Iraqi journal of biotechnology

Study the Association of IL-21 Gene Polymorphism and Some Immunological Markers with the Risk of Rheumatoid Arthritis Incidence in a Sample of Iraqi Patients

1Mais B. Mohammed , 1Jinan M. J. Al-Saffar , 2Mohammed H. Alosami — Wed, 09 Oct 2024 00:00:00 +0200

Rheumatoid arthritis (RA) is a prevalent autoimmune disorder marked by chronic inflammatory synovial joints. Throughout the last decades several autoantibody systems have been discovered that are associated with RA, anti-CCP test (which measures antibodies directed to cyclic citrullinated peptides), anti-carp test (for determination of the levels of anti-CarP antibodies in patients with RA), IL-21 and IL-22 levels. The aim of the study the association of IL-21 gene polymorphism (rs2221903) with the risk of RA. The studied group include 60 RA cases (19 males and 41 females), who diagnosed was depend on the Revised diagnostic criteria established by the American College of Rheumatology (ACR), 2010. Enzyme-linked immunosorbent assay (ELISA) has been utilized in order to estimate the IL-22, IL-21, Anti-CARP and ACPA levels in serum of studied group and result of investigations were compared with 30 healthy apparently control individuals. IL-21 gene polymorphism assessed utilizing polymerase chain reaction (PCR) and result were compared with 20 controls. The present results revealed that a higher positivity of patients sera for ACPA, Anti-CARP, IL-21 and IL-22 (308.11 ± 27.13, 3896.89 ± 343.90, 118.69 ± 13.09 and 148.70 ± 6.07) respectively in comparison with control groups (3.75 ± 0.44, 39.37 ± 5.59, 27.89 ± 3.77 and 55.86 ± 4.73) correspondingly with highly significant differences (P < 0.001), but no significant variation were recorded in the distribution of genotype, frequency of allele and frequency of haplotype for polymorphism of IL-21 gene between the RA and controls groups. It was concluded that found frequency of haplotype for polymorphism of IL-21 gene between the RA and controls groups.

Correlation of Bacteria Diversity and Drug Resistance with Colorectal Cancer Patients

1Ghofran K. Khalaf, 1Sameer A. Alash — Wed, 09 Oct 2024 00:00:00 +0200

Colorectal cancer (CRC) is the leading cause of illness and mortality worldwide. The aims of the study is investigation and detection of some bacterial interfering with CRC occurrence and progression . The study conducted between September 2022 till February 2023, a total of 50 specimens were collected from confirmed CRC patients. In addition , 50 stool specimens were collected from healty volunterrs as a control. All specimens were gathered from medical city hospital and gastro-intestinal hospital. Isolation and identification of bacteria in all collected specimens were done by using cultural and differential media ( Blood agar, macconkey agar, mannitol salt agar and Pfizer agar), as well the Vitek- 2 compect system. The bacterial species, in the specimens of control were( Escherichia coli 50 (86.20%), Klebsiella Pneumonia 3(5.17%), Salmonella typhi 2(3.44%), Staphylococcus aureus 1(1.72%), Proteus mirabilis 1(1.72%) and Pseudomonas aeruginosa 1(1.72%), while in the specimens of CRC and polyp were (Escherichia coli 30(38.69%), Streptococcus uberis 6(7.79%), Enterobacter cloacae 4(5.19%), Proteus mirabilis 11(14.28), Streptococcus constellatus pharyneis 1(1.29%), Micrococcus luteus 1(1.29%), Staphlococcus pseudintermedius 1(1.29%), Streptococcus thoraltensis 1(1.29%), Citrobacter freundii 1(1.29%), Streptoccus mutans 1(1.29%), Enterococcus faecium 5(6.49%), Enterococcus faecalis 4(5.19%), Granulicatella elegans 1(1.29%), Enterococcus gallinarum 2(2.59%), Serratia marcescens 1(1.29%), Streptococcus sangunis 1(1.29%), Staphylococcus lentus 1(1.29%), Comamons testosteroni 1(1.29%), Morganella morganii 1(1.29%), Pseudomonas aeruginosa 1(1.29%), Klebsiella pneumonia 2(2.59%). The bacteria which has been shown to be associated and more abundance in the specimense of CRC tissues are Escherichia.coli 30(38.96%), Streptococcus uberis 6(7.79%), Enterobacter cloacae 4(5.19%), Enterococcus faecium 5(6.49%), Enterococcus faecalis 4(5.19%). The relative abundance of (Escherichia coli, Streptococcus uberis, Enterobacter cloacae, Enterococcus faecium, Enterococcus faecalis) a mong CRC specimens. The biofilm formation capability for the tested isolates revealed that (Escherichia coli, Streptococcus uberis and Enterococcus faecalis) were moderate intensity production, while (Enterobacter cloacae and Enterococcus faeium) were weak intensity. The antibiotics susceptibility test (AST) showed that Streptococcus uberis. It was concluded as an Extensive drug resistance (XDR), while (Escherichia coli, Enterococcus faecalis, Enterobacter cloacae, Enterococcus faeium) were considered as Multi Drug resistance (MDR).

Antibacterial Effect of Manganese Nanoparticles Loaded on Prodigiosin against Pathogenic Pseudomonas aeruginosa

1Biotechnology Department, 1Noor J. Menshid, 1Reem W. Younis — Wed, 09 Oct 2024 00:00:00 +0200

Pseudomonas aeruginosa are opportunistic pathogens that can cause infections in the lungs, skin, and eyes of people with cystic fibrosis (CF), HIV/AIDS, and burns and abrasions. In this study, the biosynthesis method of manganese nanoparticles (MnO NPs) was performed using pigment of Serratia marcescens, called Prodigiosin, which is utilized as a stabilizing and reducing agent. The amid of study was to investigate the antimicrobial activity of manganese nanoparticles on clinical isolate of P. aeruginosa. The results, one hundred and eighty samples were collected from burn and wound infections of different patients with different ages and sexes, from Kadhimiya Hospital, Karkh General Hospital and Yarmouk Hospital during the period from September 2022 to December 2022, whereas all these samples were subjected to different examinations in order to isolate P. aeruginosa. The influences of varied concentrations (25, 50, 100 and 200 µg/ml) of MnO NPs on bacteria P. aeruginosa were demonstrated. The antibacterial action was showed to be immediately reliant on the concentration of MnO NPs. The maximum inhibition zones around P. aeruginosa isolate were 28 mm at concentration 200 µg/ml of MnO NPs, while the minimum regions of inhibition were set at 25 µg/ml MnO NPs concentrations, were 10 mm. In conclusion, manganese nanoparticles that loaded on prodigiosin showed effective antibacterial activity against P. aeruginosa.

