Untreated Control Research Articles

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.

Are predatory mites effective as biological control agents to suppress Oligonychus ilicis (Acari: Tetranychidae) in blueberry plantings?

Southern red mite, Oligonychus ilicis McGregor (Acari: Tetranychidae), is an important polyphagous spider mite pest that causes economic damage to many ornamentals, coffee, and fruit crops. Blueberry growers in the Southeastern United States, including Florida and Georgia, have experienced severe losses due to outbreaks of O. ilicis. Predatory mites are an important management tool used for controlling spider mites; however, predators have not been studied and successfully evaluated in blueberry systems. Amblyseius swirskii Athias-Henriot, Phytoseiulus persimilis Athias-Henriot, and Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) are among the most economically important arthropod agents used in augmentative biological control worldwide. To evaluate the potential of these 3 commercially available predatory mites for use in blueberry plantings, we conducted experiments under controlled laboratory conditions and in the greenhouse. In preliminary laboratory experiments, P. persimilis and N. californicus significantly reduced the number of O. ilicis motile stages below those found in the untreated control, indicating the potential for these 2 predatory mite species to suppress O. ilicis populations. Amblyseius swirskii did not perform well controlling O. ilicis motiles in the laboratory. Under greenhouse conditions, N. californicus and P. persimilis significantly reduced the number of eggs after 7 days of release and the number of motile stages after 14 days of release. This is the first report of using phytoseiid mites to suppress O. ilicis in blueberry systems in the United States. Further studies on predator behavior, feeding preferences, and acaricide compatibility with predators are required to investigate the possibility of using P. persimilis, and N. californicus as biological control agents of O. ilicis in blueberry systems.

Shelf life and quality assessment of pasteurised red dragon fruit (Hylocereus polyrhizus L.) purée: Comparative study of high-pressure and thermal processing

Red dragon fruit (RDF; Hylocereus polyrhizus L.) contains various polyphenols with potent antioxidant properties. Purée extracted from RDF has a vibrant red hue, making it a valuable natural food colouring agent suitable for a wide range of culinary applications. To preserve this valuable purée, non-thermal pasteurisation methods, such as high-pressure processing (HPP), have emerged as an alternative to thermal pasteurisation (TP), as they do not impart the adverse effects associated with heat treatment. Therefore, the primary objective of the present work was to compare the impact of HPP and TP on several key attributes of RDF purée during 60-d storage. These attributes included the total betacyanin content (TBC), total phenolic content, total flavonoid content (TFC), antioxidant activities, enzyme activities, microbial growth, and colour stability. The RDF purée samples were divided into three groups: TP-treated (65°C/20 min), HPP-treated (350 MPa/5 min), and an untreated control group. All samples were stored at a controlled temperature of 4 ± 1°C, and analysed at 15-d intervals. Results revealed that preservation method, storage duration, and their interactions, significantly influenced the various parameters studied in RDF purée. Notably, HPP demonstrated superior efficacy in extending the shelf life of RDF purée well beyond 60 d, outperforming both TP and Control. Specifically, TP was proven effective in maintaining the phenolic content, antioxidant activities, and colour stability of the purée. On the other hand, HPP was particularly efficient in suppressing microbial growth and reducing enzyme activities in RDF purée. The findings can potentially transform the way RDF purée is preserved and utilised in the food industry, benefiting both producers and consumers, and contributing to more sustainable and health-conscious food practices.

Enhanced CTLA-4 blockade anti-tumor immunity with APG-157 combination in a murine head and neck cancer.

