Angiotensin-converting Enzyme Research Articles

Characterization of novel ACE2 mRNA transcripts: The potential role of alternative splicing in SARS-CoV-2 infection.

The human angiotensin converting enzyme 2 (ACE2) gene encodes a type I transmembrane protein, which is homologous to angiotensin I-converting enzyme (ACE) and belongs to the angiotensin-converting enzyme family of dipeptidyl carboxypeptidases. As highlighted by the COVID-19 pandemic, ACE2 is not only crucial for the renin-angiotensin-aldosterone system (RAAS), but also displays great affinity with the SARS-CoV-2 spike protein, representing the major receptor of the virus. Given the significance of ACE2 in COVID-19, especially among cancer patients, the present study aims to explore the transcriptional landscape of ACE2 in human cancer and non-cancerous cell lines through the design and implementation of a custom targeted long-read sequencing approach. Bioinformatics analysis of the massive parallel sequencing data led to the identification of novel ACE2 mRNA splice variants (ACE2 sv.7-sv.12) that demonstrate previously uncharacterized exon-skipping events as well as 5' and/or 3' alternative splice sites. Demultiplexing of the sequencing data elucidated the differential expression profile of the identified splice variants in multiple human cell types, whereas in silico analysis suggests that some of the novel splice variants could produce truncated ACE2 isoforms with altered functionalities, potentially influencing their interaction with the SARS-CoV-2 spike protein. In summary, our study sheds light on the complex alternative splicing landscape of the ACE2 gene in cancer cell lines, revealing novel splice variants that could have significant implications for SARS-CoV-2 susceptibility in cancer patients. These findings contribute to the increased understanding of ACE2's role in COVID-19 and highlight the importance of considering alternative splicing as a key factor in viral pathogenesis. Undoubtably, further research is needed to explore the functional roles of these variants and their potential as therapeutic targets in the ongoing fight against COVID-19.

Dynamin independent endocytosis is an alternative cell entry mechanism for multiple animal viruses.

Mammalian receptor-mediated endocytosis (RME) often involves at least one of three isoforms of the large GTPase dynamin (Dyn). Dyn pinches-off vesicles at the plasma membrane and mediates uptake of many viruses, although some viruses directly penetrate the plasma membrane. RME is classically interrogated by genetic and pharmacological interference, but this has been hampered by undesired effects. Here we studied virus entry in conditional genetic knock-out (KO) mouse embryonic fibroblasts lacking expression of all three dynamin isoforms (Dyn-KO-MEFs). The small canine parvovirus known to use a single receptor, transferrin receptor, strictly depended on dynamin. Larger viruses or viruses known to use multiple receptors, including alphaviruses, influenza, vesicular stomatitis, bunya, adeno, vaccinia, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and rhinoviruses infected Dyn-KO-MEFs, albeit at higher dosage than wild-type MEFs. In absence of the transmembrane protease serine subtype 2 (TMPRSS2), which normally activates the SARS-CoV-2 spike protein for plasma membrane fusion, SARS-CoV-2 infected angiotensin-converting enzyme 2 (ACE2)-expressing MEFs predominantly through dynamin- and actin-dependent endocytosis. In presence of TMPRSS2 the ancestral Wuhan-strain bypassed both dynamin-dependent and -independent endocytosis, and was less sensitive to endosome maturation inhibitors than the Omicron B1 and XBB variants, supporting the notion that the Omicron variants do not efficiently use TMPRSS2. Collectively, our study suggests that dynamin function at endocytic pits can be essential for infection with single-receptor viruses, while it is not essential but increases uptake and infection efficiency of multi-receptor viruses that otherwise rely on a functional actin network for infection.

Impact of Covid-19 Infection on Incidence and Exacerbation of Overactive Bladder Symptoms: A Systematic Review

Background The COVID-19 pandemic continues to be a major global health concern. A key factor is the presence of angiotensin-converting enzyme 2 (ACE2) receptors in various organs, including the lungs, heart, bladder, and testicles. These receptors allow the SARS-CoV-2 virus to enter cells, making these organs vulnerable to damage. This vulnerability may explain why some patients experience non-respiratory symptoms. Notably, overactive bladder (OAB) symptoms and a condition called COVID-associated cystitis (CAC) have been reported to negatively affect the quality of life of COVID-19 patients. A systematic review is needed to summarize the current understanding of these urological aspects of SARS-CoV-2 infection, considering both short- and long-term effects. Methods We conducted a systematic review in accordance with PRISMA guidelines to investigate urological complications of COVID-19, with a specific focus on OAB symptoms and CAC (characterized by frequent urination, urgency, and nocturia). We searched databases, including Medline (PubMed), Embase, and Scopus. Two reviewers independently screened the studies, and the quality of each study was assessed using the Newcastle-Ottawa Scale (NOS). Results Our search identified 343 articles published up to March 2024, of which 12 were included in this review. Many of the studies utilized scoring systems such as the International Prostate Symptom Score (IPSS) and OAB symptom scores. The evidence suggests that COVID-19 may trigger or worsen lower urinary tract symptoms, OAB, and cystitis in some patients, regardless of gender or age. However, these effects appear to be uncommon. Several studies reported an increase in IPSS scores, though it remains unclear whether this increase is temporary or long-lasting. A few studies found that symptoms resolved over several months. Conclusion This systematic review suggests that COVID-19 may affect the urinary system, leading to symptoms such as frequent urination, urgency, and nocturia. These symptoms can negatively impact the quality of life in COVID-19 patients.