Variants in the Antioxidant Gene (catalase) and its Correlation with Idiopathic Male Infertility in Iraqi Population

Mayasah M. Khalid1 , Mohammed l. Nader1 , Shatha S. AL-Marayaty2 — Wed, 09 Oct 2024 00:00:00 +0200

Infertility is a multifactorial and polygenic disease. A vast majority of infertility is still unexplained despite modern diagnostic techniques. Catalase (CAT), the key enzyme for high H2O2 elimination, constitutes, along with superoxide dismutase (SOD) and gluthatione peroxidase (GPX), the main enzymatic antioxidant system. Catalase enzyme plays an important role in seminal antioxidant defense and is present in seminal plasma at high concentrations. Low concentrations of catalase enzyme in seminal plasma have been related with male infertility. The aim of the study was to investigate the status of the antioxidant enzyme; catalase in clinically diagnosed infertile males and find the potential association of CAT gene variant in the promoter region (rs1001179 [−262C/T] and rs7943316[ −21 A/T] and other CAT gene variant CAT rs1049982 [T/C], and CAT rs12270780 [G/A] and their association with the gene expression of the CAT gene. The study consisted of 100 clinically diagnosed infertile males (50 oligozoospermia and 50 asthenozoospermia) and 50 fertile volunteers. Polymerase chain reaction followed by sequencing were performed for genotyping of catalase variants. mRNA level of Catalase was analyzed by using semi quantitative technique. There was no significant difference in allele frequencies and genotype distribution for all SNPs. It was concluded that gene expression shows little increase in the level of CAT mRNA of fertile samples when compared to infertile samples.

Synergistic Effect of Staphyloxanthin and some Antibiotics on Clinical Isolates of Klebsiella pneumoniae

1Noor AL-huda A. Raja , 1Nagham Sh. Alattar — Wed, 09 Oct 2024 00:00:00 +0200

Klebsiella pneumoniae is one of the main causes of UTIs around the world. The aim of the study to examine the impact of staphyloxanthin and combination with some antibiotics on K. pneumoniae.150 urine samples obtained from Baghdad hospitals. The collected samples were streaked onto macConkey agar, followed by biochemical tests; then, using VITEK-2-compact system; thirty isolates were confirmed as K. pneumoniae. In addition, 10 pre-isolated and identified Staphylococcus aureus (producers of staphyloxanthin) were taken. The staphyloxanthin pigment extracted, purified. Antibiotic susceptibility testing of (30) K. pneumoniae isolate was assessed using KB testing. MIC of antibiotics (meropenem and ciprofloxacin) and staphyloxanthin were assessed for isolates using broth microdilution method, the effect of combination of staphyloxanthin and antibiotics was also evaluated for the same tested isolates. The result exposed that Meropenem MIC was 2 µg/ml, while MIC for staphyloxanthin pigment was 250 mg/ml. Ciprofloxacin MIC was 1000 µg/ml regarding the synergistic effect of the (ciprofloxacin and meropenem) combined with the Staphyloxanthin their MIC decreased for all, the results showed. It was concluded staphyloxanthin and antibiotics (ciprofloxacin and meropenem) have impact on K. pneumoniae.

Detection of Hematological and Biochemical Manifestations Associated with Women infected by Systemic Lupus Erythematosus

1Ghufran S. Jaber, 1Bushra J. Mohammed — Wed, 09 Oct 2024 00:00:00 +0200

Systemic lupus erythematosus (SLE) is an autoimmune disease in which the immune system attacks its own tissues. This study was aimed to investigate the hematological and chemical manifestations of SLE Iraqi women sample by focused on tests which related with SLE disease, as hematological tests that included: complete blood count (CBC) and erythrocyte sedimentation rate (ESR), also, chemical tests to evaluate renal function (blood urea and serum creatinine), and the ratio of protein calculated by urine strip only. The results (mean ± SE) of patients vs. control showed that ESR level was 40.08 vs. 10.96 mm/hour, Blood urea was 33.41 ±1.40 vs. 27.11 ±0.82 mg/dl with highly significant difference at P≤0.01, Creatinine was 0.758 ±0.04 vs. 0.651 ±0.02 mg/dl, with significant difference at P≤0.05 and protein urea percentages was 50 (83.33%) vs. 3(5.00%) with highly significant difference at P≤0.01. while hemoglobin was 11.37 ±0.20 vs. 12.90 ±0.08 g/dl, and platelets was 230.75 ±13.86 vs. 285.83 ±10.06 *10^3 / µl with highly significant difference at P≤0.01, white blood cells were 7.28 ±0.46 vs. 6.29 ±0.13cell X 10^3 / µl, with significant difference at P≤0.05. It concluded that it is evident that hematological testing and SLE disease are known to be related. And may interact with a variety of biomarker results and be used to forecast disease severity, which could be beneficial for SLE patients by expanding their options for diagnosis and treatment.

Using of mecA Gene as Genetic Marker for Staphylococcus aureus Detection Isolated from Atopic Dermatitis Patients

1Nardeen A. Qalawlus, 1Zainab H. Abood — Wed, 09 Oct 2024 00:00:00 +0200

Atopic dermatitis (AD), also identified as atopic eczema, is a long-term type of inflammation of the skin (dermatitis). The symptoms is red, swollen, itchy, and cracked skin, environmental factors such as emotions, sweating and exercise in addition to microbiological factors such as Staphylococcus aureus skin colonization which usually wide spread in these patients play an important role in the disease. The aim of the study: The present study was aimed to find the probable microorganism reasons that cause Atopic dermatitis (AD) in Iraqi individuals. During the period of study from beginning December 2022 to March 2023, A total of eighty swab specimens were collected from atopic dermatitis patients attending Medicine City/Baghdad Teaching Hospital and private laboratory. The culture results revealed 20 isolates to the S. aureus bacteria depend on culture characteristic, While the conformation by Vitek2 system showed that 20 isolates belonged to the genus S. aureus. It was concluded Staphylococcus aureus is frequently detected in patients with AD, and mecA gene appeared to be useful genetic marker for determination of S. aureus and PCR using species-specific primers could be represented rapid, sensitive and specific molecular method for detection of this bacteria in different AD patient.