A phase I clinical study for patients with locally advanced H&N cancer with a new class of botanical drug APG-157 provided hints of potential synergy with immunotherapy. We sought to evaluate the efficacy of the combination of APG-157 and immune checkpoint inhibitors. CCL23, UM-SCC1 (human), and SCCVII (HPV-), MEER (HPV+) (murine) H&N cancer cell lines were utilized for invitro and invivo studies. We measured tumor growth by treating the mice with APG-157, anti-PD-1, and anti-CTLA-4 antibody combinations (8 groups). The tumor microenvironments were assessed by multi-color flow cytometry, immunohistochemistry, and RNA-seq analysis. Fecal microbiome was analyzed by 16S rRNA sequence. Among the eight treatment groups, APG-157 + anti-CTLA-4 demonstrated the best tumor growth suppression (p = 0.0065 compared to the control), followed by anti-PD-1 + anti-CTLA-4 treatment group (p = 0.48 compared to the control). Immunophenotype showed over 30% of CD8+ T cells in APG-157 + anti-CTLA-4 group compared to 4%-5% of CD8+ T cells for the control group. Differential gene expression analysis revealed that APG-157 + anti-CTLA-4 group showed an enriched set of genes for inflammatory response and apoptotic signaling pathways. The fecal microbiome analysis showed a substantial difference of lactobacillus genus among groups, highest for APG-157 + anti-CTLA-4 treatment group. We were unable to perform correlative studies for MEER model as there was tumor growth suppression with all treatment conditions, except for the untreated control group. The results indicate that APG-157 and immune checkpoint inhibitor combination treatment could potentially lead to improved tumor control.

Control of Peach Leaf Curl with Foliar Applications of Plant Immunity Inducers and Insights in Elicitation of Defense Responses against Taphrina deformans.

Taphrina deformans is the causal agent of leaf curl, a serious peach disease which causes significant losses in peach production worldwide. Nowadays, in order to control plant diseases, it is necessary to adopt novel and low-cost alternatives to conventional chemical fungicides. These promising strategies are targeted at eliciting host defense mechanisms via priming the host through the consecutive application of plant immunity inducers prior to pathogen challenge. In this study, we investigated whether chitosan or yeast cell wall extracts could provide enhanced tolerance against leaf curl in two-season field trials. Furthermore, we addressed the possible molecular mechanisms involved beyond the priming of immune responses by monitoring the induction of key defense-related genes. The efficacy of spraying treatments against peach leaf curl with both inducers was significantly higher compared to the untreated control, showing efficacy in reducing disease severity of up to 62.6% and 73.9% for chitosan and yeast cell wall extracts, respectively. The application of chitosan in combination with copper hydroxide was more efficient in reducing disease incidence and severity, showing efficacy values in the range of 79.5-93.18%. Peach plantlets were also spray-treated with immunity inducers three times prior to leaf inoculation with T. deformans blastospores in their yeast phase. The relative expression levels of nine key defense and priming genes, including those encoding members of pathogenesis-related (PR) proteins and hub genes associated with hormone biosynthesis, were monitored by RT-qPCR across three days after inoculation (dai). The results indicate that pre-treatments with these plant immunity inducers activated the induction of genes involved in salicylic acid (SA) and jasmonate (JA) defense signaling pathways that may offer systemic resistance, coupled with the upregulation of genes conferring direct antimicrobial effects. Our experiments suggest that these two plant immunity inducers could constitute useful components towards the effective control of T. deformans in peach crops.

Measuring effects of ivermectin-treated cattle on potential malaria vectors in Vietnam: A cluster-randomized trial.

