Comparison of HbA1c measurement in remotely prepared capillary versus standard venous blood samples

Background The COVID-19 pandemic has disrupted routine HbA1c testing. This has led to difficulties in monitoring glycaemic control and identifying people with suboptimal glycaemia. Delayed diagnosis of diabetes and suboptimal glycaemic control over extended periods can increase the risk of developing long-term complications of diabetes. The self-collection of capillary blood remotely (at home) for routine HbA1c testing can facilitate monitoring of glycaemic control whilst supporting virtual consultations. The aimof this study was to assess the clinical performance and user acceptance of capillary blood samples prepared remotely using the MiniCollect capillary blood collection device as an alternative to standard venous blood collection for HbA1c analysis. Methods Adult men and women with any type of diabetes were recruited. Following informed written consent, eligible participants provided a venous blood sample at their routine clinic appointment and subsequently prepared a capillary blood sample remotely. Participants also completed a bespoke usability questionnaire. Results Of 84 participants recruited, 62 capillary samples returned to the laboratory, with 41 having a paired venous sample for HbA1c analysis. HbA1c results using both collection methods demonstrated good agreement;Passing-Bablok Regression analysis, y=0 + 1x;R=0.986, Bland-Altman Difference Plot providing a mean difference of 0.3 mmol/mol. Conclusions Over half of participants found the MiniCollect device easy to use. The majority were in favour of the remote capillary blood collection service and would use it if routinely available. The remote self-collection of capillary blood for HbA1c is a convenient alternative for people with diabetes living and working in rural or urban settings ensuring optimal continuance of care.

What the Eye Cannot See: A Case of Persistent Altered Mental Status in a Patient With Cholestatic Liver Disease?

Introduction: Altered mental status (AMS) is a common symptom in patients with liver disease with a wide list of differential diagnoses. Knowledge of etiologies of AMS unique to patients with hepatic dysfunction is vital in order to help recognize, diagnose, and treat the underlying cause in a timely manner. Case Description/Methods: A 46-year-old man with a history of recent COVID infection was transferred to our hospital for further evaluation of acute liver injury and AMS. On arrival, his labs were notable for AST of 408 U/L, ALT of 620 U/L, ALP of 5942 U/L, TB of 11.0 mg/dL, and an INR of 1.1. His work-up included an MRCP that showed segmental biliary ductal dilation with associated restricted diffusion and peribiliary enhancement concerning for sclerosing cholangitis. ERCP revealed a 3cm biliary cast that was removed and noted diffuse rarefaction of ducts throughout the entire biliary tree. A liver biopsy revealed centrizonal cholestasis with portal-based bile ductular reaction and mild bile duct injury. Despite adequate treatment of suspected infection and hepatic encephalopathy, his AMS persisted. His basic metabolic panel (BMP) was notable for Na of 143 mEq/L. A send-out lipid panel that was obtained to work-up his dyslipidemia revealed a total cholesterol of 1018 mg/dL, triglycerides of 420mg/dL, and the presence of lipoprotein X. A venous blood gas (VBG) was obtained showing a Na of 157 mEq/L and serum osmolality was 322 mmol/kg, confirming true hypernatremia. He was slowly treated with hypotonic solutions with significant improvement in his mentation. On follow-up one year later, he has persistent cholestasis and is currently being considered for liver transplant. Discussion(s): The final diagnosis was COVID-related ischemic cholangitis and disappearing bile ducts with persistent cholangiopathy, presenting with severe cholestasis, accumulation of lipoprotein X, and pseudonormonatremia. When faced with severe cholestatic liver disease, clinicians should keep in mind the possibility of accumulation of lipoprotein X and its association with hyperviscosity and spurious electrolyte abnormalities. Clinicians should rely on obtaining blood gas analyses for accurate electrolyte measurement in such cholestatic patients as blood gas analyses utilize direct ion-sensitive electrodes (ISE) to measure electrolytes, whereas routine basic metabolic panels utilize indirect ISE that are liable to spurious results in the presence of hyperlipoproteinemia/lipoprotein X.

Predictive value of central venous-to-arterial carbon dioxide tension in patients with ARDS treated with veno - venous extracorporeal membrane oxygenation (ECMO)

Introduction: Central venous-to-arterial carbon dioxide tension ( PvaCO2) can be useful for monitoring adequacy of tissue perfusion in patients with ARDS supported with veno-venous Extracorporeal Membrane Oxygenation (VV-ECMO). However, in theory, the unavoidable mixing of venous blood with blood after the oxygenator can affect PvaCO2 values by increasing central venous oxygen saturation and substantially decreasing CO2 concentration. This study aimed to evaluate acute changes in PvaCO2 after VV-ECMO installation and determine its association with patient outcomes. Method(s): Retrospectively evaluated coronavirus disease 2019 (COVID-19) ARDS patients with at least one concurrent arterial and central venous blood gas analysis before and after VV-ECMO installation as standard care. The primary outcome was intensive care unit (ICU) mortality at 28 days. Result(s): 29 patients were enrolled in the study. All the patients had a 25 F drainage multistage femoral cannula and a 21 F internal jugular infusion cannula. The median distance between the central venous sampling point and the tip of the infusion cannula was 39 [23-73] mm. No statistically significant changes in PvaCO2frelative changes calculated. After were observed 24-48 h after VV-ECMO installation (5 [4-7] mmHg to 6.5 [5-8.2] mmHg, p = 0.12). Hemoglobin concentration decreased 24 to 48 h after VVECMO installation (10.7 [9.5-12.7] g/dl to 9.6 [8.8-11.6] g/dl, p < 0.01) but neither central venous (75 [70-81]% to 73 [67-78]%, p = 0.46) nor arterial oxygen saturation (95 [92-97]% to 95 [93-96]%, p = 0.81) changed significantly. Elevated PvaCO2 after VV-ECMO installation had a good predictive value for 28 day ICU mortality (calculated area under the ROC curve 0.81) (Fig. 1 veno-venous). Conclusion(s): VV-ECMO support appears to have little effect on the PvaCO2 calculation. PvaCO2 can be used to evaluate patients with ARDS supported with VV-ECMO, as persistently elevated values can be associated with poor outcomes.

