Gul Sahika Gökdemir, Sümeyye Çakmak, Berjan Demirtas
et al.
In order to investigate the effects of acute CO poisoning and subsequent oxygen therapy on cardiac necrosis in rats, with a specific focus on adiponectin levels, twenty–one male Wistar albino rats were divided into three groups (Control, CO, CO+O2). The Control group was placed in a container and exposed to room air for 30 min. Acute CO poisoning was induced in the CO group and CO+O2 group by exposing the rats to CO gas for 30 min. Following CO exposure, the CO+O2 group received oxygen therapy for 30 min, while the CO group did not receive any additional intervention. The animals were euthanized by cardiac puncture under anesthesia, following the approved ethical procedures. Carboxyhemoglobin (COHb), serum levels of creatine kinase (CK), creatine kinase myocardial band (CK–MB), C–reactive protein (CRP) and lactate dehydrogenase (LDH), as well as cardiac and serum adiponectin levels were measured. CO poisoning caused necrosis in cardiac tissue however, oxygen therapy alleviated the negative effect of CO on cardiac injury. COHb and LDH levels in CO group were increased, whereas both cardiac and serum adiponectin levels were decreased (all, P<0.05). There were no changes in CK, CK–MB, CRP levels among groups (all, P>0.05). Oxygen therapy decreased COHb, but increased both cardiac and serum adiponectin levels (all, P<0.05). Adiponectin and LDH may serve as potential biomarkers for early diagnosis of cardiac necrosis caused by acute CO poisoning. The assessment or quantification of adiponectin can also be useful for the early prognosis of cardiac necrosis after oxygen therapy.
Ana Sredojevic, Dragan Radivojevic, Steva M. Levic
et al.
Apples are the most widely consumed temperate fruit worldwide and are often stored for long-term to ensure year-round availability. However, maintaining fruit quality during storage and subsequent shelf-life remain a significant postharvest challenge. This study investigated the combined effects of the harvest stage, cold storage duration, and shelf-life on the physico-chemical properties of Granny Smith apples. Key quality attributes including texture, maturity indices, color, and starch degradation were evaluated using instrumental methods and Raman microscopy. Fruit quality was affected differently by individual factors and their interactions. Texture parameters showed varied sensitivity: the harvest stage affected several parameters, storage duration had the strongest overall impact, shelf-life influenced a moderate number of parameters, and some were affected by combined factor interactions. Maturity indices were significantly influenced by all factors individually and combined. Color parameters were consistently affected by harvest stage and storage, with shelf-life and interactions influencing fewer parameters. These findings emphasize the complex interplay of factors shaping apple quality after harvest. The study demonstrates the importance of timing harvest and tailoring postharvest handling to maintain apple quality. It also demonstrates the potential of combining traditional and advanced techniques for effective ripeness monitoring.
Bjelanović Jasna, Nikolić Aleksandra, Aslan Mutay
et al.
Background: Altered sphingolipid levels might contribute to rectal cancer development, progression and therapy response by regulating various biological processes, including apoptosis. This study aimed to analyse the serum sphingolipid profile in rectal cancer patients and investigate its association with the apoptotic status of tumour tissue and therapy response. Methods: Ceramide (CER) and sphingomyelin (SM) serum levels were analysed in 22 patients with locally advanced rectal cancer and 24 healthy individuals by ultrafast liquid chromatography coupled with tandem mass spectrometry. The expression of pro-apoptotic BAX (BCL2 associated X, apoptosis regulator) and anti-apoptotic BCL2 (BCL2 apoptosis regulator) was analysed in tumour and corresponding healthy tissue samples of patients by quantitative real-time PCR. Results: Significantly lower serum levels of C18 CER, C22 CER, C24 CER, C18 SM and C24 SM were observed in patients than in controls (P<0.05). For C20 CER, C22 CER and C24 CER, a positive correlation with the pro-apoptotic status of tumour tissue was found (r=0.619, P=0.018; r=0.694, P=0.006 and r=0.601, P=0.023, respectively). No difference in serum sphingolipid levels was found between patients with good, moderate, and poor responses to therapy. Conclusions: These results support the involvement of sphingolipids in rectal cancer. However, further studies, including a larger cohort of subjects, are needed to clarify the association of sphingolipid metabolites with therapy response.
