GMP Manufacturing of Cryopreserved peripheral Blood Stem Cell Products

Background: Hematopoietic stem cell preparations are pharmaceuticals that are typically transfused as fresh products. Due to the travel restrictions caused by the Covid-19 pandemic since March 2020, the supply of fresh stem cell preparations to patients became significantly challenging. To maintain global patient supply, a GMP-compliant manufacturing process for cryopreservation of allogeneic stem cell donations was established and validated. Method(s): The donor leukapheresis is transported from the collection center to the manufacturing site at 2-6degreeC using qualified containers. In class D clean rooms, this material is further processed in a completely closed process. If necessary, volume reduction is performed by centrifugation. The suspension is mixed with the cryoprotectant Cryostore CS10 to achieve a final DMSO concentration of 5%. The formulated product is filled into cryobags and cryopreserved under controlled freezing conditions. The frozen products are stored and transported at <= 140 degreeC. The cell counter NC-200 is used to determine the cell count and viability of the product. 97 products in 2020 and 127 products in 2021 were successfully cryopreserved and transported worldwide. Result(s): The mean values of viability of 215 products are 99.3 % (+/- 1.1 % SD) for fresh apheresis products and 93.1 % (+/- 6.2 % SD) for the corresponding cryopreserved final product after thawing. For a good recovery after thawing, the age of the apheresis as well as the DMSO contact time are generally considered to be critical factors. The analysis of the viability of 222 cryopreserved end products after thawing shows a correlation in relation to the age of the apheresis (Fig. 1). As the age of the apheresis increases, the viability decreases. Therefore, care should be taken about the age of the apheresis. A correlation between the DMSO contact time and the viability of the end products after thawing was not observed (Fig. 2). Conclusion(s): GMP-compliant closed system manufacturing of cryopreserved allogeneic stem cell products provides safe and high quality drugs that can be used for transplantation. We have shown that with our manufacturing process, DMSO, which is potentially toxic to cells, has no effect on cell recovery after thawing. Cryopreservation is therefore a suitable and safe method to provide patients with essential therapy worldwide. (Figure Presented).

Probability of Covid-19 in Abo Blood Type during Second Wave in Southern Rajasthan, India

Objectives: Along the course human history of scientific research, the severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) is the most concerning global health problem. Second wave of COVID-19 has adversely affected India. However, India embarked on its immunization program on January 16, 2021, operating 3006 vaccination centers onset Covaxin and CoviShield. This study aimed to ascertain if there is an association amidst ABO blood type and probability of COVID-19 infection in wave. Method(s): This is analytical and observational study conducted on 713 SARS-COVID-19-positive patients of a known ABO blood type, who attended outpatient department and inpatient department during March 26-May 20, 2021, in tertiary care hospital Udaipur (Raj.) Serum inflammatory markers were evaluated by Cobas 6000. Result(s): Out of the 713 patients who were tested positive, 15.56% was blood group Type A, 19.91% was blood group Type B, 13.65% was blood group Type AB, and 46.28% was blood group Type O. On statistical analysis, there were positive association between O+ blood type and peak inflammatory marker (interleukin-6 and D-Dimer). Patients with blood Type O who received a test were more likely to test positive and blood Type B+, A+, A+, AB+, O-, A-, B-, and AB- were less likely to test positive. Conclusion(s): The present study shows an evidence for interrelation between ABO blood groups and SARS-COVID-19. Reported infection prevalence is moderately increased among O+ blood type individuals. Determination of level of inflammatory markers might prove to be helpful to clinicians so as to keep track of severity of infection and evaluate the prognosis of SARS-COVID-19 with specific ABO blood groups. Copyright © 2022 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)

Evaluation of a Propofol Dosing Capping Strategy in Patients with Hypertriglyceridemia and Covid-19

