ACE Levels Research Articles

Circulating ACE2 level and zinc/albumin ratio as potential biomarkers for a precision medicine approach to COVID-19.

Highly mutable influenza is successfully countered based on individual susceptibility and similar precision-like medicine approach should be effective against SARS-COV-2. Among predictive markers to bring precision medicine to COVID-19, circulating ACE2 has potential features being upregulated in both severe COVID-19 and predisposing comorbidities. Spike SARS-CoVs were shown to induce ADAM17-mediated shedding of enzymatic active ACE2, thus accounting for its increased activity that has also been suggested to induce positive feedback loops leading to COVID-19-like manifestations. For this reason, pre-existing ACE2 activity and inhibition of ACE2/ADAM17 zinc-metalloproteases through zinc chelating agents have been proposed to predict COVID-19 outcome before infection and to protect from COVID-19, respectively. Since most diagnostic laboratories are not equipped for enzymatic activity determination, other potential predictive markers of disease progression exploitable by diagnostic laboratories were explored. Concentrations of circulating albumin, zinc, ACE2 protein and its activity were investigated in healthy, diabetic (COVID-19-susceptible) and SARS-CoV-2-negative COVID-19 individuals. ACE2 both protein levels and activity significantly increased in COVID-19 and diabetic patients. Abnormal high levels of ACE2 characterised a subgroup (16-19%) of diabetics, while COVID-19 patients were characterised by significantly higher zinc/albumin ratios, pointing to a relative increase of albumin-unbound zinc species, such as free zinc ones. Data on circulating ACE2 levels are in line with the hypothesis that they can drive susceptibility to COVID-19 and elevated zinc/albumin ratios support the therapeutic use of zinc chelating inhibitors of ACE2/ADAM17 zinc-metalloproteases in a targeted therapy for COVID-19.

Role of the Renin-Angiotensin System in Long COVID's Cardiovascular Injuries.

The renin-angiotensin system (RAS) is one of the biggest challenges of cardiovascular medicine. The significance of the RAS in the chronic progression of SARS-CoV-2 infection and its consequences is one of the topics that are currently being mostly discussed. SARS-CoV-2 undermines the balance between beneficial and harmful RAS pathways. The level of soluble ACE2 and membrane-bound ACE2 are both upregulated by the endocytosis of the SARS-CoV-2/ACE2 complex and the tumor necrosis factor (TNF)-α-converting enzyme (ADAM17)-induced cleavage. Through the link between RAS and the processes of proliferation, the processes of fibrous remodelling of the myocardium are initiated from the acute phase of the disease, continuing into the long COVID stage. In the long term, RAS dysfunction may cause an impairment of its beneficial effects leading to thromboembolic processes and a reduction in perfusion of target organs. The main aspects of ACE2-a key pathogenic role in COVID-19 as well as the mechanisms of RAS involvement in COVID cardiovascular injuries are studied. Therapeutic directions that can be currently anticipated in relation to the various pathogenic pathways of progression of cardiovascular damage in patients with longCOVID have also been outlined.

Angiotensin I-Converting Enzyme Inhibitory Activity of Two Peptides Derived from In Vitro Digestion Products of Pork Sausage with Partial Substitution of NaCl by KCl.

This study aimed to identify angiotensin I-converting enzyme (ACE) from in vitro digestion products of pork sausage with partial substitution of NaCl by KCl (PSRK). Peptides from in vitro digestion products of PSRK were identified through liquid chromatography with tandem mass spectrometry analysis coupled with de novo sequencing. Subsequently, the ACE inhibitory peptides LIVGFPAYGH and IVGFPAYGH were screened based on PeptideRanker, in silico absorption, molecular docking, and the determination of ACE inhibitory activity. In addition, the ACE inhibitory peptides LIVGFPAYGH and IVGFPAYGH were mixed-type inhibitors; these peptides' ACE inhibitory activities were expressed as the 50% inhibitory concentration (IC50) values in vitro, which were 196.16 and 150.88 μM, respectively. After 2 h of incubation, LIVGFPAYGH and IVGFPAYGH could be transported through Caco-2 cell monolayers with paracellular passive diffusion. Furthermore, LIVGFPAYGH and IVGFPAYGH significantly increased the levels of ACE2 and nitric oxide while decreasing the levels of ACE, angiotensin II, and endothelin-1 in Ang I-treated human umbilical vein endothelial cells, indicating the ACE inhibitory effect of LIVGFPAYGH and IVGFPAYGH. In summary, LIVGFPAYGH and IVGFPAYGH from PSRK can be used as functional foods with antihypertensive activity.