José Edson Fontes Figueiredo, Gisele de Fátima Dias Diniz, Mikaely Sousa Marins
et al.
IntroductionBacillus velezensis is a ubiquitous bacterium with potent antifungal activity and a plant growth promoter. This study investigated the potential of B. velezensis CNPMS-22 as a biocontrol agent against phytopathogenic fungi under diverse experimental conditions and its potential as a plant growth promoter. Genome sequencing and analysis revealed putative genes involved in these traits.MethodsThis research performed in vitro experiments to evaluate the CNPMS-22 antagonistic activity against 10 phytopathogenic fungi using dual culture in plate (DCP) and inverted sealed plate assay (ISP). Greenhouse and field tests evaluated the ability of CNPMS-22 to control Fusarium verticillioides in maize plants in vivo. The CNPMS-22 genome was sequenced using the Illumina HiSeq 4,000 platform, and genomic analysis also included manual procedures to identify genes of interest accurately.ResultsCNPMS-22 showed antifungal activity in vitro against all fungi tested, with notable reductions in mycelial growth in both DCP and ISP experiments. In the ISP, volatile organic compounds (VOCs) produced by CNPMS-22 also altered the mycelium coloration of some fungi. Scanning electron microscopy revealed morphological alterations in the hyphae of F. verticillioides in contact with CNPMS-22, including twisted, wrinkled, and ruptured hyphae. Eight cluster candidates for synthesizing non-ribosomal lipopeptides and ribosomal genes for extracellular lytic enzymes, biofilm, VOCs, and other secondary metabolites with antifungal activity and plant growth promoters were identified by genomic analysis. The greenhouse and field experiments showed that seed treatment with CNPMS-22 reduced Fusarium symptoms in plants and increased maize productivity.ConclusionOur findings highlight the CNPMS-22’s potential as bioinoculant for fungal disease control and plant growth with valuable implications for a sustainable crop productivity.
Maria Rossello-Gelabert, Kevin Las Heras, Ainhoa Gonzalez-Pujana
et al.
Abstract Background Mesenchymal stromal cells (MSCs) exert therapeutic effects primarily through their secretome, rich in bioactive factors with immunomodulatory and regenerative properties. However, clinical application of MSC-derived secretome is hindered by donor variability, limited expansion, and replicative senescence. To address these issues, we developed immortalized human hair follicle-derived mesenchymal-like stromal cells (iHF-MSCs) as a consistent and scalable source of therapeutic secretome. Methods HF-MSCs were isolated using a minimally invasive procedure and immortalized with lentiviral SV40 large T antigen. From 576 single-cell clones, C18 and C26 were selected based on proliferation capacity, absence of senescence, and retention of a mesenchymal-like phenotype. Characterization included RT-PCR, flow cytometry for surface markers, trilineage differentiation, and CD56 expression analysis. For immunomodulatory evaluation, cells were licensed with IFN-γ and TNF-α, and the resulting conditioned media (licensed CM) were analyzed by ELISA and cytokine arrays. PBMC proliferation and Treg induction were assessed using licensed CM, while unlicensed CM was tested for its regenerative effects on dermal and epidermal cell functions, including wound repair, tube formation assay as an indicator of angiogenic potential, and oxidative stress response. Results The selected clones (C18 and C26) exhibited long-term stability, rapid expansion, and preserved mesenchymal-like phenotype and multipotency. Secretome profiling revealed an enriched composition of immunoregulatory (e.g., Gal-9, IL-1Ra, TSG-6) and pro-regenerative (e.g., VEGF, PDGF-AA, EGF) factors, with enhanced responsiveness to inflammatory licensing. Notably, iHF-MSCs maintained low immunogenicity and demonstrated superior functional performance: inhibition of PBMC proliferation, Treg induction, and promotion of skin cell proliferation, migration, tube formation, and oxidative stress protection. Conclusion These results position iHF-MSCs as a robust and scalable platform for consistent production of therapeutic secretome, paving the way for innovative, cell-free immunomodulatory and regenerative therapies.
