Iraqi Journal of Bioscience and Biomedical

Circulating cell-free tumor DNA (ctDNA) released from apoptotic or necrotic sold tumor cells, such as
breast cancer, into the bloodstream. Breast cancer patients show high levels of circulating DNA shed from
the tumor compared to healthy individuals. This case-control study involved 35 women diagnosed with
breast cancer and 50 healthy controls. Blood samples were processed for serum and plasma, utilizing
K3_EDTA tubes for ctDNA extraction and gel tubes for ELISA assays. The study assessed the evaluation
of the TP53 and BRCA2 proteins from serum using ELISA kits. CtDNA was extracted from plasma using
a special kit and real-time PCR (RT-qPCR) to quantify the levels of TP53 and BRCA2. The purpose of this
study is to explore the advancements in ctDNA detection technologies, monitor disease progression, and
provide insight into the genetic landscape of breast cancer in Iraqi patients, suggesting the ctDNA of BRCA2
and TP53 potential as valuable biomarkers for diagnosis and treatment monitoring. The results identified
several important findings regarding breast cancer patients and their healthy control counterparts, in terms
of genetic markers, serum BRCA2 levels showed (327.02) in breast cancer patients which is significantly
lower compared to controls (511.85) and serum TP53 levels followed a similar trend (198.41) in breast
cancer group and (389.57) in control group. DNA concentration analysis which is ctDNA showed (2.440)
DNA level of BRCA2 gene in breast cancer patients, compared to controls (1.000). On the other hand, DNA
level of TP53 gene was (0.191), significantly lower in breast cancer patients compared to controls (1.000).
These findings highlight the importance of using ctDNA and molecular markers in improving personalized
cancer treatment

This research study compared the levels of compounds, like phenolic, flavonoid, alkaloid, tannin, glycoside and saponine in both the leaves and roots of Cassia Glauca and Conocarpus Lancifolius collected in September 2023 locally from the local gardens in Baghdad, The aqueous plant extract was prepared using the traditional method. The leaves C. lancifolius exhibited concentrations of phenolic content (235.4 mg/g) total flavonoid content (108.9 mg/g) total tannin content (55.7 %) and glycoside content (1.9 mg/g) than C. glauca which had higher levels of total alkaloid content (2.44 mg/g). While the roots C. glauca showed levels of phenolic content (62.5 mg/g) and total tannin content (9.58 %) compared to C. lancifolius where higher amounts of total saponine content were observed (0.25%). In summary it was noted that C. lancifolius leaves generally displayed concentrations, across categories when compared to those of C. glauca.

Gout is the mainly prevalent type of inflammatory arthritis that is characterized by deposition of monosodium uric acid crystals in and around peripheral joints. This study was aimed to examine the effect of some biochemical parameters (serum uric acid, serum creatinine) and some immunological parameters (TNF-α , IL1-β) in Iraqi patients with gouty arthritis .Samples of blood were drawn from 41 gout patients and 50 healthy individuals as the control group, spanned from December2023 to march 2024, at Rheumatology Department of clinical Counselor at AL-Yarmouk Hospital. Their ages ranged from 25 to 80 years old, Clinical assessments, including imaging and laboratory tests, led to the diagnosis of acute gouty arthritis in all patients. Serum uric acid and creatinine were measured, serum cytokines (IL-1β and TNF-α) were determined using enzyme-linked immunosorbent assay (ELISA). The results indicated that serum uric acid concentration was significantly higher in gout patients (6.05±1.85 mg/dl) as compared to controls (4.40±0.97 mg/dl). No significant change was observed in the concentration of serum creatinine. Both serum cytokines (IL-1β and TNF-α) were significantly higher in gout patients as compared to controls.

The global threat of antimicrobial resistance (AMR) has driven an urgent need to explore alternative strategies to combat resistant pathogens. Metal nanoparticles (NPs), owing to their unique physicochemical properties, have emerged as promising antimicrobial agents. This review highlights the diverse mechanisms of action exhibited by metal nanoparticles, including membrane disruption, reactive oxygen species (ROS) generation, and interference with intracellular processes. Particular emphasis is placed on silver, gold, copper, zinc oxide, and titanium dioxide nanoparticles due to their broad-spectrum antimicrobial activity and synergistic potential when combined with conventional antibiotics. Furthermore, the review discusses the role of nanoparticle synthesis methods, size, shape, and surface modifications in enhancing antimicrobial efficacy and reducing toxicity. While metal nanoparticles show immense potential in overcoming AMR, challenges such as scalability, biocompatibility, and long-term safety remain key considerations for clinical translation. This review underscores the need for further multidisciplinary research to harness the full potential of metal nanoparticles in addressing the AMR crisis.