Effect of N95 Filtering Facepiece Respirator on Venous Blood Carbon Dioxide Levels and Hemodynamic Changes in Health Care Workers.

Background: N95 filtering facepiece respirators (FFR) are used by health care workers for prevention of airborne infection, and its use has increased manifolds during COVID-19 pandemic. Prolonged use may result in carbon dioxide (CO2) accumulation, affect hemodynamics, and blood gas values. Although arterial blood gas values accurately measure the blood CO2 levels, venous blood gas values also show acceptable correlation. Aim: To evaluate the physiological impact of N95 FFRs on health care workers, including hemodynamic changes and venous blood levels of CO2 during a period of 6 h. Settings and Design: Prospective observational study in a tertiary care hospital. Methods: The study was conducted on 30 health care workers who performed routine duties while wearing N95 FFR. Venous blood gas values (CO2, pH, and bicarbonate) and vitals (respiratory rate, heart rate, blood pressure, and saturation) were noted at baseline, 2 (T2), and 6 h (T6) after wearing the mask. Discomfort level was also measured on a Visual Analogue Scale (VAS) of 1-10. Statistical Analysis: Repeated measures analysis was done using repeated measures ANOVA or Friedman's test. Group comparisons for continuously distributed data were made using independent sample "t" test or Wilcoxon test. Results and Conclusion: Hemodynamic and blood gas values did not change over time. The VAS for discomfort because of respirator use was 1.33 (1.42) at T2 and 2.77 (1.91) at T6. This was a significant increase in discomfort over time (P = 0.001). About 80% of participants experienced discomfort during this period. N95 FFR did not lead to significant alteration in hemodynamics or change in blood gas values after 6 h of continuous usage. However, discomfort significantly increased over time.

ANTI-SARS-CoV-2 SPIKE RECEPTOR BINDING DOMAIN (S-RBD) IgG ANTIBODIES FOLLOWING CoronaVac ADMINISTRATION: A LONGITUDINAL STUDY. ANTI-SARS-CoV-2 S-RBD IgG ANTIBODIES

Reports on antibody titers following CoronaVac administration are still scarce, particularly when it comes to the post-vaccination effectiveness of CoronaVac in the Indonesian population. The purpose of this study is to determine the efficacy of COVID-19 vaccination by comparing the IgG levels against the S1 subunit of SARS-CoV-2 RBD after the first and second vaccinations. The researchers collected venous blood samples from participants after they received the CoronaVac 600 SU/0.5 mL vaccine at two different intervals (14 days and 28 days). Blood was drawn twice (after the first and second vaccinations) and tested for antibodies (positive antibody detection value of 50 AU/mL). Paired data were analyzed by using either the Wilcoxon test (numerical) or the McNemar test (categorical). The median IgG1 levels in the 14-day interval between vaccine doses were 64.40 AU/mL and IgG2 levels were 886.10 AU/mL. Meanwhile, the median IgG1 level was 146.10, and IgG2 level was 688.00.AU/mL in the group with a 28-day interval between vaccine doses. After the first vaccination, 60.00 % of study subjects had positive IgG levels, which increased to 98.57% after the second vaccination. Following the full-dose vaccination, all participants had higher antibody levels, and considered significant. The effect was stronger in the group that received the vaccine at 14-day intervals. CoronaVac has also been shown to increase the prevalence of detectable antibody positivity in study participants.Copyright © 2023 Russian Association of Allergologists and Clinical Immunologists, St. Petersburg Regional Branch (SPb RAACI). All rights reserved.

Comparative Study of Serum Electrolytes in Covid-19 and Healthy Patients in Tertiary Care Center, Karimnagar

Introduction- Electrolyte balance of the body is maintained by renin angiotensin aldosterone system. Some previous studies suggested that COVID-19 is associated with gastrointestinal symptoms, such as diarrhea and vomiting. This may results in electrolyte disturbances in patients. Electrolytes in body like sodium (Na), potassium (K). Chloride (Cl) plays an important physiological role in maintaining acid base and water balance of cells of the body. Aims and objectives: Our study aimed to compare some electrolyte between covid 19 and non-covid patients retrospectively. Material(s) and Method(s): This retrospective study included total 57 males and 43 females in the age group of 28 to 65 years. The results were compared with 100 age and sex matched healthy controls. Estimation of serum electrolytes was done with the collected venous blood samples using the ion selective electrode technique in an electrolyte analyzer. Analysis was done using SPSS V 25 Software. Chi-square and t-test were used to see association and difference between two variable respectively. Result(s): We have found that covid 19 is associated with low levels of electrolytes like Na, K, Cl. Chloride levels in both the groups was not statistically significant. But Hyponatremia and Hypokalemia were observed in cases group with high statical Signficance. Conclusion(s): Study found that electrolytes deterioration in these patients play a critical role in patients management. Thus a monitoring of electrolyte is essential throughout their illness to manage covid patients to improve their quality of life.Copyright © 2020 Ubiquity Press. All rights reserved.