Nuria Estévez-Gómez, Tamara Prieto, Laura Tomás
et al.
Summary: Single-cell genomics enables studying tissues and organisms at the highest resolution. However, since a cell contains a small amount of DNA, single-cell DNA sequencing (scDNA-seq) typically requires single-cell whole-genome amplification (scWGA). Unfortunately, scWGA methods introduce technical biases that complicate the interpretation of scDNA-seq data. We compared six scWGA methods, three MDA (multiple displacement amplification; GenomiPhi, REPLI-g, and TruePrime) and three non-MDA (Ampli1, MALBAC, and PicoPLEX), on 206 tumoral and 24 healthy human cells. scWGA methods performed differently depending on the parameter of interest. REPLI-g minimized regional amplification bias, while non-MDA methods showed a more uniform and reproducible amplification. Ampli1 exhibited the lowest allelic imbalance and dropout, the most accurate insertion or deletion (indel) and copy-number detection, and a low polymerase error rate. However, REPLI-g yielded higher DNA quantities, longer amplicons, and greater genome coverage. We offer a comprehensive guide for selecting a scWGA approach, outlining trade-offs that influence the interpretation of scDNA-seq data. Motivation: Single-cell whole-genome amplification (scWGA) is a critical step for the genomic study of single cells, yet its impact on downstream analysis remains poorly understood. Previous studies were not comprehensive in the number of methods tested and the metrics assessed. Besides, they have mostly been conducted or replicated by the original authors of a method rather than by independent third-party laboratories. Here, we test six commercially available scWGA methods to understand their performance and propose a guide for selecting the best scWGA approach for studies on single-cell genomics.
Omid Moztarzadeh, Arezou Taherpour, Avat Arman Taherpour
et al.
Abstract The growing concern over antibiotic resistance necessitates novel strategies to enhance antimicrobial efficacy. This study investigates the synergistic effect of fullerene C60 on antibiotic susceptibility in Escherichia coli. In a standardized modification of the disc diffusion method, the zones of inhibition were measured on Mueller–Hinton agar with and without supplementation of fullerene C60 after incubation for 10 and 24 h, respectively. Fullerene C60 was noted for strengthening the antibacterial action of several antibiotics, especially to the extent of conversion of the bacteriostatic effect to bactericidal over a period. A statistically significant enhancement in the diameter of zones of inhibition was observed with tetracycline and co-trimoxazole, whereas amikacin and gentamicin evidenced only slight increases. The findings indicate that fullerene C60 could disrupt bacterial membrane integrity or metabolic activity, thereby potentiating the antibiotics. This paves the way for introducing nanomaterials like fullerene C60 into treatment strategies to target antibiotic resistance and bring about better therapeutic outcomes in bacterial infections.
Abstract The global production of soybean, among other drought-susceptible crops, is reportedly affected by drought periods, putting more pressure on food production worldwide. Drought alters plants’ morphology, physiology and biochemistry. As a response to drought, reactive oxygen species (ROS) concentrations are elevated, causing cellular damage. However, lower concentrations of ROS were reported to have an alleviating role through up-regulating various defensive mechanisms on different levels in drought-stressed plants. This experiment was set up in a controlled environment to monitor the effects of exogenous spray of different (0, 1, 5 and 10 mM) concentrations of H2O2 on two soybean genotypes, i.e., Speeda (drought-tolerant), and Coraline (drought-susceptible) under severe drought stress conditions (induced by polyethylene glycol) during flowering stage. Furthermore, each treatment was further divided into two groups, the first group was kept under drought, whereas drought was terminated in the second group at the end of the flowering stage, and the plants were allowed to recover. After 3 days of application, drought stress significantly decreased chlorophyll-a and chlorophyll-b, total carotenoids, stomatal conductance, both optimal and actual photochemical efficiency of PSII (Fv/Fm and Df/Fm, respectively), relative water content, specific leaf area, shoot length and dry weight, and pod number and fresh weight, but significantly increased the leaf concentration of both proline and total soluble sugars, the root length, volume and dry weight of both genotypes. The foliar application of 1 mM and 5 mM H2O2 on Speeda and Coraline, respectively enhanced most of the decreased traits measurably, whereas the 10 mM concentration did not. The group of treatments where drought was maintained after flowering failed to produce pods, regardless of H2O2 application and concentration, and gradually deteriorated and died 16 and 19 days after drought application on Coraline and Speeda, respectively. Overall, Speeda showed better performance under drought conditions. Low concentrations of foliar H2O2 could help the experimented soybean genotypes better overcome the influence of severe drought during even sensitive stages, such as flowering. Furthermore, our findings suggest that chlorophyll fluorescence and the cellular content of proline and soluble sugars in the leaves can provide clear information on the influence of both drought imposition and H2O2 application on soybean plants.
