Untreated Control Research Articles

Long chain unsaturated fatty acids alter growth and reduce biofilm formation of Cronobacter sakazakii

AbstractCronobacter sakazakii is a foodborne pathogen predominately transmitted through contaminated dried foods and affects populations including neonates, infants, and the elderly. Following several recent outbreaks, it is now a notifiable infection in those under 12 months of age. Current control methods include strict manufacturing guidelines, with monitoring of this genus a legal requirement in powdered infant formula production. Fatty acids have long been known as antimicrobials, with long‐chain fatty acids increasingly identified as agents that target virulence factors. This study gives insight into the changes promoted by three long‐chain unsaturated fatty acids (oleic, linoleic, and α‐linolenic) on C. sakazakii growth, morphology, and biofilm formation. Each fatty acid was individually introduced to C. sakazakii 29544 both as a sole carbon source and as an addition to complex media. Following comparison to the untreated control, bacterial cells treated with these fatty acids showed a significant and media‐dependent impact on growth and biofilm inhibition. With further characterization, long‐chain fatty acids, including α‐linolenic acid, could be utilized as a control method with minimal safety constraints regarding their use in the food production environment.

Ameliorative Effect of Faidherbia albida Against 2.4-Dinitrophenylhydrazine Induced Hyperbilirubinemia in Wistar Albino Rats.

Faidherbia albida, popularly known as gawo in Hausa, is traditionally used to treat jaundice in Zuru emirate of Kebbi State. Herein, the ameliorative effect of F. albida against 2.4-dinitrophenylhydrazine-induced hyperbilirubinemia in Wistar albino rats was investigated. Thirty healthy rats were administered 75 mg of 2.4-dinitrophenylhydrazine to induce hyperbilirubinemia. Thereafter, groups 1-3 received 500, 750, and 1000 mg/kg body weight of the methanol stem-bark extract, and 15 mg/kg of phenobarbitone (standard drug) was administered to group 4. Groups 5 and 6 served as the untreated and normal controls, respectively. The phytochemical composition was evaluated using standard methods, and acute oral toxicity was evaluated using standard OECD 2008 guidelines. Phytochemical analysis revealed the presence of alkaloids, phenols, and a substantial amount of tannins. A significant (P<0.05) reduction of direct bilirubin, total bilirubin, and total protein levels for all the doses of the extract and standard drug compared to untreated groups was observed. Similarly, there were significant reductions in serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) levels of the group treated with the standard drug and all extract-treated groups compared to elevated levels observed in untreated controls. However, a significant (P<0.05) increase in serum albumin (ALB) levels, red blood cells, hemoglobin, and pack cell volume was observed in all extract-treated compared to the untreated control in contrast to a significant decrease in MCH levels in treated groups compared to the untreated group. F. albida ameliorated the hyperbilirubinemia induced by 2.4-dinitrophenylhydrazine in Wistar albino rats, thus providing some support for its use in traditional medicine to treat jaundice.

Evaluating the seasonal efficacy of commonly used chemical treatments on Varroa destructor (Mesostigmata: Varroidae)populationresurgence in honey bee colonies.

The purpose of this research was to determine how common chemical treatments influence Varroa destructor (Anderson and Trueman) populationresurgence rates (defined as time posttreatment for mite populations to reach 3 mites/100 adult bees) in managed honey bee (Apis mellifera L.) colonies seasonally. We conducted 2 experiments that followed the same basic protocol to address this purpose. We established 6 treatment groups in Experiment 1 in the fall of 2014: untreated control, Apivar, Apistan, CheckMite+, ApiLifeVar, and Mite Away II applied to 10 colonies per treatment. In Experiment 2, we applied 8 chemical treatments to each of 4 seasonal (spring, summer, fall, and winter) cohorts of honey bee colonies to determine how mite populations are influenced by the treatments. The treatments/formulations tested were Apivar, Apistan, Apiguard, MAQS, CheckMite+, oxalic acid (dribble), oxalic acid (shop towels), and amitraz (shop towels soaked in Bovitraz). In Experiment 1, Apivar and Mite Away II were able to delay V. destructor resurgence for 2 and 6 months, respectively. In Experiment 2, Apiguard, MAQS, oxalic acid (dribble), and Bovitraz treatments were effective at delaying V. destructor resurgence for at least 2 months during winter and spring. Only the Bovitraz and MAQS treatments were effective at controlling V. destructor in the summer and fall. Of the 2 amitraz-based treatments, the off-label Bovitraz treatment was the only treatment to reduce V. destructor populations in every season. The data gathered through this study allow for the refinement of treatment recommendations for V. destructor, especially regarding the seasonal efficacy of each miticide and the temporal efficacy posttreatment.

Transcriptomic analysis of the defense response in "Cabernet Sauvignon" grape leaf induced by Apolygus lucorum feeding.

