Untreated Control Research Articles

Effects of electrical stimulation with alternating fields on the osseointegration of titanium implants in the rabbit tibia - a pilot study.

Introduction: Electrical stimulation has been used as a promising approach in bone repair for several decades. However, the therapeutic use is hampered by inconsistent results due to a lack of standardized application protocols. Recently, electrical stimulation has been considered for the improvement of the osseointegration of dental and endoprosthetic implants. Methods: In a pilot study, the suitability of a specifically developed device for electrical stimulation in situ was assessed. Here, the impact of alternating electric fields on implant osseointegration was tested in a gap model using New Zealand White Rabbits. Stimulation parameters were transmitted to the device via a radio transceiver, thus allowing for real-time monitoring and, if required, variations of stimulation parameters. The effect of electrical stimulation on implant osseointegration was quantified by the bone-implant contact (BIC) assessed by histomorphometric (2D) and µCT (3D) analysis. Results: Direct stimulation with an alternating electric potential of 150mV and 20Hz for three times a day (45min per unit) resulted in improved osseointegration of the triangular titanium implants in the tibiae of the rabbits. The ratio of bone area in histomorphometry (2D analysis) and bone volume (3D analysis) around the implant were significantly increased after stimulation compared to the untreated controls at sacrifice 84days after implantation. Conclusion: The developed experimental design of an electrical stimulation system, which was directly located in the defect zone of rabbit tibiae, provided feedback regarding the integrity of the stimulation device throughout an experiment and would allow variations in the stimulation parameters in future studies. Within this study, electrical stimulation resulted in enhanced implant osseointegration. However, direct electrical stimulation of bone tissue requires the definition of dose-response curves and optimal duration of treatment, which should be the subject of subsequent studies.

Competitive exclusion products as an antimicrobial alternative to control Salmonella Heidelberg in broilers.

Intestinal infections caused by non-typhoidal Salmonella spp., along with antimicrobial resistance spread are a major food safety concern worldwide. Here, we evaluate the potential of competitive exclusion products developed by anaerobic or aerobic conditions to control systemic infection, cecal colonization, fecal excretion, and improve the intestinal health in broilers challenged by Salmonella Heidelberg (SH). A total of 105 day-old chickens were randomly distributed into three experimental groups: A (untreated control), B (treated with anaerobic culture), and C (treated with aerobic culture). During 21 days, morphometric parameters of the small intestine were analyzed using microscopy, fecal excretions by cloacal swabs, systemic infection, and cecal colonization by colony-forming unit counts (CFU/g). The results indicated the lowest number of positive swabs (45.33%) recovered from Group C, followed by Group B (71.8%) and Group A (85.33%). The bacterial enumeration revealed the lowest amounts in Group C at the necropsy realized in 5-, 7-, and 14-days post-infection (DPI) (P=0.0010, P=0.0048, and P=0.0094, respectively). Statistical differences between intestinal morphometrics were observed in the Group C at 21 DPI. Our results suggest that the product developed under aerobic conditions can improve intestinal health, protecting birds against SH.

Antiviral therapy in cats progressively infected with feline leukaemia virus: lessons from a series of 18 consecutive cases from Australia.