Assessment the of Interleukin-6 Level and Gene Polymorphism (rs1800795) at the Risk for Colorectal Cancer Patients

1Miriam J. Shehab, 1Dhuha S. Namaa, 1Mohammed M. AL-Zubaidi — Wed, 09 Oct 2024 00:00:00 +0200

During tumor development, proinflammatory cytokines have critical role. Interleukin -6 is pleotropic cytokine that play crucial role in inflammation, immune regulation and tumorigenesis by activating multiple carcinogenic pathways. This study aimed to demonstrate an associated between IL-6 polymorphism (rs1800795) and circulating level of IL-6 with colorectal cancer in Iraq patients. Fifty blood samples were collected from both colorectal cancer patients and healthy individuals. The level of IL-6 in the serum of the study groups was determined using a specific sandwich enzyme-linked immunosorbent test (ELISA). While, the (rs1800795) polymorphisms was screened by using DNA- Sequencing technology. In the present study results revealed that the body mass index and IL-6 were significant higher in patients group compared with control group, (P-value = 0.0371 and 0.0006, respectively). Analysis of the IL-6 -174G/C polymorphism revealed that both in the patient and control groups, persons with the G allele had considerably higher serum levels of IL-6 than those with the C allele. (25.13 ± 1.35 versus 19. 81 ± 0.98, P- value= 0.0093) and (16.42 ± 0.3 versus 11.46 ± 0.89, P- value= 0.0016), respectively. It was concluded IL-6 - 174G allele could be used as prognostic biomarker for colorectal cancer in Iraqi patients. Thereby, several therapeutics targeting IL-6 may be used as promising opportunity in the treatment of colorectal cancer.

Study Neurotoxicity Effect of Acrylamide and Amelio-rating Effect of Curcumin on Adult Male Rats

1Estabraq H. Khyoon, 1Ammar A. Abdulwahid — Wed, 09 Oct 2024 00:00:00 +0200

Acrylamide (ACR) is also present in tobacco smoke and foods heavy in carbohydrates hat have cooked at high temperatures. It was thought that ACR exposure exclusively occurred at work and was partially brought on by cigarette smoking, water, and cosmetics. The study was aimed to study the harmful and neurotoxic effect of the acrylamide exposure and the ameliorating effects of curcumin in neurotoxicity that induced by acrylamide. Thirty adult male rats were subjected in this study, animal were divided into three groups, ten rats per group, group control were intubated distilled water for 40 days, group ACR (Acrylamide), rat in this group were administered acrylamide 5mg/kg B.W. for 40 days and group ACR+CUR (Acrylamide+ curcumin), animals had acrylamide 5mg/kg B.W. and curcumin 100mg/kg B.W. for 40 days. The results showed a significant decrease in the dopamine in the brain and increase AchE in serum, moreover, acrylamide affected spatial memory and locomotor activity behavioral testing, however curcumin restored these parameters close to control values. Histopathological examination of brain sections of ACR rats revealed proliferation of microglial cells with Alzheimer type II astrocyte, whereas, section from ACR+CUR, animals showed mild congested blood vessels in the pia mater. In conclusion Acrylamide induced alteration in neurotransmitters, neurobehaviors and structure of brain and this can be protected by curcumin.

ThiopurineMethyl Transferase (TPMT) Gene Expres-sion in Imuran and Biological Response in Ulcerative Colitis in Iraqi Patients

1Noor S. Ali, 1RafidA. Abdulkareem — Wed, 09 Oct 2024 00:00:00 +0200

Ulcerative colitis (UC) is a chronic IBD that involves the rectum and colonic mucosa. The study aim to expression in biological therapy more than its expression in Imuran treatment.This study included 50 patients with UC and 22 healthy. The RNA was extracted by the triazole method and the ages ranged from 15-50 years. The response toImuran treatment was weak, so they were referred to biological therapy. The gene expression of 22 patients was lower than the control ratio, meaning low gene expression, and the folding percentage was over gene expression.Venous blood samples were collected and preserved in TRIzol tubes, RNA extraction was performed, cDNA was synthesized, PCR was performed, expression levels were calculated and T-tests used to measure gene expression.Thiopurines are immunomodulators used for UC, and TPMT gene expression see significant value in Iraqi patients with high expression in biological therapy more than its expression in Imuran treatment groups.It was concluded gene expression of TPMT have a.significanteffect in Iraqi patients with UC.

Evaluation of miRNA 34a-3p Genes Expression Level Associated with Breast Cancer in Females

1Huda M. Taher, 2Ismail H. Aziz — Wed, 09 Oct 2024 00:00:00 +0200

MicroRNAs (miRNAs) are short non-coding RNA (approximately 22 nucleotides) that plays an important role in gene regulation. The study aims to evaluate the expression of serum (miR-34a-3p) in the Iraqi population as more efficient biomarkers. Circulating serum miRNA-34a-3p expression was measured using RT-qPCR in 50 patients of breast cancer compared to 50 healthy controls. The results showed the expression of miRNA-34a-3p non-significant (p=0.38) in patients as compared to control. According to histopathological type 96 % were Invasive ductal carcinoma (IDC), 4 % Invasive Lobular Carcinoma (ILC) of all breast cancer patients. Estrogen positive receptors (ER) were in (74%) (37/50) of the cases and progesterone positive receptors (PR) in (72%) (36/50) of the cases. Also out 20 of 50 cases (20%) positive and (60%) (30/50) for her-2/neu expression. It was concluded that Luminal A molecular subtype is the most common type in Iraqi female patients with breast cancer.