To investigate the molecular mechanism of the defense response of "Cabernet Sauvignon" grapes to feeding by Apolygus lucorum, high-throughput sequencing technology was used to analyze the transcriptome of grape leaves under three different treatments: feeding by A. lucorum, puncture injury, and an untreated control. The research findings indicated that the differentially expressed genes were primarily enriched in three aspects: cellular composition, molecular function, and biological process. These genes were found to be involved in 42 metabolic pathways, particularly in plant hormone signaling metabolism, plant-pathogen interaction, MAPK signaling pathway, and other metabolic pathways associated with plant-induced insect resistance. Feeding by A. lucorum stimulated and upregulated a significant number of genes related to jasmonic acid and calcium ion pathways, suggesting their crucial role in the defense molecular mechanism of "Cabernet Sauvignon" grapes. The consistency between the gene expression and transcriptome sequencing results further supports these findings. This study provides a reference for the further exploration of the defense response in "Cabernet Sauvignon" grapes by elucidating the expression of relevant genes during feeding by A. lucorum.

Video-tracked Anopheles arabiensis entry and exit behaviour at washed and damaged pyrethroid-treated bednets.

Insecticide-treated nets (ITNs) are the most effective method for malaria prevention in Africa. Using near-infrared video tracking in a laboratory environment, we recorded and assessed bednet entry and exit by a northern Tanzanian population of Anopheles arabiensis at a human-occupied untreated net and a PermaNet® 2.0 ITN. Both had 12 holes, each 10 cm in diameter, punctured at specific locations, and the ITN was washed 20 times to further simulate the wear and tear of ageing. Washing reduced the insecticide content of ITNs by 61%, which then showed similar rates to the untreated nets for net entry (39% entered untreated net and 41% entered ITN; p = 0.84) and exit (37% and 43%, respectively; p = 0.67). Regardless of treatment, approximately 40% of mosquitoes entered nets within 20 s of first appearing in the field of view and reached the volunteer's skin within 5 s of entering the net. Mortality rates post-exposure were significantly higher (p = 0.048) at ITNs (26.6%; 95% CI 13.4%-39.7%) than at untreated controls (6.4%; 95% CI 1.8%-14.6%). The washed and aged ITN provided little additional personal protection for the sleeper over an untreated net. Simple adjustments to materials and design that could extend the effective lifespan of ITNs are discussed.

The Beneficial Effects of Soluble Silicon Fertilizer in Dendrobium Orchids: Silicon-Augmented Resistance against Damage by Insect Pests and Fungal Pathogens.

The effects of soluble silicon fertilization on monocots and dicots have been widely studied. However, little is known regarding its effects on protecting epiphytes against insect and fungal pests. The efficacy of silicon fertilizer to reduce damage by thrips pest complexes, namely: Thrips palmi Karny, Frankliniella occidentalis Pergande, Chaetanaphothrips orchidii Moulton, and Chaetanaphothrips signipennis Bagnall (Thysanoptera: Thripidae), and the fungal pathogens: Botrytis cinerea Persoon (Helotiales: Sclerotiniaceae) and Fusarium spp. Link (Hypocreales: Nectriaceae) was examined during a nine-month greenhouse trial in Hawaii. The trial assessed yield, quality, and pest damage across three common varieties of dendrobiums. All replicates received additional soluble silicon fertilizer applications alternating weekly between soil drench and foliar (50 mg Si/plant) applications. Yield, quality, and spray length, pest damage, plant vigor, SPAD, and leaf temperature were measured. Data were analyzed using a generalized linear model (glm) with repeated measures followed by post-hoc pair-wise comparisons in R, version 4.3.1. Treatment effects were significant at p < 0.001 for the majority of the explanatory variables including: marketable yield, spray length, thrips damage, and fungal damage. Overall, the lavender variety ('Uniwai Supreme') benefited the most from silicon applications with a 73.0% increase in marketable yield, compared to the white variety ('Uniwai Mist'), which had an increase of 50.6% marketable sprays in contrast to its untreated control. Si benefits conferred to the purple variety ('Uniwai Royale') were intermediate to the lavender and white varieties. Although the magnitude of Si benefits varied among the varieties, all dendrobium varieties significantly benefited from silicon fertilization.

Human plasma can modulate micronucleus frequency in TK6 and OE33 cells in vitro

In this paper, we studied the potential genotoxic effects of human plasma from healthy volunteers, as well as patients with gastro-oesophageal reflux disease, Barrett’s oesophagus (BO) and oesophageal adenocarcinoma (OAC) using the oesophageal adenocarcinoma cell line (OE33) and the lymphoblastoid cell line (TK6). Both TK6 and OE33 cells were treated with plasma (10 % volume, replacing foetal bovine serum (FBS) or horse serum (HS)) at different time points of 4 h (for the micronucleus (Mn) assay and the invasion assay) and 24 h (for the cell cycle studies). Plasma-induced effects on DNA damage levels, cell viability and the cell cycle were studied by the micronucleus assay, cytokinesis block proliferation index (CBPI) and flow cytometry respectively. The expression of IL-8 in supernatants of TK6 cells and IFN-β in OE33 cells was also analysed by enzyme-linked immunosorbent assay (ELISA). Finally, we carried out an assessment of cellular invasion of OE33 cells following plasma treatment.The results of the micronucleus assay confirmed the genotoxicity of direct plasma treatment from some participants through the increase in DNA damage in TK6 cells. Conversely, some individual patient plasma samples reduced background levels of TK6 cell Mn frequency, in an anti-genotoxic fashion. In TK6 cells, (on average) plasma samples from patients with Barrett’s oesophagus induced higher micronucleus levels than healthy volunteers (p= 0.0019). There was little difference in Mn induction when using plasma versus serum to treat the cells in vitro. Cell cycle results showed that direct plasma treatment had a marked impact on OE33 cells at 24 h (p=0.0182 for BO and p=0.0320 for OAC) by decreasing the proportion of cells in the S phase, while plasma exposure was less impactful on the cell cycle of TK6 cells. Invasion of OE33 cells was also seen to be non-significantly affected by plasma treatment of OE33 cells.The addition of N-acetyl cysteine NAC in a dose-dependent matter did not alter the formation of Mn in TK6 cells, suggesting that reactive oxygen species (ROS) are not the root cause of plasma’s genotoxicity. The concentration of IL-8 in TK6 cells and IFN-β in OE33 cells was significantly higher in cells treated with OAC-derived plasma than in the untreated negative control. Collectively, our results demonstrate that plasma-specific effects are detectable which helps us better understand some important aspects of the biology of blood-based biomarkers under development.