Antibacterial Effects of Silica Nanoparticles Loading Nano-silver and Chlorhexidine in Root Canals Infected by Enterococcus faecalis.

The persistence of microbial infection can lead to endodontic failure. Enterococcus faecalis (E. faecalis) is acknowledged to be a closely associated bacterium. This study investigated the antimicrobial effects of mesoporous silica nanoparticles (nMS) carrying nano-silver and chlorhexidine (nMS-nAg-Chx) on E. faecalis. Analyses were conducted to assess the antimicrobial efficacy of nMS-nAg-Chx towards planktonic E. faecalis, including the zone of inhibition, minimal inhibitory concentration (MIC), and growth curves. The measurement of lactic acid, scanning electron microscopy (SEM), live-dead bacteria staining, and quantitative real-time PCR (qRT-PCR) were done to further investigate its anti-biofilm effect. Colony forming unit (CFU) and SEM were used to assess its efficacy in infected root canals. The growth of planktonic E. faecalis was suppressed with a MIC value of 25 μg/mL (P＜0.05). nMS-nAg-Chx concentration-dependently inhibited biofilm formation of E. faecalis with the reduction of lactic acid (P < 0.05), sparse biofilm structure, reduced percentage of viable bacteria (P < 0.05), and suppressed expression of ebpR, gelE, ace, and efa genes (P < 0.05). The seven-day sealing of nMS-nAg-Chx resulted in a notable reduction in bacterial counts compared to the saline control group in the E. faecalis infected root canals (P < 0.05). NMS-nAg-Chx effectively inhibits E. faecalis and removes its biofilm from infected human root canals. It may be used for endodontic treatments in the control of E. faecalis bacteria as an intracanal medication.

Oat-Protein-Based Diet Lowers Blood Pressure and Prevents Cardiac Remodeling and Dysfunction in Spontaneously Hypertensive Rats

Background/Objectives: Hypertension and its associated complications, such as cardiac remodeling and dysfunction, continue to impose a significant burden on global healthcare. Nutritional interventions have been recognized as playing a crucial role in addressing this devastating condition termed a ‘silent killer’. Plant-based proteins could potentially be utilized as a non-pharmacological strategy to combat hypertension and its related risk factors. In this study, we investigated the efficacy of an oat protein diet in managing hypertension and cardiac abnormalities. Methods: Four-week-old male spontaneously hypertensive rats (SHRs) and Wistar–Kyoto (WKY) rats were fed a regular diet with casein as a protein source or an oat-protein-based diet for 16 weeks. Twenty-week-old male SHRs showed high blood pressure (BP), cardiac remodeling, cardiac dysfunction, higher levels of markers of oxidative stress [malondialdehyde (MDA)] and inflammation [tumor necrosis factor-α (TNF-α)], as well as lower levels of a marker of vascular function (nitric oxide). Results: The oat protein diet was able to significantly lower high BP, prevent cardiac remodeling and dysfunction, improve the levels of nitric oxide, and reduce the levels of TNF-α. Oat protein, after in vitro gastrointestinal digestion, also exhibited angiotensin-converting enzyme inhibition and significantly higher antioxidant activity than casein when assessed with the 2,2-diphenyl-1-picrylhydrazyl and the iron-chelating assays in vitro. Conclusions: oat protein lowers BP and prevents cardiac remodeling and dysfunction partly via improving the levels of nitric oxide and TNF-αin SHRs. Its high antioxidant potential could contribute to the observed cardiovascular effects.

AI Promoted Virtual Screening, Structure-Based Hit Optimization, and Synthesis of Novel COVID-19 S-RBD Domain Inhibitors.