ACE2 ve TMPRSS2 Genlerinin Farklı Hücre Hatlarındaki İfade Düzeyleri

Objective: ACE2 and TMPRSS2 proteins have received increased attention gained emphasis together with the pandemic COVID-19. These proteins have roles in respiratory and hypertension disorders as well as cardiovascular and renal diseases. The objective of this work was to examine the mRNA and protein levels of ACE2 and TMPRSS2 in cell lines derived from various tissue origins.
 Methods: After the growth of 14 different cell lines, protein and mRNA were isolated from the cell pellets. The amounts of mRNAs and proteins were then determined and quantified using RT-PCR and ELISA.
 Results: Findings showed that VERO, HEK293T, and VERO E6 cell lines significantly differed from others in the mRNA levels of both the ACE2 and TMPRSS2 genes. In protein levels obtained using ELISA, PNT1A cell line had the highest level of ACE2 protein expression, while for TMPRSS2, A549 had the highest level of protein expression. 
 Conclusion: It was showed in this study how the expressions of ACE2 and TMPRSS2 depend on the cell type. This may be an explanation for why virulence entrance differs in different types of tissues. It is thought that HEK293T cells with high levels of both genes may be a suitable option for studies at the RNA level by using these two genes. MCF7 may be a good option for studies at the protein level. Given the high levels of mRNA expression of both genes, it may be inferred that cells derived from the kidney were among those that were most susceptible to virus entry.

P-040 The Role of ACE2 and DNA fragmentation index on covid-19 post infected, healthy and infertile men

Abstract Study question to investigate the relationship between ACE2 and DNA fragmentation index and their impact on men’s reproductive system in covid-19 post-infected patients. Summary answer the drastic effect of covid-19 on the male reproductive system might be due to elevated DFI and ACE2 levels. What is known already In the last two years, many studies had been devoted to uncovering the depraved impact of covid-19 by measuring ACE2 concentrations or sperm DNA fragmentation but no study was applied to measure both parameters, this study performed on both parameters to figure out the role of angiotensin-converting enzyme (ACE2) with seminal fluid parameters and DNA fragmentation index in covid-19 post infected men and infertile patient in comparison to healthy fertile men. Study design, size, duration This prospective cohort study was conducted from November 2021 to June 2022. one hundred and nine men who participated in this study. Participants/materials, setting, methods The participants were divided into four groups: post-infected men with Covid-19 who recovered from Covid-19 infection with a recovery interval of no more than three months from the time of symptoms onset and all were fertile married men, healthy fertile men who haven't been infected with Covid-19 in the last three months obtain normal seminal fluid parameters as control, The third group is composed of oligospermia, while the fourth group is constituted of azoospermia volunteers Main results and the role of chance In this study, the result showed that the seminal fluid parameters for post-infected men with covid-19; were demotion in sperm concentration, total count, motility, and morphology with a p-value < 0.001 when compared with healthy men. DNA fragmentation in post-infected men was 4 times higher than in non-infected and it was found that post-infected men had 10 times more ACE2 concentrations in comparison with healthy men. The study showed that in oligospermia and azoospermia volunteers own a high ACE2 level compared to healthy volunteers and that the oligospermia subjects have a significantly high sperm DNA fragmentation index. In conclusion, the drastic effect of covid-19 on the male reproductive system might be due to elevated DNA fragmentation and ACE2 levels, and there is a statistically significant positive correlation between the DNA fragmentation index and ACE2 levels. Limitations, reasons for caution number of post-infected men with covid-19 Wider implications of the findings 1- A positive relationship exists between DNA fragmentation index percentage (DFI) and angiotensin-converting enzyme level (ACE 2). 2- Covid-19 infection alters sperm parameters such as concentrations, total sperm count, motility, and morphology. 3- The percentage of DNA fragmentation rose significantly after covid-19 infection. Trial registration number self funding

Value of Chest X-Ray and Chest Computed Tomography for Systemic Sarcoidosis Diagnosis in Undifferentiated Uveitis

ABSTRACT Background To evaluate the contribution of chest X-ray and chest CT for the diagnosis of sarcoid uveitis. Methods Retrospective study on consecutive patients with uveitis of unknown etiology, who underwent both chest X-ray and CT during uveitis diagnosis workup in a tertiary French university hospital. Results A total of 914 patients were included. Systemic sarcoidosis was identified in 23.1%. The probability of discordance between chest X-ray and CT increased with age at diagnosis (p < 0.001). In patients 30 years of age and younger, the probability of discordance was 5% or less, and 0.8% if the ACE level was normal. After 78.3 years of age, the probability of discordance was 20% or more. Conclusion We recommend not to perform CT in patients under 30 years of age with a normal chest X ray and ACE level, and suggest performing chest CT first in the elderly.