Clemens Steinek, Miguel Guirao-Ortiz, Gabriela Stumberger
et al.
Summary: The genome contains numerous regulatory elements that may undergo complex interactions and contribute to the establishment, maintenance, and change of cellular identity. Three-dimensional genome organization can be explored with fluorescence in situ hybridization (FISH) at the single-cell level, but the detection of small genomic loci remains challenging. Here, we provide a rapid and simple protocol for the generation of bright FISH probes suited for the detection of small genomic elements. We systematically optimized probe design and synthesis, screened polymerases for their ability to incorporate dye-labeled nucleotides, and streamlined purification conditions to yield nanoscopy-compatible oligonucleotides with dyes in variable arrays (NOVA probes). With these probes, we detect genomic loci ranging from genome-wide repetitive regions down to non-repetitive loci below the kilobase scale. In conclusion, we introduce a simple workflow to generate densely labeled oligonucleotide pools that facilitate detection and nanoscopic measurements of small genomic elements in single cells. Motivation: While three-dimensional chromatin conformations can be explored with fluorescence in situ hybridization (FISH), the visualization of small genomic loci with high spatial resolution remains challenging. For such applications, programmable oligonucleotides with high brightness are required. To further improve precision and sensitivity, secondary hybridization steps should be omitted. Here, we present a simple, quick, and inexpensive approach to generate labeled FISH probes that carry several fluorophores. Our workflow allows for the free choice of fluorophores, flexible adjustment of labeling density, and selective probe synthesis from large probe pools. With our probes, we reliably detect genomic loci below the kilobase level and examine their topological relationships.
Allesandra Stratigakis, Dylan Paty, Peng Zou
et al.
The development of an effective method for predicting the transfer of biologics from plasma into breast milk is important to ensure the safe use of medications during lactation. The aim of this study was to develop a regression model that could predict the transfer of monoclonal antibodies (mAbs) and Fc-fusion proteins from plasma into breast milk. By searching various databases, a list of eleven mAbs and Fc-fusion proteins with available information of presence in the breast milk was generated. Physicochemical properties such as the isoelectric point (pI), molecular weight (MW), dissociation constant (Kd), and pharmacokinetic (PK) parameters such as clearance (CL), volume of distribution (Vd), and half-life (T1/2) were collected or calculated. A two-variable non-linear regression analysis and a multivariate regression analysis were employed to establish correlation of milk-to-plasma (M/P) ratios with different combinations of two physicochemical properties. The 3D isoelectric point (pI) of the Fv region and the buried surface area (BSA) between the light and heavy chains (LC_HC) were two factors that emerged as a promising predictor of the milk-to-plasma concentration ratio (M/P). The correlation between M/P ratio, 3D pI of Fv region, and BSA_LC_HC was found to be good with R2 of 0.9058. Other combinations of the physicochemical properties did not show a statistically significant correlation. The multivariate regression model was used to predict the MP ratios for 79 different mAbs. We believe that this regression model could serve as a valuable tool to estimate the M/P ratios of mAbs and Fc-fusion proteins. Further model validation is necessary when the M/P ratios of additional biologics are available. This could inform clinical decision-making and improve the safety of large molecule drug use during lactation.
