Cellular and Molecular Biology

Genotoxic of co-codamol for human lymphocyte culture in vitro

Sada Jasim Abdulameer — 2025-07-30

Co-codamol, a combination analgesic containing paracetamol and codeine phosphate, is widely used for pain relief, but its potential genotoxic effects on human lymphocytes remain largely unknown. This in vitro study investigates the genotoxic potential of co-codamol on cultured human lymphocytes by assessing cell viability, mitotic index (MI), chromosomal aberration frequency, and micronucleus (MN) formation. Lymphocytes were exposed to varying concentrations of co-codamol (0.02-0.12 mg/mL), and cytotoxicity was determined using the MTT assay. Results showed that co-codamol significantly reduced cell viability in a dose-dependent manner, with complete cell death at 0.12 mg/mL. The mitotic index was significantly decreased at higher concentrations, and a statistically significant increase in chromosomal aberrations and micronucleus formation was observed in treated lymphocytes compared to the control group (p < 0.05). These findings provide important evidence that co-codamol exhibits genotoxic potential in vitro, suggesting a potential risk of DNA damage associated with its use. Further in vivo investigations are warranted to assess the clinical relevance of these genotoxic effects and to elucidate the underlying mechanisms of toxicity.

Protective effects of matrine on cardiomyocytes infected with coxsackievirus B₃ via modulation of the calpain-2/caspase-12 signaling pathway

Yongmei Sun — 2025-07-30

Viral myocarditis (VMC) presents a substantial threat, especially for children, often leading to cardiogenic shock and fulminant myocarditis. Our study aimed to evaluate the role of calpain-2 and caspase-12, which were involved in the endoplasmic reticulum apoptosis pathway, and the influence of Matrine on these proteins during Coxsackie virus B₃ (CVB₃)-induced acute VMC mice in vitro and in vivo, shedding light on the potential cardioprotective effects. We first performed primary cultured cardiomyocytes, which were infected with CVB₃in vitro. We observed cell viability, the beating of cardiomyocytes and cytopathic effects. And we utilized Balb/c mice to establish the VMC animal model and determined viral titers, histopathological changes, and myocardial pathological scores. Furthermore, we detected CK-MB levels and myocardial cell apoptosis in vitro and in vivo. In order to further explore the possible mechanisms, the protein expression of calpain-2 (by immunohistochemistry and Western blot) and caspase-12 activity (by fluorescence assay for substrate cleavage) were detected in vitro and in vivo. Our findings indicated that, in comparison to the normal control group, the virus-infected group exhibited increased injured myocardial cells, virus titer, CK-MB levels, and apoptotic cells (P<0.05). Matrine treatment groups significantly reduced CK-MB levels, myocardial cellular damages and apoptosis in vitro and in vivo, with Matrine notably suppressing calpain-2 protein expression and Caspase-12 activity compared to the virus-infected group (P<0.05). In conclusion, our study revealed that calpain-2 and caspase-12 played roles in CVB₃-induced myocardial cell apoptosis. Matrine effectively mitigated myocardial cell injury and reduced apoptosis, thereby providing substantial protection against CVB₃infection in vitro and in vivo, which may be related to the down-regulation of calpain-2/caspase-12 signaling pathway.

Rutin treatment alleviates obesity-related aortic endothelium dysfunction in albino rats fed a high-fat diet

Hassan A. Madkhali — 2025-07-30

Flavonoids have recently been shown to be useful to people suffering from vascular disorders caused by a high-fat diet (HFD). The flavonoid rutin (RT) exhibits numerous pharmacological effects, including antioxidant, cytoprotective, vasoprotective, and cardioprotective activities. The primary objective of this research was to assess the efficacy of RT against obesity-related vascular endothelial dysfunction (VED) in rats fed HFD. A total of 24 mature Wistar rats were blindly categorized into 4 treatment and control groups: normal control, obese control, and obese which were given RT at 50 and 100 mg/kg for the final 3 weeks of the experimental period. Animals' body mass and food consumption have been estimated periodically. In addition, liver mass and retroperitoneal fat mass per body mass, abdominal circumference (AC), LEE index, and body mass index (BMI) were estimated. Moreover, lipid profile parameters were assessed in serum. The effect on vascular endothelium reactivity was investigated in an isolated rat aorta. A histopathological investigation of the aorta was performed. The obese control group exhibited higher body, liver, and retroperitoneal fat weights. Significantly, RT intake reverses all these alterations. Furthermore, RT decreased food intake, AC, Lee index, and BMI in HFD-fed rats. The lipid profile of HFD-fed rats was also improved after RT treatment, with lower triglycerides, total cholesterol, LDL-C, and VLDL-C levels and higher HDL-C levels in the serum of HFD-fed rats. Through the ex-vivo investigation, RT groups showed improved vascular endothelium function in HFD-fed animals compared to the obese control group. Taking together, RT could be a promising option for preventing obesity-associated VED.