Functional Foods: A Promising Strategy for Restoring Gut Microbiota Diversity Impacted by SARS-CoV-2 Variants.

Natural herbs and functional foods contain bioactive molecules capable of augmenting the immune system and mediating anti-viral functions. Functional foods, such as prebiotics, probiotics, and dietary fibers, have been shown to have positive effects on gut microbiota diversity and immune function. The use of functional foods has been linked to enhanced immunity, regeneration, improved cognitive function, maintenance of gut microbiota, and significant improvement in overall health. The gut microbiota plays a critical role in maintaining overall health and immune function, and disruptions to its balance have been linked to various health problems. SARS-CoV-2 infection has been shown to affect gut microbiota diversity, and the emergence of variants poses new challenges to combat the virus. SARS-CoV-2 recognizes and infects human cells through ACE2 receptors prevalent in lung and gut epithelial cells. Humans are prone to SARS-CoV-2 infection because their respiratory and gastrointestinal tracts are rich in microbial diversity and contain high levels of ACE2 and TMPRSS2. This review article explores the potential use of functional foods in mitigating the impact of SARS-CoV-2 variants on gut microbiota diversity, and the potential use of functional foods as a strategy to combat these effects.

Single-cell analysis of adult human heart across healthy and cardiovascular disease patients reveals the cellular landscape underlying SARS-CoV-2 invasion of myocardial tissue through ACE2

BackgroundThe distribution of ACE2 and accessory proteases (ANAD17 and CTSL) in cardiovascular tissue and the host cell receptor binding of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are crucial to understanding the virus’s cell invasion, which may play a significant role in determining the viral tropism and its clinical manifestations.MethodsWe conducted a comprehensive analysis of the cell type-specific expression of ACE2, ADAM17, and CTSL in myocardial tissue from 10 patients using RNA sequencing. Our study included a meta-analysis of 2 heart single-cell RNA-sequencing studies with a total of 90,024 cells from 250 heart samples of 10 individuals. We used co-expression analysis to locate specific cell types that SARS-CoV-2 may invade.ResultsOur results revealed cell-type specific associations between male gender and the expression levels of ACE2, ADAM17, and CTSL, including pericytes and fibroblasts. AGT, CALM3, PCSK5, NRP1, and LMAN were identified as potential accessory proteases that might facilitate viral invasion. Enrichment analysis highlighted the extracellular matrix interaction pathway, adherent plaque pathway, vascular smooth muscle contraction inflammatory response, and oxidative stress as potential immune pathways involved in viral infection, providing potential molecular targets for therapeutic intervention. We also found specific high expression of IFITM3 and AGT in pericytes and differences in the IFN-II signaling pathway and PAR signaling pathway in fibroblasts from different cardiovascular comorbidities.ConclusionsOur data indicated possible high-risk groups for COVID-19 and provided emerging avenues for future investigations of its pathogenesis.Trial registration(Not applicable).

Chromatography-Spectroscopic Isolated MO11 (Moringa oleifera) and MS06 (Musa sapientum) Positively Immunomodulated ACE2 Levels in Blood, Kidney and Liver of Rats

