Available Potential Energy Research Articles

A mathematical fractal-fractional model to control tuberculosis prevalence with sensitivity, stability, and simulation under feasible circumstances

Background:Tuberculosis, a global health concern, was anticipated to grow to 10.6 million new cases by 2021, with an increase in multidrug-resistant tuberculosis. Despite 1.6 million deaths in 2021, present treatments save millions of lives, and tuberculosis may overtake COVID-19 as the greatest cause of mortality. This study provides a six-compartmental deterministic model that employs a fractal–fractional operator with a power law kernel to investigate the impact of vaccination on tuberculosis dynamics in a population. Methods:Some important characteristics, such as vaccination and infection rate, are considered. We first show that the suggested model has positive bounded solutions and a positive invariant area. We evaluate the equation for the most important threshold parameter, the basic reproduction number, and investigate the model’s equilibria. We perform sensitivity analysis to determine the elements that influence tuberculosis dynamics. Fixed-point concepts show the presence and uniqueness of a solution to the suggested model. We use the two-step Newton polynomial technique to investigate the effect of the fractional operator on the generalized form of the power law kernel. Results:The stability analysis of the fractal–fractional model has been confirmed for both Ulam–Hyers and generalized Ulam–Hyers types. Numerical simulations show the effects of different fractional order values on tuberculosis infection dynamics in society. According to numerical simulations, limiting contact with infected patients and enhancing vaccine efficacy can help reduce the tuberculosis burden. The fractal–fractional operator produces better results than the ordinary integer order in the sense of memory effect at diffract fractal and fractional order values. Conclusion:According to our findings, fractional modeling offers important insights into the dynamic behavior of tuberculosis disease, facilitating a more thorough comprehension of their epidemiology and possible means of control.

Oral pimonidazole unveils clinicopathologic and epigenetic features of hypoxic tumour aggressiveness in localized prostate cancer

BackgroundTumor hypoxia is associated with prostate cancer (PCa) treatment resistance and poor prognosis. Pimonidazole (PIMO) is an investigational hypoxia probe used in clinical trials. A better understanding of the clinical significance and molecular alterations underpinning PIMO-labeled tumor hypoxia is needed for future clinical application. Here, we investigated the clinical significance and molecular alterations underpinning PIMO-labeled tumor hypoxia in patients with localized PCa, in order to apply PIMO as a prognostic tool and to identify potential biomarkers for future clinical translation.MethodsA total of 39 patients with localized PCa were recruited and administered oral PIMO before undergoing radical prostatectomy (RadP). Immunohistochemical staining for PIMO was performed on 37 prostatectomy specimens with staining patterns evaluated and clinical association analyzed. Whole genome bisulfite sequencing was performed using laser-capture of microdissected specimen sections comparing PIMO positive and negative tumor areas. A hypoxia related methylation molecular signature was generated by integrating the differentially methylated regions with previously established RNA-seq datasets.ResultsThree PIMO staining patterns were distinguished: diffuse, focal, and comedo-like. The comedo-like staining pattern was more commonly associated with adverse pathology. PIMO-defined hypoxia intensity was positively correlated with advanced pathologic stage, tumor invasion, and cribriform and intraductal carcinoma morphology. The generated DNA methylation signature was found to be a robust hypoxia biomarker, which could risk-stratify PCa patients across multiple clinical datasets, as well as be applicable in other cancer types.ConclusionsOral PIMO unveiled clinicopathologic features of disease aggressiveness in localized PCa. The generated DNA methylation signature is a novel and robust hypoxia biomarker that has the potential for future clinical translation.

On the emergence of ecosystem decay: A critical assessment of patch area effects across spatial scales

Recent analyses suggest that positive patch area effects on biodiversity occur ubiquitously when comparing equal-effort samples from remnant habitat patches. The mechanisms underlying the emergence of this so-called “ecosystem decay” remain poorly understood despite conservation relevance. We leverage spatially-explicit information on the occurrence of plant species across the Swiss Alps (415 plots, 668 species) to test two mechanisms compatible with the emergence of patch-scale ecosystem decay: (i) plots sampled within small patches might have lower biodiversity than plots sampled within large patches (plot-scale decay hypothesis), and (ii) plots sampled within large patches might share a lower proportion of species (turnover hypothesis). We found that patch-scale ecosystem decay occurs also in our system. While plots sampled in large patches tended to be more dissimilar, supporting the turnover hypothesis, we did not find support for the plot-scale decay hypothesis. Additionally, distance between plots and elevational changes explain turnover between plots better than patch area effects. Taken together, these results indicate that applications of ecosystem decay in biodiversity conservation require a better understanding of the mechanisms that potentially underlie this pattern. Patch area effects might be less important than previously assumed when assessing landscape-scale biodiversity, because such effects can be confused with the effects of distance-decay in community similarity, environmental heterogeneity, and sampling effort. More broadly, our findings align with mounting evidence that protecting as much habitat as possible – regardless of whether such habitat exists continuous or fragmented – might be the most effective means to sustain biodiversity across human-dominated landscapes.