The study aims to compare biofilm formation quantitatively and compositionally between two selected isolates of Pseudomonas aeruginosa from soil rhizosphere zone and infection cases. Fifteen isolates were isolated out of 50 samples from the root rhizosphere zone of maize plants from the fields of the College of Agriculture / University of Baghdad during November, 2023, Ten isolates from burn injuries, and 10 isolates from 25 identified strains obtained from the isolate bank / College of Biotechnology, Al-Nahrain University. Isolates from soil and infections were identified by biochemical tests and compared with identified strains. The isolates were subjected to a qualitative biofilm formation test, and 15 isolates were selected from the soil and 15 from the infections and standard strains (10 from the specimens and 5 from the pure strains (P11-P15). Statistical analysis of qualitative biofilm formation indicated to significant differences between these selected isolates. It was found also the significant differences at P-value (0.0017) among pathogenic isolates and 0.0001 among soil isolates. So, ten soil isolates and 10 pathogenic isolates were selected for quantitative test of the biofilm using the Microtiter plat method. The results of quantitative measurement of biofilm formation showed that all selected pathogenic and the soil isolates were of the strong biofilm formation category and to the significant differences at P- value (≤0.05) among pathogenic isolates and also among soil isolates, but in comparison between soil and pathogenic isolates, the most of soil isolates have significance P- values at (≤0.05) over pathogenic isolates. Soil isolate (S2) and pathogenic isolate (P10) were selected for their high concentration of biofilm formation, but soil isolate has more biofilm concentration compared with P10. Antibiotic sensitivity test using the antibiotic disc diffusion method showed that 71% of the soil isolates were resistant to all 10 antibiotics from 8 groups of antibiotics, and 62% of the pathogenic isolates. As for the selected pathogenic isolates p10 and S2 from the soil, they were resistant to 80 % of antibiotics. The results of FTIR analysis showed that the components of the biofilm of the pathogenic isolate are more complex than those of the soil isolate and that the soil isolate contains more lipids and mixed materials. The results of HPLC analysis of polysaccharides showed to the significant differences for GIcL and GaIA in soil isolate at (P ≤0.05) and (P≤0.01), respectively compared with pathogenic isolate and the absence of D-mannuronic acid ɤ-lactone (ManL) in biofilm of pathogenic isolate.

Incubation temperature is a critical factor in measuring biochemical assays, therefore this study was designed to measure effect of temperature variations on the some important clinical assays which are C - reactive protein (CRP), vitamin D, D-Dimer, serum ferritin and rheumatoid factor (RF). Blood samples were collected from fifty different patients and only samples closer to the normal value were used in this study. All tests were incubated at room temperature (RT) and 37 °C in five replicates. Results showed that there was a significant effect of temperature variations on these biochemical assays. It could be concluded that incubation at 37 °C is the best temperature to perform chemical assays.

A series of modern compounds N,N'-(1,4-phenylene) bis (1-(1,3- diphenyl-1H-pyrazol-4-yl)
methanimine from 5a to 5e were synthesized and characterized through methods were applied to
enhance the structures for the new compounds Mass spectrometry, infrared and nuclear magnetic
resonance. These modern compounds have antimicrobial activity estimated against bacteria and fungi,
involved both Gram-positive and Gram-negative. Specific substitutions of the pyrazole rings were
conclusive in enhancing the antibacterial activity. According to these results, hybrid compounds of
pyrazole showed promising potential as potential candidates for the creation of new antimicrobial
drugs.

Colon cancer starts in the colon or rectum, usually as tiny, noncancerous cell groupings called polyps that form along the inner lining. There are several factors to consider that may influence the development, diagnosis, and treatment of the disease. Over time, some of these polyps can become cancerous. Compounds as Oxazepine derivatives of coumarin acid designed and synthesized in order to develop a lead molecule that demonstrates anti-cancer efficacy and the side effects of NSAIDs, including GI toxicity. Oxazepien derivative compounds diagnosed using FT-IR and 1HNMR. After that evaluate the cytotoxic effects of Oxazepine derivatives on investigated in vitro versus the human colon cell line CaCo-2 with normal human liver cell line WRL68. Effective of the test substances on cell growth was assessed by utilizing the MTT assay method after treating the cell line with various doses (1,5 ,10 ,15 ,20,25 ,50 and 75µM) for 24hours. The results showed that the most active compound against cancer cells 42.12±1.28 was 5b at a concentration of 75 µM and IC50 = 39.6 µM. It also has high selectivity and has a low toxicity rate when tested against normal cells WRL68 75.53±0.77. The current study showed the highly effective action of oxazepien derivative compounds and can be used in the management and treatment of cancer.

