Updates of precision medicine in type 2 diabetes

Diabetes mellitus is prevalent worldwide and affects 1 in 10 adults. Despite the successful development of glucose-lowering drugs, such as glucagon-like peptide-1 (GLP-1) receptor agonists and sodium-glucose cotransporter-2 inhibitors recently, the proportion of patients achieving satisfactory glucose control has not risen as expected. The heterogeneity of diabetes determines that a one-size-fits-all strategy is not suitable for people with diabetes. Diabetes is undoubtedly more heterogeneous than the conventional subclassification, such as type 1, type 2, monogenic and gestational diabetes. The recent progress in genetics and epigenetics of diabetes has gradually unveiled the mechanisms underlying the heterogeneity of diabetes, and cluster analysis has shown promising results in the substratification of type 2 diabetes, which accounts for 95% of diabetic patients. More recently, the rapid development of sophisticated glucose monitoring and artificial intelligence technologies further enabled comprehensive consideration of the complex individual genetic and clinical information and might ultimately realize a precision diagnosis and treatment in diabetics.

Autologous Conditioned Serum in Lumbar and Cervical Radiculopathy: A Systemic Review

Background: Intervertebral disc degeneration causing radiculopathy is driven by catabolic cytokines like IL-1β and TNFα. Autologous conditioned serum (ACS) was found to be rich in IL-1Ra (Interleukin-1 Receptor Antagonist), and thus, can impede disc degeneration. A systematic review of available literature was conducted to ascertain the potential therapeutic application of ACS in radiculopathy. Methods: Systematic literature reviews were conducted in PubMed, Scopus and Embase databases, up to September 2020. Randomised controlled trials (RCTs), prospective, retrospective studies and case series with lumbar or cervical radiculopathy and reporting use of ACS were included, with at least one of the outcome measures like VAS (Visual Analogue Scale) for pain, SF-12 (Short Form of Health Survey-12), Oswestry Disability Index, with a minimum follow up of 3 months. Animal studies, s, review articles and case reports were excluded. Results: A total of four studies, including 107 patients who received ACS were included based on the eligibility criteria. Two were RCTs and two were prospective non-comparative studies. Three studies evaluated the effect of IL-1Ra on lumbar radiculopathy and one on cervical radiculopathy. The mean age of patients in the studies ranged from 37.15 to 53.9. The dose of ACS used was 2-4 mL injection. In 1 RCT, methylprednisolone was used as control, in the other 5 mg and 10 mg triamcinolone was used. All studies reported a statistically significant reduction in pre-injection and post-injection VAS, there was also a significant difference as compared to 5 mg triamcinolone. Three studies reported significant improvement in ODI. Two studies reported statistically significant improvement in SF-12 scores post injection (p < 0.001). For cervical radiculopathy, Neck pain disability score showed a decrease of 73.76% from pre-injection to final follow up and Neck disability index showed a decrease of 74.47%. Conclusion: All of the four studies concluded that epidural perineural injection with ACS, reduced pain scores (VAS, NPDS) and improved functional scores (ODI, SF-12 and NPDS), as compared to placebo and other conventional therapeutic modalities like steroids, and analgesic-anaesthetic-steroid cocktail. Hence, ACS is a promising new therapeutic modality in both lumbar and cervical radiculopathy, and further studies can strengthen the present evidence regarding its efficacy and safety profile. © The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2022.