The inhibitory effects of monolauroyl-galactosylglycerol against spores of Bacillus cereus and its potential application in dairy

Bacillus cereus is a typical spore-forming strain that can transform into dormant spores to resist unfavorable growth conditions. Among the microorganisms that cause spoilage of dairy products, B. cereus accounts for a large proportion. In this study, we evaluated the inhibitory activity of monolauroyl-galactosylglycerol (MLGG) on B. cereus spores and explored its inhibitory mechanism. The minimum inhibitory spore concentration (MISC) and the minimum sporicidal concentration (MSC) of MLGG against B. cereus spores were 0.313 mg/mL and 2.5 mg/mL, respectively. Scanning electron microscopy was used to observe the changes in spore morphology after MLGG treatment. It was found that low concentrations of MLGG caused irregular spore shape, with a few spores slightly ruptured and high concentrations of MLGG increased the degree of spore rupture, resulting in significant leakage of spore contents. By measuring the effect of MLGG on spore structure, it could be determined that MLGG damaged the coat of the spores and weakened the resistance of the spores, but did not affect the activity of cortical lyases. Therefore, spores could not resume growth even with the addition of exogenous lytic enzymes. After propidium iodide (PI) staining of the spores, flow cytometry analysis revealed that the positive area gradually increased, indicating that the permeability of the inner membrane of the spores was disrupted, leading to dipicolinic acid (DPA) release. The amount of DPA release was proportional to the MLGG concentration, and the growth rate of the spores decreased under hypertonic conditions. These findings indicated that MLGG inactivated spores primarily by damaging the inner membrane of the spores.

PD-L1 immunohistochemistry in gastric cancer: Comparison of combined positive score and tumor area positivity across 28-8, 22C3, and SP263 assays.

2625 Background: The clinical application of programmed death-ligand 1 (PD-L1) immunohistochemistry is complicated by multiple available assays and different testing platforms, scoring algorithms, and cut-offs applied. This study assessed the analytical comparability of three commercially available PD-L1 assays (28-8, 22C3, and SP263) and two scoring algorithms used in gastric cancer (GC). Methods: Serial sections of 100 commercially procured GC samples, selected by the 28-8 assay to represent the dynamic range of PD-L1 expression, were stained with the three in vitrodiagnostic (IVD)-grade PD-L1 assays. Stained slides were blindly and independently evaluated by three trained pathologists for intra- and inter-reader assessment. Scoring was performed using the combined positive score (CPS) and tumor area positivity (TAP) methods, followed by statistical analysis. Digital image analysis (DIA) was used to objectively assess the technical performance of each assay by simulating CPS and TAP scoring methods using the HALO platform. Results: Comparable, specific, staining patterns were observed with the three PD-L1 assays. Pathologist assessment of PD-L1 positivity was reproducible in GC sample cohorts despite discernible variability in the observed staining intensity. When the same PD-L1 cut-offs were applied, inter- and intra-assay assessments of all three assays, using either CPS or TAP scoring methods, demonstrated moderate to almost-perfect (inter-assay Cohen’s kappa [κ] ranged from 0.47 to 0.83) and substantial to almost-perfect (intra-assay κ ranged from 0.77 to 1.00) agreement, respectively. Moreover, inter-pathologist evaluation showed a significant level of reproducibility (intraclass correlation coefficient (ICC) ≥0.92). DIA confirmed no difference in technical performance when specific digital algorithms were applied. Conclusions: This GC study highlights analytical concordance in PD-L1 testing among three major PD-L1 assays when TAP and CPS scoring algorithms are prospectively applied. Independently, DIA further supports the comparability of the technical performance of the assays. These observations support flexibility in cross-application of the different PD-L1 assays and scoring algorithms currently used to characterize PD-L1–positive GC samples.

4-Deoxy-ε-Pyrromycinone: A Promising Drug/Lead Compound to Treat Tumors.