Effect of Histatin 5 Peptide on ERG11 and CDR1 Gene Expression in Fluconazole-Resistant Candida albicans Isolated from Vaginitis

1Hanna B. Kareem, 2Kais K. Ghaima — Wed, 09 Oct 2024 00:00:00 +0200

Candida albicans with antifungals resistance and biofilm formation may cause complicated infections among women with Vaginitis. Therefore, the search for new drugs capable of overcoming antifungal resistance is essential. One of the alternative therapeutic agents is Histatin 5 which is an antimicrobial peptide that can be found naturally in human saliva. This study was aimed to evaluate the role of Histatin 5 as antimicrobial and antibiofilm agent against fluconazole-resistant C. albicans isolates. Also, study the effect of this peptide on the gene expression of the fluconazole-resistant genes (CDR1 and ERG11). The results of minimum inhibitory concentrations (MICs) using microtiter plate method revealed that the MICs of seven fluconazole-resistant C. albicans isolates were (62.5-250 μg/ml). The antibiofilm activity of Histatin 5 demonstrated that this peptide was able to reduce the biofilm formation to 23% at the concentration 62.5 μg/ml, and to 41% at the concentration 31.25, in comparison with the control (100%). Relative quantification (RQ) was used to calculate the fold change in gene expression using the delta delta Ct value and the gene expression of C. albicans isolates was calculated before and after treatment with the subinhibitory concentration of each isolate. Before to Histatin 5 treatment, the fold of gene expression in the CDR1 gene was slightly higher (1), and the fold of the resistant isolates after Histatin 5 treatment was low (0.07-0.38). Also for ERG11 gene, Before Histatin 5 treatment, the fold of gene expression was slightly higher (1). After Histatin 5 treatment, the fold of gene expression was low (0.11-0.31). In conclusion, the obvious activity of Histatin 5 peptide as antimicrobial and antibiofilm agent against fluconazole-resistant C. albicans isolates may be contributed in the management and control the vaginitis infections among Iraqi women.

Correlation Between FAS Gene Polymorphism (rs:1800682) and their Serum Level with Psoriasis Patients

1Zena M. Joodi, 1Basima Q. Hasan AL-Saadi — Wed, 09 Oct 2024 00:00:00 +0200

Psoriasis is a chronic, multifactorial inflammatory skin condition can be defined by keratinocyte hyper proliferation that is mediated by T cells. It is immune-mediated inflammatory skin disease with multiple phenotypically distinct subtypes it a major genetic component, with heritability estimated to be (60-90%) The goal of the current study is genotyping of the FAS (Fas cell surface receptor gene ( -671 A>G (rs:1800682), and evaluate its serum level in Iraqi patients with psoriatic.100 blood samples collected from December 2022 to February 2023. The blood samples split into two groups first 50 psorasis patients (30males and20 females) with age range (17-66years) second group 50 healthy individual (24 males 26 females ) with age range (20-60 years) visit the Dermatology Department at Al-Yarmouk Teaching Hospital in Baghdad, Iraq. Two techniques were used in the research, the first is (PCR-HRM) to investigation gene polymorphism and the second technique is EIISA to detect FAS serum level. The aimed to investigate whether FAS (Fas cell surface receptor gene ( -671 A>G (rs:1800682) with FAS serum level. the result suggest that there are three genotype (AA),(AG),(GG) in current study shows statistically significance in the (AA) genotype frequency in the FAS gene with a percentage in psoriasis patients (6%versus 24% in healthy individuals) respectively. The (AG) genotype shows non-significant differences between psoriasis patients and control the( P-value =0.612) respectively While the GG genotype shows non-significant frequency in psoriasis patients compared control group with (p-value =0.297). and findings that the FAS serum in patient high significantly different between psoriasis patients when compared to the healthy controls with (P 0.01). It was concluded that found correlation between genotyping and serum level for FAS (rs 1800682)gene (AA) genotype serum level that significant different from (GG) genotype serum level with p-value(P≤0.05).

The association of TNF-ɑ Gene Polymorphims with the Incidence of Diabetes Mellitus in Adult Patients

1Fatimah k. kadheem, 1Ismail A. Abdul-Hassan — Wed, 09 Oct 2024 00:00:00 +0200

Diabetes mellitus (DM) are defined as the most chronic metabolic disorder resulted from a complex interaction between environmental and heredity factors, Diabetes mellitus type 2 is Previously known as non-insulin dependent DM, the most common kind of diabetes is characterized by hyperglycemia, insulin resistance, and a relative lack of insulin. The TNF- ɑ gene variant ( rs 1800629) has been associated with an increased risk of developing diabetes. The aim of study is a relation between the polymorphisms in the TNF gene and the prevalence of type 2 diabetic mellitus (T2DM) in the Iraqis as a whole. Having 50 patients with diabetes and 50 subjects who appeared to be in good health as controls, The DNA was extracted and then quantification using Nanodrop,the integrity was exam through agarose gel electrophoresis were enrolled genotyping of TNF- ɑ gene SNPs (rs1799724 and rs1800629) that determined by using High Resolution Melting (HRM) ,The results showed there were an increase in serum levels of Triglyceride and VLDL in DMT2 patients compared with apparently healthy subject .the result is found that no relation was found between rs1799724 SNP of TNF-ɑ gene with the incidence of DMT2 in Iraqi patients .No effected of rs1799724 SNP of TNF-ɑ gene on Lipid profile parameters . It was concluded that related with rs1800629 SNP of TNF-ɑ ,GG genotype represent as a protective factor against DMT2 in incidence whereas ,GA genotype represent as a risk factor for DMT2 incidence .serum triglyceride and serum VLDL levels were affected by the polymorphism rs1800629 of TNF-ɑ.

Circulating Interleukin 37 and the Chemokine CXCL9 Studies on Gastroduodenal Disorders with H. pylori Infection

Wed, 09 Oct 2024 00:00:00 +0200

The ability of the bacterium Helicobacter pylori to survive in a gastric environment causes gastroduodenal disorders (GD) due to its virulence factors. The aim of the study host cell recognizes bacterial shape and pathogen-associated molecular patterns (PAMPs) by toll-like receptors (TLRs), which trigger the immune system. The study involved 165 patients diagnosed with gastroduodenal disorders and 32 healthy individuals, the patients attended the Gastroenterology Teaching Hospital and a private clinic during the period from April 2021 to January 2023. The physicians diagnosed them with gastritis, peptic ulcers, and gastric cancer by OGD. The IL37 and CXCL9 level in sera were measured by ELISA. This study aimed to find the effect of age, gender and blood group on the susceptibility of GD related to H. pylori and its effect on interleukin 37 and chemokine CXCL9. The current findings confirm that GD affects the elderly and males more than young people and females. The susceptibility increases with the ABO phenotype and the O+ blood group exhibited the highest incidence of GD. A significant increase in IL37 and CXCL9 levels in patients’ serum compared with the healthy group was reported. The IL37 concentration in gastritis, peptic ulcer, and gastric cancer, and the healthy group was as follows: 637.3 ± 125.3, 598.7 ± 158.2, 177.06 ± 67.49, and 127.08 ± 13.41, respectively. While the concentration of CXCL9 level in serum in patients suffering from gastritis, peptic ulcer, gastric cancer, and healthy individuals was as follows: 586.06 ± 701.47, 589.94 ± 778.86, 364.16 ± 215.08, and 111.85 ± 54.28, respectively. The peptic ulcer group was the highest among the others, followed by the gastritis group and gastric cancer. It was concluded that significant relationship between the biomarkers (IL-37 and CXCL9) and gastroduodenal disorder patients in patients with H. pylori-infected especially gastritis and peptic ulcers.