Low frequency of HER2 expression in colorectal cancer: A Tunisian single-center study

Khouloud Ben Lazreg — 2025-07-30

HER2 expression is a potential theranostic and prognostic marker in some cancers, particularly in breast and gastric cancers. However, published data on HER2 expression in colorectal cancer (CRC) remain controversial. This study investigates the immunohistochemical and molecular expression of HER2 in primary CRC and evaluates its clinicopathological and prognostic significance in Tunisian patients. A retrospective analysis was conducted on 144 CRC patients. HER2 status was assessed by immunohistochemistry and tissue microarray analysis, following the diagnostic criteria for gastroesophageal adenocarcinoma. CRC cases with ambiguous results underwent chromogenic in situ hybridization. The mean patient age was 61.9 years (male-to-female ratio: 1.18:1). Tumors were classified as colonic (74.3%) or rectal (25.7%), with 45.8% located in the left colon. Stage III disease was identified in 37.5% of cases, and distant metastases were present in 13.9%. HER2 expression results were as follows: negative (score 0/1+) in 142 cases (98.6%), equivocal (score 2+) in one case (0.7%), and overexpressed (score 3+) in one case (0.7%). No HER2 gene amplification was detected, and none of the metastatic CRC cases showed HER2 immunostaining. These findings suggest that HER2 overexpression and amplification in CRC are rarer than previously reported, highlighting the need for multicenter Tunisian studies to validate these results. The variability in HER2 immunostaining criteria further underscores the importance of a standardized scoring system to ensure consistency in both diagnosis and research.

Effects of abiotic factors on Zingiber officinale and Glycyrrhiza glabra to extract bioactive compounds under different time incubation and different salt concentrations

Mustafa I. Almaghasla — 2025-07-30

Zingiber officinale, commonly known as ginger, and Glycyrrhiza glabra, commonly known as licorice, are medicinal plants that are rich in bioactive compounds with various health benefits. This study aimed to investigate the effects of different growth durations and salt concentrations on the production of bioactive compounds by these plants. This experiment was conducted under natural conditions and the plants were subjected to salt stress at different stages of growth. This analysis focused on assessing the production of polysaccharides, flavonoids, ergosterol, adenine, adenosine, hypoxanthine, and guanosine by UV spectrophotometer and high-performance liquid chromatography (HPLC) in both plants. The results showed that Z. officinale exhibited the highest polysaccharide content at 20-d of growth with 3mM salt, whereas G. glabra showed slightly lower polysaccharide content. Similarly, Z. officinale had higher flavonoid content at 25-d of growth with 5 mM salt compared to G. glabra. Additionally, Z. officinale demonstrated higher concentrations of ergosterol, adenine, adenosine, hypoxanthine, and guanosine than G. glabra, particularly at 25-d of growth with 5 mM salt. This study provides valuable insights into the production of bioactive compounds in Z. officinale and G. glabra under different growth conditions, which can be beneficial for optimizing their cultivation and utilization in various applications including pharmaceuticals and functional foods.

