Adult Control Research Articles

A scoping review to determine if adverse human health effects are associated with use of organophosphates for mosquito control.

Organophosphate insecticides are widely used for adult mosquito control. Although proven effective in reducing mosquito populations and limiting arbovirus transmission, public concern exists regarding potential human health effects associated with organophosphate exposure. The aim of this scoping review was to describe any reported human health conditions associated with organophosphates during their use for adult mosquito control in the United States and Canada. Original peer-reviewed articles published in English language journals from 1 January 2000 to 22 May 2024, were obtained by searching from the databases MEDLINE, EMBASE, Agricultural & Environmental Science Collection, CAB Abstracts, and Scopus. The search identified 6,154 screened articles. Following an independent review, 10 studies were identified that described human health conditions associated with organophosphate exposure during adult mosquito control applications. Of the 10 included studies, only two articles were published within the last 11 years (2013 to 22 May 2024). Three types of study design were represented in the included studies: cohort (n = 5), case study (n = 1), and risk assessment (n = 4). The included studies could not determine causality between exposure to adulticides and development of illness or adverse impacts. Exposure to organophosphates did not contribute to an observed increase in metabolic toxicity, hospitalization rates, or self-reported symptoms and exposure. The available and limited evidence indicates that organophosphates can be used safely to control nuisance mosquitoes or mosquitoes that transmit arboviruses. Continued research regarding the human health effects associated with organophosphate applications for adult mosquito control could help evaluate the basis of the public's concerns and inform public health decision-making.

Improving Sphenophorus levis Adult Mortality Through Solid Insecticide Applications and Increased Insecticide Dose.

The sugarcane weevil (Sphenophorus levis Vaurie, 1978) is currently considered the most important sugarcane pest in Brazil, causing significant yield losses. Application methods of insecticides for S. levis control have not been effective, mostly due to the insect's cryptic behavior below the soil surface which suppresses the correct placement of insecticide active ingredients on target. Two experiments were conducted using an innovative bioassay methodology that simulates sugarcane field conditions to effectively evaluate S. levis adult mortality and insecticide residues in the soil under different treatments. The first experiment aimed to assess the efficacy of two liquid- and solid-applied insecticides, while the second aimed to examine the effect of increasing the dose of lambda-cyhalothrin + thiamethoxam on S. levis adult control. The novel bioassays simulated liquid and solid insecticide applications on sugarcane by exposing S. levis adults to chemical residuals on rhizomes and in soil after insecticide application. In the first experiment, low S. levis adult control was detected (< 53% mortality) across all treatments, where both solid and liquid applications of lambda-cyhalothrin + thiamethoxam provided greater efficacy levels than imidacloprid and control treatments, respectively. Solid applications maintained higher insecticide concentrations in the soil for longer periods than liquid insecticide applications, providing maximum insect control levels 7days after application. The second experiment revealed that solid applications at higher insecticide doses significantly improved control of S. levis adult (76.7% mortality) and resulted in greater insecticide concentrations in the soil compared to the recommended label rate (58.8% mortality).

Coconut oil affects aging-related changes in Mongolian gerbil liver morphophysiology

Aging causes changes in liver morphophysiology, altering hepatocyte morphology and organ function. Due to its antioxidant and anti-inflammatory properties, coconut oil has been used as a therapeutic agent in diets, in an attempt to attenuate alterations in the liver naturally caused by aging. Herein, we evaluated the effects of coconut oil consumption during aging on Mongolian gerbil liver morphophysiology. The animals were divided into three experimental groups: the gerbils in the Adult Control Group (AC) were euthanized at 3 months of age, the gerbils in the Old Control Group (OC) at 15 months of age, and the gerbils in the Coconut Oil Group (CO) received 0.1 ml/day of coconut oil for 12 months and were euthanized at 15 months of age. Prolonged consumption of coconut oil during aging prevented the animals and the liver from gaining mass. However, the other results showed that coconut oil intensified the morphophysiological alterations of aging, promoting an increase in the hepatocyte cytoplasm and nuclei. In addition, an increase in blood vessels, reticular fibers, lipid droplets, and lipofuscin granules were observed in the CO group. Finally, the results also demonstrated that coconut oil promotes an increase in lipid peroxidation, indicated by an increase in MDA levels. We therefore conclude that coconut oil has the potential to intensify the morphophysiological alterations that occur in the liver during aging.

Attachment Styles and Parenting Attitudes in Adults With "Pseudo-Attention-Deficit/Hyperactivity Disorder".