Synthesis and In vitro and In silico Anti-inflammatory Activity of New Thiazolidinedione-quinoline Derivatives.

Inflammation is a series of complex defense-related reactions. The inflammation cascade produces various pro-inflammatory mediators. Unregulated production of these pro-inflammatory mediators can lead to a wide range of diseases, including rheumatoid arthritis, sepsis, and inflammatory bowel disease. In the literature, the anti-inflammatory action of quinoline and thiazolidinedione nuclei are well established, alone, and associated with other nuclei. The synthesis of hybrid molecules is a strategy for obtaining more efficient molecules due to the union of pharmacophoric nuclei known to be related to pharmacological activity. Based on this, this work presents the synthesis of thiazolidinedione-quinoline molecular hybrids and their involvement in the modulation of cytokines involved in the inflammatory reaction cascade. After synthesis and characterization, the compounds were submitted to cell viability test (MTT), ELISA IFN-γ and TNF-α, adipogenic differentiation, and molecular docking assay with PPARy and COX-2 targets. LPSF/ZKD2 and LPSF/ZKD7 showed a significant decrease in the concentration of IFN- γ and TNF-α, with a dose-dependent behavior. LPSF/ZKD4 at a concentration of 50 μM significantly reduced IL-6 expression. LPSF/ZKD4 demonstrates lipid accumulation with significant differences between the untreated and negative control groups, indicating a relevant agonist action on the PPARγ receptor. Molecular docking showed that all synthesized compounds have good affinity with PPARγ e COX-2, with binding energy close to -10,000 Kcal/mol. These results demonstrate that the synthesis of quinoline-thiazolidinedione hybrids may be a useful strategy for obtaining promising candidates for new anti-inflammatory agents.

Bazi Bushen alleviates reproductive aging in aged male mice

Bazi Bushen (BZBS), a traditional Chinese medicine (TCM), has demonstrated therapeutic efficacy in testicular dysfunction within D-galactose and NaNO2 mouse models. This study aimed to ascertain if BZBS could also mitigate the decline in testicular function associated with natural aging. Therefore, male aged mice were employed to evaluate the preventive effects of BZBS on male reproductive aging. This was achieved by assessing sex hormone production, testicular histomorphology, and spermatogenesis. Relative to the untreated aged control group, BZBS administration elevated the levels of sex hormones and spermatocyte populations and preserved normal testicular structure in aged mice. Notably, spermatogenesis was maintained. Further analyses, including malondialdehyde (MDA) assays and real-time PCR, indicated that BZBS diminished testicular oxidative stress and the inflammatory burden. Corroborating these findings, mice treated with BZBS exhibited reductions in the populations of senescent and apoptotic cells within the seminiferous tubules, suggesting alleviated cellular damage. In contrast, we observed that rapamycin, a drug known for its longevity benefits, induced excessive testicular apoptosis and did not decrease lipid peroxidation. Collectively, our results highlight BZBS’s promising clinical potential in counteracting male reproductive aging, underlining its mechanisms of action.

Protein Kinase C Beta II Inhibitor Conjugated with Myristic Acid and Trans-Activator of Transcription improves intracellular delivery and cardiovascular protection in porcine myocardial ischemia/reperfusion injury