It is doubtful that any of the treatments proposed for feline leukaemia virus (FeLV) infection are effective, despite the entity being described 60 years ago. Eighteen pet cats with progressive FeLV infections were recruited in Australia. One or more antiviral drugs were trialled in 16 cats, while two FeLV-infected cats were not handleable and served as untreated controls. Six cats were administered RetroMAD1™ only (0.5 mg/kg orally twice daily), a commercially available recombinant chimeric protein with proposed antiretroviral activity. Three cats were administered the integrase inhibitor raltegravir only (10-15 mg/kg orally twice daily), a drug used as a component of highly effective antiretroviral therapy for human immunodeficiency virus (HIV-1) infection. Three cats were administered RetroMAD1™ and raltegravir concurrently, and four cats were administered raltegravir and the reverse transcriptase inhibitor zidovudine (AZT, 5 mg/kg orally twice daily) concurrently. FeLV RNA and p27 antigen loads were measured at two timepoints (T1-2 months and T3-5 months) during therapy and compared to baseline (pretreatment) levels, to assess the response to therapy using linear modelling. The median survival time (MST) of the cats from commencement of FeLV treatment to death was also determined and compared between treatments. The MST for the 16 FeLV-positive cats which received antiviral therapy was 634 days, while the MST from FeLV diagnosis to death for the two untreated control cats was 780 days. In cats treated with RetroMAD1™, FeLV viral load decreased from T0 to T1-2 months (median viral load reduced from 1339 × 106 to 705 × 106 copies/mL plasma; P = 0.012), but MST was reduced compared to cats not given RetroMAD1™ (426 days vs 1006 days; P = 0.049). Cats treated with raltegravir and AZT had no significant changes in FeLV viral load over time, but p27 antigen load was decreased from T0 to T3-5 months in cats treated with raltegravir (median p27 antigen level reduced from 50.2% to 42.7%; P = 0.005). All other results were not significantly affected by the treatment provided. Importantly, statistically significant and substantial associations were found between age at FeLV diagnosis and survival time (P = 0.046, R2 = 18.6) and between FeLV viral load at T0 and survival time (P = 0.004, R2 = 44.4). Younger cats, and cats with higher levels of pretreatment FeLV RNA, had reduced survival times. Cats treated with RetroMAD1™ were typically younger (median age 2.0 vs 8.0 years), likely explaining the observed reduction in MST. A significant association was found between FeLV viral load and p27 antigen load at T0 (P = 0.015, R2 = 32.9). Results from this small case series do not provide convincing support for the use of RetroMAD1™, raltegravir or AZT, alone or in combination, for the treatment of cats progressively infected with FeLV. The changes observed were biologically insignificant. Age and FeLV viral load at diagnosis are useful prognostic markers, and p27 antigen concentration can be used to predict viral load. Larger field trials should be performed examining antiretroviral therapy in FeLV-positive cats with progressive infections, preferably using three or more drugs from at least two classes, as is standard with human antiretroviral therapy. Future studies would be easier in countries with a higher prevalence of FeLV infections than Australia.

Potential of Sorghum Seeds in Alleviating Hyperglycemia, Oxidative Stress, and Glycation Damage.

Diabetes mellitus, characterized by dysregulated glucose metabolism, oxidative stress, and the formation of advanced glycation end products, poses a significant global health burden. In this study, we explored the potential of sorghum (Sorghum bicolor) seeds, known for their abundant phytochemical composition, as a natural remedy for diabetes and its associated damage. High-performance liquid chromatography/high-resolution mass spectrometry analysis revealed a remarkable phenolic richness in sorghum grains, including gallic acid, quercetin, and the predominant procyanidin B-1, with ecotype-specific variations in flavonoid distribution. Elemental analysis by ICP showed an abundance of macro-elements (Ca, K, Mg), trace elements (Fe, Mn, Si, Zn), and ultra-trace elements (B, Co, Cr, Cu, Mo, Se, V) essential for human health, supporting its therapeutic and nutritional potential. Additionally, the results demonstrated variable total phenolic contents (188-297 mg GAE/g dE) and total flavonoid contents (66-78 mg QE/g dE), with corresponding differences in antioxidant activities across the five ecotypes. Treatment with sorghum seed extract (SE1) significantly reduced oxidative stress markers, such as malondialdehyde (MDA)by 40% and hydrogen peroxide (H2O2) by 63%, in diabetic mice, compared to untreated diabetic controls. Moreover, sorghum extracts exhibited a remarkable increase in antioxidant enzyme activities, including a 50% increase in superoxide dismutase (SOD) activity and a 60% increase in glutathione peroxidase (GPx) activity, indicating their potential to bolster antioxidant defenses against diabetes-induced oxidative stress. These findings underscore the therapeutic potential of sorghum seeds in diabetes management and prevention, paving the way for the development of functional foods with enhanced health benefits.

Testosterone Inhibits Lipid Accumulation in Porcine Preadipocytes by Regulating ELOVL3.

Castration is commonly used to reduce stink during boar production. In porcine adipose tissue, castration reduces androgen levels resulting in metabolic disorders and excessive fat deposition. However, the underlying detailed mechanism remains unclear. In this study, we constructed porcine preadipocyte models with and without androgen by adding testosterone exogenously. The fluorescence intensity of lipid droplet (LD) staining and the fatty acid synthetase (FASN) mRNA levels were lower in the testosterone-treated cells than in the untreated control cells. In contrast, the mRNA levels of adipose triglycerides lipase (ATGL) and androgen receptor (AR) were higher than in the testosterone-treated cells than in the control cells. Subsequently, transcriptomic sequencing of porcine preadipocytes incubated with and without testosterone showed that the mRNA expression levels of very long-chain fatty acid elongase 3 (ELOVL3), a key enzyme involved in fatty acids synthesis and metabolism, were high in control cells. The siRNA-mediated knockdown of ELOVL3 reduced LD accumulation and the mRNA levels of FASN and increased the mRNA levels of ATGL. Next, we conducted dual-luciferase reporter assays using wild-type and mutant ELOVL3 promoter reporters, which showed that the ELOVL3 promoter contained an androgen response element (ARE); furthermore, its transcription was negatively regulated by AR overexpression. In conclusion, our study reveals that testosterone inhibits fat deposition in porcine preadipocytes by suppressing ELOVL3 expression. Moreover, our study provides a theoretical basis for further studies on the mechanisms of fat deposition caused by castration.