Background The number of individuals who experience the onset of attention-deficit/hyperactivity disorder (ADHD)-like symptoms after the age of 12 years has been growing, which does not meet the diagnostic criteria for adult ADHD. We designated this condition as "pseudo-ADHD" and investigated its psychological implications by comparing the parenting attitudes in childhood and attachment styles among individuals with "pseudo-ADHD," those with adult ADHD, and healthy controls. Methods We conducted an online cross-sectional survey of adults aged 18 years. Participants in the analysis included individuals with "pseudo-ADHD" (n = 46), adults with ADHD (n = 23), and healthy controls (n = 758). The Parental Bonding Instrument (PBI) was used to evaluate parenting attitudes, and the Experiences in Close Relationships Inventory for the Generalized Other (ECR-GO) was used to evaluate attachment styles. One-way analysis of variance was performed to compare the PBI and ECR-GO scores among the three groups. Results Regarding the PBI scores, a significant main effect of group was found for the paternal care scores (p = 0.002), and the "pseudo-ADHD" group did not differ significantly from the adult ADHD group or healthy control group (p = 0.378, p = 0.228, respectively). A significant main effect of the group was found for the maternal care scores (p < 0.001). The "pseudo-ADHD" group scored significantly lower than the adult ADHD group (p = 0.005), whereas there was no significant difference compared to the healthy control group (p = 1.000). A significant main effect of the group was also found for the paternal overprotection scores (p < 0.001). The "pseudo-ADHD" group scored significantly lower than the adult ADHD group (p = 0.002), whereas there was no significant difference compared to the healthy control group (p = 0.571). A significant main effect of group was found for the maternal overprotection scores (p < 0.001), and the "pseudo-ADHD" group did not differ significantly from either the adult ADHD group or the healthy control group (p = 0.060, p = 0.161, respectively). Regarding ECR-GO scores, a significant main effect of group was found for the fear of abandonment score (p < 0.001), and the "pseudo-ADHD" group scored significantly lower than the adult ADHD group (p < 0.001) and significantly higher than the healthy control group (p = 0.005). Conclusion This study suggested that the parenting attitudes of parents of individuals with "pseudo-ADHD" were not different from those of the healthy controls. For attachment styles, the fear of abandonment was higher in individuals with "pseudo-ADHD" than in healthy controls, but not as high as in patients with adult ADHD. This study highlights, for the first time, the importance of considering attachment styles in the diagnosis and treatment of individuals with ADHD symptoms in adulthood.

Effects of Life-Long Exercise on Age-Related Inflammation, Apoptosis, Oxidative Stress, Ferroptosis Markers, and NRF2/KAEP 1/Klotho Pathway in Rat Kidneys

To investigate the effects of life-long exercise (LLE) on age-related inflammatory cytokines, apoptosis, oxidative stress, ferroptosis markers, and the NRF2/KAEP 1/Klotho pathway in rats. Eight-month-old female Sprague-Dawley rats were divided into four groups: 1) LLE: 18-month LLE training starting at 8 months of age, 2) Old moderate-intensity continuous training (OMICT): 8 months of moderate-intensity continuous training starting at 18 months of age, 3) Adult sedentary (ASED): 8 month-old adult sedentary control group, and 4) Old sedentary (OSED): a 26-month-old sedentary control group. Hematoxylin eosin staining was performed to observe the pathological changes of kidney tissue injury in rats; Masson’s staining to observe the deposition of collagen fibers in rat kidney tissues; and western blotting to detect the expression levels of IL-6, IL 1β, p53, p21, TNF-α, GPX4, KAEP 1, NRF2, SLC7A11, and other proteins in kidney tissues. Results: Compared with the ASED group, the OSED group showed significant morphological changes in renal tubules and glomeruli, which were swollen and deformed, with a small number of inflammatory cells infiltrated in the tubules. Compared with the OSED group, the expression levels of inflammation-related proteins such as IL-1β, IL-6, TNF α, and MMP3 were significantly lower in the LLE group. Quantitative immunofluorescence analysis and western blotting revealed that compared with the ASED group, KAEP 1 protein fluorescence intensity and protein expression levels were significantly enhanced, while Klotho and NRF2 protein fluorescence intensity and protein expression levels were reduced in the OSED group. Compared with the OSED group, KAEP 1 protein fluorescence intensity and protein expression levels were reduced in the LLE and OMICT groups. Klotho and KAEP 1 protein expression levels and immunofluorescence intensity were higher in the LLE group than in the OSED group. The expression levels of GPX4 and SLC7A11, two negative marker proteins associated with ferroptosis, were significantly higher in the LLE group than in the OSED group, while the expression of p53 a cellular senescence-associated protein that negatively regulates SLC7A11, and the downstream protein p21 were significantly decreased. LLE may ameliorated aging-induced oxidative stress, inflammatory response, apoptosis, and ferroptosis by regulating Klotho and synergistically activating the NRF2/KAEP 1 pathway.