Coronavirus disease 2019 (COVID-19) is caused by a new, highly pathogenic severe-acute-respiratory syndrome coronavirus 2 (SARS-CoV-2) that infects human cells through its transmembrane spike (S) glycoprotein. The receptor-binding domain (RBD) of the S protein interacts with the angiotensin-converting enzyme II (ACE2) receptor of the host cells. Therefore, pharmacological targeting of this interaction might prevent infection or spread of the virus. Here, we performed a virtual screening to identify small molecules that block S-ACE2 interaction. Large compound libraries were filtered for drug-like properties, promiscuity and protein-protein interaction-targeting ability based on their ADME-Tox descriptors and also to exclude pan-assay interfering compounds. A properly designed AI-based virtual screening pipeline was applied to the remaining compounds, comprising approximately 10% of the starting data sets, searching for molecules that could bind to the RBD of the S protein. All molecules were sorted according to their screening score, grouped based on their structure and postfiltered for possible interaction patterns with the ACE2 receptor, yielding 31 hits. These hit molecules were further tested for their inhibitory effect on Spike RBD/ACE2 (19-615) interaction. Six compounds inhibited the S-ACE2 interaction in a dose-dependent manner while two of them also prevented infection of human cells from a pseudotyped virus whose entry is mediated by the S protein of SARS-CoV-2. Of the two compounds, the benzimidazole derivative CKP-22 protected Vero E6 cells from infection with SARS-CoV-2, as well. Subsequent, hit-to-lead optimization of CKP-22 was effected through the synthesis of 29 new derivatives of which compound CKP-25 suppressed the Spike RBD/ACE2 (19-615) interaction, reduced the cytopathic effect of SARS-CoV-2 in Vero E6 cells (IC50 = 3.5 μM) and reduced the viral load in cell culture supernatants. Early in vitro ADME-Tox studies showed that CKP-25 does not possess biodegradation or liver metabolism issues, while isozyme-specific CYP450 experiments revealed that CKP-25 was a weak inhibitor of the CYP450 system. Moreover, CKP-25 does not elicit mutagenic effect on Escherichia coli WP2 uvrA strain. Thus, CKP-25 is considered a lead compound against COVID-19 infection.

Inactivation of Pseudovirus Expressing the D614G Spike Protein Mutation using Nitric Oxide-Plasma Activated Water.

Variants of concern (VOCs) of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) exhibit high infectivity due to mutations, particularly in the spike protein, that facilitate enhanced binding of virus to human angiotensin-converting enzyme 2 (hACE2). The D614G mutation, situated in S1-domain, promotes the open conformation of spike protein, augmenting its interaction with hACE2. Activated water neutralizes pathogens by damaging biological molecules; however, its effect on mutated SARS-CoV-2 or VOCs requires further exploration. Here, the efficacy of nitric oxide (NOx)-plasma activated water (PAW) in inhibiting infections by SARS-CoV-2 pseudovirus expressing D614G-mutated spike protein is investigated, which serves as a model for mutated SARS-CoV-2. Results demonstrated high prevalence of D614G mutation in SARS-CoV-2 and its VOCs. NOx-PAW is non-toxic to cells at high concentration, inhibiting infection by 71%. Moreover, NOx-PAW induced structural changes in S1-domain of spike protein, reducing its binding affinity and lowering clathrin-mediated endocytosis-related gene expression. Additionally, in silico analysis revealed NOx species in NOx-PAW played key role in impairing S1-domain function of the mutated SARS-CoV-2 pseudovirus by interacting directly with it. Collectively, these findings reveal the potent inactivation ability of PAW against mutated SARS-CoV-2 and suggest its potential application in combating emerging variants of SARS-CoV-2 and other viral threats.

Guideline-directed antihypertensive medication use among young adult participants with uncontrolled hypertension at enrollment in the MyHEART study.

Hypertension is a major risk factor for heart disease, heart failure and stroke. Lifestyle changes are recommended as first-line treatment for management of high blood pressure for young adults, when 10-year atherosclerotic cardiovascular disease risk score is < 10%. If lifestyle changes alone do not control blood pressure, then providers have access to four classes of first-line blood pressure lowering agents to treat hypertension, when other contra-indications are not present. This is a cross-sectional, retrospective, secondary analysis performed of the MyHEART trial on study participants at enrollment to determine they were prescribed anti-hypertensive medication. Of those prescribed medications, we aimed to determine the frequency first-line medications including thiazide or thiazide-like diuretics, angiotensin converting-enzyme inhibitors, angiotensin receptor blockers, and calcium channel blockers were prescribed. This analysis categorized participants into four medication status categories: no antihypertensive medication, prescribed only first-line antihypertensives, prescribed only non-first-line antihypertensives, and prescribed a combination of first-line and non-first-line antihypertensives. Participant clinical and sociodemographic factors by medication use were evaluated. Linear regression models were fit to determine the association between antihypertensive medication and blood pressure. At enrollment, 157/311 (50.5%) participants were not on antihypertensives. Of the 154 on antihypertensives, reported use included monotherapy 97/154 (63.0%), combined therapy 57/154 (37.0%), only first-line antihypertensive 111/154 (72.0%), and only non-first-line antihypertensives 21/154 (13.6%), and combination of first-line and non-first-line antihypertensives 22/154 (14.2%). Antihypertension medication use varied based on age (p < 0.001), sex (p = 0.008), race (p = 0.001), body mass index (BMI) kg/m2 (p = 0.016), anxiety and/or depression (p = 0.048), diabetes (p = 0.007), and sodium intake (p = 0.042). Participants with only first-line medications had lower in-office systolic (-4.66 mmHg, CI -8.31 to -1.02, p = 0.013) and diastolic (-3.51 mmHg, CI -6.30 to -0.71, p = 0.015), and lower ambulatory diastolic (-2.12 mmHg, CI -4.15 to -0.09, p = 0.041) blood pressure than those without antihypertensives. Among MyHEART study participants, all of which had uncontrolled hypertension, 50.5% were not on an antihypertensive at enrollment. This finding supports the call to improve management of blood pressure earlier in life to potentially contribute to the reduction of long-term cardiovascular disease. Of the participants who were prescribed blood pressure medication, providers prescribed guideline-based antihypertensive therapy the majority of the time, however, this study indicates there may be an opportunity to increase the use of first-line, guideline-based antihypertensives, regardless of age, sex, or type of hypertension to lower long-term cardiovascular risk. https://www. gov Identifier: NCT03158051, registered 5-15-2017. IRB approval obtained: IRB # 2017 - 0372.

