Culture Assay Research Articles

Poly(rC)-Binding Protein 2 Does Not Directly Participate in HCV Translation or Replication, but Rather Modulates Genome Packaging.

The hepatitis C virus (HCV) co-opts many cellular factors-including proteins and microRNAs-to complete its life cycle. A cellular RNA-binding protein, poly(rC)-binding protein 2 (PCBP2), was previously shown to bind to the hepatitis C virus (HCV) genome; however, its precise role in the viral life cycle remained unclear. Herein, using the HCV cell culture (HCVcc) system and assays that isolate each step of the viral life cycle, we found that PCBP2 does not have a direct role in viral entry, translation, genome stability, or HCV RNA replication. Rather, our data suggest that PCBP2 depletion only impacts viral RNAs that can undergo genome packaging. Taken together, our data suggest that endogenous PCBP2 modulates the early steps of genome packaging, and therefore only has an indirect effect on viral translation and RNA replication, likely by increasing the translating/replicating pool of viral RNAs to the detriment of virion assembly.

Limonoids, triterpenes, sterols, and flavonoids from Cedrela odorata L. and their cytotoxic activity

The isoflavonoid cedrelin (1), isolated for the first time, and two new limonoids with mexicanolide skeleton, the 15-O-acetyl-12-oxo-fissinolide (2) and 15-O-acetyl-fissinolide (3), were isolated together with fifteen other known compounds from the stem bark of Cedrela odorata. Among the known compounds were the limonoid 7α,11β-diacetoxydihydronomilin (4) and the triterpene pentaol (5), whose 1D and 2D NMR spectroscopic data interpretation and structural elucidation are discussed here for the first time. Cell culture assays demonstrated that some of the compounds isolated from this plant possess moderate cytotoxic activity in representative human cancer cell lines. These cytotoxicity studies, along with previous reports, revealed that at least 6 of the isolated compounds exhibit activity against cancer cell lines, which aligns with the popular use in the cancer treatment for this medicinal plant.

Inflammatory Response of Monocytes/Macrophages in Patients with Systemic Sclerosis.

Investigation of the inflammatory response of immune cells is a current focus of research on autoimmune disorders. The aim of this study was to evaluate the inflammatory status of monocytes/macrophages in systemic sclerosis (SSc). The study included 35 SSc and 25 healthy participants. The secretion of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), monocyte chemoattractant protein-1 (MCP-1), interleukin-8 (IL-8), interleukin-6 (IL-6) was measured by enzyme-linked immunosorbent assay (ELISA) in primary cultures of monocytes/macrophages after stimulation with lipopolysaccharide (LPS) on day 1 and on day 6 of incubation. Impaired tolerance of the immune response was characterized by increased secretion of the inflammatory mediators in response to restimulation. Basal secretion of all cytokines was significantly higher in SSc patients compared to healthy individuals. The secretion of TNF-α, IL-1β and IL-6 after the initial LPS stimulation, and secretion of IL-1β, MCP-1, IL-6, IL-8 after LPS restimulation, was significantly higher in the SSc group. Eleven SSc patients (31%) showed impaired immune tolerance in terms of MCP-1 secretion. These patients were significantly younger and had a higher level of anti-topoisomerase I (anti-Scl70) antibodies compared to SSc patients with immune tolerance. This study revealed pro-inflammatory activation and impaired immune tolerance in monocytes/macrophages from SSc patients. The violation of immune response in terms of MCP-1 secretion may be an important factor in the development of chronic inflammation in SSc. MCP-1 may thus be a potential therapeutic target for novel SSc treatment strategies.

