Turkish journal of biology = Turk biyoloji dergisi最新文献

Background/aim: In an aging model established using male Wistar albino rats via the administration of D-galactose (D-gal), the aim of this study was to examine the effects of chelidonic acid (CA) on cognitive function and the levels of glutathione (GSH), malondialdehyde (MDA), total antioxidant status (TAS), and brain-derived neurotrophic factor (BDNF).

Materials and methods: Thirty-two, three-month-old Wistar albino male rats (n = 8) were divided into four groups, as the control (C) group, CA group (2 mg/kg of CA via oral gavage), D-gal group (150 mg/kg of D-gal, subcutaneously), and D-gal + CA group (150 mg/kg of D-gal and 2 mg/kg of CA). Following overnight fasting, the 10-week trial was concluded with intramuscular injections of anesthetic drugs xylazine (8-10 mg/kg) and ketamine (80-100 mg/kg), and subsequently, the collection of cardiac blood. The brain tissues of the rats were removed. The GSH, MDA, TAS, and BDNF levels were determined in the collected serum samples and prepared tissue homogenates. Novel object recognition and Morris water maze (MWM) experiments were also used to evaluate cognitive function.

Results: The D-gal group demonstrated a statistically significant improvement in the discrimination index for memory in both the short and long term compared to the D-gal + CA group. Further analysis of the MWM data for these two groups indicated a notable decrease in the amount of time required for finding the platform. In comparison with the D-gal group, the MDA levels decreased in the CA and D-gal + CA groups, whereas the GSH, TAS, and BDNF levels increased in both the serum and hippocampus samples.

Conclusion: CA showed positive effects on age-related neurodegenerative disorders and memory-related processes, especially by increasing TAS and BDNF levels.

MEIS1, a member of the TALE-type homeobox gene family, has emerged as a pivotal regulator of cardiomyocyte cell cycle arrest and represents a promising therapeutic target. Our study reveals that inhibition of MEIS1 using two novel small molecules, MEISi-1 and MEISi-2, significantly enhances neonatal cardiomyocyte proliferation and cytokinesis. Specifically, MEISi-1 and MEISi-2 increased the proportion of proliferating cardiomyocytes (Ph3+TnnT cells) up to 4.5-fold and the percentage of cytokinetic cardiomyocytes (AuroraB+TnnT cells) by 2-fold, compared to untreated controls. MEIS1 inhibition resulted in a notable downregulation of MEIS1 target genes and cyclin-dependent kinase inhibitors, demonstrating its effect on key regulatory pathways. Additionally, the culture and differentiation of human induced pluripotent stem cells into cardiomyocytes were studied, with MEIS1 inhibition showing no adverse effects on cell viability. Extended treatment with MEIS inhibitors led to a substantial upregulation of critical cardiac-specific genes, including a 15-fold increase in the expression of Nkx2.5. This upregulation significantly impacted both cardiac mesoderm and cardiac progenitor cells. These findings underscore the potential of MEIS1 inhibitors as effective agents in enhancing cardiac regeneration and highlight their therapeutic promise in regenerative cardiology.

Background/aim: No specific pharmacological treatment regimen for idiopathic pulmonary fibrosis (IPF) exists. Therefore, new antiinflammatory therapeutic strategies are needed. Cannabinoids (CBs), known for their inflammation-modulating and antifibrotic effects, may be potential medication candidates for treating IPF. We aim to evaluate the inflammation-modulating and antifibrotic effects of CB receptor (CBR) agonists and antagonists in lipopolysaccharide-stimulated normal human lung fibroblast, epithelial cells, IPF fibroblast cells, and monocytes.

Materials and methods: We detected CBRs in normal human lung fibroblasts (LL24) and IPF fibroblast cells (LL29), epithelial cells (A549) and monocytes (THP-1) by flow cytometry. We determined TGF-β1, IL-8, and TNF-α inflammatory cytokines in the LL24, LL29, A549, and THP-1 cell culture supernatants on days 1 and 5 by ELISA. We evaluated the cell viability in LL24, LL29, and A549 cells on days 1, 3, and 5 spectrophotometrically and detected collagen Type I (ColI) production in the LL24 and LL29 cell culture supernatants on days 1, 3, and 5 by ELISA.

