Articles in Press

Background: Aspergillus fumigatus is the most common species causing invasive aspergillosis (IA), a life-threatening infection with more than 80% mortality. Interactions between A. fumigatus and human blood platelets lead to intravascular thrombosis and localized infarcts. To better understand A. fumigatus pathogenesis, we aimed to analyze the genetic basis of interactions between the pathogen and blood platelets. Methods: A bioinformatic pipeline on microarray gene expression dataset, including analysis of differentially expressed genes (DEGs) using Limma R package and their molecular function, as well as biological pathways identification, was conducted to find the effective genes involved in IA. In the wet phase, the gene expression patterns following fungal exposure to blood platelets at 15, 30, 60, and 180 min were evaluated by quantitative reverse transcriptase-PCR analysis. Results: Three genes encoding aspartic endopeptidases including (Pep1), (Asp f 13), and (β-glucanase) were the standing candidates. The invasion-promoting fungal proteinase-encoding genes were down-regulated after 30 min of hyphal incubation with blood platelets, and then up-regulated at 60 and 180 min, although only Pep1 was greater than the control at the 60and 180 min time points. Also, the same genes were downregulated in more the clinical isolates relative to the standard strain CBS 144.89. Conclusions: Our findings delineate the possible induction of fungal-encoded proteinases by blood platelets. This provides a new research line into A. fumigatus’ molecular pathogenesis. Such insight into IA pathogenesis might also guide researchers toward novel platelet-based therapies that involve molecular interventions, especially in IA patients.

Background: Critical role of interleukin-7 (IL-7) in homeostatic proliferation and survival of T-cells has made it a promising cytokine for various clinical conditions especially those associated with lymphopenia. Methods: In the present study we reported the expression of recombinant human interleukin-7 (rhIL-7) in Chinese Hamster Ovary ( CHO)-K1 cell as a mammalian expression system. Circular and linearized forms of pBud-hIL-7 recombinant vector, stably transfected into CHO-K1 cells by electroporation. The expression of rhIL-7 molecules in CHO-K1 was confirmed by Enzyme-linked Immunosorbent Assay (ELISA), Dot blot, and Western blot. Results: In Western blot analysis a single band of expressed rhIL-7 was observed at 25KD which is consistent with the expected molecular weight of the glycosylated hIL-7. No significant difference was observed in expression level between transfected cells with circular and linear plasmids. Conclusion: In conclusion, we could establish a stable CHO-K1 cell line expressing rhIL-7 which can be considered as a promising system for producing rhIL-7 as a biopharmaceutical.

Abstract Background: Cisplatin (Cis) is a chemotherapeutic drug recognized for its hepatotoxic effects and its potential to trigger oxidative stress. This research aimed to evaluate the protective function of naringenin (NG) in mitigating Cis-induced liver toxicity by examining the inflammatory pathways and oxidative stress in mice. Methods: Adult male mice were given 50 mg/kg NG for two weeks, without Cis or with 10 mg/kg Cis on the 10th day. Histological analysis, biochemical indicators, and oxidative stress biomarkers were evaluated. The protein levels of Nrf2 and NF-κB were also measured to assess inflammation within the liver tissue. Results: The current research revealed a notable rise in the levels of Nrf2, AST, ALT, ALP, and MDA, alongside a marked reduction in the levels of NF-κB, CAT, GSH, and SOD in the Cis group. Pretreatment with NG resulted in a decrease in biochemical markers, histological alterations, and oxidative stress. NG was found to lower Nrf2 protein levels while elevating the levels of NF-κB protein in the Cis-intoxicated mice. Conclusion: The findings indicated that NG exerts a protective influence against Cis-induced hepatotoxicity by reducing histological alterations, oxidative stress, and inflammation.