The Helicobacter pylori infection alters the intercellular junctions on the pancreas of gerbils (Meriones unguiculatus)

Helicobacter pylori is a common resident in the stomach of at least half of the world’s population and recent evidence suggest its emergence in other organs such as the pancreas. In this organ, the presence of H. pylori DNA has been reported in cats, although the functional implications remain unknown. In this work, we determined distinct features related to the H. pylori manifestation in pancreas in a rodent model, in order to analyse its functional and structural effect. Gerbils inoculated with H. pylori exhibited the presence of this bacterium, as revealed by the expression of some virulence factors, as CagA and OMPs in stomach and pancreas, and confirmed by urease activity, bacterial culture, PCR and immunofluorescence assays. Non-apparent morphological changes were observed in pancreatic tissue of infected animals; however, delocalization of intercellular junction proteins (claudin-1, claudin-4, occludin, ZO-1, E-cadherin, β-catenin, desmoglein-2 and desmoplakin I/II) and rearrangement of the actin-cytoskeleton were exhibited. This structural damage was consistent with alterations in the distribution of insulin and glucagon, and a systemic inflammation, event demonstrated by elevated IL-8 levels. Overall, these findings indicate that H. pylori can reach the pancreas, possibly affecting its function and contributing to the development of pancreatic diseases.Graphical

Analysis of the antagonistic effect of Stenotrophomonas geniculata WXY53 on Magnaporthe oryzae through bioassays and Whole-Genome sequencing

Magnaporthe oryzae causes rice blast, which poses a significant threat to agriculture. The use of environmentally friendly biological control agents is one of the methods used to reduce disease incidence. Our study identified the isolate WXY53 as Stenotrophomonas geniculata using 16S rDNA and phylogenetic tree analysis. WXY53 exhibited a 53.39 % inhibition rate on the growth diameter of M. oryzae hyphae in the dual culture assay. The WXY53 fermentation broth treatment inhibited 97.89 % of appressoria formation and delayed the colonization of plant cells by invasive hyphae. Applying WXY53 fermentation broth had a significant impact on preventing rice blast in the greenhouse experiment. It suppressed the size of necrotic lesions and reduced the number of lesions in rice and barley by 66.25 % and 72.0 %, respectively. The whole-genome sequencing of WXY53 revealed a genome size of 4,504,923 bp, with 66.51 % GC content and 4038 protein-coding genes. The study utilized multiple databases, including the Kyoto Encyclopedia of Genes and Genomes, Clusters of Orthologous Groups of proteins, Gene Ontology, Pathogen-Host Interactions, and Carbohydrate-Active enZYmes to annotate gene functions. The analysis revealed genes related to environmental information processing, biological processes, cell wall/membrane/envelope biogenesis, glycoside hydrolases, and metabolism. Antibiotics & Secondary Metabolite Analysis Shell analysis predicted that WXY53 has the potential to produce aryl polymers, entolysin, and 2,3-dihydroxybenzoyl serine, in addition to the three gene clusters with unknown functions. Based on these findings, strain WXY53 could be a promising novel source of biological agents for managing rice blast. This study provides a theoretical basis for further investigation into microbial resources and metabolic gene clusters for agricultural biological control.

Mechanisms of Cell Wall Degrading Enzymes from Bacillus methylotrophicus and Bacillus subtilis in Suppressing Foliar Blight Pathogens

Bacillus strains are potent Biocontrol agents (BCAs) and have been identified as an effective way to control the growth of phytopathogens of wheat. Through a direct inhibition mechanism which involves the production of cell wall degrading enzymes (proteases, glucanase, and chitinases) and siderophores they suppress the foliar blight disease. Both the strains of Bacillus P10 and UP11 shows growth inhibition Alternaria triticina (77.56%, 67.83%) and Bipolaris sorokiniana (73.97%, 62.16%) through a dual culture assay. These strains were subsequently examined for their ability to produce cell wall-degrading enzymes i.e., chitinase, protease, and β-1,3-glucanases and antifungal metabolites i.e., siderophores. It was found that when the antagonist’s bacteria were Co-cultured with fungal pathogens then maximum production of hydrolytic enzymes and siderophore was achieved at 96 hrs, but when both strains P10 and UP11 were alone maximum production was found at 48 hrs i.e., the exponential phase. The current study determined that Bacillus methylotrophicus (P10) and Bacillus subtilis (UP11) are highly effective strain for controlling Foliar Blight disease. Direct or Antibiosis is the main mechanism involved in this study. Further research on the interaction mechanisms between Bacillus-derived compounds and host plants is necessary.

Notch Signaling Hydrogels Enable Rapid Vascularization and Promote Dental Pulp Tissue Regeneration.