Elimination of olfactory sensory neurons by zinc sulfate inoculation prevents SARS-CoV-2 infection of the brain in K18-hACE2 transgenic mice.

Coronavirus disease-2019 (COVID-19), attributed to the severe acute respiratory syndrome-related coronavirus-2 (SARS-CoV-2), has posed global health challenges since it first emerged in 2019, and its impact continues to persist. The neurotropic nature of SARS-CoV-2 remains undisclosed, though researchers are proposing hypotheses on how the virus is transmitted to the central nervous system. One of the prevailing hypotheses is that SARS-CoV-2 travels through the olfactory nerve system via the olfactory epithelium (OE). Using a K18-human angiotensin converting-enzyme 2 (hACE2) transgenic mouse model with impaired olfactory sensory neurons (OSNs) induced by zinc sulfate, we examined the role of the olfactory nerve in the brain invasion by SARS-CoV-2. Mice lacking OSNs exhibited reduced levels of viral transmission to the brain, leading to significantly improved outcomes following SARS-CoV-2 infection. Moreover, a positive correlation was observed between viral persistence in the OE and brain infection. These results indicate that early inhibition of the olfactory nerve pathway effectively prevents viral invasion of the brain in K18-hACE2 mice. Our study underscores the significance of the olfactory nerve pathway in the transmission of SARS-CoV-2 to the brain.

Effect of SARS-CoV-2 Infection on Selected Parameters of the Apelinergic System in Repeat Blood Donors

Background: SARS-CoV-2 enters cells primarily by binding to the angiotensin-converting enzyme 2 (ACE2) receptor, thereby blocking its physiological functions, affecting the apelinergic system, and inhibiting the cleavage of its peptides. The appropriate concentration of peptides in the apelinergic system influences the maintenance of homeostasis and protects against cardiovascular diseases. In our research, we determined the level of selected parameters of the apelinergic system—apelin (AP), elabela (ELA), and the apelin receptor (APJ)—in repeat blood donors. Methods: We analyzed 120 serum samples obtained from 30 repeat donors (study group) within four time periods after a SARS-CoV-2 infection: &lt;60 days, 61–90 days, 91–120 days, and &gt;120 days. We compared the results from the study groups with those of the control group, which consisted of 30 serum samples collected from donors donating blood in the years 2018–2019. Results: We observed that the AP, ELA, and APJ concentrations in the control group are higher than in any period in the study group. In the study group, the concentrations of AP and ELA increased in subsequent study periods. AP and ELA concentrations were lower shortly after SARS-CoV-2 transfection and then slowly increased in subsequent periods. APJ concentrations, on the other hand, were lowest at 61–90 days after the infection, but the decrease, relative to their level in healthy subjects, was significant in every period studied. Conclusions: The results suggest that infection with SARS-CoV-2 causes changes in the parameters of the apelinergic system, both after a short period of time has passed since the onset of the SARS-CoV-2 infection, and even up to 4 months after the infection.

Abstract 4122247: Impact of serum chloride levels for ventricular arrhythmias in patients with heart failure undergoing cardiac resynchronization therapy