Antibacterial Activity of Chitosan Nanoparticles With Ocimum basilicum oil Extract on Pseudomonas aeruginosa

1Ahmed A. Obeid, 1Emad H. Jassim — Wed, 09 Oct 2024 00:00:00 +0200

The present study aims to describe the antibacterial activity of chitosan nanoparticles (CSNPs‎‏( ‏ loaded ‎with ‎Essential Oil of Ocimum basilicum (BEO) extract ‎against Multiple drug resistance MDR ‎Pseudomonas ‎aeruginosa. The aim of the study effect antibacterial activity of chitosan nanoparticles (CSNPs‎‏( ‏ loaded ‎with ‎Essential Oil of Ocimum basilicum (BEO) extract ‎against Multiple drug resistance MDR ‎Pseudomonas ‎aeruginosa. The 40 isolates ‎of Pseudomonas ‎aeruginosa were ‎collected from ‎patients suffering ‎from ‎‎burn ‎infection‎.The isolates ‎were ‎diagnosed as ‎P. aeruginosa by classical biochemical tests ‎and PCR technique using 16S rRNA gene as a ‎diagnostic gene ‎and gel ‎electrophoresis. The sensitivity ‎of ‎bacteria to antibiotics was tested using ‎the ‎Kerby-Bauer method. The antibacterial activity ‎for This was ‎determined by using the Agar well Diffusion ‎Method (ADM) ‎for‎ Essential Oil of Ocimum basilicum‎ and ‎the Disc Diffusion Method for chitosan ‎nanoparticles ‎ loaded ‎Essential Oil of Ocimum basilicum.The results showed that both ‎the ‎test ‎substances have ‎antibacterial activity against Pseudomonas aeruginosa isolates with ‎superior ‎inhibition of ‎nanoparticles at ‎low concentrations. The current ‎study ‎concluded ‎the ‎possibility of ‎using chitosan ‎nanoparticles loaded ‎with Essential Oil of ‎Ocimum ‎basilicum extract as ‎a natural ‎alternative ‎against ‎MDR Pseudomonas ‎aeruginosa due to ‎the nanoparticles' ‎unique ‎properties that ‎increase ‎drug ‎effectiveness and ‎efficiency in ‎drug delivery.

Impact of Alcoholic Extract of Brassica olecera on Endothelial Dysfunction of Lead Acetate Exposed Rabbits

1Dina S. Dheyab, 2Majida A. J. Al-Qayim — Wed, 09 Oct 2024 00:00:00 +0200

The present study aimed to investigate the modulatory effect of alcoholic extract of broccoli on lead acetate-induced endothelial dysfunction. The aim of the study alcoholic extract of Brassica olecera on endothelial dysfunction of lead acetate exposed rabbits. Three groups of female local rabbits (6 each group) aged 5 months with a verge weight 1500 Kg were handled as following:1st as a control group (C) and 2nd lead acetate (Pb-AC) ,and 3rd lead and broccoli (Pb-Ac-B) groups were orally gavage with 10 mg/kg/Bw of lead acetate for 4 weeks. Six adult rabbits of 3rd group were treated with 300 mg/kg alcoholic extract of broccoli orally after exposures to lead acetate for another 4weeks. Blood samples collected before and after treatment. Aorta was harvested after euthanasia, for endothelial nitric oxide synthase gene expression. Vascular endothelial growth factor and antioxidant markers assayed using Eliza. The results showed the ameliorative effects of alcoholic extract in reducing the significant elevation in e NOS expression (p≤0.05) and serum VEGF and MDA caused by lead acetate, accompanied with significant modulation of TAC. It was concluded the results confirmed the superiority of the Brassica olecera in treating the pb-acetate adverse effects on the endothelial, which contributed to its phytochemical and nutrient value.

Impact of Gene Polymorphisms (Rs5046) and Serum Levels of AGT Gene Attribution in Prevalence of Hypertension Patients

1Marwah Y. Hameed, 1Rafid A. Abdulkareem — Wed, 09 Oct 2024 00:00:00 +0200

Some genes might probably contribute to the regulation of blood pressure due to their previously established effect on cardiovascular or renal function, in addition to physical and environmental factors. The aim of the study detects the relationship between the genetic polymorphism of AGT and the level of AGT in serum and its association with high blood pressure patients. The AGT levels in serum have been measured by ELISA kit. In this study, Genomic DNA was extracted by DNA extraction kit from both patients and control group and the measurement of concentration and purity was done by the use of nanodrop. (T-ARMS-PCR) was used to determine the SNPs rs5046 (G/A) in the 5′ near region of AGT gene in hypertension patients and healthy controls, by using specific Primers, The Results detected that genotype CC (34 (68.0) % vs. 50 (100.0) %) was decreased frequency in patients than the control group. In contrast, the CT genotype showed a significant increase (16 (32.0) vs. 0 (0.0) %; OR = 48.30; p<0.001) in the patient group compared with the control group. While the TT genotype does not appear in any groups of patients and controls. The results of the assessment of AGT plasma levels in AGT rs5046 SNP genotypes showed no significant differences according to the rs5046 genotypes in the level of AGT. For CC genotypes, AGT levels were higher in the hypertension patient group than in the control group and significantly (p<0.001). It was concluded investigation indicate that CT was a risk factor for hypertension (EF =31.7).