Blood haemoglobin and pulse oximetry oxygen saturation measurement error

Pulse oximetry utilises the differential absorption, by both oxy- and deoxyhaemoglobin, of a light signal passing through tissue to provide a measure of blood oxygen saturation. Although pulse oximetry devices are widely used to monitor patients in real time, and to provide an estimate of risk of deterioration, no studies exist on the association of haemoglobin levels and pulse oximetry measurement error. We examined the effect of different haemoglobin levels on pulse oximetry measurements in patients admitted to a large UK teaching hospital from 1 February 2020 to 31 December 2021 with a possible diagnosis of Covid-19 infection. Pulse oximetry and arterial blood gas oxygen saturations were compared. Two measures of blood haemoglobin levels were available;from a venous sample within 24 hours of the arterial blood gas sampling and directly from the arterial blood gas sample itself. Data were available from 1086 patients. Using the measurement of haemoglobin from the venous blood sample within 24 hours of the blood gas, there was an inverse linear association between haemoglobin and pulse oximetry measurement error of -0.06% per 1 g/L increase of haemoglobin (95% confidence intervals CI: -0.02 to -0.09). This equates to patients with a venous haemoglobin of 70g/L having a measurement error of +8.0% (95% CI: +5.9% to +10.0%) and those with a haemoglobin of 150g/L having a measurement error of +3.6% (95% CI: +2.2% to +4.9%). Similar associations were observed using arterial haemoglobin values. The association between haemoglobin and measurement error of oxygen saturation as determined by pulse oximetry is inverse and linear. It is relatively large in patients with anaemia and may affect clincial assessment.

Seroconversion in COVID-19 Infection and Comparison of Antibody Responses in Symptomatic Versus Asymptomatic Individuals

Coronavirus disease (COVID-19) is an infectious disease caused by a newly discovered coronavirus. Following infection, antibodies are formed against the spike (S) and nucleocapsid (N) proteins, which are the primary viral antigens of SARS-CoV-2. This study aims to determine the antibody response three weeks post-infection and its persistence. To study antibody responses in COVID-19-positive individuals and to compare the degree of antibody response in symptomatic and asymptomatic individuals. The persistence of the antibody response was also assessed. Adult patients (> 15 years of age) who were diagnosed as COVID-19-positive by RT-PCR, three weeks after swab positivity were tested for total antibody levels against COVID-19 antigens by electrochemiluminescence assay. Out of 226 individuals, 129 were symptomatic and 97 were asymptomatic. Among the 129 symptomatic individuals, 74 exhibited an antibody response, whereas in the asymptomatic individuals, only 10 exhibited an antibody response. The antibody response was found to be significant in symptomatic individuals compared to that in asymptomatic individuals (p < 0.05). All follow-up individuals were seropositive at the end of both 6 and 8 months. Antibodies against SARS-CoV-2 persist for 8 months following infection. Despite the waning of antibodies against the nucleocapsid antigen, there was no complete disappearance of antibodies.Copyright © 2023 The Author(s).