Background: As previous reports of ventricular arrhythmia (VA) as a prognostic marker of mortality in heart failure (HF) patients included those with implantable cardioverter defibrillator (ICD) and cardiac resynchronization therapy (CRT) devices, little is known about the risk factors of VA among patients undergoing cardiac resynchronization therapy (CRT), especially those without previous VA. The prognostic role of electrolyte abnormalities has been highlighted in HF patients. This study evaluated the impact of serum chloride levels for VA and all-cause death in patients with HF who underwent CRT without VA before CRT implantation. Methods: From the retrospective cohort of 397 HF patients (age 63+/-13, male 74%) who undergoing CRT implantation between 2010 and 2020, this study enrolled 175 patients (82% implanted CRT with a defibrillator) without a previous history of VA (sustained ventricular tachycardia (VT) of ventricular fibrillation (VF)). We studied the impact of serum chloride levels before CRT implantation. The primary endpoint was defined as VA, VA with ICD therapy and sudden cardiac death. The secondary endpoint was all-cause mortality. Results: During median follow-up of 35 months, 53 patients (30%) experienced VA. Patients with VA had a tendency toward high LVESV (172ml vs. 150ml, p&lt;0.09) and low left ventricular ejection fraction (26 vs. 29, p&lt;0.09), and significantly reduced rate of angiotensin converting enzyme inhibitors or angiotensin receptor blockers (30% vs. 70%, p&lt;0.05) at the time of CRT implantation. The rate of CRT responders was significantly higher in patients without VA than with VA (59% vs. 40%, p&lt;0.05). The cut-off value of chloride for VA was determined to be 102mEq/l using receiver operating characteristic analysis. The patients with chloride ≤102mEq/l had significantly higher rates of VA (41% vs. 25%, log-rank p&lt;0.05) and mortality (33% vs. 18%, log-rank p&lt;0.05) (Figure). By multivariate cox analysis, the level of chloride had an independent predictive value of VA in patients with CRT (hazard ratio [HR] 1.08 per 1mEq/l decrease, 95% confidence interval [CI] 1.00-1.16, p&lt;0.05) (Table). Conclusions: The chloride level had a predictive value for VA after CRT implantation in HF patients without previous VA. The management of electrolytes, including chloride, might be important for preventing fatal arrhythmia in patients with HF undergoing CRT implantation.

Abstract 4131412: Effects of Angiotensin Converting Enzyme Inhibition And Afterload Reduction in Single Ventricle Patients Prior To Fontan: Not What You Might Think

Introduction: Pts with single ventricle (SV) after bidirectional Glenn (BDG) are placed on angiotensin converting enzyme inhibitors (ACEI) to reduce afterload, improve ventricular performance and allow for decreased systemic to pulmonary collateral flow (APC). No large scale data exists to support ACEI use in BDG pts to accomplish this. We hypothesized that ACEI may not perform as predicted. Goals: To determine if ACEI use in BDG pts decreases APC and improves ventricular function. Methods: Single center retrospective study of all BDG pts who underwent cardiac magnetic resonance (CMR) from 2010 to present. Demographics, medication, ventricular function and flow data were collected. Pts included were &gt;1 year old and on ACEI for &gt;3 months prior to CMR. Wilcoxon and Fisher Exact tests were used. Significance P&lt;0.05 Results: Of the 320 BDG pts (median 2.8 years old) studied, 137 (43%) were treated with ACEI. See table for all data. There was no difference in age, sex, medications (except for ACEI) or body surface area (BSA) between both groups. APC as a % of aortic flow and APC/BSA were not different between those treated with ACEI compared to those who were not, whether right (RV) or left ventricle (LV) dominant. There was no correlation of APC with time on ACEI or ACEI dosage. Ejection fraction (EF) was lower and end-diastolic volume (EDV) was higher in BDG on ACEI for the entire group and RVs. Although LV EDV was higher with those on ACEI, LV EF was the same for those on ACEI and those who weren’t (graph). APC as a % of aortic flow and APC/BSA positively correlated with EDV (rho 0.25-0.34, P&lt;.001). There was no difference in atrioventricular valve regurgitation between those on ACEI and those who were not (median 5 vs 6% respectively). As a surrogate for cerebral blood flow (CBF), superior vena caval (SVC) flow was lower for those on ACEI compared to those who were not. Conclusion: BDG pts treated with ACEI did not demonstrate decreased APC or improved ventricular performance. There is a suggestion that ventricular performance may be worse on ACEI. CBF is decreased in BDG on ACEI using SVC flow as a surrogate. This suggests that ACEI use in SV pts at the BDG stage is not indicated and should be discouraged.

Abstract 4138024: Identifying the Medications with the Greatest Reduction in Mortality in Coronary Artery Dissection

Introduction: Coronary artery dissection (CD) is a potentially fatal complication, usually arising either spontaneously or iatrogenically. Therefore, identifying treatment methods that reduce mortality is of great importance. The American College of Cardiology recommendations stipulate that patients who have experienced CD may benefit from beta-blocker therapy, as beta-blockers have been shown to reduce the incidence of recurrent CD. Currently, there is little published data in support of beta-blockers providing mortality reduction for CD. Given the significant risk of mortality from CD, we aimed to test the veracity of these recommendations using a retrospective multicenter study as well as having a main objective of exploring means to lower mortality in CD. Methods: We conducted a retrospective, multicenter cohort that identified 2,104 patients diagnosed with CD. We extracted the data from across 16 West Florida hospitals in between the years of 2017-2022. Our patients were identified and stratified based on common lab values and cardioprotective medications they were taking. Chi-square and binary logistic regression was used to calculate odds ratios for mortality. The odds ratio estimate for mortality was our primary outcome with attention given to factors that were associated with an increased or decreased likelihood of mortality in patients with CD. Results: Patients on beta-blockers, angiotensin converting enzyme inhibitor (ACEi) or angiotensin receptor blocker (ARB) were the only variables that were statistically significant. Out of 2,104 patients, 1,562 patients were on beta-blockers (74.2%), and 921 were on an ACEi/ARB (43.8%). Our data revealed that the odds ratio for mortality in patients with CD while on beta blockers was 0.375 (p-value 0.0009, confidence interval 0.210-0.669), and while on ACEi/ARBs was 0.447 (p-value 0.0077, confidence interval 0.247-0.808). These results indicate a significant reduction in mortality in patients who are taking beta-blockers and/or ACEi/ARB medications after being diagnosed with CD. Conclusion: Our data can be viewed as supportive of the current recommendation for the primary use of beta-blockers in patients with spontaneous CD, as well as suggesting angiotensin receptor blockers and angiotensin-converting enzyme inhibitors as possible additional therapies