Anti-inflammatory effect of thyme on rheumatoid arthritis in animal model

Sana Seamand Sabir Blbas — 2025-07-30

Thyme (Thymus vulgaris) is a Mediterranean herb known for its culinary, cosmetic, and medicinal applications as it has been discovered that the plant has many clinical properties such as anti-inflammatory, anti-microbial, and antioxidant properties. Sandwich Elisa technique was used to determine the concentration of cytokines. Phenolic contents and other active compounds of thyme were analyzed by Gas Chromatography-Mass Spectrometry (GC-Mass). In this study 15 male adult albino rats were divided into 3 groups (n=5), group one (G1) was the control group which fed on basal diet. Group two (G2) was the Rheumatoid arthritis (RA) group in which the rats were inoculated with 0.1 ml of CFA (Complete Freund’s Adjuvant) yet fed on basal diet. Group three (G3) was the treatment group in which rats were inoculated with CFA along with the administration of thyme extract orally for 22 days. The results show that treatment with thyme extract significantly reduced proinflammatory cytokines IL-1 (interleukin-1), IL-6 (interleukin-6) and TNF- α (tumor-necrosis factor-alpha). Anti-inflammatory IL-10 (interleukin-10) showed a significant increase in the thyme-treated group. CD4 T (cluster of differentiation-4) cell levels showed a significant difference, while sCD14 (soluble cluster of differentiation-14) levels were non-significant in the thyme group compared to the RA group. Inflammatory markers (C-reactive proteins) CRP and (anti-cyclic citrullinated peptide) Anti-CCP antibodies were both significantly elevated in RA and significantly reduced by thyme treatment. Although body weight changes were statistically non-significant, they were visibly prominent. Paw edema was significantly decreased in the thyme-treated group. (Matrix-metalloproteinase) MMP-1 levels and neutrophil counts were both elevated in RA and significantly reduced following thyme extract treatment.

LINC01232 regulates miR-516a-5p/BCL9 axis to promote triple-negative breast cancer progression

wei liu — 2025-07-30

Triple-negative breast cancer (TNBC) is characterised by an absence of the oestrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2), for which there are few therapeutic options and the prognosis is poor. This research sought to explore the particular function of the long non-coding RNA (lncRNA) LINC01232 in TNBC and its regulatory impacts on the miR-516a-5p/BCL9 pathway. In this study, quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to determine the expression level of LINC01232 in TNBC tissues. We also examined its regulatory influences on miR-516a-5p and BCL9 via cellular function tests and a luciferase reporter experiment. Evaluated the effect of LINC01232 silencing on proliferation, migration and invasion of breast cancer cells. Results showed that LINC01232 expression was abnormally high in TNBC tissues in comparison to normal tissues. Inhibition of LINC01232 expression markedly impeded breast cancer cell proliferation, clone formation, migration and invasion. We found that LINC01232 competes with miR-516a-5p for binding, thereby reducing its expression and subsequently increasing BCL9 expression. In conclusion, our results indicate that LINC01232 facilitates the malignant development of TNBC through the miR-516a-5p/BCL9 pathway, providing fresh perspectives on the pathogenesis of TNBC and pinpointing potential therapeutic targets.

Molecular mechanisms underlying the antitumor activity of human and bovine alpha-lactalbumin-oleic acid complexes in a murine mammary adenocarcinoma model

Muataz Mohammed Al-Taee — 2025-07-30

The alpha-lactalbumin-oleic acid complexes (BAMLET and HAMLET) derivatives have shown remarkable anticancer capabilities in various preclinical studies with potential applications in oncology. The current study investigates the anti-cancer activity of synthesized human alpha-lactalbumin oleic acid (HAMLET) and bovine alpha-lactalbumin oleic acid (BAMLET) complexes on AN3 mouse mammary adenocarcinoma cancer cell line, a long-standing model for cancer research. We investigated multiple therapeutic endpoints including tumor volume reduction, survival rates, histopathological changes, as well as the molecular mechanisms allying the treatment response. A significant suppression of tumor growth was observed in both groups treated with HAMLET and BAMLET when compared with the control group, with HAMLET showing slightly better effectiveness in tumor growth inhibition. Histological examination revealed tumor necrosis, apoptosis, and decreased cell proliferation in treated mice, cancer cells were dying and also growth architecture was disrupted which indicates that both complexes cancer cell death. Thus, we investigated major molecular pathways associated with the anticancer activity of these compounds. Findings revealed the activation of supportive apoptotic pathways alongside downregulation of fundamental oncogenes linked to growth endurance and metastasis. Likewise, the safe response was augmented in the treated groups as shown by greater infiltration of immune cells into the tumor microenvironment. Survival rates were significantly higher in the HAMLET and BAMLET treatment groups compared to control, suggesting that these assemblies may prolong survival by effectively reducing cancer burden. The results highlight once again the exceptional breast cancer treatment efficacy of the alpha-lactalbumin-oleic acid complex.