A New Registry for Genetic Diagnosis of Inborn Errors of Immunity within the Military Health System

Genotypic definition of monogenic inborn errors of immunity (IEIs) continues to accelerate with broader access to next generation sequencing, underscoring this aggregated group of disorders as a major health burden impacting both civilian and military populations. At an estimated prevalence of 1 in 1200 individuals, IEIs affect ~8,000 patients within the Military Health System (MHS). Despite access to targeted gene/exome panels at military treatment facilities, most affected patients never receive a definitive genetic diagnosis that would significantly improve clinical care. To address this gap, we established the first registry of IEI patients within the MHS with the goal of identifying known and novel pathogenic genetic defects to increase diagnosis rates and enhance clinical care. Using the registry, a research protocol was opened in July 2022. Since July we have enrolled 75 IEI patients encompassing a breadth of phenotypes including severe and recurrent infections, bone marrow failure, autoimmunity/autoinflammation, atopic disease, and malignancy. Enrolled patients provide blood and bone marrow samples for whole genome, ultra-deep targeted panel and comprehensive transcriptome sequencing, plus cryopreservation of peripheral blood mononuclear cells for future functional studies. We are also implementing and developing analytical methods for identifying and interrogating non-coding and structural variants. Suspected pathogenic variants are adjudicated by a clinical molecular geneticist using state-of-the-art analysis pipelines. These analyses subsequently inform in vitro experiments to validate causative mutations using cell reporter systems and primary patient cells. Clinical variant validation and return of genetic results are planned with genetic counseling provided. As a proof of principle, this integrated genetic evaluation pipeline revealed a novel, candidate TLR7 nonsense variant in two adolescent brothers who both endured critical COVID-19 pneumonia, requiring mechanical ventilation and extracorporeal membrane oxygenation. Our protocol is therefore poised to greatly enrich clinical genetics resources available in the MHS for IEI patients, contributing to better diagnosis rates, informed family counseling, and targeted treatments that collectively improve the health and readiness of the military community. Moreover, our efforts should yield new mechanistic insights on immune pathogenesis for a broad variety of known and novel IEIs.Copyright © 2023 Elsevier Inc.

Establishment and application of triple PCR method for CPV, CCoV and CRV

Canine parvovirus (CPV), canine coronavirus (CCoV) and canine rotavirus (CRV) are the three main causative viruses of diarrhea in dogs with similar clinical symptoms;thereby it is necessary to establish a high effective molecular detection method for differentiating the above pathogens. By optimizing the primer concentration and annealing temperature, a triple PCR method was established for simultaneous detection of CPV, CCoV and CRV, and then the specificity, sensitivity and repeatability of the method were tested. The results showed that the target fragments of CPV VP2 gene (253 bp), CCoV ORF-1b gene (379 bp) and CRV VP6 gene (852 bp) could be accurately amplified by the triple PCR method with high specificity, the detection limits of CPV, CCOV and CRV were 6.44x10-1 pg/L, 8.72x10-1 pg/L and 8.35x10-1 pg/L respectively with high sensitivity, and the method had good stability. Using this triple PCR method, 135 canine diarrhea fecal samples collected in Chengdu region from 2019 to 2020 were detected, and compared with those of single PCR method. The detection rates of CPV, CCoV and CRV were 16.30%, 20.74% and 4.44%, respectively, and the total infection rate was 51.11% (65/135) with 20.00% (13/65) co-infection rate. The detection results were consistent with three single PCR methods. In conclusion, CPV/CCoV/CRV triple PCR method successfully established in this paper can be applied as an effective molecular method to detection of related pathogens and to the epidemiological investigation.

Women and Alzheimer's disease

Approximately, two-thirds of individuals with Alzheimer's disease (AD) are women. Though previously attributed to differences in lifespan, accumulating evidence suggests that the reasons for the higher prevalence of AD in women are multifactorial and related to differences in risk factors, biomarkers, and neuropathology. Sex also contributes to significant disease heterogeneity, which has important implications for prevention and treatment. This chapter discusses the evidence for sex differences in AD, with an emphasis on disease presentation, biomarkers, pathophysiology, progression, and risk. Women tend to present later in the disease course and with different clinical features, progress faster, and are disproportionately affected by the APOE-ϵ4 risk allele and AD neuropathologic changes. Lifetime estrogen exposure, pregnancy, and menopause also affect a woman's risk for cognitive decline later in life. Despite such differences, women are dramatically underrepresented in pharmacologic randomized control trials, leading to significant gaps in knowledge regarding the most effect AD treatment strategies for women. Both researchers and providers need to be aware of sex differences in AD risk, presentation, and outcomes to develop sex-specific prevention and treatment strategies, as well as provide optimum healthcare to women as they age. © 2023 Elsevier Inc. All rights reserved.