Knee muscle strength and steadiness for individuals with anterior cruciate ligament reconstruction and knee osteoarthritis

BackgroundStrength and neuromuscular decrements following knee musculoskeletal injury may accelerate knee osteoarthritis development. This study assessed isometric knee extensor and flexor strength and steadiness between individuals with knee injury, i.e., ligament reconstruction, and knee osteoarthritis to healthy age-matched controls. MethodsFour cohorts (1: knee injury and 2: age-matched controls, and 3: radiographic knee osteoarthritis and 4: age-matched controls) were recruited. Participants performed maximal voluntary isometric knee extensor and flexor contractions. Then, strength (e.g., peak and rate of torque development) and steadiness (e.g., peak power, mean, and median frequency) were derived from each raw torque-time curve and associated power spectral density. A Kruskal-Wallis H test and Spearman's rho correlation analysis assessed cohort differences and association between knee extensor and flexor strength and steadiness. FindingsThe young adult control and knee injury cohorts exhibited greater knee extensor and flexor strength than the older, knee osteoarthritis cohort (p < 0.043). The knee injury cohort, despite being as strong as their healthy counterparts, were significantly less steady with a 92% increase in peak power frequency (p = 0.046). The osteoarthritis cohort exhibited 157% less total power compared to the knee injury and young control cohorts (p < 0.019). Knee extensor and flexor peak torque, rate of torque development, and mean torque exhibit a significant, positive relation with total power (p < 0.018). InterpretationIndividuals with knee injury and disease may exhibit weaker or less steady knee musculature, predisposing them to degenerative joint disease. Clinicians may need to restore knee extensor and flexor steadiness to facilitate better joint neuromuscular control.

Dietary oxyclozanide influences antioxidant enzyme activities and damages DNA in Galleria mellonella (Lepidoptera: Pyralidae).

Oxyclozanide (OXY) is an anthelmintic widely used in the treatment of flatworm infection and fasciolosis. It also has antiadenovirus, antibiofilm, antifungal, and antibacterial activities. Various chemicals have been suggested as alternative chemicals in insect pest management. Here, the oxidative and genotoxic effects of OXY on 7th instars, pupae and adults of the model organism Galleria mellonella (Linnaeus) (Lepidoptera: Pyralidae) were examined. First-instar larvae were reared on 0.003, 0.03, 0.3, and 1.5 g OXY per 100 g artificial diets. Compared with all tested OXY concentrations and controls without OXY, dietary OXY led to increased antioxidant capacity and genotoxic effects. Concentrations of malondialdehyde, an oxidative stress marker, were significantly increased in adults of larvae reared on OXY-charged diets at 0.3 and 1.5 g/100 g compared to the adult control group. We also recorded a significant increase in the genotoxic test data (Tail length, Tail DNA %, Tail moment) at the same stages and concentrations. We recorded significant increases in glutathione-S-transferase, superoxide dismutase (SOD) and glutathione peroxidase activities in larvae fed high OXY concentrations. SOD and catalase activities were also significantly increased at the concentration of 0.03 g/100 g of OXY in the pupal and adult stages. Cytochrome P450 monooxygenase activity was significantly increased at the highest concentration of OXY in the larval and pupal stages. Also, our regression analysis indicates a correlation between the markers of oxidative stress, antioxidant enzymes and comet parameters. These data indicate that OXY induces oxidative stress and antioxidative enzyme response.

Abstract We032: FOXK1-Gli2 network regulates cardiomyocyte proliferation and heart regeneration

Unlike adult mammals, the neonatal mouse has an exceptional regenerative capacity following myocardial injury. Our goal was to decipher and define novel factors and mechanisms that have a role in regeneration of the injured adult mouse heart. FOXK1 is a transcription factor that regulates cell cycle kinetics, myogenic stem cell proliferation, and skeletal muscle regeneration. During development, its expression is restricted to progenitors, somites and the heart. In these studies, we identified a novel role for FOXK1 as a regulator of cardiomyocyte (CM) proliferation and regeneration through the regulation of Gli2 . Analysis of single nucleus RNAseq datasets of neonatal hearts revealed that FOXK1 was expressed in neonatal CMs. qPCR analysis from P1 hearts of control and Foxk1 null mice confirmed this finding and demonstrated dysregulation of the cell cycle regulators, Ccne1 and p21 , in the absence of Foxk1 . Control and Foxk1 null mice were pulsed with EdU from P3 to P7 which demonstrated that the absence of Foxk1 led to a decrease in CM proliferation in-vivo. Then, using a transgenic mouse model that we engineered to overexpress FOXK1 in muscle ( MCK-Foxk1 ), we observed CM proliferation in adult MCK-Foxk1 mice, which showed increased incorporation of EdU and increased pH3 expression compared to control. Bulk RNA-seq analysis of P1, P7, and P14 hearts indicated increased CM proliferation in the MCK-Foxk1 mouse and revealed Gli2 as a potential downstream target of FOXK1. A gel shift assay was performed, which demonstrated that FOXK1 was able to bind to a FOXK1 motif upstream of the Gli2 gene. ChIP-PCR further verified that Gli2 was a direct downstream target of FOXK1 in the embryonic stem cell-embryoid body (ES-EB) system. We then performed ligation of the left anterior descending coronary artery (LAD) in adult control and MCK-Foxk1 mice. Echocardiography following the LAD-ligation injuries showed improved cardiac function in MCK-Foxk1 transgenic mice compared with controls. These results identify FOXK1 as a regulator of CM proliferation and heart regeneration and suggest that this function is mediated through the regulation of Gli2 gene expression.