Introduction: While timely reperfusion of oxygenated blood is critical during resuscitation of ischemic myocardium, oxidative stress during reperfusion still leads to ischemia/reperfusion (I/R) injury and ultimately, cardiomyocyte death. The major sources of oxidative stress targeted in this study are NADPH oxidase (NOX-2) and mitochondria, which are both activated by protein kinase C beta II (PKCβII). In previous studies with an ex-vivo rat heart I/R model, Myristic Acid (Myr) and Trans-Activator Transcription (Tat; YGRKKRRQRRR) conjugated PKCβII inhibitor (Myr-Tat-PKCβII-; N-Myr-Tat-CC-SLNPEWNET) showed cardioprotective effects and decreased infarct size. In this study, we hypothesized that Myr-Tat-PKCβII- will offer cardioprotective effects in comparison with a scrambled peptide control (Myr-Tat-PKCβII-scram; N-Myr-Tat-CC-WNPESLNTE) in an in-vivo porcine myocardial I/R model. Methods: Regional I(1 hour)/R(3 hours) was induced in Male Yorkshire pigs (38-50kg) by inflating a balloon to occlude the Left Anterior Descending Artery (LAD) at the level of the second diagonal branch (supplies 40% of anterior left ventricle) and then deflating it to allow for reperfusion. Then, a bolus of either Myr-Tat-PKCβII- or Myr-Tat-PKCβII-scram (20ng/kg; ~100pmol in blood) was immediately administered into the LAD. Cardiac function and injury were measured by monitoring changes in ejection fraction (EF) and through routine measurements of serum creatine phosphokinase (CPK), troponin I, and myoglobin, respectively. After I/R, the hearts were stained with 1% Evans Blue dye to identify the area at risk (AR) and 1% triphenyltetrazolium chloride to determine the area of necrosis (AN). Infarct size was then quantified (AN/AR) and all data was analyzed via Student’s t-test. Results: Analysis showed that Myr-Tat-PKCβII- significantly restored EF to within 1.40.7% of baseline compared to controls which only restored EF to within 6.42.1% (p&lt;0.05) of baseline EF. Myr-Tat-PKCβII- showed a significant increase in serum myoglobin levels at 1 hr of reperfusion (1362.1 83.4 μg/mL, n=4) compared to scrambled control (202.1197.3 μg/mL, n=3 p&lt;0.05). Myr-Tat-PKCβII- reduced infarct size to 10.0±2.8%; n=4; compared to scrambled control hearts (28.5±8.3%; n=6; p&lt;0.05). CPK and Troponin I levels were comparable in both groups. These results suggest that Myr-Tat-PKCβII- can help prevent cardiac injury when given immediately after an ischemic event. Conclusions: Data from ex-vivo rat heart I/R model, coupled with data from this in-vivo porcine myocardial I/R model indicate the effcacy of Myr-Tat-PKCβII- in preventing oxidative stress-induced myocardial I/R injury when given at the beginning of reperfusion. These findings suggest that Myr-Tat-PKCβII- could be useful in the clinical setting when administered immediately after cardiac resuscitation. Future studies include the treatment of rat cardiomyocyte cells with Myr-Tat-PKCβII- prior to 24 hr hypoxia/24 hr reoxygenation, followed by cell viability assays compared to untreated control. This additional data can help determine the optimal dose to use in a 12 week survival study using the same porcine myocardial I/R protocol. Funding: This study was funded by the Philadelphia College of Osteopathic Medicine, Department of Biomedical Sciences, Division of Research, and the Center for Chronic Diseases of Aging, the National Heart, Lung &amp; Blood Institute at NIH (grant # 1R43HL160338-01) and Young Therapeutics, LLC. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Pathogens of the oak processionary moth Thaumetopoea processionea: Developing a user-friendly bioassay system and metagenome analyses for microorganisms

The oak processionary moth (OPM) Thaumetopoea processionea is a pest of oak trees and poses health risks to humans due to the urticating setae of later instar larvae. For this reason, it is difficult to rear OPM under laboratory conditions, carry out bioassays or examine larvae for pathogens. Biological control targets the early larval instars and is based primarily on commercial preparations of Bacillus thuringiensis ssp. kurstaki (Btk). To test the entomopathogenic potential of other spore-forming bacteria, a user-friendly bioassay system was developed that (i) applies bacterial spore suspensions by oak bud dipping, (ii) targets first instar larvae through feeding exposure and (iii) takes into account their group-feeding behavior. A negligible mortality in the untreated control proved the functionality of the newly established bioassay system. Whereas the commercial Btk HD-1 strain was used as a bioassay standard and confirmed as being highly efficient, a Bacillus wiedmannii strain was ineffective in killing OPM larvae. Larvae, which died during the infection experiment, were further subjected to Nanopore sequencing for a metagenomic approach for entomopathogen detection. It further corroborated that B.wiedmannii was not able to infect and establish in OPM, but identified potential insect pathogenic species from the genera Serratia and Pseudomonas.

Evaluation of Adhesion and Viability of Human Gingival Fibroblasts on Strontium-Coated Titanium Surfaces: an in vitro Study.

Applying multifunctional coatings employing strontium (Sr) ions on titanium (Ti) surfaces is a useful and biocompatible method to improve osseointegration and prevent tissue infections through antimicrobial activity. Nonetheless, the effectiveness of Sr coating on the adhesion and viability of human gingival fibroblasts (HGFs) to Ti surfaces remains unclear. The study aimed to evaluate the effect of Sr coating on the adhesion and viability of HGFs to Ti surfaces. The Ti wafers were divided into two groups based on Sr coating: uncoated Ti (control) and Sr-coated Ti. The Magnetron sputtering technique was used for Sr coating on Ti surfaces. The HGFs were seeded onto the surfaces and cultured for 48 and 96 hours before the cell adhesion and viability of the attached HGFs were assessed. The adhesion of HGFs was analyzed using the attached cell numbers at 48 h and 96 h, and the morphology at 24 h and 72 h. The cytotoxic effect on HGFs was assessed after 24 and 72 hours of incubation using cell viability assay. Student's t-test was used for statistical analysis. The number of cells attached to Sr-coated surfaces was significantly greater than those attached to uncoated Ti surfaces after 48 hours (P<0.0001) and 96 hours (P=0.0002). Sr-coated and uncoated Ti surfaces were not cytotoxic to HGFs, with the cell viability ranging from 92% to 105% of the untreated control HGFs. There were no significant differences in cell viability between Sr-coated and uncoated Ti surfaces at 24 hours (P=0.3675) and 72 hours (P=0.0982). Sr-coated Ti surfaces induce adhesion of HGFs compared to uncoated Ti surfaces. Further, Sr-coated and uncoated Ti surfaces show no cytotoxic effect on the attached HGFs.