Assessment of the therapeutic potential of Hsp70 activator against prion diseases using in vitro and in vivo models.

Prion diseases are deadly neurodegenerative disorders in both animals and humans, causing the destruction of neural tissue and inducing behavioral manifestations. Heat shock proteins (Hsps), act as molecular chaperones by supporting the appropriate folding of proteins and eliminating the misfolded proteins as well as playing a vital role in cell signaling transduction, cell cycle, and apoptosis control. SW02 is a potent activator of Hsp 70kDa (Hsp70). In the current study, the protective effects of SW02 against prion protein 106-126 (PrP106-126)-induced neurotoxicity in human neuroblastoma cells (SH-SY5Y) were investigated. In addition, the therapeutic effects of SW02 in ME7 scrapie-infected mice were evaluated. The results showed that SW02 treatment significantly increased Hsp70 mRNA expression levels and Hsp70 ATPase activity (p < 0.01). SW02 also significantly inhibited cytotoxicity and apoptosis induced by PrP106-126 (p < 0.01) and promoted neurite extension. In vivo, intraperitoneal administration of SW02 did not show a statistically significant difference in survival time (p = 0.16); however, the SW02-treated group exhibited a longer survival time of 223.6 ± 6.0days compared with the untreated control group survival time of 217.6 ± 5.4days. In addition, SW02 reduced the PrPSc accumulation in ME7 scrapie-infected mice at 5months post-injection (p < 0.05). A significant difference was not observed in GFAP expression, an astrocyte marker, between the treated and untreated groups. In conclusion, the potential therapeutic role of the Hsp70 activator SW02 was determined in the present study and may be a novel and effective drug to mitigate the pathologies of prion diseases and other neurodegenerative diseases. Further studies using a combination of two pharmacological activators of Hsp70 are required to maximize the effectiveness of each intervention.

Magnetic particles with polymeric shells bearing lithocholic and folic acid moieties: Nano-warriors to fight colon cancer

In the study, we aimed to investigate the activity of nanoformulations containing 5-fluorouracil and polymer-magnetic hybrids bearing membrane-penetrating and ligand-receptor-recognizing agents against colorectal cancer cells. The formation and characterization of iron oxide particles covered with polymeric shells comprising lithocholic acid and folic acid moieties are presented. The efficiency of nanoformulations combined by the simple mixing of low doses of 5-fluorouracil with the obtained hybrids was demonstrated against DLD-1 and HT-29 colon cancer cells. The most pronounced cytotoxic potential against HT-29 cells was observed in the cases of particles based on block and randomly arranged copolymers functionalized by FA motifs with depletion of viable cells by approximately 50 % compared to control cells and cells treated by 5-FU applied in free form. In the case of the DLD-1 cell line, the percentage of viable DLD-1 cells decreased by about 30 to 40% after treatment with the block and randomly arranged copolymer decorated by FA-moiety, when compared to 5-FU at the free form and the untreated control. The induction of apoptosis associated with PS-translocation was determined to be the main mechanism of their cytotoxic effects. Moreover, the safety profiles of the nanoformulations were established through hemolysis assay and the analysis of the viability of human colorectal fibroblasts. It was indicated that all tested nanoparticles met the compatibility requirements at the in vitro level. It should be emphasized that in many cases, there was a significant improvement in the compatibility of hybrids with the FA motif compared to non-functionalized hybrids with the addition of 5-FU. These findings suggest that the presence of FA might modulate the toxicity of chemotherapeutic agents.