The Role of Toll-Like Receptor 4 Gene Polymorphism TLR4 (Thr399Ile) and Expression in Adult Urinary Tract Infections Pathogenesis

1Esrea M. Hekmat, 2Mohammed I. Nader — Wed, 09 Oct 2024 00:00:00 +0200

The first line of protection against microbial illnesses is the immune response. Toll-like receptors (TLRs) are essential for innate immunity, and toll-like receptor 4 (TLR4) is recognized as a key pattern-recognition receptor (PRR) for identifying gram-negative bacteria's lipopolysaccharide (LPS). Numerous TLR genes have been shown to have single nucleotide polymorphisms (SNPs), which may be connected to the susceptibility or resistance to specific infections and other inflammatory diseases. The aim of this study was to determine whether there is a connection between chronic UTI and the TLR4 (Thr399Ile) gene polymorphism and TLR4 expression level. A case-control study was conducted on 52 adults This study uses HRM analysis to look for TLR4 (Thr399Ile) polymorphisms in UTI patients. TLR4 expression in the monocytes of UTI patients and healthy controls was found using real-time PCR. TLR4 (Thr399Ile) genotype and TLR4 (1196) T allele prevalence were higher in UTI patients than in controls, particularly in cases of acute cystitis and urethritis. Compared to healthy controls, chronic UTI patients had considerably greater levels of TLR4 expression. The results of the current study, show the presence of a correlation between the genotypes of the TLR4 gene and the incidence of development of Urinary Tract Infection, as the results showed the significant difference between patients and healthy controls when the genotype Heterozygous (Thr / Ile) with (OR= 0.638), while the genotype Homozygous (Ile / Ile) showed no significant difference between the patients and control group with (OR= 1: Reference). TLR4 C/T (Toll-like receptor-4 gene polymorphism). The results of gene expression of TLR4 revealed that this receptor was increased among UTI (3.50 ±0.41 fold) infected Patient. (1.20 ±0.07 fold) in comparison with the healthy control. The findings showed a connection between adult UTI, particularly severe cystitis and urethritis, and the TLR4 (Thr399Ile) allele carrier status. Chronic UTI and TLR4 expression levels are linked.

Detection of ure, emerD and znuA Virulence Genes in Brucella melitensis Isolated from Aborted Fetuses of Sheep in Thi-Qar Governorate

1Mukhtar H. Ramadan, 1Wassan A. Gharbi — Wed, 09 Oct 2024 00:00:00 +0200

Brucellosis is a major and widespread bacterial zoonosis. it is common throughout the world This has serious economic and public health consequences. Sheep, goats, and cows are examples of domestic animals.; suffer significant losses due to Brucellosis. The aim of study was to detect the ure, emerD and znuA virulence genes in Br. melitensis, samples were collected from eight fields of sheep in Thi-Qar Governorate south of Iraq during the period November 2022 - to June 2023, from animals that had abortions or certain disease symptoms, , 32 out of 67 mother sheep prove positive results for indirect Elisa ,32samples of fetal fluids fetus of sheep in Thi-Qar Governorate appeared 32 a positive for the Conventional PCR technique so 32 isolates of Brucella melitensis were determined, Generally. the result of fetal fluids was found in most prevalent gene was ure, which was found twenty isolates. (68. 75%): while the znuA gene had the lowest proportion of all analyzed genes due to its presence five isolates (15.62%). It was concluded gene were found in certain isolates, such as the emrD gene which found in 14 isolates. (43.01%).

Detection of Pyocin Multi-Drug Resistance Pseudomonas aeruginosa in Clinical Samples Collected from Patients and Study the Effects of CFSs against the Bacterium

1Aliaa G. Husain Bassi , 1Bahaa A. Laftaah Al-Rubaii — Wed, 09 Oct 2024 00:00:00 +0200

Pseudomonas aeruginosa have emerged that are resistant to all widely prescribed antimicrobial medicines, treating Pseudomonas infections has grown more challenging. The aim of this study is the detection of some resistance genes in pyocin MDR-P. aeruginosa and examine the effect of specific concentrations of Lactobacillus Acidophilus Cell-Free Supernatants (CFSs) on P. aeruginosa. The 60 P. aeruginosa isolates were gathered from clinical sources from Baghdad hospitals. the disk diffusion method was used to examine the antibiotics' sensitivity and lowest inhibitory concentration (MIC). The resistance genes were discovered by PCR, and the effects of CFSs on the growth of P. aeruginosa were studied by using a specified concentration. Obtaining Pyocin Multi-Drug Resistance isolates which carry some resistance genes, and determines the CFSs concentration effects on P. aeruginosa growth. It was concluded that use of (CSFs) as an alternative therapy for infections by Multi-Drug Resistance P. aeruginosa is thus conceivable in the future.

Molecular detection of arr2 Gene of Rifampcin Resistant Acinetobacter baumannii Isolated from Patients Referring Baghdad Hospitals

1Safaa H. Hussein, 1Mayaada S. Mahdi — Wed, 09 Oct 2024 00:00:00 +0200

The Arr enzymes that catalyze ADP-ribosylation of rifamycins to render it inactive Which contributes to increasing bacterial resistance to this antibiotics , that genes encoding predicted arr enzymes are widely distributed in the genomes of pathogenic and nonpathogenic bacteria, This study aimed to isolate A. baumannii from various clinical specimens and detection of arr2 gene in rifampcin resistant A.baumannii. A total of (300) samples were collected from patients suffering from different infections in Medical City Hospital during the period from (1/7/2022) to (30/1/2023). The Vitek 2 system was used to identify bacterial isolates, and the Kirby-Bauer method was used to test antibiotic susceptibility. Among the collected clinical samples 88 (29.3%) were identified as A. baumannii. The majority of A. baumannii isolates were found in wounds (45.4%), followed by burns (30.6%), urine (14.7%), and sputum (9.0%). Antibiotic susceptibility showed that A. baumannii isolates had varying levels of resistance to the antibiotics used. It was high resistant to cefotaxime, Ceftriaxone, Rifampin, cefepime, Amikacin, Ampicillin-sulbactam and Trimethoprim- sulfamethoxazole at a percentage of (96.5, 86.3, 85.2, 84.0, 80.6, 78.4 and 70.4 %) respectively. While exhibit a moderate level of resistance to doxycycline, meropenem and Levofloxacin at a percentage (52.2, 53.4 and 67.0 %) respectively, thirty multidrug-resistant isolates including rifampicin, were subjected to molecular study by detecting the arr2 gene, it was concluded that 9 isolates out of 30 isolates (30%) have arr2 gene

Role of DNA Isolated from Gut Microbiota Escherichia coli in Mice Joints Inflammation

1Ibtihal Fadhel Yahea , 1 Hind Hussein Obaid — Wed, 09 Oct 2024 00:00:00 +0200

The gastrointestinal microbiome is the largest and most diverse reservoir of all the human body niches. The aim of the study relationship between gut microbiota Escherichia coli in mice joints inflammation. In this study, 100 samples of stool were gathered from healthy individual and 20 urine samples from people with recurrent Urinary Tract Infection (UTI) and both types of samples were proceeded accordingly to isolate E. coli strains where 92 strains were isolated from stool samples and 10 from urine samples. The isolated strains were subjected to antibiotic sensitivity test and the results indicated that 39.2% out of the total number of the isolates were multidrug resistance while all the pathogenic strains were multidrug resistant. According to the sensitivity results three isolates were chosen to DNA isolation, two of them isolated from stool samples (Sensitive and Resistant) and one from urine samples. The extracted DNA was divided to two parts one of them was subjected to cleavage by EcoR1 restriction enzyme and the other remains without treatment as a whole DNA, both were injected directly to mice knee joints to study the histopathological effects of bacterial cell free DNA on knee joints. I was conclude that indicated no effect where all the tested tissues were similar to those of group control.