Establishment of a duplex fluorescent quantitative PCR assay for detection of Porcine circovirus type 2 and type 3

To establish a method for simultaneous detection of porcine circovirus type 2 (PCV2) and porcine circovirus type 3 (PCV3), specific primers and TaqMan probes were designed after sequence alignment according to the specific sequences of PCV2 Cap gene and PCV3 Cap gene on GenBank. By optimizing the reaction conditions, a duplex fluorescence quantitative PCR detection method for simultaneous detection of porcine circovirus type 2 and 3 was established, and the specificity, sensitivity, and reproducibility were tested. Specificity test results showed that in addition to the positive test results for PCV2 and PCV3, tests for PRRSV, CSFV, PPV, PRV, PEDV, and TGEV were all negative with no cross-reaction, indicating its good specificity. Sensitivity test results showed that the minimum detection limit for detection of PCV2 and PCV3 can both reach 10 copies.L-1, indicating its high sensitivity. The coefficient of variation within and between groups of this method was less than 2%, indicating its good stability. A total of 181 pork and whole blood samples collected from Zhejiang Province were tested using the detection method established in this article and the standard common fluorescent PCR detection method. The results showed that the positive rate of PCV2 was 50.83% (92/181), the positive rate of PCV3 was 37.57% (68/181), and the co-infection rate of PCV2 and PCV3 was 12.15% (22/181). The above detection results of ordinary fluorescent PCR were 50.28% (91/181), 36.46% (66/181), and the co-infection rate was 11.60% (21/181). The coincidence rates of the two methods for PCV2 and PCV3 can reach 98.91% and 97.06%, and the coincidence rate for PCV2 and PCV3 mixed infection were 95.45%. In summary, the duplex fluorescence quantitative PCR detection method established in this experiment can distinguish PCV2 and PCV3 rapidly, which can be used for pathogen detection and epidemiological investigation.

Silent superbug killers in a river near you how factory farms contaminate public water courses on three continents

Water downstream from factory farms harbours an invisible threat to people's health which could eclipse the COVID-19 crisis. The threat? Antibiotic Resistance Genes (ARGs) which are driving antimicrobial resistance the world's superbug crisis - projected to kill up to 10 million people annually by 2050. This publication reports the presence of ARGs in animal waste discharged from industrial farms into public waterways or onto soil (or crops) in four countries. Gauge community impact and sentiment regarding the issue was also highlighted. The water and sediment from public water courses connected to effluent discharges from 6-10 pig farms were tested in each of four countries (Canada, Spain, Thailand and the USA).

Effects of Porcine deltacoronavirus on the transduction pathway of the innate immune signal

Porcine deltacoronavirus (PDCOV) is a new type of pig intestinal coronavirus, which targets pig small intestinal epithelial cells to cause severe enteritis. After infecting the host, PDCoV finishes its proliferation in the host cell by antagonism or escape the innate immune signaling transduction pathway. In order to understand the action mechanism of PDCOV 0n the congenital immune signal transduction pathways, this paper reviews the effects of PDCOV on RLR, Jak-STAT, MAPK and mitochondrial signaling pathway to clarify the relationship between PDCOV and host innate immune signaling transduction pathways in order to provide help for the prevention and treatment of PDCOV infection.