Amphetamines, a popular psychostimulant, are causing significant health issues due to their metabolism, which generates benzoic acid derivatives. This study presents an accurate analytical technique for toxicologists, chemists, and public health professionals to ensure safety for the general population. Human benzoic acid levels from amphetamine users were quantitatively analyzed using GC-MS, which is the most effective choice for confirming drug concentrations in forensic toxicology. The study collected blood and urine samples from addicted prisoners in Wasit, dividing them into four groups: those who have not used amphetamines for less than 24-48 hours, those who have not used amphetamines for 7-8 days, those who have not used amphetamines for 15-16 days, and a control group who has never used drugs. The liquid-liquid extraction method (LLE) is a method used to extract general blood and urine components. The study found that the 24-48 hours subgroup had significantly higher levels of benzoic acid in their blood and urine compared to the control group. The 24-48 hours subgroup showed the most noticeable rise in blood benzoic acid levels, while the 24-48 days subgroup showed similar levels. The study also found that the changes made in the experiment significantly impacted the amount of benzoic acid excreted in urine, with the 24-48 hours subgroup having the most significant effect, indicating that the treatment or condition being studied may have an effect on the kidneys and liver, and it provides a precise analytical technique for toxicologists, chemists, and public health professionals to accurately analyze human benzoic acid levels from amphetamine users.

Staphylococcus aureus is a leading cause of wound infections, often exhibiting resistance to conventional antibiotics. This study investigates the antibacterial activity of immune cell secretions specifically from monocytes, macrophages, and lymphocytes against S. aureus strains isolated from wounds utilizing an in vitro tissue culture model. Clinical isolates of S. aureus were characterized and cultured in the presence of immune cell-conditioned media derived from human peripheral blood mononuclear cells (PBMCs). The antibacterial effects were assessed by evaluating bacterial viability, biofilm inhibition, and morphological changes using crystal violet staining, and microscopy. Secretions of PMNCs were collected and analyzed using high performance liquid chromatography (HPLC). Antibacterial roles of these secretions were analyzed using bacterial sensitivity assay and biofilm inhibition test compared to ten antibiotics including methicillin, doxycycline, ampicillin, gentamicin, ciprofloxacin, amoxicillin / clavulanic acid, rifampin, azithromycin, erythromycin and chloramphenicol. Results revealed that PMNCs contain IL-2, IL-6 and TGF-ᵝ and lymphocyte secretions contain IL-2, FGF-7, and IL-6. In response to immune cell secretions, the results showed a significant decrease in bacterial growth and biofilm formation, suggesting the presence of strong antimicrobial factors. These results emphasize how immune cell derived secretions may be used as a natural treatment strategy to fight S. aureus infections in wound environments.

In the field of death investigation, practical determination of the time since death, or postmortem interval (PMI), is the most important. Several approaches have been devised to allow PMI estimation including the classical methods of the developmental stages and temperature of the body, and molecular biological methods. This paper is intended to present sufficiently, perform and analyze the state of the art of such methods, discuss their strengths and weaknesses, and outline the developments’ progress in the context of forensic science, especially the prospective areas of estimating more correctly and reliably the duration of postmortem interval.

Breast cancer is one of the most prevalent malignancies worldwide, with hormone-responsive breast cancer being a significant subset of cases. Estradiol plays a crucial role in the proliferation and progression of hormone-sensitive breast cancer cells. To investigate the role of hormonal contraceptives, which commonly contain synthetic estradiol and progesterone in breast cancer development and progression. Treat MDA breast cancer cell line with Hormonal contraceptives (estradiol and progesterone and mix of both) with different concentration, replicates was 3 technical replication in one experiments was performed. The result was the estradiols have high effect on progression of cancer and increase the cell viability. Hormonal contraceptives increase the breast cancer cell and cell viability especially that have high concentration of estradiol.