Thermal quenching is described as a decrease in luminescence efficiency with increasing measurement temperature. Luminescence intensity decreases with increasing heating rates in the presence of thermal quenching. In such a case, the heating rate to be used in the measurements becomes important. Lithium fluoride (LiF) type dosimeters have been widely used in radiation dosimetry for many years. In this study, thermal quenching effect was investigated for LiF:Mg,Ti (TLD-100) and LiF:Mg,Cu,P (TLD-100H), 6LiF:Mg,Ti(TLD-600) and 7LiF:Mg,Ti (TLD-700) at two different doses (10, 1000mGy) using 90Sr/90Y beta source. TLD-100, TLD-600 and TLD-700 showed different thermal quenching behaviors according to dose values, while TLD-100H had the same characteristics at both doses. On the other hand, other dosimeters showed thermal quenching based on the total area at 10mGy, while they did not show thermal quenching when ROI was used. Again, thermal quenching was not observed at 1000mGy for all dosimeters. In conclusion, it is recommended to use ROI or low heating rate during measurements at a low dose (in the order of mGy) for TLD-100, TLD-600 and TLD-700, while desired heating rate can be used at a high dose (Gy) for all dosimeters.
Amira Awadalla, Eman T. Hamam, Fardous F. El-Senduny
et al.
Objective To investigate the renoprotective, the antioxidant, and the anti-inflammatory impact of a combination of SPL and ZnO-NPs to combat against chronic kidney disease (CKD).Methods In total, 50 males of rats were distributed into 5 groups (10 rats each); normal group, adenine sulfate (0.25% in diet for 10 days) (CKD) group. After the last dose of adenine sulfate, rats were divided into three groups: SPL + Adenine sulfate group; rats were treated orally by mixing SPL (20 mg/kg/day) into chow for 8 weeks, ZnO-NPs + Adenine sulfate group; rats were injected intraperitoneally with ZnO-NPs (5 mg/kg) three times weekly for 8 weeks, ZnO-NPs + SPL + Adenine sulfate group; rats were injected with the same previous doses for 8 weeks.Results Each of SPL and ZnO-NPs up-regulated antioxidant genes (Nrf2 and HO-1), down-regulated fibrotic and inflammatory genes (TGF-β1, Wnt7a, β-catenin, fibronectin, collagen IV, α-SMA, TNF-α, and IL-6) compared to CKD. Furthermore, a combination of SPL and ZnO-NPs resulted in a greater improvement in the measured parameters than a single treatment.Conclusion The therapeutic role of SPL was enhanced by the antioxidant and the anti-inflammatory role of ZnO-NPs, which presented a great renoprotective effect against CKD.
BackgroundThe role of circular RNAs (circRNAs) in the occurrence of gastric cancer is still unclear. Therefore, the diagnostic value and mechanisms underlying hsa_circ_0061276 in the occurrence of gastric cancer were explored.MethodsReverse transcription-droplet digital polymerase chain reaction was used to detect the copy number of hsa_circ_0061276 in plasma from healthy individuals, as well as from patients with gastric precancerous lesions or early-stage or advanced gastric cancer. Plasmids overexpressing or knocking down hsa_circ_0061276 expression were transfected into gastric cancer cells. The effects on the growth, migration, and cell cycle distribution of gastric cancer cells were then analyzed. Finally, miRanda and RNAhybrid were used to explore the binding sites between hsa_circ_0061276 and microRNAs (miRNAs). A double luciferase reporter gene assay was used to confirm the miRNA sponge effect.ResultsThe results show that plasma hsa_circ_0061276 copy number showed a trend of a gradual decrease when comparing healthy controls to the early cancer group and advanced gastric cancer group. Overexpression of hsa_circ_0061276 inhibited the growth and migration of gastric cancer cells. Through bioinformatic analyses combined with cellular experiments, it was found that hsa_circ_0061276 inhibited the growth of gastric cancer by binding to hsa-miR-7705.ConclusionHsa_circ_0061276 may be a new biomarker for gastric cancer. The tumor suppressor role of hsa_circ_0061276 on gastric cancer likely occurs through a sponge effect on miRNAs such as hsa-miR-7705.