Results: LL24, LL29, A549, and THP-1 cells exhibited CB1 (CB1R) and CB2 (CB2R) receptors. CB1R and CB2R agonists WIN55,212-2 and JWH015 inhibited fibroblastic and epithelial cell proliferation on day 5. TGF-β1 and TNF-α release increased, while IL-8 release decreased in LL24, LL29, A549, and THP-1 cells in response to the administration of WIN55,212-2 and JWH015 at a 10^-2 mM concentration. CB1R and CB2R antagonists AM251 and AM630 did not block agonistic responses, suggesting a nonclassical CBR-mediated pathway. CB2R agonist JWH015 decreased ColI expression in IPF lung fibroblasts LL29 on day 3.

Conclusion: These results suggest that CB signaling regulates the progression of pulmonary inflammation and fibrosis via CBR activation. This may offer a potential pharmacological tool for developing antifibrosis therapies.

Background/aim: The hyperglycemic environment in diabetes disrupts normal wound-healing processes, leading to chronic wounds. This study investigated whether the combination of the phenolic compounds ellagic acid and carnosic acid shows synergistic effects on diabetic wound healing and oxidative parameters in diabetic rats.

Materials and methods: Diabetic rats were divided into control, untreated, Carbopol 974P treated, topical treatment, and oral gavage treatment groups. Ellagic acid and carnosic acid in combination were applied topically and as oral gavage to the full-thickness excisional wounds of all animals except for those in the control group. We investigated oxidative events with malondialdehyde, glutathione, nitric oxide, protein carbonyl, collagen spectrophotometrically, and matrix metalloproteinase 2 and 9 and advanced oxidation protein product levels using ELISA.

Results: The combination of ellagic acid and carnosic acid decreased malondialdehyde, nitric oxide, protein carbonyl, advanced oxidation protein product, and metalloproteinase 9 levels and increased the rate of wound contraction and collagen levels in the diabetic wound healing model.

Conclusion: The combined use of ellagic acid and carnosic acid showed a synergistic effect, enhanced wound healing, and decreased oxidative stress. This combination may provide effective therapy for chronic nonhealing wounds that occur as a complication of diabetes.

Background/aim: Previous studies on general transcription factor II E (GTF2E) showed that it is associated with certain groups of diseases, such as colon cancer and trichothiodystrophy, but the global effect of GTF2E on cellular processes is still not widely characterized. This study aimed to investigate and characterize the effect of GTF2E on the transcription level of genes and identify the cellular processes and diseases associated with GTF2E.

Materials and methods: The human colorectal carcinoma cell line HCT116 used in the study was transfected at a 30 nM concentration with siGTF2E1 or nontarget negative siRNA. After 72 h, cells were harvested and prepared for further analysis. A whole transcriptome analysis was performed on the HCT116 cell line obtained from the siGTF2E1 knockdown of the HCT116 cells (n = 3) and their nontarget negative siRNA controls (n = 3) using RNA sequencing. Cell viability was tested using an MTS assay.

Results: Compared with the control group, 166 genes were identified at the time of the GTF2E1 knockdown and expressed differentially in the knockdown group, including 66 upregulated genes and 100 downregulated genes. One significantly enriched Gene Ontology term was identified, involving carbohydrate binding. One oncogene related to B cell chronic lymphocytic leukemia (B-CLL) was identified. Five genes associated with colon carcinoma were determined. Eleven genes involved in the development of atherosclerosis were identified. GTF2E1 knockdown caused a decrease in cell viability.

Conclusion: The GTF2E1 knockdown group exhibited an altered expression of multiple genes, some of which are related to the development of atherosclerosis, colon carcinoma, and B-CLL. This might shed light on the different regulatory effects of GTF2E and its association with certain diseases.

Background/aim: Melanoma arises from the uncontrolled multiplication of melanocytes, and poses an escalating global health concern. Despite the importance of early detection and surgical removal for effective treatment, metastatic melanoma poses treatment challenges, with limited options. Among optional therapies, including chemotherapy and immunotherapy, all-trans retinoic acid (ATRA), a natural metabolite of vitamin A, has shown promise in treating melanoma by inducing differentiation, apoptosis, growth arrest, and immune modulation in melanoma cells. However, ATRA treatment alone can lead to resistance and relapse. Furthermore, sphingomyelin (SM) was implicated in the inhibition of cell proliferation, differentiation, and apoptotic cell death during melanoma progression.