Effect of Drebrin protein on the proliferation of PEDV in Vero cells

Previous studies have revealed that developmental regulated brain protein (Drebrin) is involved in cell- to-cell communication, nerve transmission, tumor metastasis, spermatogenesis and other life activities, but there are few studies on viruses. The aim of the current research was therefore, to study the function of Drebrin and its effect on the proliferation of porcine epidemic diarrhea virus (PEDV). The Drebrin gene was cloned according to the Drebrin gene sequence (XM_008015438.2) of Chlorocebus sabaeus registered by GenBank, and the phylogenetic tree was constructed to analyze its homology. The results showed that the CDS region of Vero cells Drebrin gene was 2088 bp long, encoding 695 amino acids, and was relatively conserved and had high homology with all species. To investigate the effect of Drebrin on the proliferation of PEDV in Vero cells, the eukaryotic expression vector pcDNA3.1-Drebrin-Flag was constructed. After transfection of Vero cells with different concentrations of pcDNA3.1-Drebrin-Flag, cells were infected with PEDV. Our results showed that overexpression of Drebrin in Vero cells could significantly inhibit the intracellular PEDV mRNA level and N protein expression, reduce the extracellular virus titer and inhibit the proliferation of PEDV. Further study on the interaction between Drebrin and PEDV S proteins by laser confocal technique was also performed. The results showed that Drebrin and S protein were co-located in the cytoplasm, suggesting that the two proteins may interact with each other. This study demonstrated for the first time that Drebrin can inhibit PEDV proliferation in Vero cells, laying a foundation for further research in to Drebrin function and provides a valuable information for anti-PEDV research.

Development and application of a multiplex fluorescent quantitative RT-PCR for detection of Porcine epidemic diarrhea virus, porcine coronavirus and porcine acute diarrhea syndrome coronavirus

To develop a multiplex fluorescent quantitative RT-PCR for the detection of porcine epidemic diarrhea virus (PEDV), porcine deltacoronavirus (PDCoV) and swine acute diarrhea syndrome coronavirus (SADS-CoV), in this study, specific primers/probes were designed based on the conserved regions of M, M and N gene sequences of PEDV, PDCoV and SADS-CoV, respectively. After optimization of the reaction conditions, a multiplex fluorescent quantitative RT-PCR for PEDV, PDCoV and SADS-CoV was established. The results of specificity assay showed that the method was positive for detection of PEDV, PDCoV and SADS-CoV, and negative for detection of porcine transmissible gastroenteritis virus, porcine rotavirus, porcine reproductive and respiratory syndrome virus, porcine pseudorabies virus, porcine circovirus type 2, porcine parvovirus, classical swine fever virus and foot-and-mouth disease virus. The results of sensitivity assay showed that the detection limit of this method for PEDV, PDCoV, and SADS-CoV plasmids standard was 1.0x101 copies/L, and had a good linear relationship with their Ct values in the range of 101 copies/L to 106 copies/L. The results of repeatability assay showed that the coefficients of variation (CVs) of intra- and inter-assay reproducibility ranged from 0.33% to 2.53%, indicating good repeatability and stability. To evaluate the effects of the developed method, 100 clinical samples collected from different parts of Henan province were used for detection of these three viruses and compared with those of single RT-PCR and standard methods. The results of multiplex fluorescent quantitative RT-PCR showed that the positive rates of PEDV, PDCoV and SADS-CoV were 38% (38/100), 14% (14/100) and 5% (5/100), respectively. There was no mixed infection. The coincidence rate with the standard detection methods of PEDV and PDCoV was 100%, and the sensitivity was higher than that of single RT-PCR. In this study, a specific, sensitive and rapid multiplex fluorescent quantitative RTPCR method was established for the first time, which could be used for the differential detection of PEDV, PDCoV and SADS-CoV, and laid a foundation for the differential diagnosis and control of porcine diarrheal diseases.