Luminal Polyamines Disrupt Intestinal Barrier and Undergo Complex Regulation by Host and Commensal Microbiota

Background: Polyamines produced by host cells and intestinal microbiota and are critical to overall health. Studies have shown putrescine, a polyamine, can strengthen, or in some instances disrupt, gastrointestinal (GI) barrier integrity, yet the regulation of polyamine concentrations in the GI tract is unknown. Host colonocytes and microbiota both metabolize polyamines, and in GI diseases like irritable bowel syndrome (IBS) where intestinal microbiota is significantly altered, polyamine metabolism may be affected. Aim: Determine regulation of luminal polyamine concentration and its effects on intestinal barrier integrity. Hypothesis: Luminal polyamine concentration is suppressed by commensal intestinal microbiota and increasing concentration of putrescine results in intestinal barrier disruption. Methods: Microbiota effects on luminal polyamine concentrations were assessed by humanizing germ-free (GF) mice with a single oral gavage of commensal human microbiota. Polyamine effect on barrier integrity was assessed in vitro using Caco-2 cells and ex vivo on Ussing using mouse colonic tissue with 300Da fluorescein and 4kDa FITC-Dextran tracers. Changes in transepithelial resistance (TER) and rate of tracer flux was recorded. Untargeted metabolomics with shotgun metagenomics was completed on human fecal samples and sigmoid colonic biopsies were collected for bulk RNAseq. Bray-Curtis distance was determined at the species level using the PERMANOVA method and used to test relationships between beta-diversity and fecal polyamine concentration. Results: Humanization with human commensal microbiota (1.2% C. aerofaciens) lead to a 20-fold increase in cecal putrescine and a 20-fold decrease in fecal putrescine compared to GF controls. In vitro addition of putrescine (100mM) caused increased fluorescein flux (3.50±0.24 vs 1.44±0.04 mg/mL) and a rapid loss of transepithelial resistance compared to controls (% of Baseline TER, 36.6±8.2 vs 102.8±5.4, 3h postexposure). Ex vivo addition of putrescine caused a loss of TER (57% vs 89% of baseline by 3h) and an increased rate of 4kDa FITC-Dextran flux compared to untreated controls (1236±131 vs 562±70 ng/hr/cm2). Transcriptomic profiles show decreased expression of ornithine decarboxylase antienzyme (OAZ1, FDR=0.018) in IBS which inhibits the rate limiting enzyme for putrescine production. Increased lactulose excretion (2-24h), suggestive of increased distal colonic permeability, correlated with N-acetylputrescine concentration (p=0.07). N-acetylputrescine also correlated positively with increased stool frequency (p=0.009) and looser consistency (p=0.013). C. aerofaciens, a polyamine consumer, negatively correlated with n-acetylputrescine (FDR=0.02) suggesting a loss of specific taxa may lead to insuffcient polyamine metabolism. Conclusion: Polyamines, namely putrescine, disrupts intestinal barrier integrity, correlate with in vivo permeability, and are decreased in the colon by commensal microbiota. These observations identify putrescine as a negative effector of barrier function and suggest a novel role for the commensal microbiome in regulating barrier integrity through polyamine metabolism. Further work investigating host and microbiota mechanisms of polyamine metabolism, signaling and the specific taxa involved is needed to better understand the effects polyamines have on GI physiology. NIH DK 127998 (MG), support from Mayo Clinic Center for Individualized Medicine (CIM), and Mayo Clinic Center for Cell Signalling in Gastroenterology (C-Sig) P30DK084567. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Control of the plant-parasitic nematode Meloidogyne incognita in soil and on tomato roots by Clonostachys rosea.

Clonostachys rosea is a well-known mycoparasite that has recently been investigated as a bio-based alternative to chemical nematicides for the control of plant-parasitic nematodes. In the search for a promising biocontrol agent, the ability of the C. rosea strain PHP1701 to control the southern root-knot nematode Meloidogyne incognita was tested. Control of M. incognita in vitro and in soil by C. rosea strain PHP1701 was significant and concentration dependent. Small pot greenhouse trials confirmed a significant reduction in tomato root galling compared to the untreated control. In a large greenhouse trial, the control effect was confirmed in early and mid-season. Tomato yield was higher when the strain PHP1701 was applied compared to the untreated M. incognita-infected control. However, the yield of non-M. incognita-infected tomato plants was not reached. A similar reduction in root galling was also observed in a field trial. The results highlight the potential of this fungal strain as a promising biocontrol agent for root-knot nematode control in greenhouses, especially as part of an integrated pest management approach. We recommend the use of C. rosea strain PHP1701 for short-season crops and/or to reduce M. incognita populations on fallow land before planting the next crop.