Moringa oleifera (MO) and Musa sapientum (MS) are ethno-medicinal plants, while cadmium is a carcinogen. SARS-CoV-2 binds to ACE2 for host’s cell invasion and infection. This study evaluated the effects of MO11 (isolated from MO leaves) and MS06 (isolated from MS suckers) on ACE2 levels in cadmium chloride (CdCl)-induced toxicity in rats. Twentyfour adult male Wistar rats were randomly divided into six groups (n = 4). Group 1 was control. Groups 2–4 received single 1.5 mg/kg bodyweight of CdCl (i.p.) (Day 1). Groups 3 and 4 were post-treated with MO11 and MO11 + MS06 doses, respectively (Days 1–17). Groups 5 and 6 received only MO11 and olive oil (vehicle), respectively (Days 1–17). MO leaves and MS suckers were subjected to bioassay-guided fractionation and isolation procedures using chromatography and spectroscopic techniques. ACE2 levels (ELISA) were evaluated in blood, kidney and liver of rats. MO11 and MS06 were the most active antioxidant and antimicrobial compounds isolated from MO leaves and MS suckers, respectively. Liquid chromatography-mass spectrometry showed presence of therapeutic compounds and amino-acids in MO11 and MS06, which are part of 14 shared amino-acids used by SARS-CoV and SARS-CoV-2 for ACE2-binding. Post-treatments of CdCl-exposure with MO11 and MS06 showed decreased ACE2 levels in Group 4 (20.63 ± 4.33 ng/mL and 16.11 ± 0.49 ng/mL in blood and kidney samples, respectively), compared with Group 2 (39.39 ± 3.15 ng/mL and 85.39 ± 3.10 ng/mL in blood and kidney samples, respectively). In conclusion, MO11 and MS06 possess significant ethno-medicinal potentials, may compete with SARS-CoV-2 for ACE2 binding, and are recommended for evaluations as anti-SARS-CoV-2 agents.

COVID-19 and smoking: Considerations after two years

COVID-19 and smoking: Considerations after two years

The connection between systemic inflammation activity and the state of renin-angiotensin system with the course of cardiac arrhythmias

Abstract Funding Acknowledgements Type of funding sources: Public Institution(s). Main funding source(s): NSC "Institute of cardiology, clinical and regenerative medicine named after acad. Strazhesko" NAMS of Ukraine The purpose of the work is to compare the clinical course of paroxysmal and persistent forms of atrial fibrillation and flutter, depending on the systemic­ inflammation­ activity and the renin-angiotensin system state. Materials and methods The study involved 147 patients, including 103 people with rhythm­ disturbances, who werer divided into three main groups (with paroxysmal atrial fibrillation, persistant atrial fibrillation and persistant atrial flutter). Systemic inflammation activity was determined by the level of highlysensitive C-reactive protein in the serum. Renin-angiotensin activity ­ system was characterized by the angiotensin-converting factor level.The activity of ACE in blood plasma was determined by an express method using FAPGG as a substrate ("Sigma", USA), tris(oxymethyl)-aminoman, sodium chloride, EDTA ("Merck", Germany). Results The level of highlysensitive C-reactive protein in patients with Af and AFl was significantly elevated in 6. 7 times and was (8. 31±0. 42) against (1. 25±0. 01) mg/l is normal (p&amp;lt;0. 001). In the group with the persistent form of AF, the content of CRP was higher than in the group with paroxysmal form of AF, by 8. 5%, although the difference between the groups was unreliable. ACE activity in the blood plasma of the pts with arrhythmia was likely elevated, as the rateof ACE was 45. 10±2. 26 μkat/L and exceeded the nominal value by 180 %. 86 patients with rhythm disturbances, which was 83%, there was a significant increase of CRP level . Among patients with paroxysmal FP, it was 31 patients (88%), in the group with a persistent form of FP - 30 patients (78%) and in the presence of TP - 25 patients (83%, respectively). In contrast, in patients without rhythm disturbances who had hypertension and relatively healthy people, the number of patients with elevated levels is highly sensitive th CRP was only 9%, i.e. 4 patients. Comparing the data strong statistical significance was revealed. (p&amp;lt; 0.005). Conclusions An increased level of highlysensitiveC-reactive protein as a marker of systemic inflammation is observed in groups of patients with arrhythmias compared with patients with hypertension without rhythm disturbances, indicating a higher intensity of systemic inflammation. Serum ACE levels in 82 % of patients with arrhythmias were higher than normal compared to 27 % of patients without rhythm disturbances (p&amp;lt;0. 05), which may indicate hyperactivation of RAAS.