Inherited deficiency of the OAS-RNase L pathway in children with MIS-C

Multisystem inflammatory syndrome in children (MIS-C) is a rare and severe condition that follows benign COVID-19. We report autosomal recessive deficiencies of OAS1, OAS2, or RNASEL in five unrelated children with MIS-C. The cytosolic dsRNA-sensing OAS1 and OAS2 generate 2'-5'-linked oligoadenylates (2-5A) that activate the ssRNA-degrading RNase L. Consistent with the absence of pneumonia in these patients, epithelial cells and fibroblasts defective for this pathway restricted SARS-CoV-2 normally. This contrasted with IFNAR1-deficient cells from patients prone to hypoxemic pneumonia without MIS-C. Monocytic cell lines and primary myeloid cells with OAS1, OAS2, or RNASEL deficiencies produce excessive amounts of inflammatory cytokines upon dsRNA or SARS-CoV- 2 stimulation. Exogenous 2-5A suppresses cytokine production in OAS1-but not RNase L- deficient cells. Cytokine production in RNase L-deficient cells is impaired by MDA5 or RIG-I deficiency and abolished by MAVS deficiency. Recessive OAS-RNase L deficiencies in these patients unleash the production of SARS-CoV-2-triggered, MAVS-mediated inflammatory cytokines by mononuclear phagocytes, thereby underlying MIS-C.Copyright © 2023 Elsevier Inc.

Characterization of varieties biofortified in iron and zinc and local varieties of common bean

Iron and zinc are important minerals in humans in sub Saharan Africa, whose deficiency is known as "hidden hunger" due to the lack of recognised symptoms in the early stages. Although iron deficiency is the most prevalent, zinc is also involved in inhibition of replication of viruses, including the corona virus (COVID-19). In North Kivu and South Kivu provinces where more than 50% of common bean is produced and consumed in Democratic Republic of Congo, 36% and 47% of preschool children are anemic due to iron deficiency. This paradox is mainly due to insufficiency of iron-rich foods. The aim of this study is to characterise 59 iron and zinc biofortified varieties together with six local varieties of common bean for a potential selection programme in Butembo town in the Democratic Republic of Congo. We focused on 15 qualitative and five quantitative parameters. The qualitative parameters were helpful to distinguish the different morphotypes and for cluster analysis. In addition to the descriptive statistics, the quantitative data were used for Pearson correlation and for principal component analysis, PCA. Qualitative parameters enabled grouping of the study genotypes into 14 morphotypes according to the aspect and colour of the seed coat, the colour around the hilum and the size of seeds. Clustering grouped the 65 genotypes into 12 clusters with the most similar genotypes grouped in the same cluster. Quantitative parameters showed that the study genotypes were dissimilar (P=0.00). A positive correlation was obtained between the days to flowering and the days to maturity (P<0.05) and between the number of pods per plant and the days to flowering. A strong correlation was found between the number of pods per plant and seeds per pod (P<0.01). In contrast, a negative correlation was observed between the 100 seed weight and the number of seeds per pod. The PCA represented on two perpendicular axes showed 64.1% of the total variance of which the 42.3% is explained by the first axis and 21.8% by the second axis. Overall, the study genotypes are morphologically and quantitatively different and thus can be used in a selection programme.

Sensory science and chronic diseases: Clinical implications and disease management

The textbook provides an overview of the sensory science field in the context of diseases such as obesity and Coronavirus disease 2019 (COVID-19). This book brings a summary of the state of the science in key areas and provides examples of translational science from using cellular and rodent models to human clinical trials and community health. The volume structure leads the reader through the physiology of taste and smell into how sensory testing for taste and smell is studied, basic mechanisms, various protocols that are used throughout the field along with the pros/cons of the current methods used. This resource is intended for classroom teaching, for novice researchers in sensory research as well as students and postdoctoral fellows. Example of courses are nutrition, basic nursing, interdisciplinary health courses, sensory perception (psychology), neuroscience, and medical courses, dentistry, food science and others. © This is a U.S. government work and not under copyright protection in the U.S.;foreign copyright protection may apply 2021. All rights reserved.