Abstract Mo090: Altered Cardiac Cell Populations in Hypoplastic Left Heart Syndrome

Background: Congenital heart disease (CHD) affects ~1% of infants. Hypoplastic left heart syndrome (HLHS), a severe form of CHD in which the left ventricle is underdeveloped, is associated by a 15% incidence of heart failure by 6 years of age. Only 6% of HLHS patients have a genetic cause identified on exome sequencing, limiting the ability of patients to receive a diagnosis and potentially benefit from targeted treatments. Hypothesis: Novel HLHS genes are differentially expressed in HLHS cardiac tissues, or in cells with differential abundance in HLHS hearts. Goals: To use single nucleus RNA sequencing of HLHS patient cardiac tissues to identify candidate HLHS genes. Methods: Single nucleus RNA sequencing (nucSeq) was performed on paired left and right ventricular tissues (LV, RV) from 9 participants with HLHS. Left ventricular cardiac tissues from children (4) and adults (12) without CHD were used for comparison. Filtering with CellBender and Solo were used to remove low-quality nuclei. Analysis was performed in R using the Seurat package. Results: HLHS LV tissues had a higher proportion of cardiomyocytes than pediatric or adult control tissues, (78.7% vs 64.1%, p=0.005; 78.7 % vs 47.0%, p=1.8E-08, respectively), and a lower proportion of mural cells (3.1% vs 9.3%, p=3.9E-05; 3.1 % vs 23.5%, p=7.3E-15, respectively). By contrast, there were similar proportions of endothelial cells in the HLHS, pediatric and adult tissues (7.1%, 8.8% and 7.9%, respectively; p=0.56). Within HLHS tissues, the LV had a higher proportion of cardiomyocytes than the RV (78.7% vs 51.2%, p=4.8E-04) while the proportion of endothelial cells were similar in the LV and RV (7.3% and 9.1%, respectively; p=0.33). LV tissues from HLHS participants 2-18 years of age had fewer capillary endothelial cells than pediatric controls (31.2% vs 57.8%, p=0.001), and gene markers of HLHS endothelial cells included PBX1 , which is required for Hox D-3 mediated angiogenesis, as well as ARHGAP26 and RORA, inhibitors of VEGF signaling. Conclusion: HLHS cardiac tissues have a higher proportion of cardiomyocytes in the LV despite a smaller chamber size. Differences in gene expression that accompany the differences in cell proportion could identify novel HLHS genes as well as potential therapeutic targets.

Single-cell atlas of the human brain vasculature across development, adulthood and disease

A broad range of brain pathologies critically relies on the vasculature, and cerebrovascular disease is a leading cause of death worldwide. However, the cellular and molecular architecture of the human brain vasculature remains incompletely understood1. Here we performed single-cell RNA sequencing analysis of 606,380 freshly isolated endothelial cells, perivascular cells and other tissue-derived cells from 117 samples, from 68 human fetuses and adult patients to construct a molecular atlas of the developing fetal, adult control and diseased human brain vasculature. We identify extensive molecular heterogeneity of the vasculature of healthy fetal and adult human brains and across five vascular-dependent central nervous system (CNS) pathologies, including brain tumours and brain vascular malformations. We identify alteration of arteriovenous differentiation and reactivated fetal as well as conserved dysregulated genes and pathways in the diseased vasculature. Pathological endothelial cells display a loss of CNS-specific properties and reveal an upregulation of MHC class II molecules, indicating atypical features of CNS endothelial cells. Cell–cell interaction analyses predict substantial endothelial-to-perivascular cell ligand–receptor cross-talk, including immune-related and angiogenic pathways, thereby revealing a central role for the endothelium within brain neurovascular unit signalling networks. Our single-cell brain atlas provides insights into the molecular architecture and heterogeneity of the developing, adult/control and diseased human brain vasculature and serves as a powerful reference for future studies.

Microglia mediate the early-life programming of adult glucose control.

Mammalian glucose homeostasis is, in part, nutritionally programmed during early neonatal life, a critical window for the formation of synapses between hypothalamic glucoregulatory centers. Although microglia are known to prune synapses throughout the brain, their specific role in refining hypothalamic glucoregulatory circuits remains unknown. Here, we show that microglia in the mediobasal hypothalamus (MBH) of mice actively engage in synaptic pruning during early life. Microglial phagocytic activity is induced following birth, regresses upon weaning from maternal milk, and is exacerbated by feeding dams a high-fat diet while lactating. In particular, we show that microglia refine perineuronal nets (PNNs) within the neonatal MBH. Indeed, transiently depleting microglia before weaning (P6-16), but not afterward (P21-31), remarkably increased PNN abundance in the MBH. Furthermore, mice lacking microglia only from P6-16 had glucose intolerance due to impaired glucose-responsive pancreatic insulin secretion in adulthood, a phenotype not seen if microglial depletion occurred after weaning. Viral retrograde tracing revealed that this impairment is linked to a reduction in the number of neurons in specific hypothalamic glucoregulatory centers that synaptically connect to the pancreatic β-cell compartment. These findings show that microglia facilitate synaptic plasticity in the MBH during early life through a process that includes PNN refinement, to establish hypothalamic circuits that regulate adult glucose homeostasis.