Background: Infertility is a multifactorial disorder influenced by hypoxia signalling, immune modulation, and angiogenesis, which collectively determine endometrial receptivity and implantation success. Key mediators, including Hypoxia-Inducible Factor-2α (HIF-2α), Interleukin-13 (IL-13), and Vascular Endothelial Growth Factor-A (VEGF-A), play essential roles in oxygen sensing, immune tolerance, and angiogenesis, respectively. However, their coordinated expression and diagnostic potential in infertility, particularly among Middle Eastern women, remain insufficiently understood. Objective: This study aimed to estimate the expression levels of HIF-2α and IL-13 genes and serum VEGF-A, and to assess their correlations and diagnostic performance in infertile women compared with fertile controls. Materials and Methods: A case-control study was conducted involving infertile patients and age-matched fertile women. Gene expression was analysed using real-time quantitative PCR (RT-qPCR) with melt curve validation, and serum VEGF-A concentrations were determined by ELISA. Results: Findings revealed significant downregulation of HIF-2α in infertile women (fold change = 0.189, p = 0.038), while IL-13 expression showed a non-significant decrease (fold change = 0.69, p = 0.458). Serum VEGF-A levels were significantly lower in infertile women than in controls (310 ± 60 vs. 480 ± 44 pg/ml, p≤0.005). ROC analysis indicated high sensitivity but poor specificity for all three markers. A positive correlation was observed between HIF-2α and IL-13, whereas VEGF-A showed no significant correlation with either gene. Conclusions: Infertility reflects disrupted molecular interactions among hypoxia sensing, immune regulation, and angiogenic pathways. Although single markers such as HIF-2α, IL-13, and VEGF-A show limited diagnostic accuracy, integrating them into multi-marker panels could enhance predictive value and guide the development of targeted therapies to improve endometrial receptivity and implantation outcomes.

Background: Heavy metals and trace elements have a dual effect on biological systems, acting as either essential cofactors or toxicants depending on the dose. While cadmium (Cd) and cobalt (Co) are recognized environmental toxins, zinc (Zn) is an essential micronutrient with antioxidant and immunomodulatory effects. Objective: This study aimed to examine the histological and inflammatory changes in the liver and kidney tissues of rats exposed to Cd, Co, or Zn, and to evaluate possible transgenerational effects on first-generation (F1) offspring. Methods: Twenty Wistar rats were randomly divided into control, Cd-treated, Zn-treated, and Co-treated groups, with two males and three females each. Parental rats (P0) were given oral doses of cadmium nitrate (15 mg/kg), zinc (50 mg/kg), or cobalt(II) chloride (50 mg/kg), while controls received distilled water. Histopathological changes in liver and kidney tissues were examined using hematoxylin and eosin staining. Inflammatory biomarkers (NF-κβ and IL-6) were measured in both P0 and F1 generations via ELISA, adjusted for total protein content. Results: Cd and Co exposure in P0 rats caused severe liver and kidney damage, including hepatocyte degeneration, inflammatory infiltration, tubular necrosis, and glomerular alterations, along with significant increases in NF-κB and IL-6 levels (p < 0.01). Zn treatment preserved tissue structure and kept biomarker levels close to control values. In F1 offspring, Cd and Co groups showed reduced but still noticeable histological abnormalities, with moderate changes in NF-κB and IL-6, while Zn offspring mainly remained similar to controls. Conclusion: Cd and Co cause significant hepato-renal toxicity through NF-κβ/IL-6-mediated inflammatory pathways, with some effects passing to the next generation. Conversely, Zn supplementation protected against both histological and biochemical disturbances in the P0 and F1 groups. These findings highlight the dual role of trace elements and the importance of maintaining balanced Zn levels in reducing heavy metal toxicity and protecting liver and kidney health across generations.