Down regulation of IFIH1 Gene Expression in Early COVID-19 Infection

1Rand J. Sattar, 2 Hula Y. Fadhil — Wed, 09 Oct 2024 00:00:00 +0200

COVID-19 have not been completely elucidated, non-adaptive inflammatory responses appear to play a substantial part in the disease's progression. The aim of the study Down regulation of IFIH1 Gene Expression in Early COVID-19 Infection. One hundred coronaviruses infectious disease (COVID-19) patients and 58 healthy individuals were enrolled to assess the gene expression of Interferon Induced With Helicase C Domain 1 (IFIH1) by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), with some emphasis on clinical and biological markers of disease like chemokine (CCL-2), and tumor protein (P⁵³). The finding showed that CCl2 and P53 levels have a significant decrease in patients (185 ng/L and 24.5 ng/ml) from control (229 ng/L and 30 ng/ml) (p = 0.001). The ratio of relative IFIH1 gene expression in COVID-19 patients versus healthy control was 0.95. Although the 2^-∆∆Ct of IFIH1 gene expression showed no significant variation between the age group (≤45 and > 45 years): sex, and severity in patients and controls, also non-statistical increasing 2^-∆∆Ct means of IFIH1 with disease progression like critical (17.5) than mild-moderate (15.8) in patients. The findings showed the CCL2 and P53 levels were non-significantly correlated with the IFIH1 expression (p > 0.05): although there is a negative correlation (r_s= - 0.104) between CCL-2 and IFIH1 values making that prognostic indicator for reducing the development of infection. When IFIH1 expression increased, the P53 level non-significant increased (p = 0.099). It was concluded that IFIH1 expression are an impact factor correlated with P53 levels reducing the COVID-19 risk.

Phylogenetic Analysis of Pathogenic Candida Species Isolated from Iraqi Patients

Zahraa A. Al-Ameri1, Muna T. Al-Musawi1 — Wed, 09 Oct 2024 00:00:00 +0200

Pathogenic species of Candida have extensively contributed to the raising mortality, morbidity and globally increase medical prices of healthcare-related infectious diseases. The current study was focused on isolation and identification of Candida spp. from different clinical specimens and their molecular phylogenetic analyses. The study was included clinical specimens (100) from different parts of the patient’s body, such as (ear, mouth. vagina, skin, blood and urine) who attended to Gazi Al Hariri hospital, Baghdad teaching hospital in Medical City and Al-Yarmouk teaching hospital. The isolation of Candida spp. using selective media and vitek system. Then, a full inclusive tree, including the observed variant, was built by the neighbour-joining method and visualized as a circular cladogram using the iTOL suit. The findings of isolation and identification of Candida spp. in studied samples were recorded that the highest isolate was C. parapsilosis (27.7%) followed by C glabrata (22.2%), while the lowest isolates were C. tropicalis and C. krusei (5.50%) each one, Candida spp. also identified by ITS which results showed the presence of sizes ranging 550 - 625 bp depending on the difference in the lengths of the region (ITS 1 and ITS4) for yeast species, so they produce different sizes of DNA pieces. Sequencing reactions showed the accurate identity of the investigated samples and revealed that S1, S8, and S9 were homologous to C. parapsilosis, S2, S6, and S13 were similar to C. albicans, S3 was similar to C. xylopsoci while S4, S5, S7, S10, S11, and S12 were similar to C. glabrata (The presence of a total of 11 nucleic acid variants compared with the referring sequences of the Candida sequences was demonstrated. The identified variants were one deletion (g.492-494Adel) in S1, two substitutions (g.168A>G and g.233C>A) in S13, one substitution (g.83A>C) in S3, one substitution (g.116T>C) in S11, and four substitutions and one deletion (g.339C>A, g.353G>A, g.618G>A, g.356-359GGdel) in S4, S5, S7, S11, S12, one substitution in S7 (g.345C>G), one substitution (g.805T>C) in S4, S5, and S7. Meanwhile, the rest of the samples had shown a complete homology with the corresponding sequences and did not exhibit any detectable nucleic acid variations in comparison with the same reference sequences. This study suggests possible employment for these ribosomal amplicons to discriminate between the phylogenetic diversity among the other implemented tools.

Relationship of Androgen Receptor Gene Polymorphism (rs2261634) in Male Infertility and Its Correlation Effect on some Hormones Levels in Samples of Iraqi who Utilize Anabolic Steroids

1Ali S., 1Sanaa J. Kadhim — Wed, 09 Oct 2024 00:00:00 +0200

Polymorphisms that increase risk factor for male infertility. This study aimed to highlighted association between abuse anabolic steroids and male infertility by evaluate the genotyping and allele frequency of androgen receptor gene (AR gene) of (rs2261634), and found the relationship of this Single Nucleotide Polymorphism and some serum hormones levels. The samples for this study were obtained from the Kamal Al- Samara'ay IVF Hospital, Ministry of Health in Baghdad, Iraq. The study was conducted at the Institute of Genetic Engineering and Biotechnology, University of Baghdad. Genotyping of rs2261634 was done by High Resolution Melting technology. The Hormonal assay (Follicle-Stimulating-Hormone, Luteinizing Hormone, Testosterone, Estrogen and Prolactin) was performed by using cobas e 411 device. The results of genotypes and alleles frequencies at AR gene (rs2261634) SNP. in controls versus Iraqi Patients with abuse anabolic steroids showed that the percentage of wild-type TT genotype was significantly (p≤0.01) lower in Iraqi Patients with abuse anabolic steroids group than in controls group (16.6% versus 30%, respectively). The conclusion of this study found The TT genotype may represent a protective factor against the incidence of patients with abuse anabolic steroids in Iraqi patients. It was concluded that testosterone, prolactin, and estrogen levels statistically increased after using high dosages of anabolic steroids compared to control, but FSH and LH levels statistically decreased.