Malaria elimination using current tools has stalled in many areas. Ivermectin (IVM) is a broad-antiparasitic drug and mosquitocide and has been proposed as a tool for accelerating progress towards malaria elimination. Under laboratory conditions, IVM has been shown to reduce the survival of adult Anopheles populations that have fed on IVM-treated mammals. Treating cattle with IVM has been proposed as an important contribution to malaria vector management, however, the impacts of IVM in this One Health use case have been untested in field trials in Southeast Asia. Through a randomized village-based trial, this study quantified the effect of IVM-treated cattle on anopheline populations in treated vs. untreated villages in Central Vietnam. Local zebu cattle in six rural villages were included in this study. In three villages, cattle were treated with IVM at established veterinary dosages, and in three additional villages cattle were left as untreated controls. For the main study outcome, the mosquito populations in all villages were sampled using cattle-baited traps for six nights before, and six nights after a 2-day IVM-administration (intervention) period. Anopheline species were characterized using taxonomic keys. The impact of the intervention was analyzed using a difference-in-differences (DID) approach with generalized estimating equations (with negative binomial distribution and robust errors). This intervention was powered to detect a 50% reduction in total nightly Anopheles spp. vector catches from cattle-baited traps. Given the unusual diversity in anopheline populations, exploratory analyses examined taxon-level differences in the ecological population diversity. Across the treated villages, 1,112 of 1,523 censused cows (73% overall; range 67% to 83%) were treated with IVM. In both control and treated villages, there was a 30% to 40% decrease in total anophelines captured in the post-intervention period as compared to the pre-intervention period. In the control villages, there were 1,873 captured pre-intervention and 1,079 captured during the post-intervention period. In the treated villages, there were 1,594 captured pre-intervention, and 1,101 captured during the post-intervention period. The difference in differences model analysis comparing total captures between arms was not statistically significant (p = 0.61). Secondary outcomes of vector population diversity found that in three villages (one control and two treatment) Brillouin's index increased, and in three villages (two control and one treatment) Brillouin's index decreased. When examining biodiversity by trapping-night, there were no clear trends in treated or untreated vector populations. Additionally, there were no clear trends when examining the components of biodiversity: richness and evenness. The ability of this study to quantify the impacts of IVM treatment was limited due to unexpectedly large spatiotemporal variability in trapping rates; an area-wide decrease in trapping counts across all six villages post-intervention; and potential spillover effects. However, this study provides important data to directly inform future studies in the GMS and beyond for IVM-based vector control.

Retinoids in the treatment of photoageing: A histological study of topical retinoid efficacy in black skin.

Photoageing describes complex cutaneous changes that occur due to chronic exposure to solar ultraviolet radiation (UVR). The 'gold standard' for the treatment of photoaged white skin is all-trans retinoic acid (ATRA); however, cosmetic retinol (ROL) has also proven efficacious. Recent work has identified that black skin is susceptible to photoageing, characterized by disintegration of fibrillin-rich microfibrils (FRMs) at the dermal-epidermal junction (DEJ). However, the impact of topical retinoids for repair of black skin has not been well investigated. To determine the potential of retinoids to repair photoaged black skin. An exploratory intervention study was performed using an invivo, short-term patch test protocol. Healthy but photoaged black volunteers (>45 years) were recruited to the study, and participant extensor forearms were occluded with either 0.025% ATRA (n = 6; 4-day application due to irritancy) or ROL (12-day treatment protocol for a cosmetic) at concentrations of 0.3% (n = 6) or 1% (n = 6). Punch biopsies from occluded but untreated control sites and retinoid-treated sites were processed for histological analyses of epidermal characteristics, melanin distribution and dermal remodelling. Treatment with ATRA and ROL induced significant acanthosis (all p < 0.001) accompanied by a significant increase in keratinocyte proliferation (Ki67; all p < 0.01), dispersal of epidermal melanin and restoration of the FRMs at the DEJ (all p < 0.01), compared to untreated control. This study confirms that topical ATRA has utility for the repair of photoaged black skin and that ROL induces comparable effects on epidermal and dermal remodelling, albeit over a longer timeframe. The effects of topical retinoids on black photoaged skin are similar to those reported for white photoaged skin and suggest conserved biology in relation to repair of UVR-induced damage. Further investigation of topical retinoid efficacy in daily use is warranted for black skin.

Injectable biomimetic hybrid nanofibers for targeting cartilage in early osteoarthritis treatment