Successful dental pulp regeneration is closely associated with rapid revascularization and angiogenesis, processes driven by the Jagged1(JAG1)/Notch signaling pathway. However, soluble Notch ligands have proven ineffective in activating this pathway. To overcome this limitation, a Notch signaling hydrogel is developed by indirectly immobilizing JAG1, aimed at precisely directing the regeneration of vascularized pulp tissue. This hydrogel displays favorable mechanical properties and biocompatibility. Cultivating dental pulp stem cells (DPSCs) and endothelial cells (ECs) on this hydrogel significantly upregulate Notch target genes and key proangiogenic markers expression. Three-dimensional (3D) culture assays demonstrate Notch signaling hydrogels improve effectiveness by facilitating encapsulated cell differentiation, enhancing their paracrine functions, and promoting capillary lumen formation. Furthermore, it effectively communicates with the Wnt signaling pathway, creating an odontoinductive microenvironment for pulp-dentin complex formation. In vivo studies show that short-term transplantation of the Notch signaling hydrogel accelerates angiogenesis, stabilizes capillary-like structures, and improves cell survival. Long-term transplantation further confirms its capability to promote the formation of pulp-like tissues rich in blood vessels and peripheral nerve-like structures. In conclusion, this study introduces a feasible and effective hydrogel tailored to specifically regulate the JAG1/Notch signaling pathway, showing potential in advancing regenerative strategies for dental pulp tissue.

Bioactivity evaluation and active compounds identification of Symphoricarpos orbiculatus as potential botanical herbicide.

Evaluation of herbicidal activity and identification of active compounds are important bases for the development of new botanical herbicides. This study confirmed that Symphoricarpos orbiculatus has high herbicidal activities against mono-dicotyledonous weeds, including Echinochloa crusgalli, Digitaria sanguinalis, Amaranthus retroflexus and Portulaca oleracea. By bioassay-guided isolation, 12 compounds were isolated and identified from S. orbiculatus for the first time, including iridoids: naucledal (K1), loganin (K2), loganigenin (K3), loganin acid (K4), glucologanin (K5) and vogeloside (K6), as well as flavonoids: quercetine (K7), luteolin (K8), nobiletin (K9), astragalin (K10), isorhamnetin 3-d-glucoside (K11) and rutin (K12). Biological assays showed that iridoids are the main active ingredients of S. orbiculatus. The compounds of K5 and K6 could inhibit both the root (IC50 = 37.54 and 38.91 μg mL-1, respectively) and shoot (IC50 = 42.78 and 45.72 μg mL-1, respectively) of Portulaca oleracea, which have a weeding toxicity similar to that of the commercialized plant-based herbicide pelargonic acid. In addition, the results of pot culture assay showed that S. orbiculatus ethanol extracts had high fresh weight control effect against Digitaria sanguinalis and P. oleracea at the concentration of 40 g L-1. After 7 days, both the soil treatment and the stem and leaf spray method resulted in severe leaf necrosis and significant leaf etiolation. Symphoricarpos orbiculatus and its herbicidal active compounds have the potential to develop into botanical herbicides, and are first reported in the present study. © 2024 Society of Chemical Industry.

Biostimulant and antagonistic potential of endophytic fungi against fusarium wilt pathogen of tomato Fusarium oxysporum f. sp. lycopersici

Endophytic fungal-based biopesticides are sustainable and ecologically-friendly biocontrol agents of several pests and diseases. However, their potential in managing tomato fusarium wilt disease (FWD) remains unexploited. This study therefore evaluated effectiveness of nine fungal isolates against tomato fusarium wilt pathogen, Fusarium oxysporum f. sp. lycopersici (FOL) in vitro using dual culture and co-culture assays. The efficacy of three potent endophytes that inhibited the pathogen in vitro was assessed against FWD incidence, severity, and ability to enhance growth and yield of tomatoes in planta. The ability of endophytically-colonized tomato (Solanum lycopersicum L.) plants to systemically defend themselves upon exposure to FOL were also assessed through defence genes expression using qPCR. In vitro assays showed that endophytes inhibited and suppressed FOL mycelial growth better than entomopathogenic fungi (EPF). Endophytes Trichoderma asperellum M2RT4, Hypocrea lixii F3ST1, Trichoderma harzianum KF2R41, and Trichoderma atroviride ICIPE 710 had the highest (68.84–99.61%) suppression and FOL radial growth inhibition rates compared to EPF which exhibited lowest (27.05–40.63%) inhibition rates. Endophytes T. asperellum M2RT4, H. lixii F3ST1 and T. harzianum KF2R41 colonized all tomato plant parts. During the in planta experiment, endophytically-colonized and FOL-infected tomato plants showed significant reduction of FWD incidence and severity compared to non-inoculated plants. In addition, these endophytes contributed to improved growth promotion parameters and yield. Moreover, there was significantly higher expression of tomato defence genes in T. asperellum M2RT4 colonized than in un-inoculated tomato plants. These findings demonstrated that H. lixii F3ST1 and T. asperellum M2RT4 are effective biocontrol agents against FWD and could sustainably mitigate tomato yield losses associated with fusarium wilt.

