaluate the underlying cause of secondary infertility in BD patients.
MethodsAnti-sperm antibodies (ASA), testosterone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) in the sera of 39 male BD patients were measured and compared with 39 age-matched Caucasian male controls.
ResultsSix patients declared that they suffered from impotency. The ASA level was positive in 25.6% of the patients and 2.4% of the controls (p= 0.003, CC= 6.96). The mean levels of testosterone in the patients and controls were 393.12±32.9 ng/dl and 354.37±30.9 ng/dl, respectively. The mean levels of LH in the patients and controls were 0.88±0.12 mIU/r and 0.85±0.1 mIU/r, respectively. The mean levels of FSH in the patients and controls were 4.1± 0.35 mIU/r and 3.56±0.33 mIU/r, respectively. No significant difference in the serum levels of testosterone, LH, or FSH was found between the patients and controls (p> 0.05). The spermograms of three ASA-negative patients demonstrated impaired sperm motility.
ConclusionsAnti-sperm antibodies, disturbed genital circulation, autonomic dysfunction and sperm motility may be responsible for secondary infertility in Buerger’s Disease.
Keywords: Anti-sperm antibody, Buerger’s Disease, Infertility, Thromboangiitis Obliterans -
Pages 252-261Background
In the field of recombinant protein production, downstream processing, especially protein purification, is critical and often the most expensive step. Carbohydrate binding module 64 (CBM64) was shown in 2011 to bind efficiently to a broad range of cellulose materials.
MethodsIn this study, we developed a protein purification method using nanocrystalline cellulose embedded in a polyacrylamide monolith cryogel and CBM64 affinity tag linked by intein to PD1 as a model protein. The CBM64-Intein-PD1 gene cassette was expressed in E. coli. Following cell lysis, CBM64-Intein-PD1 protein bound to the monolith PA-NCC cryogel. After washing and reducing the pH from 8.0 to 6.5, the intein underwent self-cleavage, resulting in the release and elution of pure PD1 protein.
ResultsThe synthesized monolith column had a porous structure with an average pore size of 30 μm and a maximum binding capacity of 497 μg per gram of dried column. The yield of this purification method was 84%, while the yield of the His tag-acquired CBM64-Intein-PD1 method was 89%.
ConclusionsWe used cellulose as support for affinity chromatography, which can be used as a cost-effective method for protein purification.
Keywords: Affinity tag, CBM64, PD1, Protein purification -
Pages 262-269Background
The assembly and disassembly of the focal adhesions (FA) components occurs throughout life cycle of adhesion, with conservation of balance between removal and recruitment rate during temporal stages. Previous studies have demonstrated that phosphotidyilinositols play a role in regulating FA turnover. However, a little attention has been given to quantify the dynamics changes of Phosphatidylinositol 3,4,5-trisphosphate (PtdIns (3,4,5) P3) within and during fast and slow turnover rates of FA.
MethodsMDA-MB-231 breast cancer cell line was used as a model in this study due to high metastatic and motile. These cells were co-transfected with GFP- paxillin/vinculin, as FA marker, and the GFP/mCherry-Btk-PH, as a biosensor to visualize PtdIns (3,4,5) P3. Confocal time-lapse images were used
to monitor changes or differences in the local generation of PtdIns (3,4,5) P3 within and during assembly and disassembly of FA. Following transfection, immunostaining was used to examine the spatial colocalization between FA and PtdIns (3,4,5) P3.ResultsOur data demonstrated that PtdIns (3,4,5) P3 co-localized with FAs and increase during assembly and decline during disassembly of FA which exhibits slow turnover rates and was in a constant level during assembly and disassembly of FA that displays fast turnover rates.
ConclusionsOur result suggested that the dynamic changes of PtdIns (3,4,5) P3, it may depend on components undergo turnover, such that early, nascent FA displays fast turnover rates and mature FA exhibits slow turnover rates. Thus, the local enrichment of PtdIns (3,4,5) P3 enhances FA assembly and disassembly activation.