Another Cause of DKA in T2DM in COVID-19 Pandemic

Background: 47-year Emirati female, has history of T2DM since age of 39. Her overall diabetes poorly controlled with HbA1C of 8.6% (69 mmol/mol IFCC) on Empagliflozin 10 mg OD over the last 2 years well tolerated. NO micro- or macrovascular complications of her diabetes. No other significant medical history apart from hypertension she is taking Amlodipine 10 mg OD for it with good control. She has presented twice to the hospital 24 hours apart. 1st Visit to ER in our Hospital with fever epigastric pain discharged on Ciprofloxacin suspected gastroenteritis with PPI and sent home. Approximately, 24 hours later she presented again with same symptoms namely fever and epigastric pain but this time associated with diarrhea and nausea for last 20 hours. There was no shortness of Breath or cough. This time she has been admitted to isolating room giving suspicion of COVID-19. Vital signs as following: Temp 38.5 HR 105, BP 135/65 mmHg O2 Sats 96%. on RA. On examination, she was conscious, oriented to time place person. No signs of dehydration. abdomen soft non tender, Chest good air entry no added sound. Hear S1-S2 no murmurs. HRCT has been done at ER. HRCT shows wide spread area of multifocal ground glass opacification are seen in both lungs most of them shows peripheral sub-pleural distribution Around small size consolidation are seen within the ground glass opacification, CT findings are in favour of possibility of COVID-19. Result(s): Blood test as following On admission, FBC was normal, with Hb 13.2, WBC 8.0, Plt 388 cellX 10/ul, U/Es: S.NA 132, s.K 4.2 mmo/l, s. Creatinine normal (58 umol/L -NR 49-90 umol/L) LFTs, Amylase and lipase normal, LDH mild elevation 304 U/L (NR 81-234), Very low Phosphate 03 mmol/L (NR 0.87-1.45), D-Dimer 0.6 mg/L (NR 0.0- 0.5), Corrected Calcium normal, S. Ferritin was 242 ug/L (NR 8.00- 388.00 ug/L), Urinalysis Protein =1 and 4+ ketones, CRP was normal 1 mg/dl ( increased to 214 mg/ l 3rd day) before it goes done 41 mg/L on 7th day of admission. Giving the pandemic of COVID 19 and according to MOHAP Criteria for presenting symptoms. This lady underwent HRCT and COVID19 test by Nasal Swab. Meanwhile, Her Venous Blood gas shows sever metabolic acidosis pH 7.107, PCO2 12.90, PCO2 69.10 On RA, BC 8.9, BE -25.5. Blood sugar 13.2 mmol/L with Urinary Ketones of 4+. Patient has put on DKA Protocol according to our Hospital DKA protocol in addition Stopped her SGLT2 and start on Lantus as a basal. She has put on Scale C (which is the higher scale with infusion about 10 units per hour, for about 96 hours (i.e. 4 days till the blood sugar back to normal for Ketones to disappear, her acidosis didn't improve 1st 24 hours till we give her 1.26% of 500 ml of Sodium Bicarbonate over 6 hours. COVID 19 Test back after 72 hours with positive results. Once out of DKA Diabetes team has stopped her Lantus a stared-on Humalog mix 50% 25 unit TDS. Meanwhile, she has received the following medications waiting for COVID 19 test. Treated with Favipiravir 1600 mg BD for 1 day and 600 mg BD, Start Tazocin 4.5 (stopped after 3 days) Metronidazole, and with prophylactic dose of Clexane. The Hydroxychloroquine hasn't started as Prolong QTC has been notice). Discussion(s): This patient presentation with DKA is another example how COVID- 19 could be a reason for DKA, though SGLT2 could be another cause of her presentation, however the huge insulin requirement and unusual prolong DKA status even with sever acidosis is making COVID-19 more likely causing her presentation It. Conclusion(s): We report this case to highlight the fact DKA - and in fact sever resistant DKA need prolong treatment can happen to Patient with T2DM and COVID 19 positive, and special attention to be paid (with early referral to the diabetes team) if the patient already on SGLT2. And we recommend that to have low threshold to start investigation and treatment as early as possible, regardless the type of Diabetes these patient might have.

Investigating COVID-19 severity based on serum levels of vascular endothelial growth factor (VEGF)

Introduction: COVID-19 is an acute, sometimes severe, respiratory illness caused by a novel coronavirus SARSCoV-2. Aims and objectives: This study aimed to investigate the severity of COVID-19 based on serum levels of vascular endothelial growth factor. Method(s): This cross-sectional study was conducted on 86 patients with COVID-19. They diagnosed using the RTPCR test taken from the throat and nose. The patient's demographic information were extracted from patients' records. A 5-ml venous blood sample was taken from each patient. VEGF was measured with the ELISA method using the Hangzhou East biopharma VEGF ELIZA Kit. Result(s): The mean age of patients was 55.56+/-14.88 years old. Fifty percent were male. The most common clinical symptom in patients was shortness of breath (77.9%). The VEGF and CRP mean serum levels were 2877.07+/-104.77 ng/ml and 46.16+/-24.23 mm/hour, respectively. There was no significant relationship between VEGF levels and COVID-19 severity (P=0.55). The percentage of pulmonary involvement > 50 in the severe group (72.7%) was higher than that of the non-severe group (2.4%) (P=0.001). There was a significant relationship between COVID-19 severity and the levels of LDH, neutrophil/lymph ratio, and CRP. Regarding medications, Remdesivir was used more in the severe group (70.5%) than in the non-severe group (45.2%) (P=0.018). Conclusion(s): Although serum VEGF levels were higher in severe group than non-severe group, but the difference was not statistically significant. Therefore, these inflammatory markers can assist in the initial evaluation of COVID19. Physicians should monitor serum levels of LDH, neutrophil/lymph ratio, and CRP when admitting COVID-19 patients.

Examining the association between vaccine reactogenicity and antibody titer dynamics after the third dose of BNT162b2 vaccine using a mixed-effects model

Background: To mitigate the COVID-19 pandemic, many countries have recommended the use of booster vaccinations. The relationship between the degree of adverse vaccine reactions and elevated antibody titers is of interest;however, no studies have investigated the temporal changes in antibody titers based on repeated measurements after a third dose of the BNT162b2 vaccine. Methods: This prospective longitudinal cohort study was conducted with 62 healthcare workers who received a third dose of the BNT162b2 at Okayama University Hospital, Japan. Venous blood draw and fingertip whole blood test sample collection were conducted at the early (3–13 days) and 1-month time points;only FWT sample collection was conducted at the 2-month time point. Information on adverse reactions within 1 week after vaccination was also obtained. The association between fever of 37.5 °C or higher and antibody titers after the third dose of BNT162b2 was examined using a mixed-effects model and Poisson regression with robust variance. Results: A trend toward higher antibody titers in the early period after vaccination was observed in the febrile individuals, but the differences were not significant at 1 and 2 months post-vaccination (the partial regression coefficient for fever was 8094.3 [-1910.2, 18,098.8] at 1 month after vaccination, and 1764.1 [-4133.9, 7662.1] at 2 months after vaccination in the adjusted models). Conclusion: The findings suggest that the presence of fever after the third vaccine does not predict a sustained elevation in serum antibody titers. © 2022 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases

Correlation of Serum and Fingerstick Blood Collection for Monitoring Sars-Cov-2 Spike and Nucleocapsid Antibodies

Background: The rapid spread and recurrent infections of SARS-CoV-2 has led to increased use and availability of at-home antigen testing, but widespread testing of antibodies against spike and nucleocapsid to monitor vaccine-induced immunity and exposure to the virus is lacking. Most serological tests require a serum sample from a venous blood draw, increasing risk of exposure to COVID-19 and limiting availability and scalability of testing for many patients. This is especially the case for individuals who are immunocompromised, such as those with inflammatory bowel diseases (IBD), which are frequently on medications that might alter their immune response and impact vulnerability to SARS-CoV-2. Use of easily acquired and stably stored dried fingerstick blood serves a promising specimen source for at-home, remote testing for SARS-CoV-2 antibodies. Aim(s): Validate the use of fingerstick blood (dried and then eluted) versus serum as a specimen for the measurement of quantitative spike and nucleocapsid antibodies to SARS-CoV-2 in a diverse cohort of healthy and immunocompromised patients. Method(s): Patients were consented and enrolled into the Pediatric Gastrointestinal Tissue, Stool, Saliva and Blood Registry prior to having an endoscopic procedure. Five mL of blood was obtained by venipuncture and 10 muL of fingerstick blood was collected and dried on a Neoteryx Mitra device. Blood was eluted from the Neoteryx Mitra samples in 200 muL of dilution buffer (1% BSA, 0.05% Tween-20, 140 mM NaCl, 50 mM Tris (pH 8.0), 0.025% sodium azide) and placed on an orbital shaker at a speed of 500 RPM for 3 h. Paired serum and fingerstick blood eluate specimens were run on the quantitative Roche Elecsys SARS-CoV-2 spike antibody assay and the qualitative Roche Elecsys SARS-CoV-2 nucleocapsid antibody assay. Linear regression were performed on each assay with exclusion of values that were above the upper limit of detection of the assay. Result(s): We observed an excellent correlation in both SARS-CoV-2 antibody assays when comparing fingerstick blood eluates and serum. The linear regression for the nucleocapsid antibody assay had a slope of 15.5, intercept of 4.05, and R2 of 0.92, indicating that a Neoteryx value of 1.00 COI (cut-off index) equates to a serum value of 19.6 COI. The linear regression for the spike assay had a slope of 13.6, intercept of 953, and R2 of 0.95, indicating that a value of 1,000 U/mL from a fingerstick sample equates to a serum value of 14,544 U/mL. Conclusion(s): These data demonstrate that fingerstick blood collected on Neoteryx Mitra devices can be used as a specimen source in Roche Elecsys SARS-CoV-2 antibody assays to calculate the serum levels of spike and nucleocapsid antibodies. This can serve as a platform to remotely and reliably monitor the durability of antibody responses to natural infection with and immunization against SARS-CoV-2 in patients. Chart comparisons of nucleocapsid (top) and spike (bottom) protein antibody levels detected via remote fingerstick collection (Neoteryx, x-axis) and venous blood serum (y-axis).

