Archives of Biological Sciences

CORRIGENDUM: Bobinac M, Šušić N, Šijačić-Nikolić M, Kerkez Janković I, Veljović-Jovanović S. Photosynthetic insights into winter-green leaves in Quercus pubescens Willd. seedlings Arch Biol Sci. 2024;76(2):223-32

Editorial team — Fri, 20 Dec 2024 00:00:00 +0100

The authors of the article: Bobinac M, Šušić N, Šijačić-Nikolić M, Kerkez Janković I, Veljović-Jovanović S. Photosynthetic insights into winter-green leaves in Quercus pubescens Willd. seedlings. Arch Biol Sci. 2024;76(2):223-32. have notified the Editorial Office of an error in the Funding section. The name “Ministry of Education, Science and Technological Development of the Republic of Serbia” was incorrectly stated and should instead read “Ministry of Science, Technological Development and Innovation of the Republic of Serbia.” The authors have requested this correction, and as a result, the journal is publishing this Corrigendum. The correct funding source is: This research was funded by the Ministry of Science, Technological Development and Innovation of the Republic of Serbia, Contracts 451-03-66/2024-03/200053 and 451-03-65/2024-03/200169, and by the Secretariat for Environmental Protection of the City of Belgrade, Project: Identification and Monitoring of the Gene Pool of Rare, Vulnerable, and Endangered Plant Species in NM Kosutnjak Forest.

Link to the corrected article: https://doi.org/10.2298/ABS240417015B

Biochemical and toxicological evaluation of Solanum viarum fruit extract on Dugesia tigrina

Felipe Schaly, Welligton Luciano Braguini — Fri, 20 Dec 2024 00:00:00 +0100

Paper description:

The environmental risk of Solanum viarum fruit extract on planarians was examined because of its toxicity to livestock.
Dugesia tigrina served to evaluate locomotor velocity, reproduction, glycogen content, antioxidant enzyme and acetylcholinesterase activities; phytochemical analysis and toxicity assays were performed.
AEF from viarum reduced planarian locomotion, fecundity, fertility, glycogen content, and affected enzymes.
This study highlights the need for environmental risk assessments and future research on natural compounds extracted from viarum.

Abstract: Solanum viarum has alkaloid-rich fruits. We evaluated the antioxidant and chelating activities of the aqueous extract fraction (AEF) from S. viarum fruits and its effects on the brown planarian Dugesia tigrina. The AEF demonstrated an iron-chelating effect comparable to ascorbic acid but lower than EDTA and a significant reducing power. The 96-h LC₅₀ for planarians was 1.22 g/L. At a concentration of 0.50 g/L, the AEF reduced planarian locomotor velocity by 34.7%, while decreasing fecundity and fertility by 98.4% and 96.7%, respectively, leading to a significant decline in the number of cocoons and emerging planarians. Cocoon hatching time was increased at all AEF concentrations. Planarians exposed to 0.50 g/L AEF for 14 days showed a 62.3% reduction in glycogen content. After 14 days, catalase was inhibited at all concentrations, with 83.2% inhibition at 0.50 g/L. Superoxide dismutase (SOD) increased by 60.8% and 59.9% after 24 h and 14 days, respectively, at 0.50 g/L AEF. Catalase (CAT) was stimulated by 72.1% and 44% at 0.10 and 0.25 g/L AEF, respectively, but inhibited by 25.2% at 0.50 g/L after 24 h. The iron-chelating activity of the AEF reduces iron availability for catalase, an iron-dependent enzyme, leading to its inhibition, while also limiting Fenton and Haber-Weiss reactions, resulting in hydrogen peroxide accumulation and subsequent stimulation of SOD under prolonged exposure. Acetylcholinesterase (AChE) was stimulated by 40.6% and 44.8% after 24 h and 14 days, respectively, at 0.50 g/L AEF. These findings show that S. viarum is harmful to planarians and may pose environmental risks, highlighting the need for toxicity assessments in other species.

Association of fatty acid desaturase 2 gene polymorphism (rs28456) with susceptibility to bipolar disorder in the Turkish population: a case-control study

Dilek Pirim, Ceren Gümüş, Esra Boztepe, Emine Merve Akdağ, Hande Şirin — Fri, 20 Dec 2024 00:00:00 +0100

Paper description:

Variations in the genes related to fatty acid metabolism may contribute to the risk of bipolar disorder (BD).
The dysregulation of fatty acid desaturase 2 (FADS2) in BD patients indicates that it may be involved in BD development.
Limited population-specific research suggests a potential association between the regulatory gene variant rs28456 in FADS2 and BD.