Telomerase Reverse Transcriptase Prevents Doxorubicin-induced Microvascular Endothelial Dysfunction

Introduction &amp; Objective: Doxorubicin (Dox) is a frequently used chemotherapeutic known to induce cardiovascular (CV) complications by impairing microvascular- and cardiac function. We previously submitted an abstract showing that Dox-induced microvascular dysfunction preceded Dox-induced cardiac dysfunction and thus microvascular dysfunction might contribute to the damage of the heart itself.&gt;Microvascular function specifically relies upon the adequate release of the vasodilator nitric oxide (NO). Our previous work showed that Telomerase Reverse Transcriptase (TERT), the catalytic subunit of telomerase, is necessary for physiological release of NO to mediate vasodilation. NO suppresses reactive oxygen species (ROS) levels. While Dox correlates with increased ROS production and suppressed TERT activity, attenuated TERT activity may explain the elevated levels of ROS after Dox exposure. The objective of this study was to determine whether TERT transcriptional activation is suffcient to protect against Dox-induced microvascular endothelial- and cardiac dysfunction. We utilized cycloastragenol (CAG) as transcriptional activator. Hypothesis: We hypothesized that TERT transcriptional activation is suffcient to prevent or attenuate Dox-induced endothelial dysfunction. Methods: WT or TERT loss-of-function rats on a Sprague Dawley background were assigned to untreated control-, CAG-, Dox and CAG+Dox groups. Animals were ~14 weeks of age to start the six-week protocol. Dox and CAG+Dox animals received one weekly IP Dox-injection (3x 4mg/kg*BW). CAG administer via diet with 50-60mg of CAG/day*BW. Microvascular function was assessed six-weeks after the first dose of Dox via videomicroscopy. Vasodilator responses to flow (flow-mediated dilation; FMD) and mediators of dilation (NO, H2O2) via fluorescent probes were evaluated. Cardiac function was evaluated using echocardiography. Human microvessels were treated either with 100nM Dox or 1μM CAG or a combination of both. Results: Dox treatment resulted in reduction in FMD in both WT and TERT mutant animals. Smooth-muscle dependent vasodilation was not impaired indicating endothelial impairment. CAG prevented Dox-induced endothelial dysfunction in WT but not TERT mutant rats. AGS499 another activator of TERT showed similar results suggestion a TERT mediated phenotype. Dox-treated WT animals exhibited an increase in H2O2 but not NO production. This phenotype was opposite in CAG+Dox WT animals. TERT mutant animals showed an increase in H2O2 but not NO production even without Dox. To assess which vasodilator mediates dilation to flow, we used L-NAME and PEG-Catalase to block NO- and H2O2, respectively. In CAG+Dox WT animals, L-NAME blocked the vasodilatory response to flow, while PEG-Cat blocked the vasodilatory response non-significantly. Similarly, we observed this protective effect of TERT in the human microcirculation. Human adipose microvessels treated over-night with Dox showed no vasodilatory response to flow while CAG+Dox treated vessels dilated normally similar to untreated control vessels. Smooth muscle function was not affected in neither group. Conclusion: We show evidence that TERT prevents Dox-induced endothelial dysfunction in an in vivo model and using human tissue. We highlight the specificity to TERT by a) using a genetic model lacking normal TERT activity and b) showing similar results using two different compounds which have been shown to activate TERT/promote TERT transcriptional activation. Ultimately, the present data shows that CAG is a potential therapeutic to counteract the detrimental effects of Dox treatment in the CV system via TERT signaling. RO1 HL133029-01. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Abstract 3005: Transgenerational Impact Of E-cigarette Nicotine Vaping On Abdominal Aortic Aneurysm

Background: Smoking and family history are major risk factors for abdominal aortic aneurysm (AAA). Smoking is a powerful modulator of DNA methylation, and nicotine can cause heritable epigenetic alterations in animals and humans. We found that nicotine infusion and e-cigarette (e-cig) vaping augment murine AAA, and that parental nicotine infusion augments model AAA in offspring, altering tissue DNA methylation patterns. Hypothesis: We investigated the effects of maternal e-cig vaping in mice on their offspring’s experimental AAA risk, and transgenerational DNA methylation. Methods: Two treatment arms were employed: 1) pre-fertilization or 2) peri-natal. Female ApoE-/- or C57BL6 mice (F0) were exposed to e-cig nicotine (24 mg/ml) puffs, 9 sec/min, 1 hour/day (or room air). Arm 1: treated 28 days, then mated to untreated controls. Arm 2: mated after 1 week of e-cig, then maintained on treatment until gestation was complete. Aortic tissue of 10-week-old F1 offspring was evaluated using RRBS-Seq DNA methylation analysis. Parallel offspring underwent angiotensin-II infusion (1μg/kg/min; ApoE-/-) or elastase infusion (2U/ml for 5 min at 120 mmHg; C57) to induce AAA. Aneurysm growth was ultrasound tracked over 28 days. Results: Maternal vape exposure altered aortic DNA methylation in offspring, with numerous differentially methylated regions (DMRs; q&lt;0.1). Hundreds of these DMRs were located in the same genes previously identified in offspring of nicotine-infused mice (see Background). Vaping altered offspring aortic methylation patterns in AAA-related genes, including miRNAs. DMR pathway analysis was enriched in transcription factors. Further, many DMRs were located in genes harboring risk loci for human AAA from our recent collaborative publication. Maternal e-cig exposure augmented AAA size in both models, arms, and genders (p&lt;0.05). AAA incidence and mortality were increased in ApoE-/- in both genders and arms. Effects were most significant in males. Conclusions: E-cig nicotine exposure augments experimental AAA growth in multiple models across generations. These effects are accompanied by broad aortic epigenetic changes, including in key AAA modulator genes, with enrichment in transcription factors that target known AAA-related genes.