Background: Osteoarthritis (OA) is the most common type of arthritis, characterized by inflammation that can affect the joint structures, leading to pain and disability, and is directly linked to ageing. As an epigenetic risk factor, microRNAs (miRNAs), small non-coding RNAs, can play a role in OA pathogenesis by regulating genes involved in processes such as inflammation, apoptosis, and autophagy. Due to the unique therapeutic potential of curcumin, isolated from Curcuma longa, particularly its anti-inflammatory properties, the current study aimed to investigate the effect of nanomicelle formulation of curcumin, named SinaCurcumin&reg;, on the expression of miRNA-21 and miRNA-34a genes in OA patients. Methods: Thirty female patients with osteoarthritis were randomly assigned to two equal groups, who received nanocurcumin (n=15) and placebo (n=15) in a double-blind clinical trial for 3 months. The gene expression levels of miRNA-21 and miRNA-34a were evaluated using the SYBR&reg; Green Real-Time PCR method. The association between the changes in expression of miRNA-21 and miRNA-34a and the changes in clinical and laboratory indexes in OA patients was analyzed using Spearman's rank correlation. Results: In the nanocurcumin group, the gene expression of miRNA-21 and miRNA-34a was significantly reduced, while no significant changes were observed in the placebo group. Also, there was a significant positive correlation between the downregulation of miRNA-21 and miRNA-34a and a decline in clinical and laboratory indexes (Visual Analog Scale and C-reactive protein) in patients receiving nanocurcumin. Conclusion: Curcumin may provide a promising therapeutic perspective for improving inflammation in OA, through its modulatory effects on miRNA-21 and miRNA-34a.

Abstract Background: Non-Hodgkin Lymphoma (NHL) is a malignancy with a significant genetic component. The IKAROS family zinc finger 1 (IKZF1) gene is a critical regulator of lymphoid development, and its polymorphisms have been implicated in haematological cancers. This study aimed to evaluate the association of IKZF1 rs4132601 T>G and rs11978267 A>G polymorphisms, individually and as haplotypes, with NHL risk in a Southeast Iranian population. Materials and Methods: This case-control study included 195 NHL patients and 200 age- and sex-matched healthy controls. Genotyping was performed using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using logistic regression under codominant, dominant, and recessive genetic models. Haplotype analysis was also conducted. Results: The rs4132601 G allele was associated with a significantly reduced risk of NHL, with the GG genotype showing a protective effect in codominant and recessive models. Conversely, the rs11978267 G allele was associated with an increased risk, with AG and GG genotypes conferring higher risk in codominant and dominant models. Haplotype analysis revealed that the GA haplotype significantly decreased NHL risk compared to the TA reference haplotype. Conclusion: Our findings suggest that IKZF1 polymorphisms have a significant influence on NHL susceptibility. The rs4132601 G allele exerts a protective effect, while the rs11978267 G allele is a risk factor. The protective GA haplotype highlights the importance of evaluating combined genetic variants. These results emphasise the role of IKZF1 germline variation in NHL pathogenesis.

Background: The renin-angiotensin system (RAS) and advanced glycation end-products (AGEs) contribute to diabetes pathogenesis by activating AT1R and RAGE, respectively. This study investigated the effects of crocin and losartan, alone and in combination, on renal AT1R and RAGE expressions in STZ-induced diabetic rats. Methods: Diabetes was induced in 40 male Wistar rats with STZ (50 mg/kg, i.p.). Rats were assigned to five groups: healthy control, diabetic control, and three diabetic treatment groups receiving crocin (50 mg/kg), losartan (25 mg/kg), or crocin plus losartan for 8 weeks. Fasting blood glucose (FBG), serum urea, creatinine, and TNF-α levels were measured. Renal AT1R and RAGE gene expressions were analyzed by quantitative PCR. Results: Crocin significantly decreased FBG, serum urea, and creatinine levels. Combined crocin and losartan therapy further reduced urea compared to losartan alone. TNF-α levels were reduced in all treated groups, with the most pronounced decrease in the combination group. AT1R and RAGE expression were markedly downregulated, particularly in rats receiving the combined therapy, indicating enhanced anti-inflammatory and renoprotective effects. Conclusion: Crocin attenuates inflammation and downregulates RAGE and AT1R expression. Crucially, the combined administration of crocin and losartan produced a more pronounced and synergistic protective effect, suggesting its potential as an adjuvant therapy targeting the RAGE and AT1R expression axis in diabetes management.