FADS2 rs28456 may be associated with BD in a gender-specific manner and could potentially play a role in BD development as a non-modifiable risk factor.

Abstract: The FADS2 gene encodes a key, rate-limiting enzyme involved in polyunsaturated fatty acid (PUFA) metabolism. Recent studies suggest that changes in plasma PUFA levels can lead to disruptions in the neurotransmission system and increase the risk of mood disorders. FADS2 variations may contribute to the individual risk of developing bipolar disorder (BD). We investigated the association of regulatory FADS2 rs28456 with BD in the Turkish population. We performed TaqMan genotyping on 100 patients with BD and 91 healthy controls. Our results did not show significantly different genotype or allele frequencies of rs28456 in the BD cases compared to controls. However, we stratified the cases based on family history, which revealed that minor rs28456-G was observed more frequently (P=0.056) in cases without a family history of psychiatric illness compared to those with a family history of psychiatric illness. A marginally significant difference in the distribution of the “G” allele (P=0.053) between male patients and healthy males without a family history was observed. Our findings did not provide strong evidence supporting the reported association between rs28456 and BD, yet they point to its potential gender-specific effect, which requires further investigation. Future studies are necessary to explore the impact of FADS2 variations on BD risk in larger study groups, considering their potential interaction with non-inherited risk factors.

Mesenchymal stem cell-derived extracellular vesicles enriched with miR-124 exhibit anti-inflammatory effects in collagen-induced arthritis

Fri, 20 Dec 2024 00:00:00 +0100

Paper description:

Rheumatoid arthritis (RA) is a chronic inflammatory disease treated with limited success. Mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) may help by secreting anti-inflammatory factors.
We investigated whether human umbilical cord MSC-EVs overexpress miR-124 using lipofectamine 3000 could enhance their therapeutic potential.
EVs enriched with miR-124 reduced disease severity in a mouse model of arthritis, by modulating inflammatory cytokines.
MSC-EVs that overexpress specific miRNAs offer a promising cell-free approach for optimizing the treatment of inflammatory joint diseases.

Abstract: Rheumatoid arthritis (RA) is a chronic autoimmune disorder characterized by joint inflammation, leading to pain, swelling, stiffness, and joint damage. Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) hold significant promise as therapeutic agents owing to their potent paracrine activities. This study investigated the anti-inflammatory effects of human umbilical cord-derived MSC (hUCSC)-EVs overexpressing miR-124 in collagen-induced arthritis (CIA). hUCSCs were transfected with miR-124, and hUCSC-EVs were isolated and characterized. Arthritis was induced in mice by collagen injection, followed by intravenous administration of miR-124 EVs, miR-control, or vehicle. Cytokine levels, including interleukin (IL)-6, IL-1β, tumor necrosis factor (TNF)-α, IL-10, and transforming growth factor (TGF)-β, were measured in the joints using real-time PCR by enzyme-linked immunosorbent assay (ELISA). miR-124-hUCSC-EVs showed enrichment of miR-124. In arthritic mice, intravenous treatment with miR-124 EVs significantly reduced the clinical score, increased anti-inflammatory cytokines IL-10 and TGF-β, and lowered inflammatory cytokines IL-6, TNF-α, and IL-1β in the joints. This study shows that miR-124 EVs have anti-inflammatory activity in arthritis by suppressing pro-inflammatory cytokines through miR-124 overexpression. MSC-EVs overexpressed with inflammation-modulating miRNAs offer a promising cell-free approach for treating inflammatory arthritis.

ABA enhances the apoptotic effect of docetaxel in the multidrug-resistant DU145 prostate cancer cell line

Deniz Şumnulu, Zeynep Banu Doğanlar — Fri, 20 Dec 2024 00:00:00 +0100

Paper description:

Serial application of docetaxel and mitoxantrone causes drug resistance.
Development and reversal of drug resistance caused by combined treatment with docetaxel and mitoxantrone was examined in the DU145 prostate cancer cell line.
Abscisic acid increased the apoptotic effect of docetaxel on the resistant DU145 cell line via the suppressive effect on heat shock proteins.
Abscisic acid exerts a complementary effect for drugs used to treat prostate and other cancers.