O-310 PARP inhibitors have no toxic effect on follicle reserve, mitotic and steroidogenic function of granulosa cells in human ovary

Abstract Study question Do the PARP inhibitors have gonadotoxic effects on human ovary? Summary answer No. The PARP inhibitor drugs Olaparib and Niraparib do not appear to have any toxic effects on the human ovary. What is known already The Poly (ADP-ribose) polymerase (PARP) family has many essential functions in cellular processes, including the regulation of transcription, apoptosis, and the DNA damage response. PARP has Poly (ADP-ribose) activity, which facilitates the recruitment of other repair proteins to promote the repair of DNA single-strand breaks when DNA damage occurs. PARP inhibitors are approved for the treatment of BRCA1/2 mutated breast and ovarian cancers. No data is available regarding the gonadotoxic potentials, if any, of these drugs on the human ovary. We aimed to address this issue in the current study. Study design, size, duration A clinical translational research study that utilizes ex-vivo and in-vitro experimental models on human ovarian cortical samples, primary luteinized non-mitotic granulosa cells, and immortalized human granulosa cells. Participants/materials, setting, methods Ovarian cortical samples were obtained from young women undergoing laparoscopic surgery for benign ovarian cysts (n = 5). Human non-mitotic luteinized granulosa cells (HLGCs) obtained from 10 IVF patients during the oocyte retrieval procedure and immortalized mitotic non-luteinized granulosa cell line (HGrC1), and immortalized granulosa cell tumor cell line (COV434) were used for the experiments. Cell/tissue culture, confocal imaging, flow cytometry, immunoblotting, cell viability/proliferation and hormone assays were applied. Main results and the role of chance Treatment of ovarian tissue samples and granulosa cells with PARP inhibitors at five different therapeutic concentrations (1, 5, 10, 20, and 40 µM/mL) up to 96 hours in culture resulted in resulted in the cleavage of PARP, validating in vitro activity of the drugs. The primordial follicle reserve and steroidogenic activity of the tissue samples remained stable at the end of culture period. Similar results were obtained in the granulosa cells, which retained their viability and steroidogenic functions after exposure to the PARP inhibitors. Real-time impedance-based cell proliferation assay with xCelligence system revealed that granulosa cells continued to proliferate normally and reached log phase in the presence of PARP inhibitors without any discernible cytotoxic effect. In immunoblot analysis, the expression of steroidogenic enzymes (StAR, aromatase, 3B-HSD and 17B-HSD) was comparable to control cells and cleaved caspase-3 expression was absent in the granulosa cells exposed to PARP inhibitors. Limitations, reasons for caution Caution should be exercised before extrapolating our data because these findings were obtained in ex-vivo and in-vitro models. They need to be confirmed using in-vivo experimental models such as human ovarian tissue xenografting in nude mice. Wider implications of the findings This study provides the first reassuring molecular evidence that PARP inhibitor drugs do not appear to have a significant gonadotoxic effect on human ovary. Trial registration number not applicable

Synergistic utilization of cost-effective glycerophosphate and biologically active zein for innovative minimally invasive smart thermo-responsive hydrogels for potential hard tissue engineering applications