Neoplasms. Tumors. Oncology. Including cancer and carcinogens
Zemene Demelash Kifle, Agumas Alemu Alehegn, Meaza Adugna
et al.
Background: Serum lipid profile abnormalities are major predictors for coronary artery diseases. The relationship between demographic factors and dyslipidemia in Ethiopia is not completely explored. Thus, this study aimed to assess the prevalence and predictors of dyslipidemia among hypertensive patients in Lumame Primary Hospital. Methods: A cross-sectional study was conducted from June to August 30, 2020, on the hypertensive patients in Lumame Primary Hospital. All adult hypertensive patients who visited the adult hypertensive care services during the study period were included. Interview-guided self-administered questionnaire and a chart review were used for data collection. Statistical Package for the Social Sciences (SPSS) software version 24.0 was used for data analysis. Results: Out of 372 hypertensive patients, 190(51.1%) were females and the mean age of the study participants was 43.56 years (SD ± 4.31). The overall prevalence of dyslipidemia in this study was 48.4%. Besides the overall prevalence, the prevalence of TC, TG, LDL-c, and HDL-c was 73(19.6%), 91(24.5%), 60(16.1%), and 115(30.9%), respectively. Females were at higher risk for having high levels of TC (AOR = 2.31, 95% CI = 1.54–3.13), TG (AOR = 1.70, 95% CI = 1.34–3.79), LDL-c (AOR = 2.15, 95% CI = 1.56–2.86), and HDL-c (AOR = 2.67, 95% CI = 1.44–5.67) than males. Respondents who were from urban were at higher risk for having high levels of TC (AOR = 1.98, 95% CI = 1.04–6.83), TG (AOR = 1.78, 95% CI = 1.09–2.86), LDL-c (AOR = 3.01, 95% CI = 1.45–7.43), and HDL-c (AOR = 2.01, 95% CI = 1.94–4.55) than respondents who were from rural. Similarly, obese respondents were at higher risk for having high levels of TC (AOR = 2.03, 95% CI = 1.64–2.00), TG (AOR = 3.78, 95% CI = 1.06–6.42), LDL-c (AOR = 1.92, 95% CI = 1.66–2.12), and HDL-c (AOR = 4.23, 95% CI = 2.84–4.32) than to respondents who were underweight. Conclusion: The prevalence of dyslipidemia among hypertensive patients was high. Independent variables such as age, gender, residence, family history of HTN, smoking, alcohol drinking, fruit diet habits, physical activity, DM, and BMI were significant determinants of dyslipidemia.
Zulhisyam Abdul Kari, Muhammad Anamul Kabir, Khairiyah Mat
et al.
The development of plant ingredients as an alternative to fish meal (FM) has received sustained interest in the aquaculture sector. The study investigated the replacement of FM with different percentages of dietary fermented soy pulp (FSP) to assess the growth performance, haematology, blood biochemical, liver, and gut morphology of African catfish. Five isonitrogenous (32 % crude protein) diets were prepared with FSP by replacing 0 % FSP (D1), 25 % FSP (D2), 50 % FSP (D3), 75 % FSP (D4) and 100 % FSP (D5) of FM component of the diets. The results showed a significant difference (p < 0.05) in growth parameters where the fish fed D3 diet showed the highest weight gain, specific growth rate, and condition factor compared with other diets. The mean values of Red Blood Cell (RBC) and Lymphocytosis (LYM) were significantly highest (p < 0.05) in fish fed the D3 diet. The albumin (ALB), globulin (GLOB), and total protein (TP) were significantly lower (p < 0.05) in the control diet compared with the experimental diets. In addition, the D3 diet provides the highest total lactic bacteria (LAB) and total bacteria (TB) compared with other diets. FSP as a protein replacement of the FM had a significant effect (p < 0.05) on villus length, width, and crypt depth in fish's anterior and posterior gut. The histological study of the intestinal revealed that the gut of the D3 diet had an intact epithelial barrier with goblet cells arrangement and very well-organized villus structure, tunica muscularis compared with the other treatments. In conclusion, the replacement of 50 % FM with FSP could be used in the aquafeed industry for better growth and health status of African catfish and possibly for freshwater species.