Materials and methods: The combinational anticancer effects of ATRA and SM on an in vitro B16F10 melanoma model were investigated based on cell viability, apoptotic cell death, cell cycle progression, and gene expression levels; whereas the safety properties of the treatments were tested on RAW264.7 macrophages.

Results: The combination of 123 μM of ATRA + 136 μM of SM was the most effective treatment, showing a 50% reduction in cell proliferation, leading to 53.91% apoptotic cell death in 48 h, and G2/M phase-cell cycle arrest in the B16F10 cells. While 123 μM of ATRA alone did not change the caspase 3 and Bax gene expressions, the combinational ATRA + SM treatment resulted in 2- and 5-fold increases in the gene expression level, respectively. A 13-fold increase in cyclin-dependent kinase inhibitor 2A was observed with the combinational ATRA + SM treatment, while suppressing the programmed death ligand 1 (PD-L1) expression by 0.5-fold.

Conclusion: Combinational ATRA and SM therapy could be a promising therapeutic approach for melanoma, potentially improving efficacy, while reducing toxicity to healthy cells.

Background/aim: Quercetin (Q) is a compound that can inhibit the growth of cancer cells in the colon; however, to do so, a high dose is needed, requiring a drug delivery system to target cancer endothelial cells directly. This study investigates the potency of nanodiamond-conjugated quercetin (NDQ) as an anticancer drug against colon cancer in Rattus norvegicus induced by N-methyl N-Nitrosourea (MNU).

Materials and methods: This study is experimental-based and was designed using a six-group treatment method, namely normal control (KN: not treated by MNU, nanodiamond (ND), or Q); negative control (K-: treated by MNU); positive control (K+: treated by MNU and capecitabine); ND (treated by MNU and NDs); Q (treated by MNU and Q); and NDQ (treated by MNU and NDQ). To induce colon cancer in rats, MNU (10 mg/Kg BW) was administrated intrarectally three times per week for four weeks. The treatment of Q (40 mg/Kg BW) or NDQ (40 mg/Kg BW) was given intraperitoneally twice a week for 6 weeks. Cancer progression of all cohorts was evaluated by performing body and colon weight measurements, which involved the following: ELISA assay-specific to metastatic marker matrix metalloprotein-9 (MMP-9), carcinoembryonic antigen (CEA), hypoxia-inducible factor 1 α (HIF1α), vascular endothelial growth factor, protein 53 (p53) and immunohistochemistry staining of Caspase-3 and Ki-67 proteins. Observation of cancer metastasis to the lung was also performed.

Results: NDQ significantly inhibited cancer aggressiveness by causing an increment in body weight gain and the growth rate-while reducing the colon weight compared to the K- group. Moreover, decreased levels of MMP-9, CEA, HIF-1α, and Ki67 and increased levels of p53 and Caspase-3 were more significant in the NDQ group than in the Q group. The lung tumor metastases in the NDQ group were fewer than in the K- group.

Conclusion: NDQ increased Q's anticancer activity, suggesting that NDs have an effective drug delivery property.

Background/aim: The conserved phosphatase Cdc14 facilitates mitotic exit in budding yeast by counteracting mitotic cyclin-dependent kinase activity. Cdc14 is kept in the nucleolus until anaphase onset, when it is released transiently into the nucleoplasm. In late anaphase, Cdc14 is fully released into the cytoplasm upon activation of the mitotic exit network (MEN) to trigger mitotic exit. Cdc14 also localizes to yeast spindle pole bodies (SPBs) in anaphase and dephosphorylates key targets residing on SPBs to allow SPB duplication and prime the MEN. Protein phosphatase 1 (Glc7) with regulatory subunit Bud14 is another phosphatase that plays a key role in the spatiotemporal control of mitotic exit. In this study, we investigated the regulation of Cdc14 localization by Bud14-Glc7.

Materials and methods: We used fluorescence microscopy to analyze Cdc14 localization in BUD14 wildtype and BUD14 knockout cells (bud14Δ) as well as in cells expressing a mutant allele of BUD14 (bud14-F379A) that cannot bind Glc7. We also utilized a yeast two-hybrid (Y2H) system to examine the interaction of Bud14 with Cdc14.