Abstract 4117180: Hyperkalemia-Related Hospitalization Associated with Short-Term vs. Long-Term Outpatient SZC Therapy Among RAASi Users: The GALVANIZE Outcome study

Introduction: Patients receiving renin-angiotensin-aldosterone system inhibitors (RAASi) are at increased risk of developing hyperkalemia (HK). Sodium zirconium cyclosilicate (SZC) is used to treat HK, but the impact of duration of SZC on healthcare resource utilization (HRU) in RAASi users is unknown. The GALVANIZE Outcome study compared HK-related HRU among RAASi users between long-term and short-term SZC users. Methods: Adults with ≥1 outpatient prescription for SZC (index date) and ≥1 RAASi prescription spanning the index date were identified from a large US insurance claims database (7/2018-12/2022) and were stratified based on duration of SZC use. Long-term SZC users (&gt;90 days) and short-term SZC users (≤30 days) were exactly and propensity score matched on key baseline characteristics. Rates of HK-related hospitalizations or emergency department (ED) visits, HK-related ED visits, and HK-related hospitalizations were compared during follow-up from index to the earliest of 6 months post-index, end of data availability, other potassium binder use, or re-initiation of SZC post-discontinuation. Results: Among 1,586 matched pairs, the mean age was 65.5 years, 41.0% of patients were female, and most patients had any stage chronic kidney disease (91.9%), hypertension (90.8%), and diabetes (73.4%). Also, 30.0% of patients had heart failure. The most used RAASi therapies at index were angiotensin-converting enzyme inhibitors (57.3%) and angiotensin-receptor blockers (56.3%). Patients with long-term SZC use had a 44% lower rate of HK-related hospitalizations or ED visits, a 41% lower rate of HK-related hospitalizations and a 52% lower rate of HK-related ED visits than patients with short-term SZC use during follow-up (all p&lt;0.01; Figure 1). Conclusions: Among patients with RAASi and SZC therapy use, long-term SZC use was associated with significantly lower rates of HK-related HRU compared to matched patients with short-term SZC use. Funding: AstraZeneca

DeepBP: Ensemble deep learning strategy for bioactive peptide prediction.

Bioactive peptides are important bioactive molecules composed of short-chain amino acids that play various crucial roles in the body, such as regulating physiological processes and promoting immune responses and antibacterial effects. Due to their significance, bioactive peptides have broad application potential in drug development, food science, and biotechnology. Among them, understanding their biological mechanisms will contribute to new ideas for drug discovery and disease treatment. This study employs generative adversarial capsule networks (CapsuleGAN), gated recurrent units (GRU), and convolutional neural networks (CNN) as base classifiers to achieve ensemble learning through voting methods, which not only obtains high-precision prediction results on the angiotensin-converting enzyme (ACE) inhibitory peptides dataset and the anticancer peptides (ACP) dataset but also demonstrates effective model performance. For this method, we first utilized the protein language model-evolutionary scale modeling (ESM-2)-to extract relevant features for the ACE inhibitory peptides and ACP datasets. Following feature extraction, we trained three deep learning models-CapsuleGAN, GRU, and CNN-while continuously adjusting the model parameters throughout the training process. Finally, during the voting stage, different weights were assigned to the models based on their prediction accuracy, allowing full utilization of the model's performance. Experimental results show that on the ACE inhibitory peptide dataset, the balanced accuracy is 0.926, the Matthews correlation coefficient (MCC) is 0.831, and the area under the curve is 0.966; on the ACP dataset, the accuracy (ACC) is 0.779, and the MCC is 0.558. The experimental results on both datasets are superior to existing methods, demonstrating the effectiveness of the experimental approach. In this study, CapsuleGAN, GRU, and CNN were successfully employed as base classifiers to implement ensemble learning, which not only achieved good results in the prediction of two datasets but also surpassed existing methods. The ability to predict peptides with strong ACE inhibitory activity and ACPs more accurately and quickly is significant, and this work provides valuable insights for predicting other functional peptides. The source code and dataset for this experiment are publicly available at https://github.com/Zhou-Jianren/bioactive-peptides .