Anthraquinone drugs are widely used in the treatment of tumors. However, multidrug resistance and severe cardiac toxicity limit its use, which have led to the discovery of new analogues. In this paper, 4-Deoxy-ε-pyrromycinone (4-Deo), belonging to anthraquinone compounds, was first been studied with the anti-tumor effects and the safety in vitro and in vivo as a new anti-tumor drug or lead compound. The quantitative analysis of 4-Deo was established by UV methodology. The anti-cancer effect of 4-Deo in vitro was evaluated by cytotoxicity experiments of H22, HepG2 and Caco2, and the anti-cancer mechanism was explored by cell apoptosis and cycle. The tumor-bearing mouse model was established by subcutaneous inoculation of H22 cells to evaluate the anti-tumor effect of 4-Deo in vivo. The safety of 4-Deo was verified by the in vitro safety experiments of healthy cells and the in vivo safety experiments of H22 tumor-bearing mice. Tumor tissue sections were labeled with CRT, HMGB1, IL-6 and CD115 to explore the preliminary anti-cancer mechanism by immunohistochemistry. In vitro experiments demonstrated that 4-Deo could inhibit the growth of H22 by inducing cell necrosis and blocking cells in S phase, and 4-Deo has less damage to healthy cells. In vivo experiments showed that 4-Deo increased the positive area of CRT and HMGB1, which may inhibit tumor growth by triggering immunogenic cell death (ICD). In addition, 4-Deo reduced the positive area of CSF1R, and the anti-tumor effect may be achieved by blocking the transformation of tumor-associated macrophages (TAMs) to M2 phenotype. In summary, this paper demonstrated the promise of 4-Deo for cancer treatment in vitro and in vivo. This paper lays the foundation for the study of 4-Deo, which is beneficial for the further development anti-tumor drugs based on the lead compound of 4-Deo.

Shape of the membrane neck around a hole during plasma membrane repair

Plasma membrane damage and rupture occurs frequently in cells, and holes must be sealed rapidly to ensure homeostasis and cell survival. The membrane repair machinery is known to involve recruitment of curvature-inducing annexin proteins, but the connection between membrane remodeling and hole closure is poorly described. The induction of curvature by repair proteins leads to the possible formation of a membrane neck around the hole as a key intermediate structure before sealing. We formulate a theoretical model of equilibrium neck shapes to examine the potential connection to a repair mechanism. Using variational calculus, the shape equations for the membrane near a hole are formulated and solved numerically. The system is described under a condition of fixed area, and a shooting approach is applied to fulfill the boundary conditions at the free membrane edge. A state diagram of neck shapes is produced describing the variation in neck morphology with respect to the membrane area. Two distinct types of necks are predicted, one with conformations curved beyond π existing at positive excess area, whereas flat neck conformations (curved below π) have negative excess area. The results indicate that in cells, the supply of additional membrane area and a change in edge tension is linked to the formation of narrow and curved necks. Such necks may be susceptible to passive or actively induced membrane fission as a possible mechanism for hole sealing during membrane repair in cells.

Prediction of Positive Patient Health Questionnaire-2 Screening Using Area Deprivation Index in Primary Care.

Depression is recognized as a significant public health issue in the United States. The National Survey on Drug Use and Health reports that 21.0 million adults aged 18 or older had major depressive disorder in 2020, including 14.8 million experiencing a major depressive episode with severe impairment. The aim is to predict the positivity of Patient Health Questionnaire-2 (PHQ-2) outcomes among patients in primary care settings by analyzing a range of variables, including socioeconomic status, demographic characteristics, and health behaviors, thereby identifying those at increased risk for depression. Employing a machine learning approach, the study utilizes retrospective data from electronic health records across 15 primary care clinics in South Florida to explore the relationship between social determinants of health (SDoH), including area of deprivation index (ADI) and PHQ-2 positivity. The study encompasses 15 primary care clinics located in South Florida, where a diverse patient population receives care. Analysis included 94,572 patient visits; 74,636 records were included in the study. If a zip+4 was not available or an ADI score did not exist, the visit was not included in the final analysis. Screening involved the PHQ-2, assessing depressed mood and anhedonia, with a cutoff >2 indicating positive screening. ADI was used to assess SDoH by matching patients' residential postal codes to ADI national percentiles. Demographics, sexual history, tobacco use, caffeine intake, and community involvement were also evaluated in the study. Over 40 machine learning algorithms were explored for their accuracy in predicting PHQ-2 outcomes, using software tools including Scikit-learn and stats models in Python. Variables were normalized, scored, and then subjected to predictive regression models, with Random Forest showing outstanding performance. Feature engineering and correlation analysis identified ADI, age, education, visit type, coffee intake, and marital status as significant predictors of PHQ-2 positivity. The area under the curve and model accuracies varied across clinics, with specific clinics showing higher predictive accuracy and others (p > .05). The study concludes that the ADI, as a proxy for SDoH, alongside other individual factors, can predict PHQ-2 positivity. Health organizations can use this information to anticipate health needs and resource allocation.