(323) Mechanistic Insights into a Rare Genetic Cause of COVID Orchitis

Abstract Introduction To date, biomedical efforts against the SARS-CoV-2 virus have mainly focused on mitigating spread and primary infection mortality. While significant strides have been made in community response and vaccine production, post-acute COVID-19 syndromes (Long COVID) are growing in prevalence and becoming an increasing burden on public health. Current evidence suggests COVID-19 can affect the testis and cases of orchitis (testicular pain) appear to present in up to 20% of men with COVID-19. With an aim of uncovering causal mechanisms of COVID-19 sequela outside of pulmonary symptomology, a recent study from our group has identified a nonsynonymous non-frameshift mutation in the NACAD protein associated with COVID-19 orchitis and decreased ACE2 serum levels. NACAD is thought to prevent inappropriate targeting of non-secreted peptides to the endoplasmic reticulum. Objective We hypothesized that a defect in NACAD function/processing will decrease ACE2 serum levels by disrupting extracellular transport and endoplasmic reticulum interaction. We explored whether intracellular levels of ACE2 as well as TMPRSS2 were altered and if cell membrane protein deposition was increased, which may result in more severe effects to SARS-CoV-2 infection. Methods We obtained testis biopsies from men undergoing sperm retrieval for infertility. After siRNA NACAD knockdown, we analyzed total ACE2 expression using Western blot and qPCR as well as immunofluorescence/H&amp;E staining for colocalization (with NACAD). Membrane-anchored ACE2 (mACE2) levels were quantified using immunostaining and flowcytometry while secreted ACE2 (sACE2) was analyzed by ELISA and Western blot. Results NACAD and ACE2 co-localize in both the Leydig and germ cells. When NACAD is knocked down, primary testes cells show 80% decreased mRNA and 50% total protein levels of ACE2. The secreted ACE2 was also reduced by nearly 50%. However, when we specifically measured the level of mACE2 on the cell surface, we found that the knockdown of NACAD did not lead to reduction. Conclusions NACAD likely specifically affects the level of sACE2, which is speculated to act as a protection factor by retaining the virus in serum and reducing cellular entry. While the exact reason for the differential effects of NACAD on mACE2 and sACE2 remain unclear, our data is consistent to other studies, which found a reduction of total ACE2 expression and sACE2 in patients is coupled with worse outcomes in COVID-19. We hope that establishing a relationship between NACAD and ACE2 expression will prove useful not only to elucidate why some men with COVID develop orchitis but, ultimately, why some men develop multiorgan failure with COVID infection whereas most do not. Disclosure No

Prioritization of Drug Targets for Neurodegenerative Diseases by Integrating Genetic and Proteomic Data From Brain and Blood

BackgroundNeurodegenerative diseases are among the most prevalent and devastating neurological disorders, with few effective prevention and treatment strategies. We aimed to integrate genetic and proteomic data to prioritize drug targets for neurodegenerative diseases. MethodsWe screened human proteomes through Mendelian randomization to identify causal mediators of Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, multiple sclerosis, frontotemporal dementia, and Lewy body dementia. For instruments, we used brain and blood protein quantitative trait loci identified from one genome-wide association study with 376 participants and another with 3301 participants, respectively. Causal associations were subsequently validated by sensitivity analyses and colocalization. The safety and druggability of identified targets were also evaluated. ResultsOur analyses showed targeting BIN1, GRN, and RET levels in blood as well as ACE, ICA1L, MAP1S, SLC20A2, and TOM1L2 levels in brain might reduce Alzheimer’s disease risk, while ICA1L, SLC20A2, and TOM1L2 were not recommended as prioritized drugs due to the identified potential side effects. Brain CD38, DGKQ, GPNMB, and SEC23IP were candidate targets for Parkinson’s disease. Among them, GPNMB was the most promising target for Parkinson’s disease with their causal relationship evidenced by studies on both brain and blood tissues. Interventions targeting FCRL3, LMAN2, and MAPK3 in blood and DHRS11, FAM120B, SHMT1, and TSFM in brain might affect multiple sclerosis risk. The risk of amyotrophic lateral sclerosis might be reduced by medications targeting DHRS11, PSMB3, SARM1, and SCFD1 in brain. ConclusionsOur study prioritized 22 proteins as targets for neurodegenerative diseases and provided preliminary evidence for drug development. Further studies are warranted to validate these targets.

Abstract #1406508: A Rare Case of Calcitriol-mediated Hypercalcemia in a Patient with Pneumocystis Jiroveci Pneumonia and the Role of Steroids in its Management