Background: Human T-cell lymphotropic virus type 1 (HTLV-1) infects an estimated 10-20 million people worldwide and is the etiological agent of adult T-cell leukemia/lymphoma (ATLL), a highly aggressive cancer. The endoribonuclease L (RNase L) is a critical component of immune response that degrades viral and host RNA and inhibits protein synthesis, while ATP-binding cassette E1 (ABCE1) regulates its activity. Although the expression of RNase L and ABCE1 genes has been studied in various cancers and viral infections, their role in HTLV-1, particularly in ATLL context, remains unexplored. This study aims to determine the expression of RNase L and ABCE1 in ATLL patients, HTLV-1 asymptomatic carriers (ACs), and healthy controls (HCs). Methods: This study included 20 ATLL patients, 20 HTLV-1 ACs, and 10 HCs. DNA and mRNA were extracted from peripheral blood mononuclear cells (PBMCs) to assess HTLV-1 proviral load (PVL) and the expression of ABCE1 and RNase L genes using real-time PCR. Statistical analysis was performed with SPSS version 18 and GraphPad Prism 10.3.1. Results: ABCE1 and RNase L expression was significantly lower in ATLL patients compared to ACs and HCs (p 0.05). However, proviral load was significantly higher in ATLL patients compared to ACs (p>0.0001). Conclusion: The observed downregulation of ABCE1 and RNase L expression in ATLL suggest a profound dysregulation of this critical innate immune pathway, which may contribute substantially to ATLL pathogenesis. Despite this, the lack of correlation with proviral load indicates that other factors probably influence the expression of these genes. Further research is needed to explore the mechanisms of these genes in ATLL.

Abstract Background: Myeloid-derived suppressor cells (MDSCs) promote immune evasion and tumor progression in breast cancer. Pycnogenol&reg;, a standardized extract from Pinus pinaster bark, possesses antioxidant and immunomodulatory properties, but its effect on MDSCs in cancer remains unclear. Therefore, the aim of this study was to evaluate the effect of Pycnogenol on MDSC s function and tumor progression in a mouse model of breast cancer. Methods: Female BALB/c mice were inoculated with 4T1 mammary carcinoma cells and treated with Pycnogenol (50 mg/kg/day) for four weeks. MDSCs were isolated from tumor-bearing mice for analysis of nitric oxide (NO), reactive oxygen species (ROS), arginase activity, and gene expression (Arg1, iNOS, S100A8, S100A9). Tumor volume and survival were also assessed. Results: Pycnogenol significantly reduced Arg1 mRNA expression, NO production, and ROS levels in MDSCs. No significant changes were observed in arginase activity, iNOS, or S100A8/A9 expression. Pycnogenol modestly delayed tumor growth and slightly extended survival without reaching statistical significance. Conclusion: Pycnogenol attenuates key immunosuppressive functions of MDSCs in breast cancer, suggesting its potential as a supportive immunomodulatory agent. Further investigation is warranted to explore its role in combination with immunotherapies.

Background: Arsenic is widely found in the environment, and its toxicity is a significant public health issue worldwide. Syringic acid (SYRA) is a flavonoid compound with anti-inflammatory effects. Given that the effect of SYRA on sodium arsenite (As)-induced nephrotoxicity remains largely unknown, this study aimed to investigate the effect of SYRA on sodium arsenite-induced nephrotoxicity in mice by reducing oxidative stress, inflammation, and nuclear factor kappa B p-65 (NF-κB-p65) expression. Methods: Thirty male mice were split into five groups of six, based on a power analysis. The groups were set up like this: control (3 ml/kg normal saline); SYRA (25 mg/kg, administered via gavage); As (3 mg/kg, administered via gavage); As + SYRA (10 mg/kg, administered via gavage) and As + SYRA (25 mg/kg, administered via gavage). As was administered daily via gavage for a total of 30 days. SYRA was then given by gavage for 7 days, from day 23 to day 30. On the 30th day, the mice were put under anesthesia, and samples of their heart blood and kidney tissue were taken to check biochemical markers, oxidative stress, inflammatory factors, and the expression of NF-κB-p65 protein. Results: Giving SYRA lowered the levels of blood urea nitrogen, creatinine, and uric acid in the blood compared to the As group (P<0.001). In the SYRA group, levels of total thiol, catalase, superoxide dismutase, and glutathione peroxidase were higher compared to the As group (P<0.001). The SYRA group showed lower levels of thiobarbituric acid-reactive substances, nitric oxide, tumor necrosis factor alpha, and NF-κB-p65 protein expression compared to the As group (P<0.001). The histological results backed up these findings. Conclusion: Giving SYRA might help reduce kidney damage caused by As in mice by lowering oxidative stress, inflammation, and the expression of NF-κB-p65 protein.