Phytochemical screening and evaluation of antibacterial and antifungal activities of Callistemon viminalis cultivated in Iraq

Zaineb Aziz Ali — 2025-07-30

Callistemon viminalis, a member of the Myrtaceae family, is traditionally used to treat infections, respiratory, and gastrointestinal disorders. This study aimed to investigate the phytochemical profile and evaluate the antibacterial and antifungal activities of C. viminalis leaves cultivated in Iraq. Leaves were collected, dried, and extracted sequentially with hexane and 70% ethanol. Preliminary phytochemical screening revealed the presence of saponins, terpenoids, alkaloids, and flavonoids. The antimicrobial activity of the ethanol extract was assessed using the agar well diffusion method against three Gram-positive bacteria (Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus sp.), two Gram-negative bacteria (Escherichia coli, Klebsiella sp.), and one fungal strain (Candida albicans). The ethanol extract exhibited notable inhibitory effects, particularly against Gram-positive bacteria and C. albicans, with activity increasing in a concentration-dependent manner. GC-MS analysis of the hexane extract identified key bioactive compounds, including beta-sitosterol and vitamin E. These findings highlight the significant pharmacological potential of C. viminalis leaves and support their traditional use as a source of natural antimicrobial agents. Further studies are recommended to isolate and characterize the active constituents responsible for these effects.

Impact of smoking and environmental toxins on diabetic retinopathy: role of trace elements, lipid profiles, and vitamin A

Nagham Hadi Alrikabi — 2025-07-30

Diabetic retinopathy (DR), a leading cause of adult blindness, is influenced by physiological factors such as lipid profiles, vitamin A, and trace elements, as well as environmental factors like smoking. This study investigated the relationship between HbA1C, glucose levels, trace elements (lead, zinc, selenium, magnesium), vitamin A, and lipid profiles (TC, TG, HDL, LDL, VLDL) in type 2 diabetes (T2D) and DR patients, considering the impact of smoking. The study, conducted at Nasiriya General Hospital and the Center for Endocrinology and Diabetes (May 2023–March 2024), included three age groups (15–35, 35–55, and 55–75 years) divided into smokers and non-smokers. Results showed significantly higher glucose, HbA1C, lipid and selenium levels in T2D and DR patients compared to controls (p≤0.05), with smokers exhibiting greater lead levels. Zinc and vitamin A were significantly lower in DR patients, particularly among smokers. The findings highlight smoking as a source of lead and other toxins that exacerbate DR and diabetes-related complications, emphasizing the critical link between environmental health and chronic disease management.

Sequencing of TNFα and IFNϒ genes associated with Toxoplasma gondii infection among Erbil residents-Iraq

Ahmed Akil Khudhair Al-Daoody — 2025-07-30

Toxoplasmosis is a serious disease that affects all age groups and may even threaten the lives of some people. Fifty serum samples were taken from patients has Toxoplasmosis who attended Rizgary Teaching Hospital in Erbil City, and 38 serum samples were taken from healthy individuals as a control group, with ages between 18-60 years old, from the period January 2024 to March 2025. The purpose of the research is to determine the gene sequence of TNFα and IFNϒ associated with Toxoplasmosis. The results showed that the infection prevalence rate in the age group (18-27) years was 17 (56.7%) in comparison to healthy individuals 13 (43.3%), and in the age group (28-37) years was 20 (62.5%) compared to healthy control 12 (37.5%), and in the age group (38-47) years was 10 (76.9%) in comparison to the controls 3 (23.1%), while in the age group (≥47) years was 3 (23.1%) compared to the controls 10 (76.9%) with significant variations (P=0.03). The prevalence rate of Toxoplasmosis in males was 17 (48.6%) in comparison to the healthy controls 18 (51.4%), while in females it was 33 (62.3%) compared to the healthy individuals 18 (51.4%), with no significant variations (P=0.29). The mean level of Toxoplasmosis IgM was (2.28±0.18) when compared to healthy group (0.12±0.03), and the mean level of Toxoplasmosis IgG was (2.12±0.18) when compared to healthy control (0.09±0.02). The mean level of TNF-α (pg/ml) was (10.34±0.39) in comparison to the control group (4.89±0.31), and the mean level of INF-γ (pg/ml) was (10.72±0.36) in comparison to the controls (4.80±0.29) with highly significant variations (P=0.001). Moreover, direct correlation between Toxoplasma IgM with IgG was (r=.568), with TNF-α was (r=.607), with INF-γ was (r=.528), with highly significant variations P= (.000, .000, .000) respectively. Also, there were direct correlations between Toxoplasma IgG with IgM (r=1), with TNF-α (r=.550), with INF-γ (r=.576), with highly significant variations P= (.000, .000, .000) respectively. The TNFα gene sequence ID 7132 of rs767455 in the position AA was changed to AG and GG, respectively, in samples 1-10 in comparison to the control group. Also, the INFϒ gene sequence ID 3458 of rs2430561 in the position TT was changed to TA and AA, respectively, in samples 1-10 in comparison to the control group.

