Iraqi Journal of Biotechnology

This review provided a broad overview of the basic theory, methodology and applications of Genome-Wide Association Studies (GWAS) and Quantitative Trait Loci (QTL) mapping, two cornerstone technologies in plant genetics. This will be combined with an introduction of GWAS and QTL mapping, emphasizing the function of these methods to identify genetic variations underlying plant complex traits. Additionally, we explored the statistical models behind both methods, understanding the science behind regression models, including linear and logistic regression, but also addressing Type I and Type II errors and methods to minimize them, highlighting the keys such as multiple testing correction, replication and functional validation. the review also showed the practical applications of GWAS and QTL mapping in agriculture, crop improvement, livestock breeding and sustainable farming. Examples like flood resistant rice and drought tolerant maize showed the power of these technologies. Finally, the review discussed the challenges and future directions in the agriculture field including the integration of new technologies like CRISPR and high throughput phenotyping.

This review provided a broad overview of the basic theory, methodology and applications of Genome-Wide Association Studies (GWAS) and Quantitative Trait Loci (QTL) mapping, two cornerstone technologies inplant genetics. This will be combined with an introduction of GWASand QTL mapping, emphasizing the function of these methods to identify genetic variations underlying plant complex traits. Additionally, we explored the statistical models behind both methods, understanding the science behind regression models, including linear and logistic regression, but also addressing Type I andType II errors and methods to minimize them, highlighting the keys such as multiple testing correction, replication and functional validation.the review also showed the practical applications of GWAS and QTL mapping in agriculture, crop improvement, livestock breeding and sustainable farming. Examples like flood resistant rice and drought tolerant maize showed the power of these technologies. Finally, the review discussed the challenges and future directions in theagriculturefield including the integration of new technologies like CRISPR and high throughput phenotyping.

A persistent infection with the hepatitis B virus (HBV) can cause significant morbidity and mortality. Although antiviral therapy, monitoring, and liver cancer surveillance are not considered therapeutic, they can lower morbidity and mortality. The Polymorphic gene IL1B analyzed in a total of 45 individuals: 30 patients with chronic hepatitis B from different cities of Iraqi recruited the Hospital for diseases of the digestive system and liver in Baghdad between November 2022 to February 2023 and 15 controls. Genomic DNA was obtained from both patients and control and polymerase chain reaction (PCR) was perform with specific primer designed to target the promoter region of IL-1β gene, followed by sequencing. Serum concentrations of IL-1β were measured using Enzyme-Linked Immunosorbent Assay (ELISA). The results showed serum levels of IL-1B which significantly decreased (P≤0.01), from 1.10-+0.09 in hepatitis specimen in comparison with the level in healthy controls 7.58± 0.28. The results also revealed that the age group ˃ 30 years have a significant (P≤0.01) low level of IL-1B (0.83±0.06) in comparison with the age group ≤ 30 years (1.71±1.36). The molecular results suggested that IL-1B single nucleotide polymorphism (insertion/deletion) is probably associatedwith susceptibility to HBV chronic infection patients, the sequencing alignment data for 30 samples from patients with hepatitis B were compared to 15 control samples. The findings indicated frame shifts (deletion) in 124 and 125 positions in patients’ comparison with control groups. The results showed a cystosine base pair (C) in the patient group, while the control group noticed a deletion. In conclusion, the results suggest that IL-1B is probably associated with susceptibility to hepatitis B virus chronic infection