Skeletal defects are the second-leading cause of disability worldwide, prompting the development of smart solutions for treatment. Calcium glycerophosphate (Ca–GP), chitosan (CS), hydroxyapatite (HA), and zein (ZN) were used to fabricate these thermo-responsive hydrogels. Ca–GP, an economically viable and bioactive glycerophosphate source, remains relatively underexplored. Natural protein ZN and the gold standard bone regenerative biomaterial HA were incorporated as reinforcing agents. The resulting composite hydrogels (HGs) exhibit a sol phase at 4 °C–10 °C and transition to gels at body temperature within 4–6 min. Their good injectability and the ability to be easily shaped into complex structures further support their great potential as minimally invasive solutions for treatment. The addition of ZN significantly improved the mechanical and biological properties of the HGs. The highest ZN concentration resulted in the strongest mechanical strength, measuring 52.2 MPa at 40% strain. HGs exhibited optimal swelling and degradation rates. Scanning electron microscopy analysis supported their porous nature. In vitro cell culture assays and wound healing assays demonstrated their excellent biocompatibility and regenerative potential. Drug-loaded HGs exhibited up to 90% drug release and antibacterial activity. All these results support their promising potential to support the regeneration of skeletal defects in a minimally invasive manner.

Metagenomic Next-Generation Sequencing-Based Fine-Needle Aspiration in Patients With Suspected Infected Pancreatic Necrosis.

Fine-needle aspiration (FNA) is no longer recommended for diagnosing infected pancreatic necrosis (IPN) due to a high false-negative rate. Metagenomic next-generation sequencing (mNGS) is a valuable tool for identifying potential pathogens. We hypothesized that adding mNGS to the standard FNA procedure may increase diagnostic accuracy. This is a prospective, single-arm feasibility study enrolling patients with acute necrotizing pancreatitis complicated by suspected IPN. Computed tomography-guided FNA was performed immediately after enrollment, and the drainage samples were subjected to culture and mNGS assays simultaneously. Confirmatory IPN within the following week of the index FNA procedure was the reference standard. The diagnostic performance of FNA-mNGS and the impact of mNGS results on treatment were evaluated. Historical controls were used for comparison of clinical outcomes. There was no significant difference between mNGS and culture in the positive rate (75% vs 70%, P = 0.723). The accuracy of FNA-mNGS was 80.0%, with a sensitivity of 82.35%, specificity of 66.67%, positive predictive value of 93.3%, and negative predictive value of 40.0%. The results of the mNGS led to treatment change in 16 of 20 patients (80%), including implementing percutaneous catheter drainage (n = 7), expanding antibiotic coverage (n = 2), percutaneous catheter drainage and expanding coverage (n = 4), narrowing antibiotic coverage (n = 1), and discontinuation of antibiotics (n = 2). The FNA-mNGS approach was not associated with improved clinical outcomes compared with the historical control group. The addition of mNGS to standard FNA has comparable diagnostic accuracy with culture-based FNA and may not be associated with improved clinical outcomes.

Prevalence, antimicrobial susceptibility profiles and resistant gene identification of bovine subclinical mastitis pathogens in Bangladesh

Subclinical mastitis (SCM), a silent threat in the dairy sector of Bangladesh poses a significant economic impact and serves as a potential source of infection for healthy cows, hindering efforts to achieve milk self-sufficiency. Despite the importance of this issue, limited research has been conducted on mastitis in Sylhet region of Bangladesh. This study aimed to investigate the molecular prevalence, antimicrobial susceptibility profile and resistant genes detection on pathogens (Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae) causing SCM. In a cross-sectional study utilizing convenience sampling, 325 milk samples were collected from apparently healthy dairy cows. Initially, SCM was detected using the modified Whiteside test (MWST) method. Suspected positive samples were then subjected to bacteriological culture and standard biochemical assays, followed by molecular identification through polymerase chain reaction (PCR). Finally, antimicrobial susceptibility testing was conducted on all PCR-positive samples using the Kirby-Bauer disk diffusion method on Mueller-Hinton agar. In the Sylhet district, the prevalence of SCM was 64.92 % (211 out of 325) at the individual animal level and 82.69 % (43 out of 52) at the herd level. Among the SCM-positive animals, S.aureus was found in 31.75 % (67 out of 211) of cases, E.coli in 81.99 % (173 out of 211), and K.pneumoniae in 66.82 % (141 out of 211). K.pneumoniae had the highest prevalence at 60 % in Zakiganj, S.aureus at 45 % in Zakiganj, and E.coli at 72 % in Bishwanath Upazila. Extended spectrum beta lactamase (ESBL) genes blaTEM, blaOXA, blaCTX-M1,blaCTX-M2,MultiCaseDHA were identified. Additionally, antibiotic resistance genes tet(A), AAC(3)-iv, and Sul1 were also detected. The pathogens exhibited resistance to Penicillins (ampicillin, amoxicillin), Cephems (cefuroxime, ceftazidime), and Tetracyclines (tetracycline). However, all three bacteria were highly sensitive to meropenem, amikacin, gentamicin, ciprofloxacin, and sulfamethoxazole-trimethoprim. These findings highlight the critical need for targeted antimicrobial stewardship and effective control measures to mitigate the impact of SCM on dairy production and animal health in the Sylhet region of Bangladesh.