Molecular evaluation of quercetin effects in a murine model of giant cell tumor of bone: an in vivo pilot study

Dalia Lizbeth Monroy-Quiroz — 2025-07-30

Quercetin, a flavonoid derived from plant sources, has been extensively studied for its numerous biological properties, particularly its potential antitumor action against various malignant neoplasms. In our experience with a giant cell tumor of bone cell line (TIB-223), we demonstrated that quercetin has the ability to induce apoptosis via caspase-3. Therefore, this study aimed to evaluate molecular markers for apoptosis, necrosis, and cell proliferation in a murine model of giant cell tumor of bone, to determine whether the behavior reported for quercetin in 2D remains consistent in a 3D in vivo tumor model. Tumor constructs based on TIB-223 cells were implanted into athymic mice, and two weeks post-implantation, the mice were orally administered quercetin at a concentration of 100 mg/kg body weight once a day for two weeks. The control group received only 200 µL of the vehicle. Our results demonstrate the activation of two cell death pathways in the implanted tumors: apoptosis, via Caspase-8 to Caspase-3 activation, and necroptosis, via RIPK1. No significant effect on cell proliferation was observed, as PCNA expression remained unchanged. Our results suggest that quercetin may induce specific mechanisms of cell death without significantly altering cell proliferation in the tumor model induced in mice.

Modern methods of detecting mitophagy

Alexander V. Blagov — 2025-07-30

Inhibition of mitophagy is one of the signs of chronic disease pathogenesis. Detection and measurement of mitophagy levels under in vitro and in vivo models provide a better understanding of the role of mitophagy disorder in disease development and serve as prerequisites for creating a clinically applicable system test. The development of such a system is potentially feasible, but taking into account a number of factors that will be discussed in detail in this article. Here it is considered the main models of mitophagy-based test systems and an analysis is carried out showing their advantages and disadvantages. The future potential for the development of mitophagy-based diagnostic test systems is also discussed here.

Analysis of the prospects of new therapeutic agents for the treatment of rheumatoid arthritis

Alexander V. Blagov — 2025-07-30

Rheumatoid arthritis (RA), a chronic autoimmune disease, is one of the major research themes in medicine. The current therapies have their limitations and cannot completely cure RA, but new therapeutic strategies are being proposed to reduce the shortcomings of approved drugs. This review will consider new potential treatment strategies for RA, including T-cell therapy, genetic editing and epigenetic regulation, what advantages and disadvantages they have and to what pathological target in RA they are directed.

The use of monoclonal antibodies for the treatment of atherosclerosis: current status and prospects

Alexander V. Blagov — 2025-07-30

Atherosclerosis remains a leading cause of cardiovascular morbidity and mortality worldwide, underlying major conditions such as coronary heart disease and stroke. The pathogenesis of atherosclerosis is tightly linked to chronic inflammation and dysregulated lipid metabolism, processes that are also implicated in other inflammatory diseases like rheumatoid arthritis and psoriasis. Monoclonal antibodies (mAbs) have emerged as a promising therapeutic strategy, offering targeted intervention against key molecular drivers of atherosclerosis. This review summarizes recent advances in the development and clinical application of mAbs targeting both lipid-lowering pathways—such as low-density lipoprotein (LDL) and proprotein convertase subtilisin/kexin type 9 (PCSK9)—and inflammatory mediators, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-17 (IL-17). Notably, anti-PCSK9 antibodies like alirocumab and evolocumab have demonstrated significant reductions in LDL-C levels and cardiovascular events in large-scale clinical trials. Similarly, antibodies targeting inflammatory cytokines have shown efficacy in reducing vascular inflammation and associated risks. The review also discusses the advantages and limitations of therapeutic mAbs, such as their high specificity, potential for adverse immune responses, and challenges related to tissue penetration and cost. Overall, monoclonal antibody therapy represents a significant advancement in the management of atherosclerosis, with ongoing research aimed at optimizing efficacy, safety, and accessibility. Future directions include the development of novel mAbs and combination therapies to further improve cardiovascular outcomes in patients with atherosclerotic disease.