The coronavirus pandemic of 2020 was caused by the coronavirus disease 2019 (COVID-19), an infectious disease with severe acute respiratory syndrome that was first discovered in Wuhan, China. Since then, it has spread throughout the world.Co-infections between viruses and bacteria are among the deadliest medical conditions, with greater fatality rate.Research on bacterial superinfections in patients with coronavirus disease 2019 (COVID-19) is scarce. COVID-19 patients are at risk for colonization with K. pneumoniaand hospital-acquired infections. Therefore,Infections with K. pneumonia, particularly those caused by extremely virulent strains, may make COVID-19 more difficult to treat. Fifty sample of COVID-19 infection collected and detected positive by (RT)PCR with 50 apparentlycontrol healthy samplethan we cultured 50 sputum and nasal pharyngeal swabs for all COVID19 positive samples ,fifty sample of coinfection bacterial in the COVID19 patients identified by A-Vietk-2system,B-confirmed by molecular detection PCR,The fifty-isolate distributed as 27 (54 %) k. pneumonia, 6 (12 %) E. coli, P. aeruginosa 15 (30%),other bacteria 2(4%)the k. pneumoniais the largest number of coinfection.The results of the current study demonstrated a positive correlation between the presence of certain virulence genes and the ability of bacterial isolates to resist antibiotics. Certain virulence genes (rmpA, mrkA, and iuCc, ybts, magA,) have been found bymolecular means. our study was done by conventional PCR technique.wefound Klebsiella pneumoniaisolates which was found rmpA18(66%) ofthis isolates at bp 535, then 7(25%)) isolate were positive for mrkA gene at 115bp,whereas iucC gene was recorded in 15(55%) isolates at bp 1075 ,magA found in 3 (11%) at 1283, and ybts found in 20(74%) at 242 bp

Hyperglycemia, or abnormally high blood sugar levels, is a hallmark of diabetes mellitus (DM), a serious metabolic illness caused by the body's incapacity to either create or use insulin as intended. Thisstudy wasdesignedinvestigated the relationship between the expression levels of the long noncoding RNA GAS5(LNCRNA GAS5)and various biochemical parameters in individuals with type 2 diabetes mellitus (T2DM) compared to a control group of apparently healthy individuals. The studyincluded the collection of bloodsamples from 50 suspected T2DMpatients who attendedthe AL-Furat General Hospital in Baghdad after medical investigation and 50 apparently healthy individuals as a control group for the period from July 2023 to November 2023.The Serum Biochemical parameters were analyzedafter the collection of the sample to observe the changes among T2DM cases and healthy controls. In the present study,the age was classified into three groups 40–50 years, 50-60 yearsand > 60 yearswith highly significant differences(p<0.001)in biochemicalparameters (RBS, HbA1c and C. peptide).the GAS5levels were decreased in T2DM calculated ratios for GAS5gene fold expression in the patients compared tohealthy groups were 0.73, and 1.00, respectivelywerehighly significant differencesp<0.001. GAS5expression profiles did not show a significant correlation with clinical parameters, including random blood sugar (RBS) and HbA1c.The ROC analysis revealed thecutoff GAS5 value ofthe predictive cut-off value of (0.16) Analysis indicated an area under the curve (AUC) of ROC of 0.81(95% CI: 72.3%, 92.6%)with 74% sensitivity and 88% specificitywith. (p≤0.001). in distinguishing non-diabetic from diabetic subjects. The positive predictive value is 71.4%.In conclusion, the results of this studyindicate thatthe potential circulating LNCRNA GAS5expression level isimportant in the diagnosis and progression pathogenesis of T2DM.The findings highlight the potential utility of GAS5as a biomarker for distinguishing between diabetic and non-diabetic individuals

Abstract: The purpose of this studyto extract phenolic compounds from Camellia sinensis,investigate their antibacterial and anti-biofilm properties, and apply them to certain virulence genes that are involved in the production of biofilm in multidrug-resistant Pseudomonas aeruginosa.Twenty-five isolates ofP. aeruginosawere collected from the Institute of Genetic Engineeringand Biotechnologylabs atBaghdad University. Using the VITEK-2 technology and growing the isolates on cetrimide agar, the diagnosis was verified. Extracts of Camellia sinensisleaves are created methanolic and aqueous,respectively, using the maceration technique and the Soxhlet equipment. The results indicated that, at concentrations of 128 and 256 mg/ml, As compared to the aqueous extract, the methanolic extract demonstrated greater efficacy, giving the highest inhibition zone values of 19.33 and 23.67 mm, respectively, when compared to the aqueous extract, which gave inhibition zones of 14.33 and 19.00mm, respectively. All P. aeruginosaisolates had MIC of 4 mg/ml for the methanolic extract, with the exception of isolates No. 1, 2, and 10, which had MICs of 8 mg/ml and 16 mg/ml, respectively, the MIC of an aqueous extract for five P. aeruginosaisolates was 16 mg/ml, for four isolates itwas 32 mg/ml, and for isolate No. 10, it was 64 mg/ml. The methanolic leaf extract of C. sinensiscompletely prevented P. aeruginosafrom forming biofilms at a concentration of 8 mg/ml, whereas the aqueous extract completely inhibited P. aeruginosaisolates' ability to produce biofilms at a concentration of 16 mg/ml.Investigating the virulence gene pelA, which is responsible for P. aeruginosabiofilm formation, the gene expression data revealed reduced pelAgene levels following treating with the methanolic extract’s sub-MIC in contrast to the untreated isolates.