Assessing functional properties of diet protein hydrolysate and oil from fish waste on canine immune parameters, cardiac biomarkers, and fecal microbiota

Locally produced fish hydrolysate and oil from the agrifood sector comprises a sustainable solution both to the problem of fish waste disposal and to the petfood sector with potential benefits for the animal’s health. This study evaluated the effects of the dietary replacement of mainly imported shrimp hydrolysate (5%) and salmon oil (3%; control diet) with locally produced fish hydrolysate (5%) and oil (3.2%) obtained from fish waste (experimental diet) on systemic inflammation markers, adipokines levels, cardiac function and fecal microbiota of adult dogs. Samples and measurements were taken from a feeding trial conducted according to a crossover design with two diets (control and experimental diets), six adult Beagle dogs per diet and two periods of 6 weeks each. The experimental diet, with higher docosahexaenoic (DHA) and eicosapentaenoic (EPA) acids contents, decreased plasmatic triglycerides and the activity of angiotensin converting enzyme, also tending to decrease total cholesterol. No effects of diet were observed on serum levels of the pro-inflammatory cytokines interleukin (IL)-1β, IL-8, and IL-12/IL-23 p40, and of the serum levels of the anti-inflammatory adipokine adiponectin. Blood pressure, heart rate and echocardiographic measurements were similar between diets with the only exception of left atrial to aorta diameter ratio that was higher in dogs fed the experimental diet, but without clinical relevance. Diet did not significantly affect fecal immunoglobulin A concentration. Regarding fecal microbiome, Megasphaera was the most abundant genus, followed by Bifidobacterium, Fusobacterium, and Prevotella, being the relative abundances of Fusobacterium and Ileibacterium genera positively affected by the experimental diet. Overall, results from the performed short term trial suggest that shrimp hydrolysate and salmon oil can be replaced by protein hydrolysate and oil from fish by-products without affecting systemic inflammatory markers, cardiac structure and function, but potentially benefiting bacterial genera associated with healthy microbiome. Considering the high DHA and EPA contents and the antioxidant properties of fish oil and hydrolysate, it would be worthwhile in the future to assess their long-term effects on inflammatory markers and their role in spontaneous canine cardiac diseases and to perform metabolomic and metagenomics analysis to elucidate the relevance of microbiota changes in the gut.

Persistently low tricuspid annular plane systolic excursion and its prognosis in Japanese hospitalized patients with heart failure with reduced ejection fraction.

In patients with heart failure (HF) with reduced left ventricular ejection fraction (HFrEF), low tricuspid annular plane systolic excursion (TAPSE) on echocardiography is associated with poor prognosis. The significance of TAPSE changes post-HF treatment among HFrEF patients remains unclear. We evaluated the factors associated with persistently low TAPSE and its prognostic impact in Japanese hospitalized patients with HFrEF. We prospectively examined 260 HFrEF patients from the prospective observational HIJ-HF III study of HF patients hospitalized at Tokyo Women's Medical University between 2015 and 2019. Persistently low TAPSE was defined as TAPSE < 17mm on both pre- and 1-year post-discharge echocardiography. The primary endpoint of the study was all-cause mortality or re-hospitalization due to HF. Prognosis and characteristics were compared between patients with and without persistently low TAPSE. Using characteristics and echocardiography data, factors associated with persistently low TAPSE were assessed using logistic regression analysis. We identified the prognostic impact of persistently low TAPSE in HFrEF patients using Cox proportional hazards models. Seventy-eight (30%) of the 260 patients had persistently low TAPSE. They had higher New York Heart Association functional class; lower baseline TAPSE and left ventricular ejection fraction; and fewer angiotensin-converting enzyme inhibitors or angiotensin receptor blockers. Significant factors associated with persistently low TAPSE included higher brain natriuretic peptide level at 1year after discharge, lower baseline levels of TAPSE and septal s'. Over a follow-up period of 32months (range 12-69months) after the 1-year echocardiography, the rate of the primary endpoint was significantly higher among patients with persistently low TAPSE than that among others (n = 31 (40%) vs. n = 39 (21%), respectively, log-rank p < 0.001). Cox multivariate analysis revealed that persistently low TAPSE was independently associated with adverse events (Hazard ratio, 1.975; 95% confidence interval 1.183-3.295; p = 0.009). Exactly 30% of hospitalized patients with HFrEF had low TAPSE both pre- and 1-year post-discharge. Persistently low TAPSE had independent predictive value of prognosis in these patients.