Keywords: Cancer cell migration, Focal adhesion turnover, MDA-MB-231 cell line, PtdIns (3, 4, 5) P3 -
Pages 270-281Background
Focal adhesions (FAs) are highly dynamic complex structures that assembled and disassembled on an ongoing basis. The balance between the two processes mediates various aspects of cell behavior, ranging from cell adhesion to cell migration. Assembly and disassembly processes of FAs are
regulated by a variety of cellular signaling proteins and adaptors. We previously demonstrated that local levels of Phosphatidylinositol 4,5‐bisphosphate (PtdIns(4,5)P2) in MDA-MB-231 cells increases during FA assembly and declines during disassembly. In this study we aimed to investigate whether PtdIns(4,5)P2 regulates FA turnover.MethodsMDA-MB-231 cells were co-transfected with a labeling vinculin (or zyxin) and the PLC𝛅1-PH biosensor to visualize FA localization and PtdIns(4,5)P2 in the cell membrane. We also used pharmacological inhibitors to determine the mechanism underlying the changes of PtdIns(4,5)P2 level
during FA turnover and cell migration. Immunostaining, immunoprecipitation, and Western blotting were used to examine the localization and interaction between phospholipase C (PLC)/phosphatidylinositol 3-kinase (PI3K) FA proteins.ResultsWe showed that inhibition of PLC, PI3K significantly reduced the decline of PtdIns(4,5)P2 levels within FA disassembly and the slowdown rate of FA turnover and cell migration. We also showed that the inhibition of enzymes implicated in the downstream pathway of PtdIns(4,5)P2, such as diacylglycerol kinase (DAGK) and protein kinase C (PKC) significantly reduced FA turnover time and the speed of cell migration. Additionally, we demonstrated that PLC but not PI3K interact with FAs. In conclusion,
ConclusionsThis study suggests that dynamical changes of PtdIns(4,5)P2 might regulate FA turnover and facilitate cell migration.
Keywords: Cell Migration, Focal Adhesion Turnover, MDA-MB-231 Breast Cancer Cell Line, Ptdins(4, 5)P2, PLC, PI3K -
Pages 282-288Background
Receptor tyrosine kinase-like orphan receptor 1 (ROR1) is an oncofetal antigen expressed on many types of cancer cells, but not normal adult cells. ROR1 antigen contributes to cancer development and progression by several signaling pathways. ROR1 expression has been associated with tumor growth, survival, and metastasis. In this study specific human recombinant antibodies were selected against ROR1 antigen for their use in cancer immunotherapy.
MethodsPhage display technology was used to produce phage antibody from a human scFv library. Phage concentration was determined to confirm the phage rescue process. Panning procedure was performed to isolate specific scFv clones against ROR1 epitope. Phage ELISA was done to evaluate the reactivity of the selected scFvs.
ResultsTwo specific human scFvs with frequencies of 20% and 25% were selected against ROR1 peptide. The antibodies showed specific reaction to the corresponding epitopes in phage ELISA.
ConclusionsCancer targeted therapy using human specific antibodies is a new strategy, which is used in cancer therapy. The selected specific scFvs that target ROR1 epitope are human antibodies that originated from a human library and have the potential to be used in clinic in cancer immunotherapy of ROR1 positive tumors without induction of human anti mouse antibody (HAMA) response.
Keywords: ROR1, Phage display, scFV library, Cancer -
Pages 289-298Background
Cytoglobin (Cygb) is a relatively newly identified globin protein that acts as an oxygen transporter in tissues like hemoglobin (Hb) in erythrocytes and myoglobin (Mb) in muscles. The natural oxidation of the Fe2+ ion in its heme group into metglobin (globin-Fe3+) made the loses of oxygen binding functions. It is known metHb and metMb can be reduced enzymatically using diaphorase or cyb5r3. However, metCygb reductase had not been previously identified. This study aims to analyze the reducing activity of bovine diaphorase on metCygb.
MethodsDiaphorase was isolated from bovine erythrocyte and purified using gel filtration and cationicexchanger chromatography. Its purity was verified by SDS-PAGE and western blot (WB). The metCygb was obtained from Cygb oxidation with potassium ferrocyanide and its reducing activity was determined by spectroscopy.
ResultsThe diaphorase (MW=30.09 kDa) was purified 10.77-fold from crude enzyme with specific activity against metHb 8.479 U/mg. The purity was confirmed by WB using primary antibody anti-cyb5r3. The purified enzyme reduced metCygb at 0.785 μgmin-1, which was 13.7 times less than the Vmax of metHb.
ConclusionsIn conclusion, the purified diaphorase from bovine erythrocytes did not significantly reduce metCygb rather than metHb, a natural substrate in cells.