This study aims to detect the qualitative detection of Npband Apbgenes responsible for producing neutral proteases (Bea16) and alkaline serine proteases (Bace16), which are among the main proteases produced by Bacillus subtilisthat affect nematodes, and determine the extent of their effect on nematode cuticle degradation with increasing exposure time. B. subtilisis a wild type of bacteria present in the soil and can produce various proteases, some of which may be effective against nematodes. In the current study, B. subtiliswas isolated from the soil and identified molecularly based on a conserved region in the 16S rRNA. Real-time PCR amplified the Npband Apbgenes extracted from B. subtilisfor the qualitative detection of their genes, and the results of the qualitative detection showed the presence of genes at Ct27.91, 27.33 for Npb, and Ct33.51, 35.09 for Apb, respectively. After nematodes (second-stage juvenile) isolates were treated with 10, 20, 30, 40, and 100% crude extracts at 24, 48, and 72 h, the in vitro experiments after 72 h of exposure showed that 30, 40, and 100% crude dilutions had the highest event mortality rates, with averages of 90.7%, 97.3%, and 96.7%, respectively. This is due to the crude extract containing some proteases produced by B. subtilisthat can affect nematodes, including neutral and alkaline serine. This study found that among thevarious proteases produced by B. subtilisthat can impact nematodes, B. subtiliscan make two intriguing ones: Bae16and Bace16. These proteases, along with others, may degrade the nematode's cuticle

Abstract:Hepatitis A virus infectionis a health threat with multiple transmission patterns across areas. This study aimed to use HAV diagnostic methods, including reverse transcription polymerase chain reaction (RT-PCR) test, anti-HAV IgM antibodies, and biochemical analysis to assess liver function. A total of 110 Iraqi individuals participated in this study, comprising 60 HAV-infected patients (Patient group) and 50 healthy controls. The study was conducted from December 2022 to July 2023. Blood samples were collected for HAV antibody titers and RT-PCR confirmation of infection. Liver function tests measured alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST)and total serum bilirubin(TSB)in serum.Our results showed that HAV RNA was detected in samples by RT-PCR, demonstrating the effectiveness of molecular methods in diagnosing HAV infection. Higher levels of HAV IgM antibodies in patients compared to healthy individuals indicated acute infection. Biochemical testing revealed abnormalities in liver function markers in HAV-infected individuals, including elevated ALT, AST, ALP, and total serum bilirubin.This study highlights the benefits of liver function tests in assessing liver damage and diagnosing hepatitis. It emphasizes the need for accurate diagnosis and management of HAV infection to better understand its prevalence and health impact

Abstract:Staphylococcus aureusis the most frequent ocular infection that irritates the cornea and conjunctivitis. Determining the function of the biofilm gene (icaB) in S. aureusisolated from eye infection is the goal of the study. Through the identification of the mecAgene, the study assessed the presence of the icaBgene. Between the beginning of September 2023 and the end of January 2024, 125 patients with a clinical diagnosis of bacterial eye infections were seen in Ibn Al-Haitham Teaching Hospital, Baghdad city. The result of study of 125 swap specimen, 55 (44%) specimen were given positive results for infections caused by pathogenic bacteria and/or fungus, or other species of staphylococci, while 70 (56%) specimen were given negative results, which mean the infection caused by other agents. However among 55 positive bacterial isolation swap was 42isolate (76%) were given positive results for bacterial infections, as (Pseudomonas, Streptococcusspp, Aspergillusspp, Escherichiacoli, Proteusspp and others), while 13 (24%) specimen were S. aureus,depending on the morphologic characteristics of this bacterium on the culture media and biochemical tests included manual conventional biochemicaltests and automated biochemical tests.Molecular detection was conducted using Polymerase Chain Reaction technique of icaBand mecAgene with 117bp, and 533bp respectively,conclusion: 100% of isolates had the icaBgene, while 69% had the mecAgene.