Resurgence of Clinical Malaria in Ethiopia in the Era of Anopheles stephensi Invasion.

The invasion of Anopheles stephensi into Africa poses a potential threat to malaria control and elimination on the continent. However, it is not clear if the recent malaria resurgence in Ethiopia has linked to the expansion of An. stephensi. We aimed to summarize the major achievements and lesson learnt in malaria control in Ethiopia from 2001 to 2022, to assess the new challenges and prospects for the control of An. stephensi. We obtained the clinical malaria case reports, antimalarial drug treatment records, insecticide-treated and long-lasting insecticidal net (ITN/LLIN) distribution and utilization records, and indoor residual spraying (IRS) coverage data from the Ethiopian Ministry of Health (MoH) for the period 2001-2022. We analyzed clinical malaria hotspots using spatially optimized hotspot analysis. We investigated malaria outbreaks in 2022 and examined the potential role of An. stephensi in the outbreaks.Clinical malaria cases in Ethiopia decreased by 80%, from 5.2 million cases (11% confirmed) in 2004 to 1.0 million cases (92% confirmed) in 2018; however, cases increased steadily to 2.6 million confirmed cases (98% confirmed) in 2022. Plasmodium vivax cases and proportion have increased significantly in the past 5 years. Clinical malaria hotspots are concentrated along the western Ethiopian border areas and have grown significantly from 2017 to 2022. Major malaria outbreaks in 2022/23 were detected in multiple sites across Ethiopia, and An. stephensi was the predominant vector in some of these sites, however, it was absence from many of the outbreak sites. The malaria burden has been significantly reduced in Ethiopia in the past two decades, but in recent years it has increased substantially, and the cause of such increase is a subject of further investigation. Major gaps exist in An. stephensi research, including vector ecology, surveillance, and control tools, especially for adult mosquito control.

The inactive X chromosome drives sex differences in microglial inflammatory activity in human glioblastoma.

Biological sex is an important risk factor in cancer, but the underlying cell types and mechanisms remain obscure. Since tumor development is regulated by the immune system, we hypothesize that sex-biased immune interactions underpin sex differences in cancer. The male-biased glioblastoma multiforme (GBM) is an aggressive and treatment-refractory tumor in urgent need of more innovative approaches, such as considering sex differences, to improve outcomes. GBM arises in the specialized brain immune environment dominated by microglia, so we explored sex differences in this immune cell type. We isolated adult human TAM-MGs (tumor-associated macrophages enriched for microglia) and control microglia and found sex-biased inflammatory signatures in GBM and lower-grade tumors associated with pro-tumorigenic activity in males and anti-tumorigenic activity in females. We demonstrated that genes expressed or modulated by the inactive X chromosome facilitate this bias. Together, our results implicate TAM-MGs, specifically their sex chromosomes, as drivers of male bias in GBM.

Colonic MYO5B Expression Retains Fecal Formation but not Weight Loss in OLFM4+ Stem Cell-Specific MYO5B Deletion in Mice

Background and aim: Microvillus inclusion disease (MVID) is a congenital disorder characterized by severe diarrhea and is caused by inactivating mutations in motor protein myosin Vb (MYO5B). Mice lacking MYO5B in intestinal and colonic epithelia ( Vil-CreERT2;Myo5bfl/fl) display an MVID phenotype characterized by significant weight loss, lack of solid feces, and brush border defects. However, previous models have not discerned if this diarrheal phenotype is due to MYO5B loss in the small intestine specifically or both in the small intestine and colon. Using the small intestinal stem cell specific Cre driver under Olfm4, we phenotyped the mice lacking MYO5B only in the small intestine, while normal MYO5B expression is maintained in the colon. Methods: Olfm4-GFP-CreERT2;R26R-LSL-tdTom and Myo5bfl/fl mice were crossbred to generate Olfm4-GFP-CreERT2;R26R-LSL-tdTom;Myo5bfl/fl (Olfm4DMYO5B) mice. Adult Olfm4DMYO5B mice and control littermates received tamoxifen injection at day 0. GI tissues were collected 5 and 7 days post tamoxifen. Results: Following Cre induction, Olfm4DMYO5B mice exhibited progressive weight loss starting at day 5 and on average lost 20.6% on day 7. At day 7, the small intestine contained clear luminal contents, similar to that of the previous MVID model mice. However, the colon of day 7 Olfm4DMYO5B mice contained small, solid feces. An elongation of PCNA+ crypts was identified on day 5 throughout the small intestine. Villus blunting and microvillus inclusions were prominent in the small intestinal enterocytes on day 7, but not on day 5. The day 7 Olfm4DMYO5B small intestine demonstrated a decrease in tuft cell numbers, suggesting differentiation deficits in the MYO5B-lacking stem cells. However, the colon of Olfm4DMYO5B mice showed intact morphology. Colonic crypt depth and tuft cell numbers at both days 5 and 7 were similar to those of control mice. Immunostaining confirmed the maintenance of MYO5B expression in the colon and progressive loss of MYO5B in the small intestine of Olfm4DMYO5B mice. At day 5, MYO5B was specifically lost in small intestinal crypts, and the loss of MYO5B expanded to the villus tips on day 7. These MYO5B-lacking cells were tdTom+, including enterocytes, goblet, and rare tuft cells. Following the loss of MYO5B, small intestinal enterocytes lacked proper apical localization of numerous transporters and enzymes, including Na+-glucose cotransporter 1 and dipeptidyl peptidase 4. Conclusion: Recently established Olfm4DMYO5B mice demonstrated that the specific MYO5B deletion in the small intestinal stem cells causes hyperproliferation and differentiation deficits. Colonic MYO5B function is suffcient for the formation of solid feces, however, these mice experience prominent weight loss, likely due to the MVID-like malabsorption in the small intestine. NIH R01DK128190, RC2DK118640. 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.