Abstract: This study aimed to induce drug resistance in DU145 prostate cancer cells by exposing them to docetaxel and mitoxantrone, and to examine the effects of combining docetaxel and abscisic acid (ABA). The IC₅₀ values for docetaxel and mitoxantrone in non-resistant cells were 54.57 nM and 6.25 nM, respectively, rising to 808.53 nM and 50.07 nM after resistance had developed. RT-PCR analysis showed that treatment of resistant cells with 50.07 nM docetaxel and 500 μM ABA (ABA) resulted in the following changes in gene expression: heat shock protein (HSP) 70 (0.63-fold), glucose-regulated protein 94 (GRP94) 0.33-fold, inositol-requiring transmembrane kinase endoribonuclease-1α (IRE1α) 1.62-fold, ER degradation-enhancing alpha-mannosidase-like 1 (EDEM1) 1.77-fold, X-box binding protein 1 (XBP1) 1.53-fold, p21 (2.53-fold), cellular tumor antigen p53 (p53) 2.49-fold, bcl-2-like protein 4 (Bax) 2.7-fold, and tumor necrosis factor (TNF-α) 6.35-fold. Tali™ cytometry analysis showed a 47% increase in apoptotic/necrotic cells with the combined treatment of docetaxel and ABA, compared to a 26% increase with docetaxel alone. Fluorescent staining revealed that co-administration of docetaxel and ABA increases apoptosis in resistant DU145 cells compared to treatment with docetaxel alone. This study suggests that combining ABA with docetaxel could be effective in drug-resistant prostate cancer.

miR-146a expression and the miR-146a gene variant rs2910164 as diagnostic and prognostic biomarkers of oral cancer

Miodrag Vukovic, Goran Stojković, Katarina Zeljić — Fri, 20 Dec 2024 00:00:00 +0100

Paper description:

There is a growing interest in identifying new molecular biomarkers among miRNAs for diagnosis and prognosis of oral cancer.
The study involved 35 oral cancer patients. The expression of miR-146a-5p in cancerous and adjacent non-cancerous tissue and the genotype of the rs2910164 variant were determined using real-time PCR.
Oral cancer patients with high and low miR-146a-5p expression differed significantly in overall survival. Patients with the GG genotype and high miR-146a-5p expression had poor survival.
A high miR-146a-5p level is a potential biomarker for poor survival in oral cancer patients.

Abstract: This study aimed to analyze the diagnostic and prognostic significance of miR-146a-5p in oral cancer and the association of the rs2910164 gene variant with miR-146a-5p expression. Thirty-five oral cancer patients participated in the study. Expression of miR-146a-5p in oral cancer, adjacent non-cancerous tissue, and the rs2910164 genotype variant were examined by real-time PCR. No difference was observed in miR-146a-5p expression between cancerous and non-cancerous tissues (P=0.272). It was concluded that miR-146a-5p is not a good diagnostic biomarker. There was no difference in miR-146a-5p expression depending on the rs2910164 genotype. The rs2910164 variant was associated with tumor histological grade (P=0.037), stage (P=0.036), and lymph node status (P=0.025). There was a difference in survival between oral cancer patients with high and low miR-146a-5p expression (P=0.026), but not between patients with the GG and GC genotypes (P=0.400). Oral cancer patients with the wild-type GG genotype and high miR-146a-5p expression had significantly shorter survival when compared to patients with miR-146a-5p expression below the optimal cut-off and the wild-type genotype (P=0.035). A high miR-146a-5p level could be considered a potential biomarker of poor survival in patients with oral cancer, especially in those with the wild-type GG genotype of the rs2910164 variant.

Involvement of the TGF-β pathway in epithelial-mesenchymal transition promoted by the pulmonary microenvironment in Mycoplasma pneumoniae pneumonia

Lu Fan, Huixia Wang, Nuo Xu, Yun Guo, Ling li — Fri, 20 Dec 2024 00:00:00 +0100

Paper description:

Mycoplasma pneumoniae pneumonia (MPP) can lead to atelectasis and even pulmonary fibrosis, the rationale for which is unclear.
Transcriptome sequencing, immunohistochemistry, Western blotting, immunofluorescence, and ELISA were used to assess whether the pulmonary microenvironment in MPP patients with atelectasis induces epithelial-mesenchymal transition (EMT), and to investigate the signaling pathways involved.
The TGF-β, p53, Hippo, and Rap1 pathways were upregulated; the differentially expressed genes were enriched in the TGF-β signaling pathway.
EMT induced by BALF from MPP patients with atelectasis is closely related to the TGF-β signaling pathway, which may be one of the mechanisms leading to long-term pulmonary fibrosis.