Keywords: Bovine Erythrocyte, Cytochrome B5 Reductase, Diaphorase, Metcytoglobin, Reduction -
Pages 299-309Background
The fluctuation in serum caveolin-1 (Cav-1) concentrations is an important indicator of many diseases. Irrespective of the actual cause, a significant reduction of serum Cav-1 is associated with a significant increase in insulin secretion and hyperinsulinemia. The aim of the current study was to evaluate the relationship between serum Cav-1, serum vaspin and visfatin in newly diagnosed men with T2DM.
MethodsEighty-two newly diagnosed men with T2DM were matched for age and body mass indexes (BMIs) with a similar number of non-diabetic men. Serum Cav-1, vaspin and visfatin were assessed through enzyme-linked immunosorbent assay. Fasting serum glucose (FSG), glycohaemoglobin A1C (HbA1c) were both measured using automated method. In addition, waist-circumferences, waist-hip ratio, systolic (SBP), and diastolic blood pressure (DBP) were also obtained.
ResultsSerum concentration of Cav-1(ng/mL) was significantly low in men newly diagnosed with T2DM, (2.334±0.7627) compared with non-diabetic controls (4.321±1.143), p< 0.0001. In contrast, patients with T2DM exhibited significantly higher serum concentrations of vaspin and visfatin (ng/mL), 142.4±60.53) and 2.99±1.091), than controls, 81.53±39.32) and 1.456±0.654), respectively, p< 0.0001. Expectedly, patients with T2DM have significantly higher FSG, HbA1c, systolic blood pressure (SBP), and diastolic blood pressure (DBP).
ConclusionsThere was an inverse significant relationship between Cav-1 and vaspin, visfatin, HbA1c, FSG, and hypertension. This study suggests that serum Cav-1 can be used as a diagnostic marker to predict T2DM in individuals and families under high risk.
Keywords: Caveolin-1, HbA1c, Insulin resistance, T2DM, Vaspin, Visfatin -
Pages 310-319Background
The use of chimeric proteins that selectively interact with various immune cell receptors to treat oncology patients has increased. One effective way to obtain recombinant proteins is to use the E. coli expression system. However, in eukaryotic protein production in E. coli, severaldifficulties arise that can be solved by fusing the target protein with thioredoxin. Thioredoxin can enhance solubility, but its large size can lead to an erroneous assessment of protein solubility, folding, and activity. The present study examined the ligand-binding activity of PD-L1, and CTLA-4 receptors
fused with thioredoxin.MethodsThe de novo synthesized genes of the extracellular domains of the PD-L1 and CTLA-4 were cloned into the pET28 and pET32 expression plasmids and used to transform E. coli BL21 cells. Purified recombinant proteins were characterized by western blotting, LC-MS/MS spectrometry, and
ELISA.ResultsAmino acid sequence comparisons of the recombinant proteins obtained by LC-MS/MS with the SwissProt database resulted in the highest comparison scores from 4950 to 13396. The binding efficiencies of recombinant human B7-1 Fc to rCTLA-4 and rTrx-CTLA-4 proteins in ELISA did not
differ significantly. Similar results were obtained with recombinant rhesus monkey PD-1 hFc against rPD-L1 and rTrx-PD-L1.ConclusionsRecombinant proteins specifically reacted with recombinant human B7-1 Fc and recombinant rhesus monkey PD-1 hFc. The fusion of thioredoxin with recombinant proteins through linkers slightly affected the activity of the extracellular domains of CTLA-4 and PD-L1.
Keywords: Chimeric Protein, CTLA-4, PD-L1, Protein Refolding, Recombinant Protein, Thioredoxin -
Pages 320-326Background
Recent advancement on experiment concluded that etiology of pre-eclampsia (PE) could be explained by the "two-stage" theory. The theory of which explained that pre-eclampsia occurs due to abnormalities in spiral arteries development and release of inflammatory response. Failure of spiral arteries development, the lesion of damage could be due to ischemia-reperfusion or hypoxia-reoxygenation. Hypoxia in pre-eclamptic placenta leads to metabolic change to anaerobic in glycolysis. Lactate dehydrogenase (LDH) has important role in anaerobic glycolysis that catalyzes the conversion of lactate to pyruvate during hypoxia. On the other hand, phosphoenolpyruvate carboxy kinase (PEPCK) is merely an enzyme of gluconeogenesis. This research conduct to reveal that in early onset pre-eclampsia the placenta still hypoxic and undergoes gluconeogenesis even after delivery, through metabolic enzyme of LDH and PEPCK level.