Populationwide Screening for Chronic Kidney Disease

Sodium-glucose cotransporter-2 (SGLT2) inhibitors have changed clinical management of chronic kidney disease (CKD) and made populationwide screening for CKD a viable strategy. Optimal age of screening initiation has yet to be evaluated. To compare the clinical benefits, costs, and cost-effectiveness of population-wide CKD screening at different initiation ages and screening frequencies. This cost-effectiveness study used a previously published decision-analytic Markov cohort model that simulated progression of CKD among US adults from age 35 years and older and was calibrated to population-level data from the National Health and Nutrition Examination Survey (NHANES). Effectiveness of SGLT2 inhibitors was derived from the Dapagliflozin and Prevention of Adverse Outcomes in Chronic Kidney Disease (DAPA-CKD) trial. Mortality, quality-of-life weights, and cost estimates were obtained from published cohort studies, randomized clinical trials, and US Centers for Medicare & Medicaid Services data. Analyses were performed from June 2023 through September 2024. One-time or periodic (every 10 or 5 years) screening for albuminuria, initiated at ages between 35 and 75 years, with and without addition of SGLT2 inhibitors to conventional CKD therapy (angiotensin-converting enzyme inhibitors/angiotensin receptor blockers). Cumulative incidence of kidney failure requiring kidney replacement therapy (KRT); life years, quality-adjusted life years (QALYs), lifetime health care costs (2024 US currency), and incremental cost-effectiveness ratios discounted at 3% annually. For those aged 35 years, starting screening at age 55 years, and continuing every 5 years through age 75 years, combined with SGLT2 inhibitors, decreased the cumulative incidence of kidney failure requiring KRT from 2.4% to 1.9%, increased life expectancy by 0.13 years, and cost $128 400 per QALY gained. Although initiation of screening every 5 years at age 35 or 45 years yielded greater gains in population-wide health benefits, these strategies cost more than $200 000 per additional QALY gained. The comparative values of starting screening at different ages were sensitive to the cost and effectiveness of SGLT2 inhibitors; if SGLT2 inhibitor prices drop due to patent expirations, screening at age 55 years continued to be cost-effective even if SGLT2 inhibitor effectiveness were 30% lower than in the base case. This study found that, based on conventional benchmarks for cost-effectiveness in medicine, initiating population-wide CKD screening with SGLT2 inhibitors at age 55 years would be cost-effective.

Glycemic properties of noodles produced from acha (Digitaria exilis), fig leaves (Ficus exasperata) and wheat (Triticum aestivum) and effect on biochemical and hemodynamic parameters in diabetic-hypertensive rats

In this study, the anti-diabetic and anti-hypertensive properties of wheat (Triticum aestivum) noodles was improved by inclusion of acha (Digitaria exilis) and fig (Ficus exasperata) leaf, which are plants established to have anti-hyperglycemic and anti-hypertensive properties respectively. The glycemic indices and the effects of the formulated noodles on fasting blood glucose levels, hemodynamic blood pressures and enzymes linked to type-2 diabetes (α-amylase and α-glucosidase) and hypertension [arginase and angiotensin 1 converting enzyme (ACE)] were assessed. The results revealed that inclusion of acha and fig leaf in the noodles caused significant reduction in glycemic indices, fasting blood glucose and systolic and diastolic blood pressure and modulated the activities of enzymes linked to type-2 diabetes and hypertension in the rats. Conclusively, the results revealed that wheat noodles fortified with acha flour and fig leaf could be suitable meal choice for diabetic and hypertensive patients.

A Novel Cell- and Virus-Free SARS-CoV-2 Neutralizing Antibody ELISA Based on Site-Specific Labeling Technology.

The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) led to the global spread of coronavirus disease 2019 (COVID-19), creating an urgent need for updated methods to evaluate immune responses to vaccines and therapeutic strategies. In this study, we introduce a novel cell-free, virus-free SARS-CoV-2 neutralizing antibody ELISA (NAb-ELISA), which is based on competitive inhibition of the receptor binding domain (RBD) of spike protein binding to the angiotensin-converting enzyme 2 (ACE2) receptor. In this method, site-specific biotinylated hACE2-Fc-Avi recombinant protein is immobilized onto a 96-well plate for capture, and the RBD-Fc-vHRP recombinant proteins serve as detection probes. Evaluation of sera from wild type (WT) or Delta RBD-immunized mice using the NAb-ELISA and pseudovirus neutralization tests (pVNTs) demonstrated strong correlations between assays (R2 = 0.91 and 0.90 for the WT and Delta groups, respectively). Additionally, the NAb-ELISA successfully detected cross-neutralizing activity in sera, though with slightly lower correlation to pVNT (R2 = 0.70-0.83). By employing NAb-ELISA instead of an indirect ELISA for hybridoma screening, five monoclonal antibodies (mAbs) with neutralizing activities against WT, Delta, and BA.2 pseudoviruses were obtained. This assay offers a straightforward, rapid, and safe approach to characterizing vaccine-induced antibody responses and mAb neutralization activity. Notably, the NAb-ELISA platform can be quickly adapted to assess neutralizing antibody responses against emerging mutant strains, addressing the rapid mutation of the virus.