Osteoarthritis (OA) is a degenerative joint disease characterized by cartilage degradation and limited effective treatment options for regeneration. To address this clinical challenge, an innovative injectable biomimetic hybrid nanofiber, namely hyaluronic acid-graft-poly(2-methacryloyloxyethyl phosphorylcholine)-chondrocyte affinity peptides (HA-PMPC-CAP), was designed and synthesized through a ultraviolet light (UV)-initiated reaction process for intervention of early osteoarthritis (OA). Comprehensive in vitro assessments and molecular docking experiments have confirmed its non-cytotoxicity, exceptional lubrication properties, favorable biocompatibility and high binding affinity to cartilage proteins. Utilizing an OA rat model, the therapeutic potential of HA-PMPC-CAP was explored through intra-articular injection, with a focus on its impact on cartilage regeneration. During an 8-week period, macroscopic, histological, immunohistochemical, and microscopic CT evaluations were conducted on the treated joints. The findings were remarkable: HA-PMPC-CAP established a durable lubricating layer on the surface of cartilage, significantly reduced cartilage deterioration, replenished glycosaminoglycan (GAG) levels, lowered the scores of Osteoarthritis Research Society International (OARSI), and curtailed the development of osteophytes, which was superior to the untreated OA control groups. These results position HA-PMPC-CAP as an effective biomimetic therapy, offering a novel strategy for cartilage regeneration and lubrication in the early stages of OA, which has promising implications for future clinical applications.

Maximizing Zygopetalum maculatum orchid yield through calcium nutrition: Unveiling calcium content and dynamics across different plant parts at different growth stages of the orchid

This study explores the pivotal yet often overlooked role of calcium (Ca) in Zygopetalum maculatum orchid cultivation. Six different treatments, featuring graded doses of Ca (ranging from 0 to 100 mg Ca L−1 nutrient solution), were used to assess the impact of Ca on growth, development, and physiological responses of the orchids. Notably, the Ca untreated control treatment (Ca0) exhibited a rapid decline in potting media pH (became acidic in reaction with time), whereas under Ca100 treatment potting media pH declined slowly and remained near neutral. Dehydrogenase activity in the potting media significantly increased under Ca100 treatment. Significant improvements in vegetative growth parameters, like, bulb length, bulb width, and leaf characteristics, were evident under Ca100 treatment compared to the Ca0 treatment. The economic yield, measured by the number of flower spikes and various floral attributes (i.e. number of florets per spike, spike length, florets size), displayed a remarkable enhancement under Ca100 treatment. The Ca100 treatment produced 100 % more number of flower spikes as compared to Ca0 treatment. The number of new shoot initiation, and overall flower biomass also substantially increased under Ca100 treatment. Ca content analysis in different plant parts revealed presence of elevated Ca concentrations in leaves, back bulbs, new bulbs, and roots under Ca100 treatment. The Ca content in back bulbs and roots decreased after flowering phase of the orchid, but the Ca levels in leaves and new bulbs remained statistically similar during both pre and post-flowering phases.These findings underscore the positive influence of Ca application on Zygopetalum maculatum orchids for enhancing its vegetative growth and economic yield. The study contributes valuable insights into Ca dynamics in different plant parts of the orchid, and recommends a 100 mg L−1 Ca application at 7-day intervals for improving Zygopetalum maculatum cultivation practices.

Release from aboveground enemies increases seedling survival in grasslands

Abstract Plant enemies can influence plant community assembly and structure. However, it is unclear how insect herbivores and fungal pathogens affect seedling recruitment. Complex interactions with competition and resource availability make it difficult to isolate the effect of enemies. This uncertainty can impede understanding of community assembly drivers, species coexistence and trophic interactions; and limits hypothesis testing such as of the enemy release hypothesis, a key hypothesis in invasion biology. Using a novel species‐specific approach, we examine how enemies affect seedling survival and recruitment of 16 grassland species. We planted seedlings of 16 native species from two functional groups (C4 grasses and non‐legume forbs) into two grassland sites (early and mid‐succession). We hand‐painted 1512 individual seedlings with pesticides (insecticide and fungicide) over the course of one growing season to enforce aboveground species‐specific release from enemies and tested whether it enhanced survival relative to untreated controls. Applying treatments to individuals allowed us to test for the effect of enemies on plant performance while controlling for contexts such as competition from the resident community and resource levels. Release from insects increased seedling survival by 56% on average, with no additional benefit of release from fungal pathogens. This effect was observed for both forbs and C4 grasses across both sites and was strongest in resource‐acquisitive species. There was no effect of the mean phylogenetic distance between our target species and the resident plant community, or of light availability or soil moisture. Synthesis. The significant positive effect of release from insect herbivores on survival early in colonisation—a trend that held across functional groups and types of resident community—suggests insects play an important regulatory role in community assembly, especially for resource‐acquisitive species. As variation between species could be explained by traits but not by phylogeny, we emphasise the importance of trait‐based approaches to plant community ecology. Our results also support the key mechanism (increased performance following release from enemies) underlying the enemy release hypothesis. Enemy release may therefore aid initial recruitment of plants during the invasion process.