Telemedicine versus traditional consultations in clinical genetics: comparison of patient satisfaction rates

Background/Objectives: Telemedicine is a service delivery model in which credentialed specialists provide care remotely to clients. Data regarding patient satisfaction with telemedicine consultations in the field of clinical genetics, and specifically paediatric genetics is lacking. We aimed to compare patient satisfaction rates from telemedicine versus traditional, face-to-face genetics consultations during the COVID-19 pandemic. Method(s): A cross-sectional survey, sent to 1672 parents of minors (patients<18years), or adult patients, who received counselling through the Tel-Aviv Souraski Medical Center Genetics Institute between 1/1/2020-1/6/2020. Data were collected through REDCap and converted to Microsoft EXCEL Database Program( v16.0) and STATA(v14.1). Result(s): Full responses were collected from 457 patients (27.3%). Of them, 330 patients (72.2%) had face-to-face consultations, 80 (17.5%) were counselled through telemedicine, and 47 had both (10.3%). Satisfaction or high satisfaction were reported in 82.1% in the face-to-face consultation group, while 6.3% were unsatisfied or unsatisfied at all, compared with 82.5% and 11.2% in the telemedicine group, respectively. Differences were insignificant statistically between the two groups. Data were further stratified according to subspecialties. Of total consults, 58 (12.7%) were in paediatric genetics. None of the patients who received paediatric genetics counselling solely through telemedicine were unsatisfied. Seventy-six percent of all patients who were counselled through telemedicine would want to use telemedicine services in the future, while 18.7% are undecided, and 5% do not. Conclusion(s): Telemedicine consultations in the genetics clinic during the COVID-19 pandemic, and specifically in paediatric genetics, were associated with high satisfaction rates, non-inferior to traditional consultations satisfaction rates.

Using Multiple Models to Learn about COVID-19

Furthermore, multiple scientific disciplines, such as immunology, genetics, epidemiology, and microbiology, contribute to our understanding of the pandemic. [...]COVID-19 is a complex socioscientific issue (SSI), meaning that science concepts related to the virus have real-world implications for problems in society (Zeidler 2014). SSI-based teaching and learning creates opportunities for students to grapple with real-world problems relevant to their own lives and that require consideration and evaluation of multiple, sometimes competing, factors associated with the issue. The modeling activities were embedded in a broader unit designed for high school biology classes;descriptions of the full unit and the individual modeling activities can be accessed online at https:// epiclearning.web.unc.edu/covid (Sadler et al. 2021). For this aspect of the work, we chose a Netlogo computer simulation (www.jacobkelter.com/infection-model) that allowed students to identify patterns and make sense of underlying cause-and-effect relationships associated with social distancing-two of the NGSS crosscutting concepts.

Association of Alpha 1 Antitrypsin Deficiency with COVID-19 Mortality: Basis for Clinical Trials

Coronavirus disease 2019 (COVID-19) has emerged as a pandemic leading to unprecedented disruption of global health and economy. Countries with a large population of European/Hispanic ancestry have been found to have the highest COVID-19 related case fatality rates. This prompted us with an interesting question that whether host immune programming and host genetic modifiers might be responsible for the higher mortality rate in these ethnicities. Transmembrane protease serine 2 (TMPRSS2) is critical in priming the viral spike protein and the host ACE2 receptor before the virus enters into the host cell. Recent results from the COVID-19 Host Genetics Initiative identified ELF5 rs766826 as a protective factor to severe COVID-19 which decreases the expression of TMPRSS2. Moreover, multiple studies have experimentally demonstrated that alpha 1 antitrypsin (A1AT) (encoded by SERPINA1 gene) is an inhibitor of TMPRSS2 and provided support to the already approved therapy as a candidate for COVID-19. Interestingly A1AT deficiency is common among Europeans and Latinos. We have also analyzed the gnomAD dataset to show that Europeans and Latinos have a substantially higher carrier frequency of AlAT deficiency (~12%) compared to other large ethnicities. A1AT has the dual role of an antiviral and anti-inflammatory molecule for treating COVID-19. To date, eight clinical trials have been started to find out the effectiveness of A1AT in COVID-19. Low A1AT level in severe COVID-19 has also been found to be a poor prognostic marker. © The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Switzerland AG 2022.