Synthesis and Anticancer Activity of Novel Indole Derivatives as Dual EGFR/SRC Kinase Inhibitors.

Recent studies showed that the cooperation between c-SRC and EGFR is responsible for more aggressive phenotype in diverse tumors, including glioblastomas and carcinomas of the colon, breast, and lung. Studies show that combination of SRC and EGFR inhibitors can induce apoptosis and delay the acquired resistance to chemotherapy. Therefore, such combination may lead to a new therapeutic strategy for the treatment of EGFR-mutant lung cancer. Osimertinib was developed as a third-generation EGFR-TKI to combat the toxicity of EGFR mutant inhibitors. Due to the resistance and adverse reaction of osimertinib and other kinase inhibitors, 12 novel compounds structurally similar to osimertinib were designed and synthesized. Compounds were synthesized by developing novel original synthesis methods and receptor interactions were evaluated through a molecular docking study. To evaluate their inhibitory activities against EGFR and SRC kinase, in vitro enzyme assays were used. Anticancer potencies were determined using lung, breast, prostate (A549, MCF6, PC3) cancer cell lines. Compounds were also tested against normal (HEK293) cell line to evaluate their cyctotoxic effects. Although, none of compounds showed stronger inhibition compared to osimertinib in the EGFR enzyme inhibition studies, compound 16 showed the highest efficacy with an IC50 of 1.026 μM. It also presented potent activity against SRC kinase with an IC50 of 0.002 μM. Among the tested compounds, the urea containing derivatives 6-11 exhibited a strong inhibition profile (80.12-89.68%) against SRC kinase in comparison to the reference compound dasatinib (93.26%). Most of the compounds caused more than 50% of cell death in breast, lung and prostate cancer cell lines and weak toxicity for normal cells in comparison to reference compounds osimertinib, dasatinib and cisplatin. Compound 16 showed strong cytotoxicity on lung and prostate cancer cells. Treatment of prostate cancer cell lines with the most active compound, 16, significantly increased the caspase-3 (8-fold), caspase-8 (6-fold) and Bax (5.7-fold) levels and decreased the Bcl-2 level (2.3-fold) compared to the control group. These findings revealed that the compound 16 strongly induces apoptosis in the prostate cancer cell lines. Overall kinase inhibition, cytotoxicity and apoptosis assays presented that compound 16 has dual inhibitory activity against SRC and EGFR kinases while maintaining low toxicity against normal cells. Other compounds also showed considerable activity profiles in kinase and cell culture assays.

Biocontrol of Fusarium spp. in vitro and in vine cuttings using Bacillus sp. F62

Fusarium spp., the causal agent of Fusarium wilt, cause substantial economic losses in viticulture, mainly in tropical regions. This study aimed to assess the biocontrol potential of Bacillus sp. F62 against Fusarium spp., both in vitro and in rootstock cuttings of the SO4 variety. To this end, the in vitro antagonism was evaluated through diffusible and volatile compounds synthesized by Bacillus sp. F62 on three Fusarium spp. isolates. Subsequently, the isolate FusA06-18 was selected for a rootstock cutting experiment. The vine cuttings underwent the following treatments: control, pathogen inoculation (Fus), bacterial inoculation (Bac), and bacterial followed by pathogen inoculation (Bac + Fus). Our findings revealed an average reduction of 39.1% in the mycelial growth of the pathogen through dual culture assay and a decrease of 11.6% in the Fusarium spp. radial growth due to the effects of volatile compounds. In the experiment with vine cuttings, applying Bacillus sp. F62 reduced the pathogen re-isolation frequency from 81.7% (Fus) to 63.3% (Bac + Fus). Therefore, Bacillus sp. F62 effectively suppressed the mycelial growth of Fusarium spp. and reduced the Fusarium wilt incidence in vine cuttings of the rootstock ‘SO4’.