Convergence of ensemble forecast distributions in weak and strong forcing convective weather regimes

AbstractThe constraint of computational power and the huge number of degrees of freedom of the atmosphere means a sampling uncertainty exists in probabilistic ensemble forecasts. In our previous study, the uncertainty could be quantified, creating a convergence measure which converges proportional to in the limit of large ensemble size . This power law can then be extrapolated to determine how sampling uncertainty would decrease with larger ensemble sizes and hence find the necessary ensemble size. It is unknown, however, how the sampling uncertainty depends on different weather regimes. This study extends the previous idealised ensemble developed, by including weak and strong forcing convective weather regimes, to look at how sampling uncertainty convergence differs in each. Two ‐member ensembles were run, with weak and strong forcing respectively. Comparisons with a kilometre‐scale weather prediction model ensured realistic weak and strong forcing regimes by comparing the rain, convective available potential energy (CAPE), convective adjustment timescale, and distribution shapes throughout the diurnal cycle. Differences in distribution shape between the regimes led to differences in the convergence measure. Large differences in spread between weak and strong forcing runs throughout the hr period led to large differences in sampling uncertainty of the mean and standard deviation, which could be quantified according to well‐known equations. The timing of these differences was case‐dependent. For extreme statistics such as the quantile and for cases where there was precipitation, the moisture variables for the weak forcing case had the largest sampling uncertainty and required the most members for convergence proportional to . This was due to the tails of the weak forcing moisture variables containing the least amount of density. Different ensemble sizes will hence be required depending on whether one is in the weak or strong forcing convective weather regime.

#1593 ARGDRR, a novel disintegrin, attenuates AKI to CKD-related renal fibrosis

Abstract Background and Aims Acute kidney injury (AKI) can lead to progress renal fibrosis, resulting in chronic kidney disease (CKD) and end stage renal disease (ESRD). Nevertheless, the underlying pathological mechanisms remain unclear. Platelet activation has been demonstrated as one of the important factors involved in the pathophysiology of AKI, and studies show the therapeutic potential for using platelet inhibitors or antagonists in ischemia-reperfusion (IR)-induced AKI models. However, drug-induced side effects, including thrombotic microangiopathies (TMA)-related kidney damage, thrombocytopenia and bleeding, hinder clinical application in AKI or CKD treatment. Our newly developed dual effect disintegrin, ARGDRR, is derived from snake venom exert a high affinity to αIIbβ3 and αvβ3 binding affinity without causing bleeding and other thrombosis-related side effects. Method We performed a unilateral ischemia-reperfusion injury (UIRI) model to investigate integrin αIIb(Itga2b) and β3(Itgb3) mRNA levels in the injured kidney for 15 days. For exploring the therapeutic efficiency of ARGDRR in AKI, we administered ARGDRR post-ischemic injury and measured platelet activation after 24 hours using quantitative polymerase chain reaction (qPCR) and immunohistochemistry (IHC) for Itgb3, Lcn2, and CD42b. For exploring the therapeutic efficiency of ARGDRR in CKD progression, we administrated ARGDRR daily for 15 days after IRI with nephrectomy, and examined the difference of renal dysfunction, renal fibrosis, and cell death compared to sham group. Results We found that platelet activation with progressively increased mRNA level of platelet glycoprotein IIb-GPIIIa (Itga2b and Itgb3) in the UIRI kidney over time. After 24 hr of ischemic AKI, ARGDRR administration reduced Lcn2 and Itgb3 mRNA level and CD42b positive area in the kidney, indicating its efficacy in protecting the kidneys by inhibiting platelet activation and aggregation. In AKI to CKD transition, serum creatine and blood urea nitrogen (BUN) were significantly lower both in efficacious and high dose of ARGDRR administration group. ARGDRR significantly reduced CD42b expression and increased LTL positive area. Also, ARGDRR administration alleviated renal fibrosis, cell apoptosis and senescence-related molecules p21 and Ctgf expression in the AKI to CKD transition. Conclusion Our results have successfully proved our hypothesis that disintegrin-ARGDRR has the potential for renal protection in the transition of AKI to CKD through inhibition of platelet activation and aggregation during acute injury.

The tardigrade-derived mitochondrial abundant heat soluble protein improves adipose-derived stem cell survival against representative stressors

Human adipose-derived stem cell (ASC) grafts have emerged as a powerful tool in regenerative medicine. However, ASC therapeutic potential is hindered by stressors throughout their use. Here we demonstrate the transgenic expression of the tardigrade-derived mitochondrial abundant heat soluble (MAHS) protein for improved ASC resistance to metabolic, mitochondrial, and injection shear stress. In vitro, MAHS-expressing ASCs demonstrate up to 61% increased cell survival following 72 h of incubation in phosphate buffered saline containing 20% media. Following up to 3.5% DMSO exposure for up to 72 h, a 14–49% increase in MAHS-expressing ASC survival was observed. Further, MAHS expression in ASCs is associated with up to 39% improved cell viability following injection through clinically relevant 27-, 32-, and 34-gauge needles. Our results reveal that MAHS expression in ASCs supports survival in response to a variety of common stressors associated with regenerative therapies, thereby motivating further investigation into MAHS as an agent for stem cell stress resistance. However, differentiation capacity in MAHS-expressing ASCs appears to be skewed in favor of osteogenesis over adipogenesis. Specifically, activity of the early bone formation marker alkaline phosphatase is increased by 74% in MAHS-expressing ASCs following 14 days in osteogenic media. Conversely, positive area of the neutral lipid droplet marker BODIPY is decreased by up to 10% in MAHS-transgenic ASCs following 14 days in adipogenic media. Interestingly, media supplementation with up to 40 mM glucose is sufficient to restore adipogenic differentiation within 14 days, prompting further analysis of mechanisms underlying interference between MAHS and differentiation processes.