Current Advances and Future Avenues in Endocrinology

Endocrinology is a dynamic science with numerous advances in the field of diagnosis, prognosis and management. Newer diagnostic modalities in the field have not only revolutionised the manner glycaemic status in diabetes is assessed but have provided newer metrics of evaluation, including ‘time in range' and the importance of glycaemic variability as an independent association with vascular complications. The focus on lifestyle management for weight and glycaemic optimisation is at an all-time high, especially in terms of time-restricted feeding, intermittent fasting and chrononutrition. Precision and personalised medicine is also foraying into mainstream endocrinology, with potential applications in diabetes mellitus as well as other disorders such as acromegaly and adrenal diseases (phaeochromocytoma/paraganglioma). Genetic testing for clinical and predictive endocrinology is another rapidly advancing domain with use in disease gene identification and discerning the genetic and molecular basis of various endocrine disorders. Avenues for the future implicate improved genetics, epigenetics and environmental factors to understand the intricacies of disease as well as design more effective therapeutic options. © The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2022.

Experiences and preferences of Healthcare Professionals using telegenetics in a UK Clinical Genetics centre during the COVID-19 pandemic

Background/Objectives: The changes and restrictions precipitated by the COVID-19 pandemic have led to innovation in Clinical Genetics service delivery worldwide. At the Guy's and St Thomas' (GSTT) Clinical Genetics Service, telegenetics was implemented at the beginning of the pandemic using the AttendAnywhere videoconferencing platform. We subsequently designed a qualitative study to capture experiences and preferences of Healthcare Professional's (HCP) using this service delivery model. Method(s): We conducted semi-structured interviews with seven HCPs working at the GSTT Clinical Genetics Service, including Genetic Counsellors, Clinical Geneticists and a Clinical Psychologist. Interview content was analysed using a thematic analysis approach. Result(s): We present HCPs' experiences of transitioning between virtual and in-person appointments and their appraisal of the technical and practical aspects of telegenetics. We also present themes that emerged about how HCPs' clinical practice has changed to adapt to telegenetics, as well as differences in both patients' and HCPs' attitudes towards virtual appointments when compared to in-person encounters. Future considerations will be shared regarding the suitability of telegenetics for Clinical Genetics appointments. Conclusion(s): Based on their experience at GSTT, HCPs interviewed would welcome the addition of telegenetics to the Clinical Genetics toolkit beyond the COVID-19 pandemic, and we will provide considerations for future guidelines.

SINGLE CELL MULTIOMIC PROFILING OF DISRUPTED REGULATORY NETWORKS IN COVID19 ASSOCIATED MULTISYSTEM INFLAMMATORY SYNDROME IN CHILDREN