Maintaining skin integrity and controlling infection depend on effective wound healing; nonetheless, poor healing still presents a major clinical and public health issue. The purpose of this research was to find the correlation between the expression levels of microRNAs-155 (miRNAs) and the prevalence of burn and wound injuries aggravated by bacterial infection in Iraqi patients. Three hundred blood samples in all—one hundred from healthy controls and two hundred from burn and wound injured patients. Patients had notably higher clinical parameters than controls (p < 0.05: white blood cell count, red blood cell count, neutrophils, lymphocytes, platelets, ESR, C-reactive protein). Reverse transcription quantitative polymerase chains reaction (RT-qPCR) was used to examine miRNA-155 expression. The findings showed a significant downregulation of miRNA-155 in the sick group (0.62 ± 0.027) relative to the control group (1.0 ± 0.0), therefore approximating a 38% drop. These results suggest that miRNA-155 could be a useful molecular biomarketer for burn and wound injuries, especially those aggravated by bacterial infection. Its lower expression emphasizes its possible function in the inflammatory response and tissue damage, therefore stressing its importance for further diagnostic and therapeutic uses.

This study explores the synthesis and biological evaluation of novel imine derivatives (L1 and L2) derived from 6-aminopenicillin acid. These compounds were synthesized through condensation reactions with p-methoxybenzaldehyde and 2-hydroxybenzaldehyde, then structurally characterized via FT-IR and ¹H-NMR spectroscopy. Antibacterial assays revealed strong inhibition zones, particularly for derivative L1, which showed a 31 mm inhibition zone against E. coli at 0.1 M, outperforming both L2 and unmodified penicillin (29 mm each). At 0.001 M, L1 maintained superior efficacy with a 24 mm zone. Antifungal testing against Pleurotus species confirmed notable activity. The anticancer potential was examined through MTT assays on the PC-3 cell line. L1 significantly reduced cell viability in a dose- and time-dependent manner, with IC₅₀ values of 44.47 ppm at 24 hours and 19.75 ppm at 48 hours, while L2 exhibited IC₅₀ values of 40.05 ppm and 16.86 ppm at respective intervals. Molecular docking against androgen receptor-associated proteins (1FVT, 1MOQ) showed strong ligand-receptor binding, with L1 achieving a binding affinity of -6.91 kcal/mol (RMSD 1.95 Å) and L2 scoring -6.84 kcal/mol (RMSD 2.07 Å), demonstrating structural compatibility and interaction potential. The synthesized derivatives, especially L1, showed promise in inhibiting microbial growth and reducing viability in prostate cancer cells, with supportive molecular interactions revealed through docking studies. These findings indicate that modified penicillin derivatives could serve as dual-function agents with applications in antimicrobial and anticancer therapies.

PCOS is a multifactorial and heterogeneous endocrine disorder with a complex pathophysiology. This infertility disorder is increasingly prevalent worldwide, impacting a substantial percentage of women in reproductive age, with an estimated prevalence of approximately 8-13%. This search aimed to investigate the association of (CCR5Δ32) polymorphisms in chemokine receptor gene (CCR5) with PCOS.A case control study was organized at Baghdad Teaching Hospital in Baghdad Medical City included 150 women which were divided into three groups, Group 1 Includes (50) normal weight PCOS patients, Group 2 Includes (50) obese PCOS patients and Group 3 Includes (50) healthy women (Control). polymerase chain reaction (PCR)was used to detect and determine the presence of the gene variant (rs333) in the search subjects. The search’s findings demonstrated that the majority of the biochemical and endocrine serum biomarkers analyzed including Lipid profile by checking the amounts of Total cholesterol, triglycerides, HDL, LDL and VLDL and hormonal analysis of (LH, FSH, Testosterone, E2 and AMH) exhibited statistically significant variations in PCOS patients. Specifically, PCOS patients’ groups showed elevated levels of triglycerides, total cholesterol, LDL, and VLDL, alongside reduced levels of HDL. Furthermore, the hormonal analysis confirmed a significant increase in LH: FSH ratio, LH, total testosterone, and AMH, whereas lower levels of E2, with no significant difference in FSH values between PCOS patient groups and control group. This search found no significant association between the CCR5-Δ32 (rs333) mutation and polycystic ovary syndrome (PCOS). The wild-type CCR5 genotype was predominant among all participants with a slightly higher frequency in in heterozygous form, in the control group (4%) compared to the obese PCOS group (2%), with no individuals carrying the homozygous Δ32/Δ32 genotype in the studied population.