#2599 Adam17 inhibition moderates interstitial fibrosis and macrophage infiltration in TD1 mouse model

Abstract Background and Aims ADAM17 participates in the release into circulation of inflammatory and fibrotic molecules, such as TNF-α actively involved in the progression of Diabetic Nephropathy. We studied the effect of specific deletion of ADAM17 in the endothelium and renal tubule in a T1 diabetic mouse model demonstrating its participation in the progression of kidney damage. As ADAM17 is also expressed in other cell types we wanted to study the effect of the complete deletion of Adam17 in type 1 diabetic mouse. Method We studied total inducible (by TAM) Adam17 knockout male mice (ADAM17KO) and their control littermates. Animals were rendered diabetic by STZ injection (DB group). Blood glucose, mesangial index (PAS staining), number of podocytes and positive area of α-SMA (by immunohistochemistry) were determined. We also studied the protein expression of the cytokine MCP-1 in renal cortex by Western Blot. Results The mesangial index value observed in the 20wk-DB group was compared with the diabetic ADAM17KO group. In the latter, the value significantly decreased. Podocyte loss detected in DB group was not observed in the KO-DB group. For interstitial fibrosis, the intensity and location of α-SMA was significantly lower in the ADAM17KO group. In addition, complete deletion of Adam17 partly prevented the infiltration of macrophages assessed by the expression of MCP-1 in the protein extract of the renal cortex. Conclusion Complete inhibition of Adam17 expression in T1DM mice prevents fibrosis progression and protects the glomerulus from hypertrophy. At the same time, it reduces macrophage infiltration, limiting the inflammatory response induced by diabetes. We demonstrate the influence of Adam17 on macrophages to reduce the expression of TNF-a partly responsible for the progression of diabetic nephropathy.

#2349 Lysyl oxidase-like 2 inhibitor ameliorates tubulointerstitial fibrosis and albuminuria in cyclosporine induced nephropathy

Abstract Background and Aims Lysyl oxidase-like 2 (LOXL2), an amine oxidase, contributes to fibrotic scarring in the tissue by facilitating collagen cross-linking. Recent data demonstrated upregulation of lysyl oxidase and LOXL2 in the fibrotic kidneys after administration of cylosporine A (CsA). Thus, inhibition of LOXL2 may render therapeutic effects against CsA-induced nephropathy by ameliorating tubulointerstitial fibrosis. Method Low salt fed CD-1 mice were administered saline or CsA (15 mg/kg/day, intraperitoneally) for 8 weeks. At 4 weeks, CsA-treated animals were divided into 2 groups respectively and treated with: (1) vehicle, (2) LOXL2 inhibitor (10 mg/kg/day, gavage feeding) for another 4 weeks up to 8 weeks. We explored the reduction of tubulointerstitial fibrosis as well as albuminuria with administration of LOXL2 inhibitor in CsA nephropathy mouse model. Results CsA administration significantly increased the levels of serum creatinine (10.0 ± 1.2 vs. 28.4 ± 4.3, p = 0.02), tubular injury scores (0.1 ± 0.2 vs. 3.6 ± 1.3, p = 0.03), tubulointerstitial fibrosis (Sirius red positive area, 14 ± 6% vs. 204 ± 43%, p = 0.01), F4/80 positive inflammatory cell infiltration in mouse kidney (2 ± 1 vs. 21 ± 4, p = 0.001). These changes were associated with upregulated mRNA expression of LOXL2, TGF-β1, and monocyte chemoattractant protein-1 (MCP-1). Administration of LOXL2 inhibitor significantly suppressed the expression of alpha-SMA and collagen 1A, intrarenal F4/80 positive inflammatory cell infiltrations (21 ± 4 vs. 14 ± 2, p = 0.001) and improved tubular injury scores (3.6 ± 1.3 vs. 1.6 ± 1.1, p = 0.02). Moreover, LOXL2 inhibitor effectively reduced albuminuria excretion (46 ± 4 vs. 22 ± 6, p = 0.02). Conclusion Our results suggest that LOXL2 inhibitor treatment can attenuate CsA nephropathy progression and LOXL2 may be a potential therapeutic target in CsA nephropathy.