Methods to Induce Sprouting in Dormant Potato Tubers for Direct Tuber Testing of Potato Virus Y

The ability to initiate sprouting soon after harvest to enable direct tuber testing for potato virus Y (PVY) could aid in acquiring more rapid results compared to the traditional winter grow out tests currently used. Methods to break dormancy for PVY detection using laboratory direct tuber testing by ELISA in commercially produced Ranger Russet, Clearwater Russet, and Umatilla Russet seed lots were tested over two years and compared to leaf testing results obtained from the winter grow out and spring grow out. At harvest, three 400 tuber samples from each cultivar were obtained for the trial and included (1) untreated control (UTC), (2) application of cold aerosol smoke, or (3) application of Rindite. Tuber samples were held at 18.3 C and sprout development was monitored weekly. Treatments were direct tuber tested for PVY when one treatment of that cultivar achieved three sprouts elongating to six millimeters. A fourth 400 tuber sample was collected, treated with Rindite, and included in the Idaho winter grow out plots in Waialua, Hawaii and leaves were sampled and evaluated for PVY using ELISA. Laboratory tested seed was stored and planted in a spring grow out (Kimberly, Idaho) and leaf samples were analyzed for PVY by ELISA. Rindite treated tubers had greater sprout rating and number of sprouts elongating compared to UTC tubers and tubers receiving the smoke treatment at time of PVY testing. Smoke had a greater sprout rating but did not always significantly differ in the number of sprouts elongating compared to the untreated tubers. Overall, estimates of PVY prevalence from direct tuber testing showed limited significant differences to those obtained in the winter grow out for each cultivar, year, and PVY incidence. However, in year two, the incidence of PVY in the winter grow out (7% PVY) significantly differed from direct tuber testing (16% PVY) in Ranger Russet. In both years, the spring grow out PVY results for all cultivars were not significantly different than the direct tuber testing, except in year one the Ranger Russet direct tuber tested UTC showed 10% lower PVY detection compared to the spring grow out. This study identified a novel dormancy breaking treatment to promote earlier and accurate PVY detection by direct tuber testing using ELISA and provided data to support direct tuber testing for post-harvest evaluation of PVY in seed certification.

Whitening toothpastes with hydrogen peroxide concentrations vs. at-home bleaching.

To evaluate the effect of whitening toothpastes with different hydrogen peroxide (HP) concentrations on HP permeability, color change, and physicochemical properties, compared to at-home bleaching treatment. Forty-nine premolars were randomized into seven groups (n = 7): untreated (control); at-home bleaching with 10% carbamide peroxide gel (AH; 10% CP) with 14 and 28 applications of 180min each (AH [14 × 180min] and AH [28 × 180min]); three whitening toothpastes (3% HP; 4% HP and 5% HP) and 10% CP brushed 28 times for 90s each (TB [28 × 90s]). HP permeability was measured using a UV-VIS spectrophotometer and color change by a digital spectrophotometer (ΔEab, ΔE00, and ΔWID). Initial concentration, pH, and viscosity were measured through titration, digital pH meter, and rheometer, respectively. Statistical analysis included one-way ANOVA, Tukey's test, and Dunnett's test (α = 0.05). 4% HP group showed acidic pH, the lowest viscosity and the highest HP concentration into the pulp chamber (p < 0.05). The 10% CP groups had lower HP in the pulp chamber and greater color change than other groups (p < 0.05), except the 5% HP group in ΔEab and ΔE00. For ΔWID, the 10% CP AH groups showed greater whitening than other groups (p < 0.05). Whitening toothpaste with up to 5% HP resulted in higher HP permeability and less color change compared to 10% CP. Higher HP commercial concentrations in toothpaste increased whitening effect; however, acidic pH toothpastes exhibited greater HP permeability. Whitening toothpastes with high hydrogen peroxide concentrations were less effective than at-home bleaching, resulting in less color change and greater permeability of hydrogen peroxide, potentially increasing the risk of tooth sensitivity.

Blue-Emissive Antioxidant Carbon Dots Enhance Drought Resistance of Pea (Pisum sativum L.).

Prolonged drought conditions are a critical challenge for agricultural advancement, threatening food security and environmental equilibrium. To overcome these issues, enhancing plant resilience to drought is essential for plant growth and sustainable agriculture. In this study, blue-emitting antioxidant carbon dots (B-CDs), synthesized from citric acid and ascorbic acid, emerged as a promising solution to enhance the drought resistance of peas (Pisum sativum L.). B-CDs can efficiently scavenge reactive oxygen species (ROS), which are harmful in excess to plants under stress conditions. Through detailed experimental analyses and density functional theory (DFT) studies, it is found that these B-CDs possess structures featuring eight-membered aromatic rings with abundant oxygen-containing functional groups, providing active sites for reactions with ROS. The practical benefits of the B-CDs are evident in tests with pea plants exposed to drought conditions. These plants show a remarkable reduction in ROS accumulation, an increase in photosynthetic efficiency due to improved electron transfer rates, and significant growth enhancement. Compared to untreated controls under drought stress, the application of B-CDs results in an impressive increase in the fresh and dry weights of both the shoots and roots of pea seedlings by 39.5 and 43.2% for fresh weights and 121.0 and 73.7% for dry weights, respectively. This suggests that B-CDs can significantly mitigate the negative effects of drought on plants. Thus, leveraging B-CDs opens a novel avenue for enhancing plant resilience to abiotic stressors through nanotechnology, thereby offering a sustainable pathway to counter the challenges of drought in agriculture.