MethodsThis cross-sectional study compared early onset PE (< 34 weeks) with normal term placenta. We measured LDH enzyme activity using colorimetric assay and PEPCK protein using ELISA method.
ResultsResult show that placental LDH specific activity was increased significantly in PE with median 2.750 (0.030 - 5.680) U/mg compared to normal term placenta 0.255 (0.032 – 1.194) U/mg (Mann-Whitney, p< 0.001). PEPCK level was significantly increased in PE 8.998 (1.737-44.914) ng/mg compared to normal term placenta 1.552 (0.741-8.832) ng/mg (Mann-Whitney, p< 0.001).
ConclusionsWe conclude that anaerobic glycolysis and gluconeogenesis pathway are increased in early onset PE placenta as adaptation mechanism to hypoxic condition.
Keywords: Early onset preeclampsia, LDH, PEPCK, Placenta -
Pages 327-335Background
Sepsis is one of the most common causes of multiorgan failure. Sepsis requires the presence of infection with a resultant systemic inflammatory state. Organ dysfunction occurs from the combination of the two processes. Sepsis is the main cause of mortality at intensive care units, with 30-50% mortality rate for all septic episodes. MicroRNA (miRNA) profile data could be beneficial as a specific diagnostic biomarker for sepsis and systemic inflammatory response syndrome (SIRS).
MethodsExpression of miRNAs -122, -181b, -223 and -146a levels were assayed by quantitative real time polymerase chain reaction (qRT-PCR) in a prospective case control study, where forty septic cases were compared to 40 healthy controls of matched age and gender.
ResultsmiRNAs -122 and -181b were significantly upregulated during early septic conditions, indicating that they could be sensitive and specific biomarkers for diagnosing sepsis. miRNA-223 and miRNA-146a could also represent highly specific and sensitive diagnostic biomarkers, as they were found to be significantly down-regulated. Serum levels of miRNA-223 could be used to predict poor prognosis with 70% sensitivity and 75% specificity, whereas the other three miRNAs could not predict prognosis.
ConclusionsOur study shows that all tested miRNAs can be used for early detection of sepsis, with miRNA-223 being predictive of mortality, hence preventing multi-organ failure and reducing mortality, and predicting poor outcomes, thereby assisting in early categorization of ICU patients for rapid appropriate treatment and medico legal aspects.
Keywords: Systemic inflammatory response syndrome (SIRS), Sepsis, Mortality, Biomarker, micRNA-223, Prognostic factor -
Pages 336-343Background
Pancreatic cancer (PC) is among the most aggressive tumors with a poor prognosis, indicating the need for the identification of a novel prognostic biomarker for risk stratifications. Recent genome-wide association studies have demonstrated common genetic variants in a region on chromosome 9p21 associated with an increased risk of different malignancies.
MethodsIn the present study, we explore the possible relationship between genetic variant, rs10811661, and gene expression of CDKN2B in 75 pancreatic cancer patients, and 188 healthy individuals. DNAs were extracted and genotyping and gene expression were performed by TaqMan real-time PCR and RT-PCR, respectively. Logistic regression was used to assess the association between risk and genotypes, while the significant prognostic variables in the univariate analysis were included in multivariate analyses.
ResultsThe patients with PDAC had a higher frequency of a TT genotype for rs10811661 than the control group. Also, PDAC patients with dominant genetic model, (TT + TC), was associated with increased risk of developing PDAC (OR= 14.71, 95% CI [1.96-110.35], p= 0.009). Moreover, patients with CC genotype had a higher expression of CDKN2B, in comparison with TT genotype.
ConclusionsOur findings demonstrated that CDKN2A/B was associated with the risk of developing PDAC, supporting further investigations in the larger and multicenter setting to validate the potential value of this gene as an emerging marker for PDAC.
Keywords: CDKN2A, B, Rs10811661, Pancreatic cancer, Prognostic biomarker -
Pages 344-349Background
Bronchial asthma has a complicated genetic history. Changes in gene expression may be caused by gene polymorphism, cytokines play a central role. IL-13 is an interleukin that has been shown to play a role in the disease's immunopathogenesis. The current study investigated the relationship between rs20541 of the IL-13 gene and Bronchial asthma in Iraqi patients.