Abstract: Mycoplasma pneumoniae (MP), one of the smallest prokaryotic microorganisms capable of independent survival, causes respiratory tract infections and various extrapulmonary diseases. Mycoplasma pneumoniae pneumonia (MPP) is the most significant clinical manifestation, often leading to complications such as atelectasis and pulmonary fibrosis. We explored the role of the pulmonary microenvironment in regulating epithelial-mesenchymal transition (EMT) in MPP patients with atelectasis. Transcriptome sequencing revealed significant upregulation of pathways including transforming growth factor beta (TGF-β), tumor protein 53 (P53), protein kinase Hippo, Ras-proximate-1 or Ras-related protein 1 (Rap1), and members of class O forkhead box proteins (FoxO) in cells exposed to bronchoalveolar lavage fluid (BALF) from MPP patients with atelectasis. Among these, the TGF-β pathway exhibited the most pronounced changes in gene expression. Further analysis confirmed that BALF from these patients induced EMT in human bronchial epithelial cells and mouse lung tissues and that TGF-β receptor kinase inhibitor (TRKI) effectively reversed this process. In conclusion, the pulmonary microenvironment in MPP patients with atelectasis promotes EMT in the lungs, with TGF-β playing a key role in this process. This may represent a crucial mechanism contributing to pulmonary fibrosis, underscoring the need to focus on the pulmonary microenvironment and TGF-β-targeted therapies for the prevention and management of pulmonary fibrosis in these patients.

Antimicrobial activity of chitosan-silver nanoparticles made from jewelry industry silver waste

Fri, 20 Dec 2024 00:00:00 +0100

Paper description:

Novel chitosan-silver nanoparticles (CS-AgNPs) have been synthesized from recycled silver.
CS-AgNPs were examined as antimicrobial agents based on the inhibition zone, MIC₅₀, and microbial growth inhibition.
CS-AgNP effectively inhibited the growth of coli, S. aureus, and C. albicans.
CS-AgNP-based products may be promising in local antibacterial therapy.

Abstract: Local management of bacterial infections is challenging. The antimicrobial effect of silver has long been recognized, but its use is limited due to its expensive nature and reduced applicability in liquids. This study aimed to synthesize chitosan-silver nanoparticles (CS-AgNPs) from reusable silver waste of the jewelry industry and investigate their antimicrobial properties against pathogenic microorganisms. X-Ray diffraction (XRD) analysis was used to confirm the crystalline structure of the recycled silver, with a strong diffraction peak observed at 2θ=38.60°. Agar disk diffusion showed inhibitory effects for CS-AgNPs on the growth of Escherichia coli, Staphylococcus aureus, and Candida albicans that depended on the concentration of AgNO₃ solution used for preparation. In these tests, S. aureus was more susceptible to the treatment than E. coli and C. albicans. The CS-AgNP inhibited the growth of tested microorganisms with minimum inhibitory concentration (MIC₅₀) values between 1.7 and 4.25 mg/mL. These findings highlight the potential of CS-AgNPs as effective antimicrobial agents. The use of waste materials in nanoparticle synthesis in this research offers a promising approach for sustainable and eco-friendly nanotechnology.

Methylenetetrahydrofolate reductase gene polymorphisms, lipid profiles, and basic renal functional markers as risk for myocardial infarction: a case-control study and haplotype analysis

Fri, 20 Dec 2024 00:00:00 +0100

Paper description:

The interplay of genetic and environmental factors causes myocardial infarction (MI), characterized by gradual worsening of atherosclerosis.
The association of methylenetetrahydrofolate reductase (MTHFR) C677T and A1298C gene variants and different haplotypes, and the risk of MI were examined in the Montenegrin population.
The MTHFR C677T polymorphism is associated with a higher risk of MI (P=0.006) and the A-T haplotype (P=0.004). C677T and A1298C MTHFR polymorphisms are associated with elevated creatinine levels in MI patients (P=0.003 and P=0.002, respectively).
The MTHFR C677T polymorphism is associated with MI and is a potential genetic risk factor.

Abstract: Myocardial infarction (MI) is a serious cardiovascular disease and the primary cause of mortality, with a complex etiopathology. Identifying the genetic basis of myocardial infarction (MI) is essential for developing personalized medical treatments. This study examined the possible association between polymorphisms in the methylenetetrahydrofolate reductase (MTHFR) gene and MI. In the study, 120 patients with MI and 120 age-and-sex-matched controls were genotyped for C677T and A1298C MTHFR polymorphisms by the allele-specific or amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). In the case of the C677T polymorphism, the T/T and C/T genotypes were associated with a significantly increased risk of MI under the dominant genetic model (odds ratio (OR)=2.060; P=0.006). Although there was no significant association between the A1298C variant and MI, this polymorphism was linked to a higher level of creatinine in MI patients (P<0.002). A similar association was observed for the C677T polymorphism (P=0.003). An A-T haplotype represented an increased risk for MI (OR=1.630; P=0.014), whereas the A-C haplotype had a protective role (R=0.517; P=0.002). These findings indicate that C677T MTHFR polymorphism is strongly associated with and increased risk of MI, making it a potential genetic risk factor and a possible predictor of MI.