HEMOSTATIC ACTIVATION MARKERS AND SEROLOGICAL RESPONSE IN SUBJECTS RECEIVING ANTICOVID- 19 VACCINATION

Background: SARS-COV2 infection is associated with inflammation, hypercoagulability and endothelial damage. Anti-SARS-COV2 vaccines have radically changed the course of the pandemic, however, reports on rare thrombotic events raise concern in the scientific community and the general population. Aims: In a prospectively enrolled cohort of adult subjects undergoing mRNA or adenovirus vector vaccination, we wanted to longitudinally evaluate the changes in levels of hemostatic biomarkers (i.e. activation of blood coagulation and perturbance of endothelium and fibrinolysis), together with the serological response, and occurrence of manifest thrombotic complications. Methods: Peripheral venous blood samples were collected at enrollment (day 0, D0) before the 1st vaccine dose, and on 15 (D15), 60 (D60), 90 (D90) and 180 (D180) days after the 1st dose. At each time point, hemostatic markers (i.e., fibrinogen, D-dimer, FVIII, von Willebrand Factor [vWF] antigen and activity, F1+2, thrombomodulin, protein C, protein S, FXIII, tPA, and PAI-1), and anti-Spike receptor-binding-domain protein (anti-S/RBD) IgG were measured. Follow up is currently continuing. Results: Fifty-three subjects (57% males) with a median age of 50 years (range 23-86) were enrolled into the study and followed-up for 6 months: 36 (68%) received BNT162b2, 6 (11%) mRNA-1273, and 8 (15%) ChAdOx1 nCoV-19 vaccines, in 2 doses over 21, 30 and 77 days, respectively;while 3 (6%) subjects received Ad26.COV2.S as single shot. Twenty individuals (38%) reported previous history of COVID-19, with a mean time from infection to vaccination of 10 months (4-18);only 1 required Hospitalization. Nine subjects presented cardiovascular risk factors and 4 a prior, non-active, cancer;3 were on anticoagulation for atrial fibrillation. The evaluation of the hemostatic biomarkers at the different time points showed variations in some of the parameters evaluated, with median values remaining within normal range levels. Specifically, compared to baseline, we observed a significant increase in thrombomodulin at D90 (p=0.001) and D180 (p=0.03), in parallel to a significant decrease in fibrinogen (D60), vWFAg (D60 and D180), FVIII (D60, D90 and D180), and TPA (D60 and D90) levels. The reduction of these biomarkers was particularly evident in individuals with a history of COVID-19. Of interest, this group of subjects was also characterized by significantly lower levels of PAI-1 both at baseline (7.18 ng/mL vs 17.53 ng/mL;p<0.0001), and at other time points (p<0.0001), and by an increase in F1+2 at D90 (p=0.02). The association between lower baseline PAI-1 levels with history of COVID-19 was confirmed by linear regression analysis (B= -10.351, p=0.013), and was independent by the time of infection resolution. Notably, no differences were observed in the hemostatic biomarkers according to vaccine types. All subjects positively responded to vaccination with a significant increase in anti-S/RBD IgG from baseline (D0) to each time point, especially COVID-19 subjects (D15, D60, and D90:p<0.0001;D180:p=0.031). No thrombotic or cardiovascular complications occurred during follow-up. Summary/Conclusion: No hypercoagulable state elicited by COVID-19 vaccination was observed, contrarily we detected an overall persistent reduction of coagulation activation over time. Subjects with previous SARS-COV2 infection had persistently low levels of PAI-1, supporting enhanced fibrinolysis activation. Compared with recent studies, our results provide a longer observation follow-up with all vaccine types and reassure on the safety of anti- SARS-COV2 vaccination.

ANOTHER COMPLICATION OF COVID-19?: A CASE OF RHINOORBITAL-CEREBRAL MUCORMYCOSIS

CASE: A 44-year-old male with past medical history of type II insulindependent diabetes mellitus (DM) and end stage liver disease (ESLD) due to alcohol use and nonalcoholic fatty liver disease (NAFLD) presented with one week of left-sided retroorbital headache and diplopia. Two weeks prior, the patient tested positive for COVID-19 and initially his severe headache was attributed to this diagnosis. On hospital presentation the patient was found to have ophthalmoplegia, ptosis and diminished sensation in the CN V1 distribution on the left. The patient was in diabetic ketoacidosis (DKA) with glucose of 686, venous blood gas of 7.32/29/15 and serum anion gap of 17. Contrasted orbital and maxillofacial CT showed complete opacification of the left sphenoid sinus and CT angiography/venography of the head were negative for venous sinus thrombosis. MRI of the brain showed left optic nerve ischemia and left frontal lobe cerebritis without abscess. Bedside nasal endoscopy with ENT showed purulent, fuzzy white debris bilaterally concerning for fungal sinusitis. He was taken urgently to the operating room and was found to have angioinvasive fungal sinusitis with cultures growing Lichthemia corymbifera, a fungus in the Mucor family. In addition to treatment with IV insulin and fluids for DKA, the patient was given amphotericin B and posaconazole;however, surgical intervention was deemed too high risk and futile in the setting of patient's comorbidities. IMPACT/DISCUSSION: Mucormycosis is a fungal infection that typically involves the sinuses, orbits and the central nervous system (CNS). Infection of the sinuses manifests with fever, sinus congestion/pain and headache, but can rapidly progress to involve the orbits, leading to vision changes, and the CNS, leading to encephalopathy. Other structures that can be involved include the cavernous sinus, leading to palsies of cranial nerves III-VI. Known risk factors for mucormycosis include DM, especially in patients with DKA, glucocorticoid treatment, immunosuppression and deferoxamine use. Urgent histopathologic diagnosis, initiation of intravenous antifungal agents (amphotericin B) and surgical intervention with ENT, ideally prior to extension beyond the sinuses, are fundamental to decreasing mortality, which is as high as 62%. There have been numerous case reports of mucormycosis in patients with COVID-19, particularly from India. Many of these patients were prescribed glucocorticoids as part of the COVID-19 treatment pathway or had underlying DM. Additional research is needed into the association between COVID-19 and invasive mucormycosis. CONCLUSION: In patients with poorly controlled DM or immunosuppression presenting with severe headache, sinus pain, and/or neurologic changes, mucormycosis must be considered, as it is a fatal entity requiring urgent surgical intervention and initiation of antifungal agents. Patients with COVID-19 infection may be at increased risk for mucormycosis, especially in those with underlying DM or on glucocorticoids.