Regulatory Effects of the Silymarin on Expression of OCT4, NANOG, and P53 in MCF7 Cell Lines

Background:: Breast cancer was known as the second most common cause of death in the world, natural sources compound derived from milk thistle called silymarin had already shown anticancer properties. Objective: In the present study, silymarin was used to treat MCF7 cells and inhibition of stem cell pluripotency genes, as well as cell proliferation. Methods: MCF7 cells were cultured in the presence of RPMI-1640 medium consisting of various silymarin extract concentrations (10, 100, 500, 1000, 2000, 3000, 4000, and 5000 µg/mL) for 24, 48, and 72 hours. The inhibitory effects of the compound on cellular proliferation were assessed via employing MTT assay techniques. Following confirming apoptosis, the fold changes of OCT4, NANOG and P53 expression were determined by quantitative Real-Time PCR. Results: There was a significant difference (p value&lt;0.05) in cell viability when various concentrations of silymarin extract were used for 24, 48, and 72 h in comparison to the control. Real-Time- PCR analysis indicated that the expression of OCT4 and NANOG was downregulated while P53 upregulated in compare to untreated control cells (p value &lt;0.05). Conclusion: According to these findings, the silymarin effects on MCF7 cell line and act via modulating OCT4, NANOG, and P53 pathway mediators. Silymarin may introduce this compound as a promising therapeutic compound against MCF7.

Naltrindole attenuates irreversible Hypercontracture during Myocardial Ischemia by a Novel Mechanism and elicits robust cardioprotective effects in Myocardial Ischemia/Reperfusion Injury

Introduction: Previous studies have shown that naltrindole (NTI) produced robust cardioprotective effects (e.g.~80% reduction of infarct size) in both ex-vivo (2.5-5μM) and in-vivo (3.75-7.5 mg/kg) rat myocardial ischemia-reperfusion (MIR) injury models. We recently showed that NTI 100-200μM attenuated phorbol 12-myristate 13-acetate (PMA) induced polymorphonuclear leukocytes (PMNs) superoxide (SO) release by a novel mechanism of action (MOA) devoid of opioid receptors. We hypothesize NTI’s novel MOA is a reduction of intracellular calcium (Ca2+). To test this hypothesis, we conducted PMN SO release and isolated perfused MIR assays using KB-R7943(KB). KB is known to reduce intracellular Ca2+ via inhibition of reverse mode Na+/Ca2+ exchanger and therefore should attenuate PMN SO release, myocardial ischemic hypercontracture, infarct size, and left ventricular end diastolic pressure (LVEDP). Methods: PMNs and hearts isolated from anesthetized male Sprague Dawley (SD) rats (~400g SO release assay, ~300g for perfused hearts). PMN SO release was stimulated using PMA (100nM) ± NTI 10-200 μM, KB 5-20 μM, or vehicle (0.5% DMSO; controls) and measured spectrophotometrically as a change in absorbance @ 550nm over 390 secs and cell viability was determined using 0.2% Trypan blue exclusion. We then infused NTI 1.25-5μM, KB 10-20μM, or naloxone (NX; negative control) 10μM for 5 min into perfused hearts, just prior to global I (30min)/R(45min). Cardiac LV function was measured via a pressure transducer and infarct size using 1% triphenyltetrazolium chloride staining in MIR assay. Data were analyzed using ANOVA and Fischers post-hoc analysis, with p&lt;0.05 considered statistically significant. Results: PMN SO release assay: KB 10 μM, KB 20 μM, NTI 100 μM (~3.75 mg/kg), and NTI 200 μM(~7.5 mg/kg) significantly attenuated PMA induced PMN SO release by ~40% (KB 10μM, n=7, p&lt;0.05), ~50% ((KB 20μM, n=7, p&lt;0.05), ~30% (NTI 100 μM, n=7, p&lt;0.05), 75% (NTI 200μM, n=7, p&lt;0.05) respectively when compared to untreated control (n=10). MIR assay: NTI 5μM, 2.5μM, 1.25μM and KB 20 μM reduced infarct size (4.8±3%, n=5, p&lt;0.05), (6.8±0.8%, n=6, p&lt;0.05), (12.2±3%, n=5), and (2.9±2%, n=5, p&lt;0.05) respectively compared to vehicle (14.1±2%, n=9) and NX 10μM controls (18±3%, n=3). NTI 5 μM and KB 20 μM had significantly improved LVEDP to (32.4±11%, n=5, p&lt;0.05) and (22.8±4%, n=5, p&lt;0.05) respectively compared to vehicle control (72.4±6%, n=9) and NX 10μM controls(54.2±8%, n=3) Conclusion: NTI and KB showed concentration-dependent effects on reducing PMN SO release. Both NTI (2.5μM, 5μM) and KB (20μM) reduced infarct size and improved LVEDP. The novel MOA of NTI is most likely mediated via a reduction in intracellular Ca2+ and subsequent hypercontracture during myocardial ischemia. The infarct reducing effect of NTI can benefit patients who will undergo elective PCI, CABG, and transplants. Funding: This study was funded by the Philadelphia College of Osteopathic Medicine, Department of Biomedical Sciences, Division of Research, and the Center for Chronic Diseases of Aging and Young Therapeutics, LLC. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Effect of oxytocin use during colostrum harvest and association of cow characteristics with colostrum yield and IgG concentration in Holstein dairy cows