Age influences therapeutic ultrasound effects: From increased inflammation in young to decreased inflammaging in adults

BACKGROUND: Therapeutic ultrasound is a controversial topic in physical therapy because outcomes varied due to different experimental designs. At the cellular level, acoustic radiation force (i.e., pressure) from ultrasound previously showed activation of sodium/calcium channels, hyper-proliferation and increased cytokine signaling. However, few have studied the immune response after ultrasound despite the tight coupling between muscle inflammation and regeneration. Additionally, limited work has assessed ultrasound in aged populations. Therefore, a deeper understanding of how therapeutic ultrasound influences the muscle microenvironment is required to use it as a regenerative therapy. The aim of this study was to evaluate inflammatory cytokine signaling and immune cell infiltration after ultrasound in young and aging murine muscle. We hypothesized that the immune response will increase with ultrasound pressure but decrease with age. METHODS: Young (6-weeks-old) and Adult (52-weeks-old) male and female non-injured gastrocnemii were treated with a single sonication of either low intensity pulsed ultrasound (LIPUS) at ~0.243 MPa or pulsed focused ultrasound (pFUS) at ~5.96 MPa. After sonication, muscles were harvested at 1-, 8-, and 24-hours for cytokine analysis using an immunology multiplex assay (138 mice) and at 1- and 24-hours for flow cytometry (120 mice). Control mice were not treated. Sexes were combined due to minimal, if any, differences. Statistical analyses were performed using an ANOVA with post-hoc Bonferroni comparisons ( p &lt; 0.05); only differences compared to control are described. RESULTS: In Young post-LIPUS, IL-15 was upregulated at 1-hour and likely caused the increased M1 and M2 macrophages (M2 &gt; M1) at 1-hour. In Young post-pFUS, the upregulation of VEGF at 1-hour was counteracted by MIG at 8-hours resulting in no change in endothelial cells. Increased MCP-1 at 24-hours post-pFUS corresponded with increased neutrophils, monocytes, and M1 and M2 macrophages (M1 &gt; M2) at 24-hours. In Adult, cytokine expression throughout the 24-hour period was downregulated after LIPUS (IL-9/-15) and pFUS (IL-1α/-2/-6/-9, M-CSF, MIP-1α). Thus, there were no changes in the immune cell response over time post-LIPUS or -pFUS. Additionally, for most of these cytokines, Adult control expression was at least two times greater than Young control. CONCLUSIONS: In Young, LIPUS caused an early, temporary pro- and anti-inflammatory response while pFUS caused a late, extended pro-inflammatory response. However, this response was lost in Adult where age-induced inflammaging was decreased after sonication. In younger muscle, ultrasound could enhance regeneration by augmenting the immune response while in aging muscle it could be used to decrease inflammation induced atrophy. Overall, therapeutic ultrasound caused a pressure dependent inflammatory response that can increase or decrease intrinsic signaling and cell recruitment in adolescent or aged muscles, respectively. This work was funded by the Intramural Research Program of the National Institutes of Health, Clinical Center and National Institutes of Biomedical Imaging and Bioengineering. 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 2036: Prognostic Impact Of Pfkfb3 Expression In Endothelial Cells Insights From Peripheral Arterial Disease Patients And The Effects Of Supervised Exercise