Abstract:A varicocele is an anomalous enlargement and twisting of the veins in the spermatic cord. Varicoceles are a prevalent condition in the general population and are often discovered during normalmedical examinations. However, they are the most frequent correctable cause of male factor infertility. The aim of the current study was design to investigatethe immunological parameters, gene expression profiles, and their correlations in varicocele and non-varicocele oligospermic patients compared to healthy controls, providing insights into the molecular mechanisms underlying male infertility. A total of 120 human male personals were involved in this study over the period from October 2023 to January 2024. Seminalfluid samples were collected from patients who were diagnosed with varicocele confirmed oligospermia (40 samples) and non-varicocele oligospermia (40 samples), in addition to 40 healthy males as control.Demographically, the distribution of the study groups according to age (20-35 and 36-50) among different groups showed no significant variation. The mean age was 34.97±6.5, 31.23±6.7 and 32.0±7.5 for control, varicocele, and non-varicocele oligospermia patients, respectively. According to body mass index, no significant differences in mean of BMI among groups was observed. With respect to alcohol consumption, all participants were non-alcohol consumer, while the distribution of smoking habit among groups revealed a significant difference for varicocele smoking patients (73%) and non-varicocele oligospermic smokers (77%) compared with the proportion of smoking control (22%). The level of IL-18 and Il-37 was estimated in the seminal plasma of tested groups using Enzyme Linked Immunosorbant Assay (ELISA) technique. The seminal plasma levels of the pro-inflammatory cytokine IL-18 were significantly elevated in both varicocele and non-varicocele oligospermic patients compared with control, suggesting its potential role in the pathogenesis of these conditions. Interestingly, the anti-inflammatory cytokine IL-37 was also significantly increased, possibly as a compensatory mechanism to counterbalance the pro-inflammatory state. Notably, the level of seminal anti-sperm antibodies (ASAbs) was significantly higher in varicocele patients compared to the other groups, indicating an autoimmune component in varicocele-related male infertility.The role of androgen receptor (AR)gene expression in varicocele disease was detected using reverse transcriptase polymerase chain reaction(RT-PCR). The fold of gene expression results by RT-PCR technique revealed a significant downregulation of (AR) gene expression in varicocele patients compared with control. In conclusion,no significant correlation was found between the expression of ARand the levels of IL-18, IL-37, or ASAbs, indicating that the alterations in these parameters may involve distinct molecular pathway

Abstract: Salmonella typhimuriumisthe main cause of gastrointestinal diseases for the human population and one of the most dangerous foodborne bacteria. This study aims to detect the occurrence of different important resistance genes in Salmonella typhimurium. Stool samples were collectedfrom some hospitals in Baghdad-Iraq from different patients for bacterial isolation. 110 samples aggregate and identified by morphological tests and affirmed using the vitek-2 system. Antibiotic sensitivity tests of fourteen isolates belonging to clinicalsamples were spread by agar diffusion method.The result showed that Salmonella showed the diameters of the inhibition zones for 7 antibiotics and the antimicrobial Susceptibility rate of 14 SalmonellaIsolates while a genomic DNA kit was used to extract bacterial genome from isolates then screened for virulence and resistance genes mainly InvAand Sopperformed by polymerase chain reaction (PCR).In addition, the study showed that InvAandSopgenes are found in allthe 24 isolate which means even asymptomatic patients had resistance genes.

Abstract: Fourty samples were obtainedfrom burns and wounds, from various ages.specimenswere collected through the period extending from Februaryto December 2022, from patients were admitted in AL-kindy hospital,Alnumaan teaching hospital,Ibn-albaladi for women and children and Imam Ali hospital in Baghdad.Identification of isolates were performed bybiochemical and cultural tests, the results revealed thatall isolates were P. aeruginosa.Conventional PCR(Polymerase Chain Reactin) was also used to confirm these isolates as P.aeruginosaby detection of 16SrRNAbut the results showed that this gene was located in only 30(75%) out of 40 biochemically P. aeruginosa isolates.PCR was also used to screen furgene and iron responsive geneswhich are (pvd,pch,exoS and exoA) of the thirty isolates ofP.aeruginosaand results revealed thatexoSgene was located in only 14(46.6%) out of 40 isolates, whilefor pvdgeneresults revealed only 21(70%) of isolates were consist of this gene out of 30 P. aeruginosa isolates, for pchgene 15(50%) of isolates were consist ofthisgene out of 30 P. aeruginosa isolates, 20 (66.6%) were appeared to have for exoAgene out of 30 P. aeruginosa and 27(90%) of isolateswere positive for fur gene.The source of these isolates which was burn swab n=20 (66.6%) and wound swab n =10 (33.3%)