Evaluation of treatment with human chorionic gonadotropin at transfer of in vitro produced beef embryos on reproductive outcomes in lactating multiparous Jersey cows after a synchronized ovulation

Our objective was to evaluate the effect of treatment with human chorionic gonadotropin (hCG) at the time of transfer of in vitro produced (IVP) beef embryos on pregnancy outcomes in lactating multiparous Jersey cows. Grade 1, Stage 7 (expanded blastocyst), IVP beef embryos were produced from black Angus-based dams using 3 proven high fertility Angus sires and were frozen for direct transfer. In a preliminary experiment, lactating multiparous Jersey cows were randomized to a 2x2 factorial arrangement of treatments to test the main effect of recipient synchronization protocol (Double-Ovsynch; DO; n = 169 vs. a synchronized estrus; ED; n = 180) and were randomly assigned within recipient protocol to serve as untreated controls (DO-CON, n = 78; ED-CON, n = 44) or to receive i.m. treatment with 2,500 IU of hCG (DO-hCG, n = 79; ED-hCG, n = 46) at the time of embryo transfer (ET). The recipient utilization rate was greater for DO (93%) than for ED (50%) cows, and there was an interaction between recipient synchronization protocol and hCG treatment in which DO-hCG cows had more pregnancies per embryo transfer (P/ET) at 26, 33, and 61 d than DO-CON, ED-hCG, and ED-CON cows. Based on a partial budget analysis, the cost per pregnancy for DO cows was $135.35 less than for ED cows. In Experiment 2, lactating multiparous Jersey cows were submitted to a Double-Ovsynch protocol (DO, n = 386) and were randomly assigned to serve as untreated controls (CON, n = 192) or were treated with 2,500 IU hCG (hCG, n = 194) at ET. Progesterone concentrations and total luteal volume 7 d after ET were greater for hCG than for CON cows. In contrast to the preliminary experiment, treatment with hCG did not affect P/ET at 26, 33, or 61 d, and treatment with hCG did not affect pregnancy loss from 26 to 61 d. In conclusion, treatment with 2,500 IU of hCG at ET increased P4 concentrations and total luteal volume 7 d after ET but did not increase pregnancy outcomes or decrease pregnancy loss in lactating multiparous Jersey cows receiving frozen/thawed IVP beef embryos.

Bactericidal Effect of Different Photochemical-Based Therapy Options on Implant Surfaces-An In Vitro Study.

Objectives: Photochemical systems are frequently recommended as an adjuvant treatment option in peri-implantitis therapy. The aim of the present study was to evaluate the efficacy of these treatment options, as well as a novel curcumin-based option, in a biofilm model on implants. Methods: Eighty dental implants were inoculated with an artificial biofilm of periodontal pathogens and placed in peri-implant pocket models. The following groups were analyzed: I, photodynamic therapy (PDT); II, PDT dye; III, curcumin/DMSO + laser; IV, curcumin/DMSO only; V, dimethyl sulfoxide (DMSO) only; VI, photothermal therapy (PTT); VII, PTT dye; VIII, control. After treatment, remaining bacterial loads were assessed microbiologically using quantitative real-time polymerase chain reaction analysis. Results: The PDT, PTT, and DMSO treatment methods were associated with statistically significant (p < 0.05) improvements in germ reduction in comparison with the other methods and the untreated control group. The mean percentage reductions were as follows: I (PDT) 93.9%, II (PDT dye) 62.9%, III (curcumin/DMSO + laser) 74.8%, IV (curcumin/DMSO only) 67.9%, V (DMSO) 89.4%, VI (PTT) 86.8%, and VII (PTT dye) 66.3%. Conclusions: The commercially available PDT and PTT adjuvant treatment systems were associated with the largest statistically significant reduction in periopathogenic bacteria on implant surfaces. However, activation with laser light at a suitable wavelength is necessary to achieve the bactericidal effects. The use of curcumin as a photosensitizer for 445 nm laser irradiation did not lead to any improvement in antibacterial efficacy in comparison with rinsing with DMSO solution alone.