Results: We found that Cdc14 remains at the SPBs longer in bud14Δ and bud14-F379A compared to wildtype cells. This effect is limited to the SPB that has migrated to the daughter cell (dSPB). Cdc14 localizes to both SPBs shortly after anaphase onset. In mid-to-late anaphase, levels of Cdc14 increase at the dSPB in both wildtype and bud14Δ cells. With mitotic exit, Cdc14 disappears from the dSPB in wildtype cells but not in bud14Δ cells. Accordingly, 50% of bud14Δ cells in G1 have Cdc14 at their SPBs. We also found that Cdc14 localization at the dSPB was largely, but not entirely, dependent on Bfa1 in bud14Δ cells. Furthermore, Bud14 interacted with Cdc14 in the Y2H system.

Conclusion: Our results suggest that Glc7-Bud14 is part of a mechanism that promotes Cdc14 disappearance from the dSPB.

Breast cancer is one of the most common cancers and a significant cause of death in females worldwide. For effective breast cancer treatment, using systems with a promising delivery of anticancer agents is an important strategy. Peptide 18 (P18), a tumor-homing peptide, shows a high binding affinity toward breast cancer cells. Nanoliposomes are known to have enhanced accumulation ability in tumors with longer systemic circulation. In this study, Poly (2-ethyl-2-oxazoline) (PEtOx) polymers conjugated with DOPE are used to prepare PEtOx-DOPE nanoliposomes. BikDD, a mutant form of the Bik gene and a member of the BH3-only proapoptotic genes, mimics the constitutively phosphorylated form of the gene. To the best of our knowledge, this study presents a novel approach by investigating P18-conjugated PEtOx-DOPE nanoliposomes (P18-PEtOx-DOPE) for the targeted delivery of BikDD to the AU565 breast cancer model. A site-directed mutated BikDD was loaded into P18-PEtOx-DOPE nanoliposomes, and the targeted drug delivery system was assessed in in vitro and in vivo breast cancer models for efficiency, safety, and efficacy. The increased Bik mRNA expression levels in AU565 cells suggest a high effectiveness of the targeting PEtOx-DOPE nanoliposomes. Following the in vitro studies, the delivery of BikDD by P18-PEtOx-DOPE nanoliposomes was analyzed in CD-1 nude mice models. The animal study showed no significant difference in the tumor volume of the CD-1 nude mice treated with P18-PEtOx-DOPE-BikDD nanoliposomes compared to the free delivery of BikDD. Our preclinical studies suggest that P18-PEtOx-DOPE-BikDD nanoliposomes may be promising gene carriers for targeted breast cancer therapy. Thus, further studies should be carried out to determine the prolonged use of this drug delivery system in breast cancer therapy.

Background/aim: Sugarcane mosaic virus (SCMV; genus Potyvirus and family Potyviridae), poses a significant threat to global sugarcane cultivars, including those in Pakistan. The aim of this study was to develop a rapid and effective diagnostic tool for detection of SCMV, enabling timely implementation of control measures to mitigate potential yield losses.

Materials and methods: The study focused on the in silico analysis, physicochemical properties, immunogenicity, and subcellular localization of the SCMV coat protein (CP). The SCMV CP gene was synthesized, cloned, and expressed in Escherichia coli. The recombinant fusion CP (rFCP-SCMV) was purified and used to generate polyclonal antibodies (pABs) in mice. The immunogenicity of the antibodies was evaluated through indirect ELISA and RT-PCR.

Results: Epitope prediction using tools like the OptimumAntigen design tool from GenScript, BepiPred, and IEDB identified key B-cell epitopes on the SCMV CP, enhancing the specificity of the antibodies. Structural modeling with SWISS-MODEL and PyMOL provided insights into the 3D structures of viral proteins and their epitopes, aiding in the design of high-affinity antibodies. Molecular docking studies simulated the interaction between antibodies and viral epitopes, enabling the selection of optimal antibody candidates. The synthesized recombinant fusion CP (rFCP-SCMV) was used to produce pAbs in mice. These antibodies exhibited high sensitivity, detecting as low as 100 pg of SCMV protein in indirect ELISA. They also effectively identified SCMV in infected sugarcane field samples, confirmed by RT-PCR. The antibodies maintained high specificity and sensitivity even at a 1:10,000 dilution, proving their efficacy in recognizing both the recombinant protein and virus particles in plant sap.

Conclusion: The study reveals a rapid, effective immunodiagnostic technique for detecting SCMV in sugarcane cultivars, offering an accurate alternative to conventional virology methods, reducing contamination risk, and providing a valuable tool for mitigating yield losses.