Calcitriol-mediated hypercalcemia is an uncommon complication of Pneumocystis jiroveci pneumonia (PJP). Some cases of PJP have been reported in patients with solid organ transplants and AIDS. Here we present a case of calcitriol-mediated hypercalcemia due to granulomatous Pneumocystis jiroveci pneumonia in a patient with a history of multiple sclerosis. A 45-year-old man with a medical history of multiple sclerosis, spondyloarthritis, smoking, and bipolar disorder was admitted for the evaluation of fever, fatigue, constipation, confusion, and weight loss of 10 Lbs over three months. His medications include rituximab, methotrexate, vitamin D-4000 IU, lithium, and calcium carbonate-600 mg. Laboratory evaluation showed elevated serum calcium (15 mg/dl) with a suppressed parathyroid hormone (9 pg/mL). Other pertinent findings include mildly elevated 25-hydroxyvitamin D (87 ng/ml), significantly elevated 1,25-dihydroxyvitamin D (381 pg/ml), and normal PTHrP (<2 pmol/L), vitamin A (0.82 mg/L) and TSH (1.31 mU/L). Extensive workup including AFB smear for tuberculosis, ACE level for sarcoidosis, SPEP for multiple myeloma, leukemia-lymphoma flow cytometry, and CT chest and PET-CT for malignancy were unremarkable. Treatment with large-volume intravenous fluids, calcitonin, and zoledronic acid followed by denosumab showed a transient reduction in serum calcium level to 10.5 mg/dl in 2 days and a rebound to a peak of 15.4 mg/dl in a week. Bronchoscopy with bronchoalveolar lavage which was done for persistent fever and new onset hypoxia revealed PJP confirmed by PCR. With this new finding, the diagnosis of PJP-induced calcitriol-mediated hypercalcemia was considered. He was then treated with Bactrim and prednisone 60 mg daily. Due to the severity of hypercalcemia, he required a hemodialysis session while waiting for the medications to take effect. His calcium normalized a week after the initiation of steroids and antibiotics and he was discharged on a slow prednisone taper. Clinic follow-up 4 weeks after discharge showed normal calcium, 1,25-dihydroxyvitamin D, and 25-hydroxyvitamin D levels. Discontinuation of prednisone was attempted but resulted in rebound elevation of 1,25-dihydroxyvitamin D requiring continuation of low-dose prednisone 5 mg. Our immunocompromised patient developed hypercalcemia induced by PJP infection due to granulomatous production of 1α-hydroxylase resulting in increased calcitriol formation. Steroids unsurprisingly appear to have the best impact on PJP-induced hypercalcemia because of their anti-inflammatory property. In this clinical entity, it inhibits inflammation and granuloma formation leading to a reduction in the release of 1α-hydroxylase and 1,25-dihydroxyvitamin D levels. It is essential to consider PJP as an etiology of hypercalcemia, especially in immunosuppressed patients. Steroid appears to be effective medical therapy in this setting. The timing and duration of treatment, and the efficacy of bisphosphonates and denosumab, require further studies.

Curious Cases of Painful Ophthalmoplegia (P1-9.012)

<h3>Objective:</h3> Importance of various investigations for diagnoses by exclusion and importance of starting treatment, which might be common for specific pathologies irrespective of different etiologies. (in a resource-limited environment) <h3>Background:</h3> There were two cases of adult women (Patient G and Patient V) residing in the same neighborhood. They presented with similar complaints of right-sided orbital pain, headache, and sixth cranial nerve palsy. <h3>Design/Methods:</h3> On individual evaluation, Patient G had prominent right-sided dull orbital pain for 3 weeks with right-sided headache and diplopia for 1 week. Examination showed ipsilateral sixth nerve paresis. Investigations like Complete Blood Count, Complete Metabolic Panel, Chest X-ray, ANA, CSF Analysis (including Adenosine deaminase), ESR, and CRP were negative. MRI showed a single hypointense extra-axial lesion in right anteromedial temporal region, anterolateral to ipsilateral cavernous sinus. The lesion appeared to partially encase right sixth cranial nerve and approaching right superior orbital fissure. Patient V had similar complaints of right-sided headache for 4 weeks and diplopia for 1 week. Examination showed complete paralysis of ipsilateral sixth nerve palsy. Similarly, investigations were negative (including serum ACE levels). MRI revealed a single extra-axial hypointense lesion along right medial temporal region along with partial invasion of ipsilateral cavernous sinus. Mild asymmetric thickening of right fifth and sixth cranial nerves was seen. <h3>Results:</h3> Investigations were necessary to rule out differential diagnoses like tuberculous meningoencephalitis, thyroid ophthalmopathy, and other infectious, autoimmune, or neoplastic etiologies. By diagnosis of exclusion, patient G was diagnosed with <b>Tolosa-Hunt syndrome</b>. Though biopsy results were unavailable for patient V, she was provisionally diagnosed with <b>Neurosarcoidosis</b>. Both were started on Corticosteroids. On respective follow-up visits, both patients reported significant improvement in their symptoms. This further solidified the respective diagnoses in these patients. <h3>Conclusions:</h3> In a resource-limited environment, response to treatment is as important as other investigations to diagnose rare diseases by exclusion. <b>Disclosure:</b> Mr. Shah has nothing to disclose.