Abstract: Among the phytochemicals extracted from Cinnamomum cassiabark using hexane, which revealed the presence of terpenoids, alkaloids, flavonoids and phenols. Once 100 Klebsiella isolates from 170 UTI samples were morphologically cultured, only 60 Klebsiella pneumoniaeisolates could be identified using conventional bacteriological and morphological methods. A highly pure RNA sample was collected. The three K. pneumoniaeisolates were tested for fimA andmrkAgene mRNA expression compared with rrsEgene (housekeeping) using RT-qPCR. The RT-qPCR results show that when exposed to subMIC CHEx and CHCsNPs, respectively, the expression of the fimAgene significantly decreased (p≥ 0.05) as compared to the control group. This study utilizes quantitative RT-qPCR screening of the mrkAgene to uncover variations in gene expression among three distinct K. pneumoniaeisolates. The mrkAgene Ct value ranging from 6.0 to 8, and did not differ statistically according to the analysis. K. pneumoniaebacteria secrete a many toxin, because the mrkAgene was not strongly expressed

Abstract:Type 2 diabetes mellitus (T2DM) is a multifactorial not-communicable disease that is characterized by insulin resistance and chronic sub-clinical inflammation. Thisresearch aimed to study therole of biochemical and immunological marker (IL23) levelits effect on the geneexpression of JAKs in T2DM.The study divided participants into two groups: 60 with T2DM and 60 control aged 35-74 who attended Sheikh Zayed Hospital from Nov 2023 to Jan 2024. Participants underwent blood sampling to extract RNA for JAK2 transcript analysis using cDNA synthesis in RT-PCR,additionally theresearch also assessed the clinicopathological traits of diabetic patients and IL-23 concentration using ELISA. Type2 diabetic patients showed significantly high JAK2expression. The study found a correlation between IL-23 concentration, JAK2expression level, and they may act as clinicopathological aspects like HBA1C and FBS levels. In conclusion, T2 DM had elevated JAK2and IL-23 levelsTherapeutic targetsof the diseaselinked to disease activity and pathophysiology may exist.

Abstract:diabetes mellitus, a metabolic disorder distinguished by chronically high blood glucose levels, includes thirst, weight loss, and excessive urination. In diabetic patients, there is a correlation between the overexpression of the cellular signaling inhibitor SOCS3and both insulin resistance and glucose levels. The present research aims to examine the impact of IL-23 on the expression of the SOCS3gene. In the present study, 120 individuals were Involved between November 2023 and January 2024, 60 of whom were diagnosed with Type 2 Diabetes Mellitus (T2DM) treated with Glucophage and Amaryl, excluding patients who use sitagliptin as a treatment`and the remaining 60 were apparently healthy. To evaluate the level of SOCS3Each and every individual had blood samples taken and RNA was extracted. cDNA synthesis was performed, and real-time PCR was used for analysis. Compared to healthy individuals, SOCS3gene expression was upregulated in T2DM patients (fold change: 3.75). Moreover, T2DM patients had elevated IL-23 levels, suggesting a correlation with disease activity.in conclusion, our findings indicate that SOCS3expressionand IL-23 are positively linked with insulin resistance indicators in a diabetic environment.