MethodsSeventy-five patient and fifty healthy individuals as a control. The DNA was extracted from blood samples. Detection of genotype IL-13SNP (rs20541) were achieved by RFLP-PCR.
Resultsindicated a highly significant the levels of the IgE, and IL-13 in the patients compared to control at (p value≤ 0.01), (456.45±290.106 vs. 30.08±24.414), (59.5980±20.93750 vs.6.7034±4.10547) pg/ml respectively. Result shows no significant differences in the frequency distributions of IL-13 SNP (rs20541) for all genotypes in cases and controls. A protective role of asthma, (OR: 0.62; CI.95%: 0.23 - 1.6) and (OR 0.89; CI.95%:0.42 - 1.89) were observed for wild type homozygous and heterozygous genotype respectively. Whereas the AA genotype (42.7%) in cases and (34.0%) in control, that (OR:1.44; CI.95%:( 0.66 - 3.07) mutant homozygous were risk factors of asthma among individuals. The genotypes of IL-13 rs20541 (GG, AG, AA) among patients and controls were significantly correlated with IgE and IL-13 results at (p≤ 0.05).
ConclusionsAA genotype in case and control mutant homozygous were risk factors of asthma among individuals. It’s possible that this has a predisposing impact on the development of asthma.
Keywords: Bronchial Asthma, RFLP, IL-13, SNP -
Pages 350-357Background
Globally, cardiovascular diseases (CVDs) are the leading cause of death and disability. Elevated low-density lipoprotein-cholesterol (LDL-C) and more specifically, elevation of its small, dense phenotype (sdLDL-C) has been regarded as the key modifiable risk factors associated with atherogenesis. This study aimed to determine the association of LDL-C and sdLDL-C with the development of CVDs in the next six months to establish their predictive efficacy.
MethodsA batch of 162 anonymized serum samples sent for analysis of lipid profile parameters, were classified into tests and controls based on the calculated LDL-C values obtained by Fried Ewald formula. Direct LDL-C was also estimated automatically using assay kits. Using the formula provided by Srisawasdi et al., sdLDL-C was then computed for all samples. Six months later, samples were deanonymized, and the lipid profiles were compared with cardiovascular outcomes of these patients, to determine which parameter had the greatest correlation.
ResultsFour control group patients and three test group patients developed the outcome (any cardiovascular event) during the 6-month follow-up period. Binary logistic regression analysis showed that none of the lipid profile parameters: calculated LDL-C (OR= 0.99; 95% CI= 0.97-1.01; p= 0.826), direct LDL-C (OR= 0.99; 95% CI= 0.97-1.01; p= 0.818) or sdLDL-C (OR= 0.99; 95% CI= 0.93-1.04; p= 0.734), were significantly associated with the occurrence of outcome. The median % sdLDL-C both with respect to direct and calculated LDL-C was slightly higher in patients with the outcome.
ConclusionsThe levels of LDL-C or its individual phenotypes may not be used singly as indicator of cardiovascular morbidity in the next six months.
Keywords: Biomarkers, Cardiovascular diseases, Cholesterol, LDL, Lipoprotein, Myocardial infarction -
Pages 358-366Background
We set out to explore the effect of intrauterine human chorionic gonadotropin (hCG) instillation by intrauterine insemination (IUI) catheter before embryo transfer (ET) on assisted reproductive technologies (ART) outcomes of infertile women.
MethodsOne hundred women with infertility who were scheduled for in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles were included in the study. They were randomly devoted to two groups: experimental (n= 50) and control (n= 50). In the experimental group, 500 IU hCG passed into the internal cervical orifice via IUI catheter within 15 minutes before the transfer of fresh or vitrified cleavage-stage embryos. The control group underwent the same ET procedure without prior injection of hCG.
ResultsNone of the outcomes showed a statistically significant difference between the two groups. In the intervention and control groups, respectively, biochemical pregnancies rates were 26% and 18%, implantation rates were 13.5% and 8.6%, clinical pregnancies rates were 22% and 14%, ongoing pregnancies rates were 18% and 14%, and live birth rates were 14% and 12%.
ConclusionsIntrauterine injection of hCG via IUI catheter is not recommended in a clinical routine setting at this stage. Future efforts are warranted to further refine the applicability of this modality.
Keywords: Assisted reproductive technologies, Embryo transfer, Human chorionic gonadotropin, Intrauterine insemination catheter, Randomized clinical trial