P166 Parotid benign lymphoepithelial cyst in connective tissue disease: a rare case series and literature review

Abstract Background/Aims Benign lymphoepithelial lesions are swellings of salivary glands caused by a reactive lymphoid infiltrate with follicular hyperplasia, obliterating the acinar glands and causing proliferation and disruption of the ductal epithelium. These have been described in HIV, lymphoma and Sjögren's syndrome. We report a case series with mixed connective tissue disease/Sjögren's syndrome and reviewed the literature with similar presentations in order to highlight this unusual manifestation. Methods Index patient: A 25-year-old lady presented with a history of arthralgia, fatigue and Raynaud's 3 years ago and was diagnosed by rheumatologist to have MCTD with ANA showing U1RNP, Ro positive and histones antibody positive. She was managed with mycophenolate and improved. A year ago she developed parotitis which improved with antibiotics. She recently came to the outpatient department with painful swelling in the parotid region. Blood counts remain normal, ESR 36 mm/hr , CRP 6 mg/dl, ACE level, HIV screening normal. MRI parotid showed bilateral multiple lymphoepitheial cysts in the parotid glands. Opinion was obtained from oncology and the ENT surgeon. No evidence of malignancy was found. In view of this, consensus opinion among rheumatologists was to stop mycophenolate and give 2gm rituximab. We also reviewed 10 cases of benign lymphoepithelial cyst arising in the parotid region in acquired immunodeficiency syndrome negative patients. Cases were from 8 published articles plus the three cases of benign lymphoepithelial cyst in a mixed connective tissue disease/Sjögren's syndrome in AIDS negative patients reported in Table 1. Most of the patients had been treated surgically, however, immune therapy is sufficient. Results: Conclusion Benign lymphoepitheial cysts have been described in patients with sarcoidosis, HIV, cancer and CTD. Our patients had features of MCTD with Sjögren's syndrome and manifested with benign lymphoepithelial cysts in the parotids, which is very unusual. Parotid swelling in connective tissue disease requires evaluation and multidisciplinary input. Benign lymphoepithelial cysts will not require any surgery; once malignancy is excluded, they will require modification of immune therapy. Disclosure N. Palaniappan: None. S. Nallasivan: None.

Determining ACE-2 Level and Some Relevant Biochemical Parameters and studying the effect of Gender in Iraqi Diabetic Patients with Glomeruli and Renal Tubules Fibrosis as Early Prediction Marker

Diabetic kidney disease is an illness of the glomerulus that interferes with the glomerular filtration barrier (GFB), which is worked to enable kidney to selective purification of water and solutes in addition to limiting the movement of large macromolecules such as albumin. In the glomerular endothelium, mesangial cells, foot cells, and the brush border of the proximal tubules, ACE-2 is expressed and that the kidneys represent the highest-expressing region of this enzyme. Thus, the current study aimed to evaluate ACE-2 level in this case compared to healthy condition. The study Conducted with 120 male and female ranging in age (30-65) years old. Ninety patients with type 2 diabetes subdivided into three groups on the basis of ACR criteria including normoalbuminuria, microalbuminuria, macroalbuminuria (30 patients for each group) and 30 healthy people served as the control group, all visited Baghdad Teaching Hospital / Medical City and Al-Yarmouk Teaching Hospital, at the period between December 2021 and May 2022. ACE-2 levels were determined using the ELISA technique. Urea results showed significant differences between diabetic nephropathy in patient and control group in female cases but no significant differences in male patients with diabetic nephropathy and control group. Similar results were obtained in K ion. Also the results revealed significant differences in Na ion, ACR, eGFR, Urea, FBS, creatinine between diabetic nephropathy groups and healthy group.ACE-2 represents a good marker for early prediction in diabetic nephropathy case. ROC data analysis support the importance of ACE-2 in diagnosis of the studied disease case.

The effect of omega-3 supplements on the serum levels of ACE/ACE2 ratio as a potential key in cardiovascular disease: A randomized clinical trial in participants with vitamin D deficiency.