Abstract:Vulvovaginitis caused via Candidaspecies is a common fungal infection among adult and pregnant femals especially withC.albicans.Fluconazole resistance has been reported more frequently by researchers from around the world. The goals of this study were to determine the prevalence of Candida albicans among suspected vaginitis women, evaluate their ability to form biofilm, and the susceptibility profiles of isolates to fluconazole, as well as the molecular detection of some fluconazole resistance genes.The study included 250 women ages 18 to 45 who were Candida attending specialist hospitals in Baghdad, Iraq.Candidaspp. was identified using standard methods and biochemical testsafter Vaginal smears were cultured on selective HiChrom Candida Differential agar and (SDA).Out of the 250 vaginal smears collected, 150 (60%) isolates of fungal were isolated, with 50 (33.3%) were Candida albicansand 100 (66.6%). Out of 50 Candida albicansisolates which tested by microtiter plate assay, 21(42%) isolates form a strong biofilm, while 19 (38%) isolates were the moderate producer, and only 7(14%) isolates were weak a biofilm formation. The identification of the Candidaspp. by PCR confirmed the primary identification, using the primers from 18S rRNAgene, the PCR assays exhibited the detection of fluconazole resistancegenes(CaERG11, andCaCDR1), where these genes were found in all flucanazole resistant isolates with strong biofilm formation ability.It was concluded that highprevalence of Candidaisolates with high antifungal resistance among Iraqi females patients with Vulvovaginitisindicates the importance of regular screening and routine examination for candidiasis in Iraqi hospital

Abstract: Rheumatoid arthritis is an inflammatory autoimmune disorder that affects large and small joints. Ixifi, a biosimilar medication, is produced from infliximab, a protein that targets Tumor Necrosis Factor-α exclusively.The study determines the effect of Ixifi trough level on disease activity, rheumatoid factor, and anti-cyclic citrullinated peptide antibody. A cross-sectional observational study, which included forty-two patients, who had a diagnosis of Rheumatoid arthritis. Three months after initiation of the Ixifi therapy, Ixifilevel, CDAI, Anti-CCP, and RF biomarkers were determined. The Ixifi trough level was )5.45 ± 0.28μg/ml) inremission and )3.57 ± 0.14μg/ml), (2.2 ± 0.17μg/ml), and (0.66 ± 0.14μg/ml)in the mild, moderate, and severe groups, respectively. The Rheumatoid Factor serum level for the remission was (24.94 ± 0.92IU/ml), mild was (35 ± 1.93IU/ml), moderate and severe groups were(60.55 ±1.89IU/ml)and (70 ±3.18IU/ml), respectively. Anti-Cyclic Citrullinated Peptide, the serum level for the remission was (22.25 ±1.35IU/ml), for the mild was (31±1.62IU/ml), and for the moderate and severe groups was (57.18±2.54IU/ml, and 67 ±2.3IU/ml)respectively.Thedecrease in the Ixifi levels leads to an increase in disease severity and inflammation, whereashigh concentrations of Ixifi decrease the disease activity, Rheumatoid Factor, and Anti-Cyclic Citrullinated Peptide Antibody Serum

Abstract:Interleukin 6 (IL-6) is a well-established indicator of inflammation and is one of the first immune responses to occur in cases of sepsis. It is possible to assess serum levels quickly, typically within a few hours. The precise clinical importance of IL-6 in the initial phase of sepsis in burn individuals remains unverified. This study aimed to investigate the prognostic significance of IL-6 in burn patients with suspected sepsis, specifically in relation to bacterial infection. 44 patients with age (1.5–75 years), and 19 healthy controls with the same age range, were involved in this study during their attendance to Specialized Burns Hospital in the Medical City, Al-Kindi Teaching Hospital in Baghdad. The study was conducted from 1st December 2023 to 1st April 2024 and approved by ethical committees of Institute of Genetic Engineering and Biotechnology for Postgraduate Studies, University of Baghdad. Blood samples and data were collected to evaluate the level of IL-6 associated with bacterial infection for burn patients by RT-qPCR System. Results appeared that mean IL-6 percentages were significantly (P≤0.01) increased in burn patients as compared with control. The current study is consistent with other research as it shows no significant differences in IL-6 levels among patients with gram-positive, gram-negative, and mixed growth

Abstract:This study aimed todetermine which antibiotic is the most vulnerable and resistant to pathogen and to study the change of expression fold after antibiotic-treatment in compare to isolate without treatment by Reverse Transcription Quantitative Polymerase Chain ReactionRT-qPCR technique. A total of 55 samples with omphalitis symptoms inchicksin Baghdad province. These samples were cultivation, molecular analysis and antimicrobial susceptibility. Fifty isolates showed blue-black colonies on cultureEosin Methylene Blueand pink colonies on MacConkey agarand five isolates showed negative results. of Identification by PCR analysis for detection the 16S rRNAgene. The results of nucleotide sequencing were submitted in GenBank/NCBI database through accession number (OR142657.1). Antimicrobial susceptibility tested showed that the isolates were resistant to ampicillin, amikacin.Four E. coliout 50 isolate chosen for the minimum inhibitory concentration (MIC) exhibited by the ampicilinagainst tested E. coli bacteria,neomycinand combination of ampicillin and neomycinIn particular, the result of gene expressionthe change in expression fold was analyzed using the Livak formula. The results showed that treatment with neomycin had a higher suppression effectiveness on the studied genes compared to ampicillin. Additionally, the combination treatment (Amp/Neo) did not exhibit a clear suppression effect on the genes, which could be attributed to the antagonistic action of ampicillin when used with neomycin.