Background: Glycolysis constitutes 85% of ATP production in endothelial cells (ECs). Although upregulating PFKFB3 (6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3), the rate-controlling enzyme for EC glycolysis, has been considered a mean to induce angiogenesis, our group found this approach ineffective or even harmful in pre-clinical peripheral arterial disease (PAD) models. However, no EC specific PFKFB3 data in human PAD patients exists. Hypothesis: EC PFKFB3 upregulation is associated with poor prognosis in PAD patients. Goal: Assess PFKFB3 in ECs and correlate its’ expression with PAD clinical presentation and outcome. Methods: CD31 (EC marker) and PFKFB3 immunostaining was performed on age-matched gastrocnemius (GA) muscle sections from healthy adult control (HA), PAD patients with intermittent claudication (IC; patients experience pain during activity), and patients with chronic limb threatening ischemia (CLTI; the most severe presentation of PAD -patients experience pain at rest with increased risk of ulcers, gangrene, and amputation). A similar approach was used on GA muscle sections from PAD patients before and after 3 weeks of supervised exercise. qPCR was done on flow sorted live CD56-CD31+ EC population from HA and CLTI GA muscle biopsies, for CD31 and PFKFB3 expression. Results: The IC presentation did not increase the %PFKFB3 + ECs (17.08 ± 1.19 vs. 14.40 ± 2.32 SEM, n=6-7/group, p= 0.69) from HA. However, in CLTI tissues, the %PFKFB3 + ECs increased vs both HA (42.01 ± 4.32 SEM vs. 14.40 ± 2.327 SEM, n=7/group, p= 0.0025) and IC (42.01 ± 4.32 SEM vs. 17.08 ± 1.19 SEM, n=6-7/group, p= 0.01). Furthermore, qPCR analysis of PFKFB3 expression in flow sorted CD56-CD31+ EC fraction from CLTI vs. HA (1.855 ± 0.084 SEM vs. 1 ± 0.1768 SEM, n=4/group, P= 0.0052). Three weeks of supervised exercise in PAD patients, with tolerance for activity, decreased the %PFKFB3 + ECs compared vs. pre-exercise values (7.02 ± 1.74 SEM vs. 18.24 ± 2.15 SEM, n= 4 /group, p= 0.0077) respectively. Conclusion: In humans with PAD, higher EC expression of PFKFB3 + uniquely associated with CLTI, but not the IC presentation. Supervised exercise, the only non-surgical approved method to improve functional performance and angiogenesis in PAD patients, reduced EC PFKFB3 expression.

Long-Term Outcomes of Arthroscopic Labral Treatment of Femoroacetabular Impingement in Adolescents: A Nested Propensity-Matched Analysis.

Hip arthroscopy has demonstrated effectiveness as a treatment for femoroacetabular impingement (FAI) in adult patients, with promising long-term outcomes. However, there is a paucity of literature regarding the adolescent population. The purposes of our study were to report on survivorship and patient-reported outcomes (PROs) at a minimum 10-year follow-up in adolescent patients who underwent hip arthroscopy for FAI and labral tears and to compare the survivorship and outcomes of this population with those of a nested, propensity-matched adult control group. Data regarding adolescent patients who underwent primary hip arthroscopy between February 2008 and January 2012 were reviewed. Patients with a minimum 10-year follow-up for the modified Harris hip score (mHHS), Non-Arthritic Hip Score (NAHS), Hip Outcome Score Sport-Specific Subscale (HOS-SSS), International Hip Outcome Tool-12 (iHOT-12), and visual analog scale (VAS) for pain were eligible. The exclusion criteria were previous ipsilateral hip conditions or surgical procedures, a Tönnis grade of >1, or dysplasia. In the subanalysis, adolescent patients were matched to young adult patients with use of a 1:1 ratio on the basis of sex, body mass index, Tönnis grade, lateral center-edge angle (LCEA), labral treatment, capsular treatment, and additional procedures performed during the surgery. A total of 74 patients (61 female and 13 male; 74 hips) with a mean (and standard deviation) age of 16.7 ± 1.4 years and a minimum follow-up of 10 years were included. The latest follow-up occurred at a mean of 125.4 ± 5.3 months (range, 120.0 to 144.1 months). Significant improvement in all PROs from baseline and 100% survivorship were demonstrated at the 10-year follow-up. In the subanalysis, 58 adolescent hips were propensity-matched to 58 young adult hips. The adolescent group had higher postoperative scores for the HOS-SSS (p = 0.021), NAHS (p = 0.021), and iHOT-12 (p = 0.042) than the young adult group. Patient satisfaction at the latest follow-up was also significantly higher in the adolescent group (p = 0.00061). The rate of survivorship free from conversion to THA was similar between the adolescent and control groups (100% versus 96.6%; p = 0.200). Adolescents undergoing hip arthroscopy for the treatment of FAI and labral tears demonstrated excellent outcomes and a high rate of survivorship. These outcomes were superior to those seen in a matched adult group. The results of this study indicate that hip arthroscopy in adolescents is a safe procedure that leads to improvement in outcomes at long-term follow-up. Therapeutic Level IV . See Instructions for Authors for a complete description of levels of evidence.

Differential DNA methylation in iPSC-derived dopaminergic neurons: a step forward on the role of SNORD116 microdeletion in the pathophysiology of addictive behavior in Prader-Willi syndrome.