Assessing hurricane impact on vegetation and endangered deer habitat using airborne lidar and multispectral images

In the lower Florida Keys, the endangered Florida Key deer and numerous other wildlife species inhabit a vulnerable island environment susceptible to storm surges and rising seawater due to low elevation and flat terrain. Timely and reliable assessment of vegetation damage from natural disasters, such as Hurricane Irma, is crucial for effective habitat management. The study’s overall objective is to examine Hurricane Irma’s impact on vegetation on No Name Key, Florida, using remote sensing. The study relates the area change in vegetation obtained from remote sensing analysis to Florida Key deer population changes following the storm. The methodology involved performing a thematic change detection analysis using the following data sources: (1) aerial multispectral images (for pre- and post-Hurricane), (2) airborne lidar data (for pre- and post-Hurricane), (3) an existing vegetation map, and (4) soil data. A Support Vector Machine (SVM) image classification algorithm was applied to pre- and post-storm input image stacks to create pre- and post-Hurricane Irma vegetation maps. We were then able to obtain the area change information (for various vegetation categories) by performing the change detection analysis of the 2 SVM-classified images. The differences in areas following the storm were calculated for 7 affected vegetation types. Using the area change information following Hurricane Irma, we estimated the number of deer supported by the storm-affected vegetation. These estimated deer numbers, based on the area differences in post-Hurricane Irma vegetation types, were compared to observed deer numbers collected during the post-Hurricane Irma Texas A&M Natural Resources Institute (NRI) deer field survey. The results showed the following: mangroves had the largest negative area changes (area loss), followed by pinelands, hardwoods/hammocks, developed areas, and buttonwoods. Freshwater marshes had the largest positive area changes (area gain). The deer's preferred vegetation areas had decreased post-Hurricane Irma, resulting in a reduced deer population compared to pre-storm numbers. The predicted number of the Key deer post-Hurricane Irma fell within a 95% confidence interval of the observed deer population from the post-storm field survey. The study findings and techniques could be applied to study climate change impact, especially sea level rise. This methodology can be valuable in assessing the impact of storms on other wildlife species in similar environments. The applications and methodology are especially relevant considering the increasing frequency and intensity of storm surges and the accelerating rate of sea level rise.

#2385 Inhibition of histone lysine demethylases 5 attenuates acute kidney injury but not acute kidney disease or chronic kidney disease

Abstract Background and Aims The histone lysine demethylases 5 (KDM5) are members of the family of Jumonji C domain-containing histone demethylases and catalyze the removal of di- and tri-methyl moieties from the fourth lysine of histone 3 (H3K4me2/3). It has been reported that the KDM5 family mediates a range of physiological and pathological events including cell differentiation, motility, senescence and epithelial-mesenchymal transition (EMT) via activating or repressing transcription in demethylase-dependent or independent manners in both homeostasis and disease. This study investigated the potential role of KDM5 in the development and progression of renal fibrosis in acute kidney injury (AKI), chronic kidney disease (CKD) or AKI to CKD transition (acute kidney disease, AKD). Method We evaluated therapeutic effects of KDM5 inhibition in AKI model of unilateral nephrectomy plus contralateral ischemia-reperfusion (IR) injury, AKI to CKD transition model of unilateral IR injury plus removal of contralateral uninjured kidney after 8 days of IR injury, and CKD model of unilateral ureteral obstruction (UUO) in mice. Results KDM5 C70 inhibitor prevented the increase of blood urea nitrogen and positive areas of trichrome, Sirius red, F4/80 and α-smooth muscle actin in injured kidneys 7 days after IR injury. Significant reductions in renal mRNA levels of pro-inflammatory cytokine/chemokine, EMT and pro-fibrotic markers (IL-1β, IL-6, TNF-α, CCL2, CCL3, GM-CSF, MMP9, fibronectin, vimentin, TGF-β1, COL4A1) were also observed with attenuated mRNA levels of KDM5a-c. In addition, renal protein levels of antioxidants such as catalase, SOD1 and SOD2 were increased in injured kidneys of KDM5 inhibitor-treated mice. However, parameters of renal fibrosis and inflammation were similar between mice without and with KDM5 inhibitor in both AKD and CKD models. Conclusion These data demonstrate that inhibition of KDM5 could prevent renal inflammation and fibrosis after acute and severe IR injury. The activation of KDM5 might be an early step in the development of renal fibrosis after AKI insult.

#2663 Assessment of in situ alternative and classical convertases as indicators of ongoing glomerular complement activation in IgA nephropathy