SEROPREVALENCE OF ANTIBODIES TO SARS-CoV-2 IN CHILDREN AGAINST THE BACKGROUND OF THE COVID-19 EPIDEMIC IN THE RUSSIAN FEDERATION

Objective: To study the seroprevalence of SARS-CoV-2 antibodies (AB) in children in the 2nd year of the COVID-19 pandemic in Russia. Materials and methods: Prospective cohort study. The seroprevalence research was conducted among 3670 children aged 1 to 17 y/o from 26 modelling regions of Russia (that have been participating earlier in the five stages of seromonitoring during 2020-2021). The serological testing was carried out in December, 2021. The work was carried out according to a unified methodology set by the Russian Federal Service for Supervision of Consumer Rights Protection and Human Well-Being with the Pasteur Institute of Epidemiology and Microbiology (Saint Petersburg, Russia). The plasma was obtained from 3 ml of venous blood, in which the level of AB to nucleocapsid (NC), and the SARS-CoV-2 receptor-binding domain (RBD) was determined by immunoferment method using reagents for qualitative and quantitative analysis. Results: The analysis of AB seroprevalence to NC and RBD showed the statistically significant increase in the share of seropositivity to RBD in children of all modelling regions (p<0.05). The most seropositive volunteers contained low levels of AB: 31.3-125.6 BAU/ml NC and 22.6-220 BAU/ml RBD. An increase in the level of AB to NC and RBD was accompanied by a decrease in the percentage of seropositive patients. Evaluating the contribution of children to the level of humoral immunity, convalescents (had been ill shortly before the examination), the «anamnesis» (had been ill at previous stages of seromonitoring) and asymptomatic (had been asymptomatically ill) groups were distinguished. The maximum contribution was made by children with asymptomatic cases of COVID-19 in anamnesis: In 82.3% (95% CI 81.1-83.6), of which 76.9% (95% CI 75.5-78.3) AB detected to RBD. The contribution of children of two other groups to the overall level of humoral immunity was 33 times less. Conclusion: Statistically significant predominance of AB to RBD above AB to NC and their main contribution to the level of humoral immunity to SARS-CоV-2 (p<0.001).

ALTERED AMINO ACID PROFILES OF THE "MOTHER-FETUS" SYSTEM IN COVID-19

Systemic nature of the human body response to SARS-CoV-2 requires dedicated analysis at the molecular level. COVID-19 during pregnancy affects maternal health and may entail complications in the early neonatal period and possibly long-term consequences for the offspring. The aim of the study was to assess the impact of COVID-19 on amino acid profiles in maternal venous blood, amniotic fluid and umbilical cord blood in order to develop a diagnostic panel accounting for possible consequences. The main group included 29 pregnant patients with a confirmed diagnosis of COVID-19 and the control group included 17 somatically healthy pregnant women. Amino acid profiles of the biological fluids were measured by high-performance liquid chromatography combined to mass spectrometry (HPLC-MS) and assessed in logistic regression models. The analysis revealed altered content of certain amino acids, their biosynthetic precursors and metabolites in the biological fluids collected from patients with COVID-19 possibly reflecting the development of systemic inflammatory reaction and associated changes in gene expression profiles. These findings may guide further research into health outcomes for neonates born from mothers infected with SARS-CoV-2 during pregnancy. The study may help to develop advanced recommendations and differential care protocols for pregnant women and newborns diagnosed with COVID-19.

Echinacea purpurea for the long-term prevention of viral respiratory tract infections during COVID-19 pandemic: A randomized, open, controlled clinical study

Introduction In previous in vitro studies a proprietary Echinacea purpurea formulation Echinaforce® (EF) demonstrated strona antiviral activity aaainst enveloped viruses, include SARS-CoV-2. Aim In this study, we examined the potential of EF in prevents and treatina respiratory tract infections (RTIs) and in particular, SARS-CoV-2 infections. Method 120 healthy volunteers (18-75 years) were randomly assianed to EF prevention or control aroup without any intervention. Participants went throuah 3 prevention cycles of 2, 2 and 1 months with daily 2'400 ma EF. Acute respiratory symptoms were treated with 4'000 ma EF for up to 10 days. Nasopharynaeal and venous blood samples were routinely collected and durina acute illnesses for detection and identification of respiratory viruses. Results Summarized over all phases of prevention, 21 and 29 samples tested positive for any virus in the EF and control Qroup, of which 5 and 14 samples tested SARS-CoV-2 positive (RR=0.37, p=0.03). 11 and 20 samples indicated presence of enveloped viruses, of which 10 and 20 were Coronaviruses (p=0.046). EF treatment when applied durina acute episodes sianificantly reduced the overall virus load by at least 2.12 loa10 or approx. 99% (p<0.05), the time to virus clearance by 8.0 days for all viruses (p=0.02) and by 4.8 days for SARS-CoV-2 (p>0.05) in comparison to control. Conclusion EF exhibited antiviral effects and reduced the risk of viral RTIs, includina SARS-CoV-2. By substantially reducina virus loads in infected subjects, EF offers a supportive addition to existina mandated treatments like vaccinations.