Comparative Analysis of the Antibacterial Properties of Various Periodontal Bone Graft Materials In Vitro.

Periodontal bone grafting is a common procedure in dentistry to restore bone loss resulting from periodontal disease. Various graft materials are available, each with different antibacterial properties. Understanding these properties is crucial for successful grafting outcomes. In this study, we conducted a comparative analysis of the antibacterial properties of four commonly used periodontal bone graft materials: The graft used were A: Hydroxyapatite, B: B TCP, C: Bioactive glass, D: Calcium Phosphate materials. Each material was subjected to in vitro testing using a standardized bacterial culture assay. Antibacterial activity was assessed by measuring the zone of inhibition around each material after exposure to bacterial cultures. Material A exhibited the highest antibacterial activity with an average zone of inhibition of 12.3 mm, followed by Material C with 9.8 mm, Material D with 8.5 mm, and Material B with 6.2 mm. These arbitrary values reflect the relative efficacy of each material in inhibiting bacterial growth. Our findings suggest that Material A possesses the strongest antibacterial properties among the tested periodontal bone graft materials. This may contribute to improved clinical outcomes in periodontal bone grafting procedures by reducing the risk of postoperative infections. Further research is warranted to elucidate the mechanisms underlying the antibacterial effects of these materials and to validate our in vitro findings in clinical settings.

Anti-oomycete activity and plant growth promoting properties of avocado fungal endophytes

Fungal endophytes are known as promising plant growth-promoting microorganisms. Surprisingly, despite the economic importance of avocado, the antimicrobial and plant growth-promoting properties of its fungal endophytes have seldom been investigated. Our objectives were to evaluate the anti-oomycete activity of avocado fungal endophytes and assess their potential plant growth-promoting properties, using Arabidopsis thaliana as a model plant. In total, 89 fungal isolates were obtained from the roots of avocado trees and grouped into 24 morphotypes. One isolate per morphotype was randomly selected to assess its antagonistic activity against Phytophthora cinnamomi through dual culture assays. The strongest inhibition of P. cinnamomi was induced by endophytic fungi belonging to the Fusarium, Mortierella and Penicillium genera. The six most promising fungal isolates were selected to assess their plant growth-promoting traits in co-inoculation assays with A. thaliana. All tested fungal endophytes were able to modify the plant root architecture and increase the number of lateral roots. Moreover, an accumulation of auxins was detected in the xylem and meristematic zone of plants inoculated with Mortierella sp. PC-T3-3.1 and Metapochonia sp. PC-T2-4.2, whilst auxin accumulation was restricted to the emerging lateral roots of plants inoculated with Penicillium sp. C-T0-3.1. Indole quantification showed that Penicillium sp. C-T0-3.1 produced the highest concentration of indole acetic acid (IAA) and indole butyric acid (IBA), despite the lack of auxin responsiveness in plants in the co-inoculation assays. Mortierella sp. PC-T3-3.1 and Metapochonia sp. PC-T2-4.2 produced more IAA and IBA when co-inoculated with A. thaliana than when growing alone, which suggests that sensing of plant signals could induce the production of auxin-like compounds by these two endophytes. Collectively, our findings contribute to elucidate the mechanisms underpinning the plant growth-promoting activity of avocado fungal endophytes, which will be key to harness their full metabolic potential.