In vitro activity of Western Australian honeys and Manuka honey against clinically important yeasts.

With the steady rise in antifungal resistance amongst clinically important yeasts, antifungal drug discovery remains of the utmost importance. To determine the potential of some honeys as alternative antifungal agents, we quantified the antifungal activity of 12 Western Australian honey samples, two Manuka honey samples and an artificial honey against 10 yeast isolates including clinical and reference strains. Results showed that the tested honeys varied in activity, and yeasts species also differed in susceptibility, with minimum inhibitory concentrations (MICs) determined by broth microdilution ranging from 8% to >44% w/v honey. Honeys with the highest overall activity were derived from Blackbutt (Eucalyptus patens), Jarrah (E. marginata), and Karri (E. diversicolor). The optical density of each MIC microtitre plate was determined after incubation and showed that at relatively low concentrations of honey the growth of all yeasts was enhanced compared to the untreated control, whereas at and above approximately 12% w/v, honeys exerted a dose-dependent growth inhibitory effect, the extent of which varied by honey type. Time-kill studies with 64% w/v honey showed that all eight of the natural honeys tested had greater fungicidal activity than the comparator artificial honey. Our findings suggest that the specific nectar-derived phytochemicals present within each honey play an important role in antifungal activity, and support the notion that activity is due to a combination of factors including osmotic activity, hydrogen peroxide and phytochemical compounds. These data indicate that honey is worthy of further investigation as a potential therapeutic agent for superficial yeast infections.

Inhibition of CYP1A1 expression enhances diabetic wound healing by modulating inflammation and oxidative stress in a rat model

ObjectiveWound therapies utilizing gene delivery to the skin offer considerable promise owing to their localized treatment benefits and straightforward application. This study investigated the impact of skin electroporation of CYP1A1 shRNA lentiviral particles on diabetic wound healing in a streptozotocin (STZ)-induced rat model. MethodsMale Sprague Dawley (SD) rats were made diabetic by injecting STZ and subsequently creating foot skin wounds. The rats were randomly divided into four groups: normal, diabetic foot ulcers (DFU), DFU + control shRNA (electroporation of control shRNA lentiviral particles), and DFU + CYP1A1 shRNA (electroporation of CYP1A1 shRNA lentiviral particles). Wound healing progress was monitored at multiple time points (0, 1, 3, 5, 7, 10, 14 days). On day 14, wound tissue specimens were collected for histological examination. Wound samples collected at days 7 and 14 were used for gene expression analysis via qRT-PCR, assessment of CYP1A1 protein levels using western blotting, and evaluation of oxidative stress markers. ResultsTreatment with CYP1A1 shRNA significantly enhanced diabetic wound healing rates compared to untreated controls over the observation period. Histological analysis revealed improved wound characteristics in the CYP1A1 shRNA-treated group, including enhanced epithelial regeneration, reduced inflammation, and increased collagen deposition, indicative of improved tissue repair. Furthermore, suppression of CYP1A1 corresponded with decreased expression levels of pro-inflammatory cytokines (interleukin-1β, tumor necrosis factor-α, and interleukin-6) and diminished oxidative stress markers (malondialdehyde, superoxide dismutase) within wound tissues. ConclusionTargeted suppression of CYP1A1 represents a promising therapeutic strategy to enhance diabetic wound healing by modulating inflammation and oxidative stress.

Inhibition of neutrophil rolling and migration by caADAMTS13 in vitro and in mouse models of thrombosis and inflammation