A preliminary study on the impact of nutrient stress induction on drug cytotoxicity in glioblastoma cells and fibroblasts

Abstract Background Nutrient stress (NS), one of the hallmarks of the tumour microenvironment, can render cancer cells tolerant to cytotoxicity. Fibroblasts, on the other hand, have cancer cell-like traits, such as plasticity and resiliency. Hence, this study aimed to evaluate the cytotoxicity of the drug on reseeded human U87 glioblastoma (GBM) cells as well as on mouse L929 fibroblasts in the form of monolayer and colonies that grew after NS induction. Results No treatment for 48 h showed a statistically significant difference in U87 cell viability when compared to the 50% hypothetical value. However, temozolomide (TMZ) (151.0 µg/ml) and azithromycin (AZI) (92.0 µg/ml) significantly diminished the number of U87 cell colonies compared to the untreated control, and AZI also outperformed doxycycline (DOXY) (147.0 µg/ml). L929 fibroblasts survived NS, but the cytotoxicity of AZI, DOXY, and AZI + DOXY (92.0 + 147.0 µg/ml) substantially increased than in L929 fibroblasts without NS induction. Conclusions The present findings suggest that NS does not inevitably contribute to cytotoxic drug tolerance in GBM cells. In addition, although fibroblasts can withstand NS, they can also become susceptible to cytotoxic drug-induced death; nevertheless, the type of drug may play a role. Graphical Abstract

Evaluating the effects of mefenoxam on taxonomic and functional dynamics of nontarget fungal communities during carrot cultivation

Ridomil Gold SL (45.3% a.i. mefenoxam) is a widely used chemical fungicide for the control of oomycetes. However, its impact on fungal communities remains unexplored. Therefore, the goal of this study was to examine the effects of mefenoxam on the temporal dynamics of fungal taxonomic and functional diversities during carrot cultivation under four treatment groups: mefenoxam application with and without Pythium inoculation, and untreated control groups with and without Pythium inoculation. Our in vitro sensitivity assay showed that the maximum recommended concentration of mefenoxam, 0.24 ppm, did not suppress the mycelial growth of P. irregulare. At 100 ppm, mycelial growth was only reduced by 11.4%, indicating that the isolate was resistant to mefenoxam. MiSeq sequencing data revealed transient taxonomic variations among treatments 2 weeks post-treatment. Mortierella dominated the fungal community in the mefenoxam-Pythium combination treatment, as confirmed through PCR using our newly designed Mortierella-specific primers. Conversely, mefenoxam-Pythium combination had adverse effects on Penicillium, Trichoderma, and Fusarium, and decrease the overall alpha diversity. However, these compositional changes gradually reverted to those observed in the control by the 12th week. The predicted ecological functions of fungal communities in all Pythium and mefenoxam treatments shifted, leading to a decrease in symbiotrophs and plant pathogen functional groups. Moreover, the community-level physiological profiling approach, utilizing 96-well Biolog FF microplates, showed discernible variations in the utilization of 95 diverse carbon sources among the treatments. Notably, arbutin, l-arabinose, Tween 80, and succinamic acid demonstrated a strong positive association with Mortierella. Our findings demonstrate that a single application of mefenoxam at its recommended rate triggers substantial taxonomic and functional shifts in the soil fungal community. Considering this impact, the conventional agricultural practice of repeated mefenoxam application is likely to exert considerable shifts on the soil ecosystem that may affect agricultural sustainability.