Abstract Background and Aims Complement activation is involved in IgA nephropathy (IgAN), opening up new avenues for complement-targeting therapies. However, precise patient selection and optimal timing for these therapies are essential. Current diagnostic methods of complement activation in IgAN, based on immunostaining, lack precision in identifying specific pathway(s) involved and distinguishing ongoing from past complement activation. Method Complement deposition analysis was conducted on kidney biopsy samples from 53 patients with primary IgAN at diagnosis (NCT05234463). Glomerular complement deposition was assessed through immunohistochemistry for C4d and C5b-9 and immunofluorescence for C3b/iC3b and C1q. In situ detection of C3/C5 convertases complexes from complement classical/lectin (C4b2b) and alternative pathways (C3bBb) was performed on paraffin-embedded kidney biopsies from 33 patients using a proximity ligation assay (Duolink®, Sigma). Kidney biopsies from patients with lupus nephritis (LN) and C3 glomerulopathy (C3G) were used as positive controls for classical/lectin and alternative convertases staining, respectively. The signals corresponding to the convertases were quantified using QuPath® Software and correlated with clinical parameters, MEST-C score and C5b-9 staining. Results At diagnosis, all 53 IgAN patients (100%) exhibited C3 mesangial deposition. Additionally, 51.2% were positive for C4d, 16.5% had C1q deposition and 62.7% had C5b-9 deposition. In situ analysis of C3/C5 convertases revealed the presence of glomerular C3bBb convertases in 20/33 (60.6%) patients, with a median positive area of 14.9 [2.5-26.5]%o of glomerular surface. C4b2b convertases were detected in 21/33 patients (63.6%), with a median glomerular positive area of 8.1 [3.4-12.1]%o. Both C3bBb and C4b2b convertases were detected in 13/33 (39.4%) patients (Fig. 1). By contrast, no convertases were detected in 5/33 (15.2%) patients despite positive C3b/iC3b deposition. The intensity of C3bBb staining positively correlated with terminal pathway C5b-9 glomerular deposits (p = 0.0011, r2 0.29). Patients with MEST-C score S1 lesions and/or interstitial fibrosis/tubular atrophy (T1-2) exhibited a more pronounced intensity of C3bBb at diagnosis (p = 0.0093). Conclusion Our study, for the first time, presents evidence of ongoing complement activation at the tissue level in IgA nephropathy (IgAN), showing variable intensities among patients. The visualization of in situ C3/C5 convertases could enhance the functional aspect of pathological examination, helping identify patients with ongoing complement activation, making them potential candidates for complement inhibitors. Further research is necessary to investigate the kinetics of this staining over the natural course of IgAN and its implications for IgAN progression.

Molecular mechanism of Danxiong Tongmai Granules in treatment of coronary heart disease.

Danxiong Tongmai Granules (DXTMG) are widely utilized in treating coronary heart disease (CHD) in China. This study aims to explore the molecular mechanisms underlying the therapeutic effects of DXTMG on CHD by employing a network pharmacology approach, complemented with experimental validation. Traditional Chinese Medicine (TCM) compounds and targets were identified via searches in the BATMAN-TCM database, and the GeneCards database was used to obtain the main target genes involved in CHD. We combined disease targets with the drug targets to identify common targets. The "TCM-compound-target" network was plotted using Cytoscape 3.7.2 software and a protein-protein interaction (PPI) network was constructed using the STRING database from which core targets were obtained. Gene ontology (GO) function analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed for common drug-disease targets using R Version 4.0.4 (64 bit) software. Molecular docking of core protein-small molecule ligand interaction was modeled using AutoDock software. A molecular dynamics simulation was conducted on the optimal protein-small molecule complex identified through molecular docking, using Amber18 software. The rat model for myocardial ischemia was established through pre-gavage administration of DXTMG, followed by dorsal hypodermic injection of isoprenaline. Myocardial tissues from the rats were analyzed using hematoxylin and eosin (HE) staining and Masson's trichrome staining. Relevant targets were validated by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry. 162 potential targets of DXTMG involved in CHD were identified. These included INS, ALB, IL-6 and TNF according to PPI network studies. GO enrichment analysis identified a total of 3365 GO pathways, including 3049 biological process pathways (BP) concerned with the heart and circulatory system; 109 cellular component (CC) pathways, including cation channels and membrane rafts and 207 molecular function (MF) pathways related to receptor ligands and activators. KEGG analysis revealed a total of 137 pathways (P < 0.05), including those related to AGE-RAGE signaling associated with diabetic complications, fluid shear stress and atherosclerosis. The results of molecular docking and molecular dynamics simulations demonstrated the robust binding affinity between the compounds and target proteins. Animal experiment findings indicated that, compared with the model group, the DXTMG group effectively ameliorated inflammation and fibrosis in rat myocardial tissues, reduced LDH, cTn-I, and MDA levels (P < 0.05, P < 0.01), elevated SOD and GSH-PX levels (P < 0.05), and reduced the percentage of positive area for IL-6 and TNF-α (P < 0.05). This study preliminarily suggests that DXTMG can modulate oxidative stress, inflammation response, and cardiomyocyte regulation, thereby mitigating the onset and progression of CHD.