Our objectives were to determine the effect of oxytocin use during colostrum harvest on colostrum yield (CY) and IgG concentration in Holstein dairy cows on a commercial dairy in New York and to describe associations of cow characteristics with these outcomes. Animals were enrolled between July and October 2023 using a randomized block design, with day of enrollment as the unit of randomization. A median (range) of 10 (3 to 19) cows were enrolled/d. Treatments were 1) 40 IU Oxytocin (OXY40), 2) 20 IU Oxytocin (OXY20), and 3) an untreated control group (CNTR). Oxytocin was administered intramuscularly (IM) approximately 45 s before unit attachment in a rotary parlor. Colostrum weight was measured using the colostrum bucket-embedded scale or a platform scale when the yield was less than the smallest bucket scale. The concentration of IgG ([IgG]) in colostrum was determined using radial immunodiffusion (RID) and used to calculate total IgG, and dry matter (%) was determined by oven drying. Individual cow characteristics such as parity, calf sex, weight, dystocia score, stillbirth, milk production in wk 4 of lactation, and for multiparous cows, dry period length, previous lactation dry off linear score (LS) were collected. Data were analyzed separately for primiparous and multiparous cows using backward stepwise elimination to produce final mixed effects ANOVA models Primiparous cows (n = 201) were randomized to 35.8% (n = 72) OXY40, 32.8% (n = 66) OXY20, and 31.3% (n = 63) CNTR. Multiparous groups (n = 435) were randomized to 34.7% (n = 151) OXY40, 29.7% (n = 129) OXY20, and 35.6% (n = 155) CNTR. The median (range) CY was 6.0 (0 to 20.6) kg and [IgG] was 98.5 (0.1 to 293.6) g/L in the study population. In primiparous cows, OXY40 had a higher colostrum yield (LSM [95% CI]) of 5.4 (4.9 to 5.9) kg compared with both OXY20 (4.1 [3.5 to 4.7] kg) and CNTR (3.8 [3.3 to 4.3] kg) (P < 0.001). In multiparous cows, OXY40, OXY20, and CTNR did not differ in CY (5.9 [5.3 to 6.5], 5.7 [6.3 to 5.1], and 5.4 [6.0 to 4.8] kg, respectively, P = 0.43), but colostrum yield was greater in parity 2 compared with all other parities, cows giving birth to male calves, cows with the highest milk production at wk 4 of lactation, and with a dry period of >65 d. Oxytocin use did not affect [IgG] in either primiparous or multiparous cows (P > 0.56), but [IgG] was highest in cows in parity ≥4 and lowest in cows dry >65 d. In summary, oxytocin use at 40 IU IM in primiparous was associated with a higher CY but not [IgG]. Oxytocin use likely addressed disturbed milk ejection and therefore increased CY in heifers milked for the first time in a rotary parlor. This study confirms cow characteristics associated with colostrum production within a single herd.

Impact of sonication time in nanoparticle synthesis on the nutrition and growth of wheat (Triticum aestivum L.) plant

Nanotechnology in agricultural applications is promising in improving plant nutrition and yield, pest control, and gene delivery. However, the method to synthesize nanoparticles or nanocomposites (NCs) can play a crucial role in determining the characteristics of NCs, such as size and morphology, which may be critical factors affecting plant nutrition and NCs` potential toxicity. This study elucidates the effect of sonication time in synthesizing NCs on its characteristics and plant use efficiency. For this purpose, a hard/soft nanocomposite (NC) (CoFe2O4/Ni0.8Cu0.1Zn0.1Fe2O4) was sonochemically synthesized at different sonication times (20 and 60 minutes) and comprehensively characterized. They were hydroponically applied to wheat seedlings (50, 100, 200, 400, and 800 mg/L). The physiological, morphological, and nutritional status of the seedlings were determined. The results showed that an increase in sonication time decreased the mean NC size: 26.7 nm (20 minutes) and 17.4 nm (60 minutes). Photosynthetic parameters, growth, and biomass were gradually reduced with the increasing NC concentrations, revealing their toxic effect. However, treating NCs at 60 min significantly improved the average root length, suggesting its beneficial role for plant growth at the germination stage. The content of elements in the composition of the NCs (Fe, Zn, Co, Ni, and Cu) was remarkably higher in the NC-treated roots compared to the untreated controls. In addition, 60 minutes of preparation showed better plant uptake than 20 minutes. This is the first study to evaluate the effect of sonication time in the preparation of NC on plant nutrition and their fate in plants.