Recent investigation of a constitutively active ADAMTS13 variant (caADAMTS13) in murine models of acute ischaemic stroke (AIS) have revealed a potential anti-inflammatory mechanism of action contributing to its protective effect. However, it remains unclear whether these observations are a direct result of VWF proteolysis by caADAMTS13. We have implemented state of the art in vitro assays of neutrophil rolling and transmigration to quantify the impact of caADAMTS13 on these processes. Moreover, we have tested caADAMTS13 in two in vivo assays of neutrophil migration to confirm the impact of the treatment on the neutrophil response to sterile inflammation. Neutrophil rolling, over an interleukin-1β stimulated hCMEC/D3 monolayer, is directly inhibited by caADAMTS13, reducing the proportion of neutrophils rolling to 9.5 ± 3.8 % compared to 18.0 ± 4.5 % in untreated controls. Similarly, neutrophil transmigration recorded in real-time, was significantly suppressed in the presence of caADAMTS13 which reduced the number of migration events to a level like that in unstimulated controls (18.0 ± 4.5 and 15.8 ± 7.5 cells/mm2/h, respectively). Brain tissue from mice undergoing experimental focal cerebral ischaemia has indicated the inhibition of this process by caADAMTS13. This is supported by caADAMTS13’s ability to reduce neutrophil migration into the peritoneal cavity in an ischaemia-independent model of sterile inflammation, with the VWF-dependent mechanism by which this occurs being confirmed using a second experimental stroke model. These findings will be an important consideration in the further development of caADAMTS13 as a potential therapy for AIS and other thromboinflammatory pathologies, including cardiovascular disease.

Fungal pathogen: A game changer for insect herbivore in sequential attack on tomato

Plants have several means of coping with biotic stress, which involves physiological, cellular and molecular mechanisms. The interplay of two principal phytohormonal pathways, Jasmonic acid (JA) and Salicylic acid (SA) play a crucial role in transmitting stress-induced signals throughout the plant. However, the interplay of these hormonal pathways is not clear when plants encounter biotic agents that trigger antagonistic hormonal signals. To address this, we conducted biochemical and gene expression studies on tomato, Solanum lycopersicum plants, exposed to insect alone (IA), fungus alone (FA), or sequential dual stress with fungi followed by insect (FI) and insect followed by fungi (IF), with untreated control. We also assessed the performance of late-arriving conspecific herbivore on these plant sets. The total phenol, and non-tannin phenols content in IF and FI plants showed similar increase or decrease as FA and IA plants, respectively. Gene expression analysis revealed that JA-related genes (MYC2) were dominant in FI plants, while SA-related genes (PR1) were upregulated in IF plants. Moreover, late-arriving herbivores showed significantly decreased relative growth rate, efficiency of conversion of ingested food (ECI) and digested food (ECD) when feeding on FA and IF plants, indicating that sequential treatments substantially induce plant responses. However, herbivory did not affect the late-arriving conspecific herbivore. Thus, biochemical responses and hormonal interplay of the plant are influenced by both sequence as well as the duration of the biotic stress. Furthermore, while the late-arriving conspecific herbivore is negatively affected by the presence of a pathogen, it remains unaffected by previous herbivory.

Fertilization Type Differentially Affects Barley Grain Yield and Nutrient Content, Soil and Microbial Properties.

The use of artificial fertilizers follows the intensification of agricultural production as a consequence of population growth, which leads to soil depletion, loss of organic matter, and pollution of the environment and production. This can be overcome by increasing the use of organic fertilizers in agriculture. In the present study, we investigated the effect of using vermicompost, biochar, mineral fertilizer, a combination of vermicompost and mineral fertilizer, and an untreated control on alluvial-meadow soil on the development of fodder winter barley Hordeum vulgare L., Zemela cultivar. We used a randomized complete block design of four replications per treatment. Barley grain yield, number of plants, and soil and microbiological parameters were studied. We found statistically proven highest grain yield and grain protein values when applying vermicompost alone, followed by the combined treatment and mineral fertilizer. The total organic carbon was increased by 70.2% in the case of vermicompost and by 44% in the case of combined treatment, both compared to the control. Thus, soil microbiome activity and enzyme activities were higher in vermicompost treatment, where the activity of β-glucosidase was 29.4% higher in respect to the control, 37.5% to the mineral fertilizer, and 24.5% to the combined treatments. In conclusion, our study found the best overall performance of vermicompost compared to the rest of the soil amendments.

Multistress‐tolerant Pseudomonas aeruginosa SSVP3 uses multiple strategies to control Meloidogyne incognita in tomato