Role of Calcium Sensing Receptor Gene Polymorphism r1801725 in the Evaluation of kidney Disease

1Rana A. AL-Jumaili, 1Essam F. Al-Jumaili, 2Omar M. Ramadhan — Wed, 09 Oct 2024 00:00:00 +0200

"kidney disease" refers to a diverse range of conditions that affect the structure and operation of the kidneys. It is now understood that even minor deviations in kidney structure and function are linked to a higher risk of mortality as well as consequences in other organ systems. In the kidney it has an inhibitory effect on the reabsorption of calcium, potassium, sodium, and water depending on which segment of the tubule is being activated. In this study we aimed to investigate the genotypes prevalent for the (CASR) rs1801725 variant is responsible for a non-conservative amino-acid change (A986S) in the calcium-sensing receptor cytoplasmic tail. This study involved 50 blood samples were collected as patient with kidney disease and (50) apparently healthy controls. Molecular study was performed in order to investigate the single nucleotide polymorphisms (SNPs) for CASER gene in chromosome 3 (rs1801725) for studied groups which have determined using high resolution method (HRM) genotyping assay by real time polymerase chain reaction (RT-PCR) using the Qiagen rotor gene Q real-time PCR system. . Data obtained was subjected to statistical analysis using the SPSS 25 software. The rs1801725 G/T genotype was associated with increased risk for kidney disease in Iraqi patients with highly significant difference. The average age of a patient with kidney disease is between 21 and 69 years old The Genotypes and allele frequencies of (rs1801725, G>T, A986S) in patient with kidney disease groups 48% (n=24) wild (TT), and 14% (n=7) heterozygous (TG) and 38% (n=19) homozygous (GG). Allele frequencies for T and G were 55% and 45% respectively It was concluded that the variants of CASR SNP, namely, rs1801725 might be involved in susceptibility to kidney disease.

Creating a Model for Ancient DNA Isolation from Contaminated Archaeological Materials Using Sodium Hypochlorite Pretreatment and Enzymatic Predigestion.

1Baneen A. Abdullah, 1Salwa J. Al-Awadei, 2Dheaa Sh. Zageer — Wed, 09 Oct 2024 00:00:00 +0200

The retrieval of DNA from archaeological or forensic skeletal remains provides valuable data for analysis, but it faces specific challenges. The study aims to develop a standardized method for isolating ancient DNA, with applications in archaeology, genetic ages, disasters, and human remains. One crucial challenge is the presence of contemporary contamination on bone and teeth surfaces, which can lead to misleading outcomes and false positives if not addressed before DNA extraction. To mitigate this contamination, Ancient DNA (aDNA) researchers and forensic scientists have utilized various techniques, including using bleach (sodium hypochlorite - NaOCl) to "eliminate" the contaminating DNA. However, caution is advised, as this treatment has the potential to damage a significant portion of the sample's native molecules, as indicated by some earlier research.To further investigate this subject, the study employed samples of both human and animal remains and subjected them to sodium hypochlorite treatment to eliminate contamination. The findings align with prior research, showing a notable decrease in human contamination after applying the bleach pretreatment. However, it's essential to note that this reduction in contaminant DNA comes with a trade-off, resulting in a decrease in the complexity of endogenous DNA.

Determination of Viral load of Hepatitis C Virus and Evaluation of Some Liver Function Markers and Interlukin-27 in Hepatitis C Patients

1Ramy Z. Nasser, 1Saife D. Al-Ahmer — Wed, 09 Oct 2024 00:00:00 +0200

The Hepatitis C virus (HCV) is the main contributor to chronic liver diseases and affects over 71 million people worldwide that primarily leads to significant morbidity and mortality through its predisposition to liver fibrosis, cirrhosis, and liver cancer. The present study aim to early detection of HCV and determine the viral load that can be contributed in the viral hepatitis infection, as well as estimation of some liver function markers include TSB, ALT, AST and ALP, and evaluation of IL-27 role in immune response against the hepatitis C virus. The present study was include 50 patients infected with the HCV and 50 apparently healthy individuals. The RNA was extracted from serum samples of the HCV patients and the viral load of HCV were determined using one-step qRT-PCR. The biochemical tests include TSB, ALT, AST and ALP, were done for patients and control groups. In addition, the IL-27 levels were estimated using ELISA. The detection of HCV viral load showed that 31(62%) of HCV patients were negative, while 19(38%) out of 50(100%) of HCV patients were positive for the assay. Also, the results of TSB, ALT, AST and ALP tests of showed significant differences between the patients and control groups. In addition, the results of interlukine-27 revealed that there were significant differences between the patients and control groups. The HCV viral load in acute HCV males patients were range from (126-2310000 IU/ml) while the viral load in acute HCV females patients were range from (137-2180000 IU/ml). It was concluded that the results of liver function tests showed that there was a non-significant correlation between these results and viral hepatitis infections. In addition, the estimation of IL-27 levels showed there was significant correlation between these results and viral hepatitis infections.

miR-199a-5p Confers Tumor-Suppressive Role in MCF-7 Breast Cancer Cell Line

1Arwa M. Salih, 2Ismail H. Aziz, 3Forat Y. Mohsin — Wed, 09 Oct 2024 00:00:00 +0200

Breast cancer is a leading cause of cancer death among women worldwide. Abnormal expression of microRNAs has been observed in various types of cancers, including breast cancer. This study aimed was to evaluate and suggest the optimum quantities of lipoplex components to enhance the transfection efficiency of breast cancer cell line (MCF-7). Human breast cancer cell lines MCF-7 were grown through a transfection experiment. After RNA extraction and cDNA creation, real-time PCR was used to quantitatively measure miRNA expression levels. MCF-7 cells were transfected through a series of tests. The tests involved modifying the amount of Lipofectamine used. As a result, the optimal transgene expression time point and concentration were determined to be 1 μl (with a folding value of 1.82675). This was found to achieve the highest transfection efficiency while causing minimal cytotoxicity. In conclusion, the results show that transfected MCF-7 cells exhibit increasing transgenic expression over time. 72 hours after transfection is a suitable period for further study or therapeutic application, which is more than 24 hours.