Abstract: The World Health Organization has identified antibiotic resistance among bacteria as one of the most serious dangers to human health. This present study focuses on the isolation and identification of antibiotic-resistant Escherichia coliusing 16S rRNAgene sequencing based on molecular techniques from water samples.Between September and December 2023, we collected fifty aseptic water samples from various water treatment plants in Baghdad city. A number of biochemical and molecular tests confirmed the isolation of Escherichia colifrom the samples. Isolates has been placedintonutrient broth and cultured them on various agar media, such as MacConkey agar and EMB agar. For biochemical analysis, we used Gram\\'s staining, string techniques, and Vitek.Using the Kirby-Bauer disk diffusion technique, we tested the antibiotic sensitivity of 45 Escherichia coliisolates to fourteen commonly prescribed antibiotics. The data indicates a significant variation in the percentage of antibiotic resistance among E. coli isolates across different regions. In some regions, such as medical cities, the resistance rate was very high for almost all antibiotics. Other regions, like Al-Rustaimiya Station, also exhibit a high resistance rate.We amplified and compared 16S rRNA gene sequences to the NCBI sequence database using primers. A study of the partial 16S rRNA sequence revealed that two new strains, HHWW and HS, were 100% similar to E. coliand had a lot in common with species in the genus Escherichia

The Effect of Oxidative Stress and Some Biochemical Parameter in a Sample of Iraqi Chronic Renal Failure Patients in Baghdad Governorate

Abstract: The present study was conducted to investigate the antioxidant and cytotoxicity of purify flavonoids that isolated from Cuscuta campestris.Three solvent (methanol, Dichloromethane, ethyalacetate)were used to extracted flavonoid.were measured(128.5,85.1,98.3 mg Rutin / 100 gm) respectively, the type and quantity of flavonoid were detected by HPLC .The result showed that the plant extract contains flavonoids (Rutin ,Qurcetine, Kaempferol,Apigenin, Luteolin)(124.0,130.5,80,9,58.7,51.5)μg/gm respectively.The antioxidantactivities of purify flavonoids ofCuscuta campestriswere screenedusingDPPH (2,2-diphenyl-1-picrylhydrazyl). the results revealed that the radical scavenging activity is effective and activity increase when the concentrations increase.The highest percentage of free radical scavenging is at of 200μg/mL (82.37)a concentrationcompared with control (13.20). Some species of Cuscutapossess anticancer activity on various cell lines (colon HT29, MCF10). Using MTT assay. The cytotoxic activity purify flavonoids extracted fromCuscuta campestris. The results showed that the administration of different doses (12.5,25,50,100,200) μg/mLafter 72h of Cuscuta campestrisresulted in decreased viability of(HT29) cells which have higher in inhibition rate compared with control (88.7,13.20) for HT29, MCF10 respectively. This plant may represent promising source of herbal treatment for colon cancer and is thus a safe and promising anticancer drug candidate.

Abstract:Genetic Stability analysis of in vitrocultivated GAC fruit (Momordica cochinchinensis Spreng)from the Horticulture Department stationusing ISSR markers. These are difficult to apply for breeding, resulting in limited availability.Resultingin a lack of seed availability. However, tissue culture allows the developmentof micropropagate variations, especially in plantlets undergoing several subculture stages. This study aimed to identify somaclonal variations in GAC fruit seedlings generated from in vitro culture using Inter Simple Sequence Repeat (ISSR) markers. Two accessions were used in this study consisting of GAC fruit grown in its original environment and in vitro cultured. Total of five primers, of which four primers detected polymorphisms amongthe population. This study resulted in a somewhat similargenetic similarity among the GAC fruit. The genetic diversity of GAC fruit plants propagated by in vitroculture had a similarity level of 92%, showing somaclonal variations in vitroculture.