The aim of this randomized controlled clinical trial was to determine the effect of the omega-3 fatty acid supplementations 300 mg per day for 8 weeks on the serum levels of ACE/ACE2 ratio in Jordanian participants with vitamin D deficiency (VDD). The physical and clinical characteristic of individuals in both intervention and control randomized controlled clinical trial were measured and analyzed. The comparisons between the two groups and the changes in each group before and after taking omega-3 doses were studied through independent t test and paired t test, respectively. Possible factors that have a role in the changes were determined by multivariate stepwise regression. Follow-up period lasted 10 weeks. The sample consisted of 82 participants with VDD and a mean age of 37.85 ± 9.85 years. Omega-3 Supplements resulted in a significant decrease in serum ACE levels, ACE/ACE2 ratio and serum 25-hydroxy vitamin D (25OHD). While the change in serum ACE2 levels and serum triglycerides levels were insignificant. Also, a significant increase in serum LDL levels were observed. It is possible that taking high doses of omega-3 fatty acid supplementations have positive effects on the heart and circulatory system and could protect from COVID-19 or decrease disease severity, in connection with a decrease in the ACE/ACE 2 ratio. On the other hand, omega-3 supplement may have negative effect on cardiovascular system due to the significant increase in serum LDL levels.

Cutaneous susceptibility to SARS-CoV-2 infection according to the expression of viral entry factors in the skin

Introduction: Reports of dermatological manifestations in patients with COVID-19 suggest a possible cutaneous tropism of SARS-CoV-2; however, the capacity of this virus to infect the skin is unknown. Objective: To determine the susceptibility of the skin to SARS-CoV-2 infection based on the expression of viral entry factors ACE2 and TMPRSS2 in this organ. Method: A comprehensive analysis of human tissue gene expression databases was carried out looking for the presence of the ACE2 and TMPRSS2 genes in the skin. mRNA expression of these genes in skin-derived human cell lines was also assessed. Results: The analyses showed high co-expression of ACE2 and TMPRSS2 in the gastrointestinal tract and kidney, but not in the skin. Only the human immortalized keratinocyte HaCaT cell line expressed detectable levels of ACE2, and no cell line originating in the skin expressed TMPRSS2. Conclusions: Our results suggest that cutaneous manifestations in patients with COVID-19 cannot be directly attributed to the virus. It is possible that cutaneous blood vessels endothelial damage, as well as the effect of circulating inflammatory mediators produced in response to the virus, are the cause of skin involvement.

THE ACTIVITY OF ANGIOTENSIN-CONVERTING ENZYME 2 AND OTHER COMPONENTS OF THE RENIN-ANGIOTENSIN SYSTEM (RAAS) DURING PROGRESSIVE AGE IN HYPERTENSIVE MEN

Background: ACE2 and renin-angiotensin-aldosterone system (RAAS) are complex physiological systems that contribute to blood pressure control, whereas the activity and responsiveness of the ACE2 and RAAS be changed due to advanced age and hypertension. Aim: This study is to investigate the ACE2, Renin, Ang I, Ang II, and aldosterone levels in various ages of hypertensive men in order to provide insights into the effect of aging and high blood pressure on some body organs. Methods: It was compared ACE2, Renin, Ang I, Ang II, and aldosterone levels across 80 hypertensive men with ages between 30 years - 69 years. The persons were divided into four groups (20 men/group) according to their ages. The first group was 30-39 years, the second group was 40-49 years, the third group was 50-59 years, and the fourth group was 60-69. It used a one-way Analysis of Variance (ANOVA), followed by Duncan s test for the groups. Results: It was observed that the concentration of ACE2 decreased significantly in different groups with advanced age (p ≤ 0.01), except in the fourth sample group vs. the third group. The renin was reduced considerably in other groups with aging (p ≤ 0.01), except in the second sample group vs. the first group. The peptides Angiotensin I (Ang I) and Angiotensin II (Ang II) had opposite growth directions, decreasing and increasing, respectively, it was statistically significant (p ≤ 0.01). Also, aldosterone increased significantly in different groups with advanced age (p ≤ 0.01), not including the fourth group vs. the third group). Discussion: The physiological impact of these results is discussed according to the effects of hypertension and advanced age on all the studied parameters, particularly ACE2 deficiency and high levels of Ang II, pointed out an apparent dysfunction in blood pressure regulation during advanced age. Conclusions: ACE2 deficiency may be mediated by a series of dysfunctional events caused by the present changes in RAAS and other parameters in response to hypertension and advanced ages. Low renin levels and other following changes might cause an imbalance of Ang I and Ang II ratios. In addition, high levels of Ang II might reflect the unfavorable and negative changes that contributed to the disruption of many organs and systems during hypertension and advanced age.