Climatology of thermodynamic indices and background synoptic conditions responsible for severe convection during pre‐ to post‐monsoon seasons over Indian region

AbstractTo investigate the mechanisms responsible for severe convection linked with a variety of mesoscale, synoptic systems over the Indian region, a preliminary analysis is conducted. Using IMD RSRW/Reanalysis (ERA5)/Lightning (TRMM LIS) products both local‐scale thermodynamics and large‐scale background conditions responsible for the major lightning hot spots in different seasons are investigated. In the pre‐monsoon season, high lightning activity over NE Indian region is due to moisture intrusion by south‐westerly at lower levels (warm air advection), along with mid‐level north westerlies (cold air advection) which favour the low‐level convergence and upper‐level divergence. SCS over south India are mainly attributed to wind discontinuity over the region. During monsoon season major lightning activity in NW India is due to the presence of heat low and the rest of the country less intense SCS activity is evident due to high wind shear conditions in connection with low‐level (westerly) and upper‐level (easterly jet stream). In the post‐monsoon season, major lightning in south India are due to the prevailing north‐easterlies and in north India, the activity is attributed to the passage of western disturbance (WD). Station plot analysis suggests the varied distribution of thermodynamic indices season/region‐wise due to the diverse background forcing. Correlation analysis between the lightning and thermodynamic indices suggests that more skillful indices are KI (~0.7 to 0.8), precipitable water (PPW) (~0.8) (pre‐monsoon, monsoon) and (PPW) (~0.8), convective available potential energy (CAPE) (~0.6) in post monsoon over respective lightning hot‐spot regions. Independent verification using Wyoming data/reanalysis for different case studies suggests that monitoring the monthly thresholds of highly correlated indices provides a helpful proxy to operationally forecast the severe convective systems (season/region). Mean Equivalent potential temperature, and mixing ratio in mixed layer has shown reasonable association with the lightning occurrence over all lightning hot spot regions which are implemented in the IMD GFS and IMD WRF models.

Glucagon-like peptide-1 receptor agonist exendin 4 ameliorates diabetes-associated vascular calcification by regulating mitophagy through the AMPK signaling pathway

BackgroundVascular calcification (VC) is a complication in diabetes mellitus (DM) patients. Osteogenic phenotype switching of vascular smooth muscle cells (VSMCs) plays a critical role in diabetes-related VC. Mitophagy can inhibit phenotype switching in VSMCs. This study aimed to investigate the role of the glucagon-like peptide-1 receptor (GLP-1R) agonist exendin 4 (EX4) in mitophagy-induced phenotype switching.Materials and methodsThe status of VC in T2DM mice was monitored using Von Kossa and Alizarin Red S (ARS) staining in mouse aortic tissue. Human aortic smooth muscle cells were cultured in high glucose (HG) and β-glycerophosphate (β-GP) conditioned medium. Accumulation of LC3B and p62 was detected in the mitochondrial fraction. The effect of EX4 in vitro and in vivo was investigated by knocking down AMPKα1.ResultsIn diabetic VC mice, EX4 decreased the percentage of von Kossa/ARS positive area. EX4 inhibited osteogenic differentiation of HG/β-GP-induced VSMCs. In HG/β-GP-induced VSMCs, the number of mitophagosomes was increased, whereas the addition of EX4 restored mitochondrial function, increased the number of mitophagosome-lysosome fusions, and reduced p62 in mitochondrial frictions. EX4 increased the phosphorylation of AMPKα (Thr172) and ULK1 (Ser555) in HG/β-GP-induced VSMCs. After knockdown of AMPKα1, ULK1 could not be activated by EX4. The accumulation of LC3B and p62 could not be reduced after AMPKα1 knockdown. Knockdown of AMPKα1 negated the therapeutic effects of EX4 on VC of diabetic mice.ConclusionEX4 could promote mitophagy by activating the AMPK signaling pathway, attenuate insufficient mitophagy, and thus inhibit the osteogenic phenotype switching of VSMCs.

A hypothesis test for detecting spatial patterns in categorical areal data

The vast growth of spatial datasets in recent decades has fueled the development of many statistical methods for detecting spatial patterns. Two of the most commonly studied spatial patterns are clustering, loosely defined as datapoints with similar attributes existing close together, and dispersion, loosely defined as the semi-regular placement of datapoints with similar attributes. In this work, we develop a hypothesis test to detect spatial clustering or dispersion at specific distances in categorical areal data. Such data consists of a set of spatial regions whose boundaries are fixed and known (e.g., counties) associated with a categorical random variable (e.g. whether the county is rural, micropolitan, or metropolitan). We propose a method to extend the positive area proportion function (developed for detecting spatial clustering in binary areal data) to the categorical case. This proposal, referred to as the categorical positive areal proportion function test, can detect various spatial patterns, including homogeneous clusters, heterogeneous clusters, and dispersion. Our approach is the first method capable of distinguishing between different types of clustering in categorical areal data. After validating our method using an extensive simulation study, we use the categorical positive area proportion function test to detect spatial patterns in Boulder County, Colorado USA biological, agricultural, built and open conservation easements.