Abstract: With the widespread increase of bacterial resistance to available antibiotics and the lack of re-sources for the discovery of new classes of antibiotics, the multidrug-resistant S. aureus has become a global concern. The methicillin-resistant Staphylococcusaureus(MRSA) has been reported as a serious threat in health settings. There is now a strong demand for new antibacterial agents as only a few antibiotics can combat MRSA infections. This articleaims to in vitro investigatethe antibacterial properties ofnew (phos-phanegold(I) thiolate) compoundsagainst S. aureusstrains. The disc diffusion methodwasapplied as a preliminary screeningmethodof thegoldcompoundsagainst different Gram-positive and Gram-negative bacteria. In contrast,minimum inhibitory concentration(MIC), minimum bactericidal concentration(MBC),and time-killingassay methods were carried out to investigate the antibacterial activity of selected compoundsagainst different Methicillin-resistant Staphylococcus aureus(MRSA) species.Moreover, com-binations of available antibiotics and selected (phosphanegold(I) thiolate) compound were investigated us-ing the checkerboard method.Two out of five compoundsbelong to phosphangold(I) thiolate named 3F2 and 3F3were showing antibacterial activity against Gram-positive bacteria,predominantly Staphylococcus aureusstrains with MIC values within the rangefrom 4 μg/mL-8 μg/mL and30 μg/mL –60 μg/mL, respectively. Only 3F2 was exhibited goodand promisingantibacterial activity against different S.aureusstrains including MRSA strains.Additionally, the 3F2 was exhibited synergistic activity and positive drug interaction when combine with ciprofloxacin at sub-inhibitory concentration. These findingssuggested that the new investigatedphosphanegold(I) thiolate,mainly 3F2 compound, may have great potential to act as an antibacterial agent with action against Gram-positive bacteria that mainly prevent or control MRSA infection's growth. Thus,further investigations are neededto fully understand the mechanismof action of tested (phosphanegold(I) thiolateagainst S.aureus.

Abstract:Quinine, a classic antimalarial agent, in recent years has attracted attention for its anticancer activity. In this study, we examined the cytotoxic and anti-proliferative effects of Quinine against the Rhabdomyosarcoma (RD) cell line and an exploration ofits interaction with the major proteins overexpressed in this cancer. Cytotoxic activity was evaluated usingthe MTT assay, which indicated a dose-dependent, significant inhibition of RD cell growth, as represented by an IC₅₀ value of 39.3μg/ml at a 24-hour treatment time. Morphological changes in line with cytotoxic effect were also observed. Additional molecular docking analysis was also conducted to estimate the binding affinity of Quinine with some proteins implicated in the pathogenesis of Rhabdomyosarcoma. The in silico data showed that Quinine could inhibitthe activity of these proteins, and it could be the reason behind its antiproliferative activity. The observation is preliminary evidence of the therapeutic potentialof Quinine against Rhabdomyosarcoma and needs more mechanistic and in vivo studies

Abstract: The rising incidence of Type 2 diabetes mellitus (T2DM) globally attaining epidemic levels and is emerging as a significant public health concern. Toll-like receptors (TLRs) are innate immune system receptors that facilitate the inflammation in diabetic mellitusdisorder. The study evaluatesthe relationship betweenrs352139 SNP in TLR9 gene in patients with T2DM in Iraq,and its effect on TLR9 level in those patients. This study included 60 T2DM cases and 40 apparently healthy controls. The SNP were identified using Sanger sequencing for rs352139.Current findingsindicated a significant elevation in TLR9 levels in T2DM patients compared to healthypeople(1.182± 0.76 versus0.836 ± 0.74ng /ml, p ≤ 0.05). T2DM risk factors include AA, GG genotypesin comparison with controls(40% versus22.5%; P = 0.009; OR = 2.30 and 40% versus20%; p = 0.005; OR 2.67). Patients carrying the AG and GG genotypes, had higher Serum concentrations of TLR9 in comparison withthe control group(1.13 ± 0.26 versus0.79 ± 0.18 ng/ml; 1.22 ± 0.18 versus0.71 ± 0.16 ng/ml) respectively.This study indicate a risk association between rs352139 and T2DM. Also, rs352139 increase TLR9 serum level in patientswith T2DM