A microdeletion including the SNORD116 gene (SNORD116 MD) has been shown to drive the Prader-Willi syndrome (PWS) features. PWS is a neurodevelopmental disorder clinically characterized by endocrine impairment, intellectual disability and psychiatric symptoms such as a lack of emotional regulation, impulsivity, and intense temper tantrums with outbursts. In addition, this syndrome is associated with a nutritional trajectory characterized by addiction-like behavior around food in adulthood. PWS is related to the genetic loss of expression of a minimal region that plays a potential role in epigenetic regulation. Nevertheless, the role of the SNORD116 MD in DNA methylation, as well as the impact of the oxytocin (OXT) on it, have never been investigated in human neurons. We studied the methylation marks in induced pluripotent stem-derived dopaminergic neurons carrying a SNORD116 MD in comparison with those from an age-matched adult healthy control. We also performed identical neuron differentiation in the presence of OXT. We performed a genome-wide DNA methylation analysis from the iPSC-derived dopaminergic neurons by reduced-representation bisulfite sequencing. In addition, we performed RNA sequencing analysis in these iPSC-derived dopaminergic neurons differentiated with or without OXT. The analysis revealed that 153,826 cytosines were differentially methylated between SNORD116 MD neurons and control neurons. Among the differentially methylated genes, we determined a list of genes also differentially expressed. Enrichment analysis of this list encompassed the dopaminergic system with COMT and SLC6A3. COMT displayed hypermethylation and under-expression in SNORD116 MD, and SLC6A3 displayed hypomethylation and over-expression in SNORD116 MD. RT-qPCR confirmed significant over-expression of SLC6A3 in SNORD116 MD neurons. Moreover, the expression of this gene was significantly decreased in the case of OXT adjunction during the differentiation. SNORD116 MD dopaminergic neurons displayed differential methylation and expression in the COMT and SLC6A3 genes, which are related to dopaminergic clearance.

EVALUATION OF THERMAL FOGGER FOR EFFECTIVENESS OF LARVICIDE-ADULTICIDE MIXTURE AGAINST AEDES AEGYPTI

A hand-held thermal fogger was evaluated to spray a mixture of adulticide (Aqualuer 20-20® containing permethrin and piperonyl butoxide) and a biological larvicide (VectoBac®12AS, containing Bacillus thuringiensis subspecies israelensis). Adulticide was applied at maximum label rate of 37 mL/ha while larvicide rates were 0.58, 1.76, 2.64, and 3.50 L/ha (Maximum label rate 2.34 L/ha). Two rows, 7.6 m apart, of bioassay cages containing susceptible 25, 5-7 days old female Aedes aegypti were used to assess effectiveness of the adulticide. Along with each cage were two 473 mL empty plastic cups at 0 and 1.3 m above ground to collect the larvicide. Two control cages and two empty cups were placed upwind in the field for five minutes and removed before the spray. The study was conducted in the evening with five separated replications of a rate. The exposed cages and cups were retrieved from the field and brought back to the laboratory for further observations and tests to determine mortality at 24 and 48 h, respectively. All the adults in all treatment cages and all tests died within 24 h of the spray application. In the lab, larval mortality was assessed by introducing Ae. aegypti larvae, where mortality decreased with increasing distance from the spray line and increased with increasing application rate. The larval mortality at all distances was strongly correlated with application rate (R2 = 0.98). The highest rate used produced 64 -95% larval mortality at all distances. The results indicate that Aqualuer 20-20 and VectoBac 12AS can be applied simultaneously as a mixture achieving complete adult control and considerable larval control.

Discriminant Potential of the Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-Cog) in Greek Older Adults with Subjective Cognitive Decline and Mild Cognitive Impairment.

Alzheimer's Disease Assessment Scale Cognitive Subscale (ADAS-Cog) is a widely used screening tool for detecting older adults with Alzheimer's disease among their cognitively healthy peers. A previous study in Greek population showed that ADAS-Cog-Greek (G) is a valid tool and can identify people with Alzheimer's disease from older adult control group; however, there is no current data about whether ADAS-Cog can differentiate older adults with mild cognitive impairment (MCI) from those who have subjective cognitive decline (SCD). The current study aimed to examine the discriminant potential of ADAS-Cog-G in Greek older adults who meet the criteria for SCD or MCI. Four hundred eighty-two community-dwelling older adults, visitors of the Greek Alzheimer Association and Related Disorders, were enrolled in the current study. One hundred seventy-six of them met the criteria for SCD and three hundred six had MCI. Path analysis applied to the data showed that age, as well as educational level affected ADAS-Cog-G performance. Results showed that the cut-off scores, which better discriminate people with SCD from MCI as well as their sensitivity and specificity values, were extracted in participants with high educational level (13 educational years<) and mainly under the age of 75 years. The current study provided evidence concerning the discriminant potential of ADAS-Cog-G to differentiate older adults with SCD from those with MCI in the Greek population, and therefore contributes to the relevant literature on the field.