Inhibiting METTL3-ATG5 axis-mediated harmful autophagy in macrophages could help reduce airway epithelial inflammation and remodeling in COPD

Guiyang Chen, Zhiwei Xia, Xianyou Zeng, Hanying Liu — Fri, 20 Dec 2024 00:00:00 +0100

Pape description:

Methyltransferase-like 3 (METTL3) and autophagy-related protein 5 (ATG5) are increased in chronic obstructive pulmonary disease (COPD).
Mice exposed to cigarette smoke were used to establish the in vivo COPD model; mouse mononuclear macrophages RAW264.7 exposed to cigarette smoke extract served as the in vitro Knockdown of METTL3 was used to investigate its regulatory mechanisms.
In COPD, the inhibition of METTL3-ATG5 axis-mediated macrophage detrimental autophagy alleviated bronchial epithelial cell inflammation and reduced airway remodeling.
Our work provides insight into molecular mechanisms for intervention and treatment of COPD.

Abstract: Cigarette smoke exposure leads to chronic obstructive pulmonary disease (COPD). We investigated the role and underlying mechanisms of methyltransferase-like 3 (METTL3) and autophagy-related protein 5 (ATG5) in the progression of COPD. In a COPD mouse model exposed to cigarette smoke, lung tissues showed increased levels of METTL3, p-p65/p65, autophagy markers (LC3 and ATG5), inflammatory factors (interleukin-6, IL-8, and TNF-α), and airway remodeling markers (N-cadherin, α-SMA, and Tn-C), while p62 and E-cadherin levels were decreased. Expression of METTL3 and ATG5 was positively correlated. These findings are consistent with observations in RAW264.7 mouse mononuclear macrophages exposed to cigarette smoke extract (CSE). CSE inhibited cell viability while promoting autophagy. METTL3 knockdown counteracted CSE effects, and ATG5 overexpression reversed METTL3 knockdown outcomes. Methylated RNA immunoprecipitation-qPCR showed that METTL3 knockdown reduced m6A, and the actinomycin D assay suggested that METTL3 knockdown reduced ATG5 mRNA levels and lowered ATG5 mRNA stability. METTL3-knockdown RAW264.7 reduced the inflammation and airway remodeling markers in the co-cultured mouse bronchial epithelial cells. In conclusion, inhibition of the METTL3-ATG5 axis-mediated macrophage detrimental autophagy in COPD could alleviate bronchial epithelial cell inflammation and reduce airway remodeling.

Evaluation of the cytogenetic and genotoxic effects of an abamectin-based pesticide on Allium cepa roots

Pinar Ili, Fikret Sari — Fri, 20 Dec 2024 00:00:00 +0100

Paper description:

Abamectin, with insecticidal, anthelmintic, and acaricidal activities, one of the most intensively used pesticides, has toxic effects on some non-target organisms, raising concerns regarding its safety.
Toxicity of abamectin, the active ingredient of commercial pesticide formulation Alopec® EC, was investigated by cytogenetic and comet assays in Allium cepa root tips.
Abamectin was found to be cytogenotoxic for cepa.
To protect public health and the environment, abamectin should be carefully regulated, with particular attention given to its concentration, especially in agroecosystems.

Abstract: Abamectin, a widely used pesticide with insecticidal, anthelmintic, and acaricidal properties, has raised safety concerns due to its toxic effects on certain non-target organisms. The toxicity of abamectin, the active ingredient in the commercial pesticide formulation Alopec® EC, was evaluated using cytogenetic and comet assays on Allium cepa root tips. Mitotic index (MI) and phase index (PI) values were used for cytotoxicity assessment. Chromosomal aberration (CA) frequencies in the dividing cells and comet data were used for genotoxicity assessment. The root growth test showed a significant concentration-based decline in root growth after abamectin exposure, with a median effective concentration (EC₅₀) of 2.50 mg/L. Following 96-hour exposure to three concentrations of abamectin (1.25, 2.50, and 5.00 mg/L), cytogenetic and comet analyses indicated a significant concentration- and time-dependent decrease in the MI, alongside an increase in DNA damage. Additionally, there was a significant concentration-dependent rise in the total frequency of CAs. These findings show that abamectin is a pesticide with lethal effects on A. cepa root tip meristematic cells, even at lower concentrations, over prolonged exposure times, with CA-forming and DNA-damaging effects, and that it is highly cytotoxic and genotoxic.