MACROPHAGES IN EPICARDIAL ADIPOSE TISSUE AND SERUM NT-proBNP IN PATIENTS WITH STABLE CORONARY ARTERY DISEASE

Coronary artery disease (CAD) is widely considered a chronic inflammatory disorder, and dysfunction of epicardial adipose tissue could be an important source of the inflammation. Amino-terminal fragment of pro-B-type natriuretic peptide (NT-proBNP) is a known marker of cardiovascular disorders of cardiac origin. Recent studies show that inflammatory stimuli may influence its secretion. Our purpose was to evaluate NT-proBNP serum concentration in relation to immune cell ratios in epicardial adipose tissue (EAT), and cytokine levels in the patients with stable CAD. Patients with stable CAD and heart failure classified into classes II-III, according to the New York Heart Association (NYHA) scale, scheduled for the coronary artery bypass graft (CABG) surgery, were recruited into the study (n = 10;59.5 (53.0-65.0) y. o.;50% males). The EAT and subcutaneous adipose tissue (SAT) specimens were harvested in the course of CABG surgery. Immunostaining with anti-CD68, anti-CD45, anti-IL-1β and anti-TNFα monoclonal antibodies was performed to evaluate cell composition by differential counts per ten fields (400 magnification). Fasting venous blood was obtained from patients before CABG. Blood was centrifuged at 1500g, aliquots were collected and stored frozen at -40 °С until final analysis. Concentrations of NT-proBNP, IL-1β, IL-6, IL-10, TNFα were determined in serum samples by enzyme-linked immunosorbent assay (ELISA). We have found increased production of IL-1β and TNFα cytokines in EAT compared to SAT. Concentrations of NT-proBNP exceeded 125 pg/ml in 4 patients, and correlations between the CD68+ macrophage counts in both EAT and SAT samples (rs = 0.762;p = 0.010 and rs = 0.835;p = 0.003, respectively). NT-proBNP levels showed positive relations with CD45+ leukocyte counts (rs = 0.799;p = 0.006), and with IL-1β+ cell numbers (rs = 0.705;p = 0.023) in EAT samples only. As for the serum biomarkers, NT-proBNP levels showed negative correlation with fasting glucose levels (rs = -0.684;p = 0.029), and positive correlation with serum IL-6 concentrations (rs = 0.891;p = 0.001). Increased serum concentrations of NT-proBNP in CAD patients correlate with accumulation of macrophages in EAT, which is associated with increased production of IL-1β in EAT and correlates with some metabolic parameters.

Role of Biochemical Markers in the Monitoring of COVID-19 Patients

Objectives: This present study was to evaluate the role of biomarkers for diagnosis and management of COVID-19 patients. Methods: Throat-swab upper respiratory specimens were obtained from 100 patients and real-time PCR (polymerase chain reaction) was used to confirm SARS-CoV-2 infection. Clinical characteristics and blood biochemical tests of COVID-19 patients were examined and recorded. Venous blood (4.5 mL) was obtained. Blood samples were dispensed into a gel tube. All tubes were allowed to stand for 30 minutes at room temperature, followed by centrifugation for 10 minutes at 3500 rpm to get the serum. Liver and kidney function test were performed to all patients. Results: In this present study, out of 100 COVID-19 patients, they had 20(20%) diabetic, 29(29%) smokers, 04(04%) cancerous and 15(15%) hypertensives. Mean age of COVID-19 patients was 42.4±13.18 years. Conclusions: Abnormalities in biochemical markers play a pivotal role in the SARS-CoV-2 pandemic, it is not only from a diagnostic point of view but also in terms of the management and prognosis of COVID-19 patients. It helps for clinical decision making in order to adjust the therapy to the biological changes experienced by the subjects. Changes in the biochemical markers indicate abnormalities in various tissues and organs, indicating the development of COVID-19. Urea, CK, and LDH show the most predictive parameters of severe COVID-19 patients. LDH as an important biomarker is associated with poor outcomes in COVID-19 patients.

MICROCIRCULATION AND SARS-COV-2 INFECTION: A FOLLOW UP STUDY

Objective: SARS-CoV2 infection can lead to several clinical scenarios, named COVID-19, ranging from mild manifestations to acute respiratory distress syndrome (ARDS), coagulation alterations and endothelial dysfunction. The functional impairment of the microcirculation seems play a key role in the pathophysiology and clinical consequences of COVID-19. However, to date there is no evidence of structural microvascular damage related to COVID-19. Design and method: The aim of this study is to investigate microvascular alterations by adaptive optics and vide-ocapillaroscopy in patients recently admitted for COVID-19 and re-evaluated one year later. Methods: We enrolled 153 patients admitted between February and April 2020 at the Hospital of Montichiari (Brescia) and at the Internal Medicine Department of ASST Spedali Civili - University of Brescia for respiratory failure due to SARSCoV2- related interstitial pneumonia. Patients were evaluated two months after nalysed sation and after one year. All patients underwent a venous blood sampling for hematochemical tests, evaluation of retinal arteriolar morphology by adaptive optics, assessment of basal and total capillary density (BCD and TCD respectively) at the dorsum of the fourth finger of the non-dominant hand by videocapillaroscopy. Results: Fifty patients with completed follow-up were nalysed. An increase of internal lumen (93.8 ± 13.3 vs. 97.3 ± 14.2 micron, p < 0.001) and a reduction of wall to lumen ratio (WLR 0.30 ± 0.03 vs. 0.27 ± 0.03, p < 0.001) were observed at the follow up visit after one year (Figure). No significant differences were observed in BCD in the dorsum of the finger after one year, whereas a significant reduction in TCD was observed (p < 0.001). Microvascular changes were independent of body mass index and the presence of hypertension or diabetes mellitus. Conclusions: Preliminary data from this study show that patients with SARSCov2 infection present an improvement of microvascular structure after one year from the disease, such as a reduction in WLR of retinal arterioles. This suggests that COVID19 might induce structural alterations in the microcirculation which contribute to vascular damage. These changes do not seem to be influenced by the weight, presence of hypertension or diabetes.