AbstractTomato is the most important horticultural crop, and India is its second largest producer; however, the plant‐pathogenic nematode Meloidogyne incognita is a serious pest of tomato, causing detrimental losses in its production. This study is focused on the use of multistress‐tolerant Pseudomonas aeruginosa SSVP3 to control this devastating nematode and attempts to decipher the roles of its metabolites and volatile organic compounds (VOCs) during this interaction. The supernatant of P. aeruginosa SSVP3 caused 58% mortality in M. incognita J2 juveniles. The major nematicidal metabolites, identified using liquid chromatography‐mass spectrometry, were pyocyanin, pyoluteorin, pyochelin, benthocyanin and phenazines. The VOCs secreted by P. aeruginosa SSVP3, which were identified via gas chromatography–mass spectrometry (HS‐SPME‐GC–MS) in a mixture, caused 98% mortality in M. incognita J2 juveniles. In a pot experiment, seedling root treatment with P. aeruginosa SSVP3 activated the induced systemic response in tomato plants to M. incognita by increasing the activity of defence and antioxidant enzymes as well as the proline and phenolic contents. The malondialdehyde content in P. aeruginosa SSVP3‐treated plants decreased as compared to those in the other treatment groups. The number of galls formed and the number of endoparasitic stages of M. incognita in the untreated nematode control plants were much greater (19 and 35) than those in the P. aeruginosa SSVP3‐treated (0.33 and 1) and chemically treated groups (0.67 and 1.33). These results indicate that P. aeruginosa and its secreted metabolites and VOCs have a high potential for controlling nematodes.

Synergic Effect of the Antimicrobial Peptide ToAP2 and Fluconazole on Candida albicans Biofilms.

Candida albicans is one of the agents of invasive candidiasis, a life-threatening disease strongly associated with hospitalization, particularly among patients in intensive care units with central venous catheters. This study aimed to evaluate the synergistic activity of the antifungal peptide ToAP2 combined with fluconazole against C. albicans biofilms grown on various materials. We tested combinations of different concentrations of the peptide ToAP2 with fluconazole on C. albicans biofilms. These biofilms were generated on 96-well plates, intravenous catheters, and infusion tubes in RPMI medium at two maturation stages. Scanning electron microscopy and atomic force microscopy were employed to assess the biofilm structure. We also evaluated the expression of genes previously proven to be involved in C. albicans biofilm formation in planktonic and biofilm cells after treatment with the peptide ToAP2 using qPCR. ToAP2 demonstrated a synergistic effect with fluconazole at concentrations up to 25 µM during both the early and mature stages of biofilm formation in 96-well plates and on medical devices. Combinations of 50, 25, and 12.5 µM of ToAP2 with 52 µM of fluconazole significantly reduced the biofilm viability compared to individual treatments and untreated controls. These results were supported by substantial structural changes in the biofilms observed through both scanning and atomic force microscopy. The gene expression analysis of C. albicans cells treated with 25 µM of ToAP2 revealed a decrease in the expression of genes associated with membrane synthesis, along with an increase in the expression of genes involved in efflux pumps, adhesins, and filamentation. Our results highlight the efficacy of the combined ToAP2 and fluconazole treatment against C. albicans biofilms. This combination not only shows therapeutic potential but also suggests its utility in developing preventive biofilm tools for intravenous catheters.

Effect of Novel and Traditional Intracanal Medicaments on Biofilm Viability and Composition

IntroductionThe aim of this study was to test the hypothesis that a combination of D-amino acids (DAAs) and trans-cinnamaldehyde (TC) demonstrates superior antibiofilm activity to calcium hydroxide (CH) and untreated controls. MethodsIn this 3-part in vitro study, the concentration of DAAs (D-methionine, D-leucine, D-tyrosine, and D-tryptophan) that would significantly decrease Enterococcus faecalis and Actinomyces naeslundii biofilm biomass was first determined. Then, the effect of TC + selected DAAs on polymicrobial biofilms was characterized by quantifying the biomass and biofilm viability. Finally, the antibiofilm effects of TC + DAA was compared with CH and untreated controls by (i) determining bacterial viability and (ii) quantifying biofilm matrix composition using selective fluorescence-binding analysis. Statistical analysis was performed using one-way ANOVA and appropriate multiple comparisons test, with P < .05 considered as statistically significant. ResultsTC (0.06%) + D-tyrosine (1 mM) + D-tryptophan (25 mM) significantly reduced the biomass and biofilm viability compared to the control (P < .05). While no significant difference was observed between TC + DAA and CH in the cultivable bacterial counts (P > .05), confocal microscopy demonstrated a significantly greater percentage of dead bacteria in TC + DAA-treated biofilms compared to CH and the control (P < .05). TC + DAA significantly decreased the biovolume and all the examined components of the biofilm matrix quantity compared to the control, while CH significantly reduced only the exopolysaccharide quantity (P < .05). ConclusionThe combination of TC + D-tyrosine + D-tryptophan demonstrated superior antibiofilm activity (biofilm bacterial killing and reduction of matrix quantity) to CH and has potential to be developed as an intracanal medicament.