Background: This study aimed to investigate the harmful effects of paraquat (PQ) on sperm and testicular tissue and the protective effect of geraniol against the toxin.   Methods: In this study, 28 small white male mice were gavaged with PQ and geraniol for one month. Testicular tissue was removed to evaluate gene expression, sperm parameters and histopathological changes in testicular tissue. Results: The relative gene expression results revealed that CAT and CYP17 gene expression significantly increased in groups 3, 4, 5, 6, and 7 (P<0.001). Additionally, the activity of the catalase enzyme increased. The expression level of the USP9Y gene increased in response to low concentrations of PQ. The sperm parameters, such as number, motility, and morphology, decreased after treatment with PQ. In the groups treated with PQ, the number of germinal layer cells was reduced and separated from the basement membrane. Malformed seminiferous tubules, thick myoepithelial cells, atrophy of Leydig cells, vascular congestion and fibrosis were observed. In the testicular tissue under the influence of two doses of PQ, the number and thickness of the epithelium of the seminiferous tubules, as well as the number of spermatozoa, were significantly reduced compared to the control. Additionally, the area of the seminiferous tubules and the number of spermatogonia and primary spermatocytes decreased slightly.  Following geraniol treatment, a degree of improvement was observed in the levels of certain histopathological factors and all testicular tissue parameters. Conclusion: PQ has adverse effects on testicular tissue and sperm, and geraniol has a protective effect against oxidants .

Abstract Background : Methotrexate (MTX) is a medication widely used in the treatment of various conditions, including cancer. One potential side effect of MTX is nephrotoxicity. The purpose of the present study was to evaluate the effects of betulinic acid (BA) on MTX-induced nephrotoxicity in mice through the TLR4/NF-κB signaling pathway. Methods : Four groups– control, BA 25 mg/kg, MTX 20 mg/kg, and BA+MTX- were considered for seven days to evaluate the effect of BA on MTX nephrotoxicity. Factors such as blood urea nitrogen  (BUN ) , serum creatinine, antioxidant enzymes, oxidative stress markers, and pro-inflammatory cytokines were investigated. Toll-like receptor 4 (TLR4) and nuclear factor-kappa B (NF-κB) expression levels were measured using Western blot analysis. Results: BUN and creatinine serum levels were significantly reduced in the BA-treated group compared to the MTX group. Furthermore, BA administration inhibited the activation of the TLR4/NF-κB signaling pathway, resulting in reduced expression of  TLR4 and NF-κB. This decreased the release of pro-inflammatory cytokines linked to kidney inflammation, such as interleukin-1β and tumor necrosis factor-alpha. Moreover, BA exhibited notable antioxidant properties by enhancing the activity of enzymes like catalase, superoxide dismutase, and glutathione peroxidase. This antioxidant effect was evident by decreasing the levels of oxidative stress markers, as measured by  the lipid peroxidation assay, and increasing the total thiol level. Conclusion: These findings suggest that BA can ameliorate MTX-induced nephrotoxicity by reducing oxidative stress and enhancing the antioxidant defense system. It effectively reduces renal damage, inflammation, and oxidative stress. Our results demonstrate that BA's anti-inflammatory and antioxidant qualities could protect the kidneys from MTX's nephrotoxic effects.