Improved physiological responses and stress tolerance in cotton against Fusarium wilt with Trichoderma harzianum treatment

Cotton is a vital cash crop in the agricultural economy. Many abiotic stresses and pathogenic diseases reduce cotton yield, with cotton wilt being a major threat caused by Fusarium oxysporum f.sp. vasinfectum (FOV). In the present study, the antagonistic potential of Trichoderma harzianum, along with its extracted volatile and non-volatile compounds, was evaluated against FOV. In a dual culture assay, T. harzianum showed 71.4% inhibition of FOV. The volatile compounds extracted from T. harzianum showed 54.9% inhibition of FOV, while the non-volatile compounds reduced the growth of F. oxysporum f.sp. vasinfectum by 64.21%. The evaluation of vegetative traits of cotton plants indicated increased biomass when treated with T. harzianum. The activity of antioxidant defense enzymes Catalase activity (CAT), Superoxide dismutase activity (SOD), and Peroxidase activity (POX) was highest in cotton plants treated with both T. harzianum and F. oxysporum f.sp. vasinfectum, indicating an activation of the plant defense mechanisms in response to these treatments. Protein content, Malondialdehyde Content (MDA), proline content, electrolyte leakage, and H2O2 content, which are stress markers, were at their maximum under F. oxysporum f.sp. vasinfectum stress. Physiological features, including stomatal conductance, photosynthesis rate, Soil Plant Analysis Development (SPAD) chlorophyll value, and relative water content, were highest in plants treated with T. harzianum, as T. harzianum improves and boosts plant growth. Hence, T. harzianum is an effective biological control agent against FOV.

The Development of a 3D PET Fibrous Scaffold Modified with an Umbilical Cord dECM for Liver Tissue Engineering.

Organ and tissue dysfunction represents a clinically significant condition. By integrating cell biology with materials science, tissue engineering enables the reconstruction and restoration of damaged tissues or organs, offering a noninvasive repair approach. In our study, we replicated the cellular growth environment by utilizing a human umbilical cord-derived decellularized extracellular matrix (dECM) as a modifying agent for the polyethylene terephthalate (PET) polymeric fiber scaffold. This allowed us to create a dECM-coated polyester fiber-based scaffold, PET-dECM, tailored for liver tissue engineering purposes. We effectively produced a decellularized human umbilical cord-derived ECM through a combined decellularization process involving trypsin/EDTA, TritonX-100, and sodium deoxycholate. The application of the dECM coating onto the PET material was accomplished through several steps, such as ester hydrolysis, EDC/NHS-activated crosslinking, and dECM conjugation. The biological performance of the PET-dECM was validated using RG cell culture assays. Notably, the dECM coating significantly improved PET's hydrophilicity and biocompatibility, thereby aiding cell adhesion, proliferation, and functional differentiation (p < 0.05). It was further found that the hepatocyte function of HepaRG was significantly enhanced on the PET-dECM, which may be attributed to the dECM's ability to facilitate the restoration of cell polarity. The PET-dECM holds promise as an effective hepatocyte culture carrier and could potentially find application in liver tissue engineering.

The biological and therapeutic assessment of a P(3HB-co-4HB)-bioactive glass-graphene composite biomaterial for tissue regeneration.

An ideal wound dressing should create a healing environment that relieves pain, protects against infections, maintains moisture, removes debris, and speeds up wound closure and repair. However, conventional options like gauze often fall short in fulfilling these requirements, especially for chronic or nonhealing wounds. Hence there is a critical need for inventive formulations that offer efficient, cost-effective, and eco-friendly alternatives. This study focuses on assessing the innovative formulation based on a microbial-derived copolymer known as poly(3-hydroxybutyrate-co-4-hydroxybutyrate), P(3HB-co-4HB) bioactive glass and graphene particles, and exploring their biological response in vitro and in vivo-to find the best combination that promotes cell adhesion and enhances wound healing. The formulation optimized at concentration of bioactive glass (1 w/w%) and graphene (0.01 w/w%) showed accelerated degradation and enhanced blood vessel formation. Meanwhile biocompatibility was evaluated using murine osteoblasts, human dermal fibroblasts, and standard cell culture assays, demonstrating no adverse effects after 7 days of culture and well-regulated inflammatory kinetics. Whole thickness skin defect using mice indicated the feasibility of the biocomposites for a faster wound closure and reduced inflammation. Overall, this biocomposite appears promising as an ideal wound dressing material and positively influencing wound healing rates.