BackgroundChildren infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) usually present minimal symptoms or are asymptomatic. Nevertheless, a subset of children 2-6 weeks after the initial SARS-CoV-2 infection develops a postinfectious SARS-CoV-2-related multisystem inflammatory syndrome in (MIS-C). Recently, transient expansion of TRBV11-2 T cell clonotypes in MIS-C was associated with signatures of inflammation and T cell activation, however, the underlying pathophysiology of MIS-C is not fully understood [1].ObjectivesThe purpose of our project is to characterize the complexity of cell populations and capture cellular heterogeneity to uncover the regulatory networks and interactions that are disrupted during MIS-C flare with simultaneous profiling of gene expression and open chromatin regions from the same nuclei.MethodsSamples of peripheral blood mononuclear cells from patients with MIS-C diagnosed at the University Children's Hospital, University Medical Center Ljubljana, were collected during the initial presentation before any treatment and at 6-12 months in remission. The primary aim is to identify which regulatory networks are driving inflammation in MIS-C flare, for which we are performing single cell Multiome ATAC + Gene Expression Sequencing. To enable simultaneous profiling of epigenomic landscape and gene expression from the same nuclei, we are using Chromium Next GEM Single Cell Multiome ATAC + Gene Expression kit from 10X Genomics.ResultsWe included 32 patients with MIS-C from whom we collected paired blood samples during the initial presentation before treatment and at 6-12 months in remission. In single cell multiomic experiment we included 10 patients with paired samples, with the most viable cell count prior cryopreservation. All samples that are included into multiomic single cell analysis have 75% - 99% viability prior cryopreservation. In the protocol the key is to remove remaining granulocytes causing high mitochondrial RNA burden and extensively optimize the dilution factor of lysis buffer and the length of cell lysis step in order to get intact nuclei with no significant blebbing. Afterward, the single cell ATAC libraries as well as single-cell gene expression libraries are constructed and sequenced. Data are undergoing pairwise analysis to compare the cell population heterogeneity, expression profile and open chromatin landscape in the time of the initial presentation of MIS-C and in the remission, with Cell ranger software as well as with R package scREG [2], and custom scripting. In the second step we will inspect if the resulting altered transcriptomic signature from single-cell experiment is present on larger cohort. In that regard, we will perform bulk transcriptomic profiling on all paired collected samples during the initial presentation of MIS-C before treatment and at 6-12 months in remission.ConclusionThe results of this project are expected to enlighten the underlying pathophysiology of MIS-C flare and thus support clinical decision on more targeted treatment. The identified disrupted networks during MIS-C flare could lead the way to establish an early diagnosis and improve long-term outcome, including prevention of myocardial and neuropsychological impairment. Moreover, a better understanding of the disrupted regulatory networks that are driving inflammation in MIS-C, could lead to new insights into diseases with similar clinical presentations as is Kawasaki Disease.References[1]Sacco, K., Castagnoli, R., Vakkilainen, S. et al. Immunopathological signatures in multisystem inflammatory syndrome in children and pediatric COVID-19. Nat Med 28, 1050–1062 (2022).[2]Duren, Z., Chang, F., Naqing, F. et al. Regulatory analysis of single cell multiome gene expression and chromatin accessibility data with scREG. Genome Biol 23, 114 (2022).AcknowledgementsThis research was supported by Slovenian research agency grant J3-3061 and Interreg ITA-SLO project Cattedra.Disclosure of InterestsNone Declared.

A Tale of Two Counties With the Same Telehealth Story: Experience with Cancer Genetics Testing During COVID-19 at a Community Hospital

Introduction: With the onset of the COVID-19 pandemic in 2020, the utilization of telemedicine now offered an alternative diagnostic and treatment resource to providers in many areas of medicine including oncology and cancer genetics. This care option paired with genetic testing labs' ability to send saliva-based DNA collection kits to patients, enabled our community hospital in Detroit to offer diagnostic testing without the patient coming to a healthcare setting for a host of reasons. Social determinants of health have been found to influence success with telehealth, and this study sought to analyze how successful telehealth cancer genetics care was throughout the Detroit Metro area. Methods: Patient demographics for in person visits six months before COVID were analyzed, and then compared with demographics of patients during the 2020-2021 pandemic period where visits were telehealth. Results: Pre-pandemic there were , 192 unique patients seen in person with the top three cities patients were from were Detroit (12.1%), Clinton Township (8.3%), and Saint Clair Shores (10.4%). During the pandemic, with telehealth as the major modality, the top three cities were Macomb (7.2%), Detroit (7%), and Clinton Township (7%). Detroit is in Wayne County, while St.Clair Shores and Clinton Township are in Macomb County. Per the US Census Bureau Macomb county has a median income of $64,641 and Wayne county has a median income of $49,359, and poverty level in Macomb county is 9.2% versus in Wayne the level is 20%. Conclusions: This paper outlines the challenges of initiating a telemedicine program in an urban community area and highlights the benefits of a concierge service in serving cancer patients who may have economic and historically poor perceived technologic abilities.