INTRODUCTION: Propofol can be associated with hypertriglyceridemia (HTG), which may lead clinicians to change to non-preferred sedatives such as benzodiazepines (BZD) to avoid HTG complications. Patients with COVID-19 have been observed to require increased doses and durations of sedatives, which may increase the risk of HTG development. The purpose of this study was to evaluate a dose capping strategy of propofol versus discontinuation on minimizing BZD exposure in ventilated patients. METHOD(S): This retrospective study included patients with COVID-19 and HTG after receiving propofol between March 1, 2020 and April 30, 2021. HTG was defined as serum triglycerides (TG) greater than or equal to 500 mg/dL. The propofol dose cap used in select patients was a reduced maximum dose of 30 to 40 mcg/kg/min. The remainder of patients had propofol discontinued (standard of care). Descriptive statistics were used to evaluate differences in propofol duration, BZD doses, and intensive care unit (ICU) length of stay (LOS) between groups. Incidence of pancreatitis was also evaluated. RESULT(S): Seventy-one patients were included. The propofol dose cap was used in twenty patients. The mean baseline TG were similar in both groups, 256 mg/dL (standard deviation (SD)=125) in the propofol dose cap group compared to 252 mg/dL (SD=104) in the standard group (P=0.9). Propofol was continued four days longer after dose capping, with a median duration of 11.7 vs. 7.4 days, P=0.255. Cumulative intermittent doses of BZDs were higher in the dose cap cohort (average cumulative dose 28mg vs. 20mg, P=0.245), however BZD continuous infusion cumulative doses were lower (1214mg vs. 1377mg, P=0.778). Median ICU LOS was similar between groups (20.4 vs. 20.9 days, P=0.451). No patients in either group developed pancreatitis. CONCLUSION(S): The dose capping strategy allowed for clinicians to continue using propofol for sedation in mechanically ventilated patients with COVID-19 and HTG. This in turn helped minimize BZD exposure with no evidence of HTG associated adverse effects such as pancreatitis. Future investigation in a larger population is warranted to determine the dose cap impact on clinical outcomes, such as incidence of delirium and length of mechanical ventilation.

Effect of polymerized type I collagen in hyperinflammation of adult outpatients with symptomatic COVID-19: a double blind, randomised, placebo-controlled clinical trial

Background. Currently, therapeutic options for outpatients with COVID-19 are limited, in Mexico Polymerized Collagen type I (PCTI) has been tested as a useful option. Methods. Double-blind, randomised, placebo-controlled clinical trial of PTIC vs placebo. To evaluate the safety, efficacy and effect of the intramuscular administration of polymerized type I collagen (PTIC) on hyperinflammation, oxygen saturation and symptom improvement in adult outpatients with symptomatic COVID-19. Eighty-nine adult participants with a confirmed COVID-19 diagnosis and symptom onset within the 7 days preceding recruitment were included from August 31, 2020 to November 7, 2020 and followed for 12 weeks. Final date of follow-up was February 4, 2021. Patients were randomly assigned to receive either 1.5 ml of PTIC intramuscularly every 12 h for 3 days and then every 24 h for 4 days (n=45), or matching placebo (n=44). Results. Of 89 patients who were randomised, 87 (97.8%) were included in an intention-to-treat analysis;37 (41.6%) were male and mean age was 48.5+/-14.0 years. The IP-10 levels decreased 75% in the PTIC group and 40% in the placebo group vs baseline. The comparison between treatment vs placebo was also statistically significant (P=0.0047). The IL-8 (44%, P=0.045), M-CSF (25%, P=0.041) and IL-1Ra (36%, P=0.05) levels were also decreased in the PTIC group vs baseline. Mean oxygen saturation >=92% was achieved by 40/44 (90%), 41/42 (98%) and 40/40 (100%) of participants that received PTIC at 8, 15 and 97 days of follow-up vs 29/43 (67%), 31/39 (80%) and 33/37 (89%) of patients treated with placebo (P=0.001). The unadjusted accelerated failure time model showed that patients treated with PTIC achieved the primary outcome 2.70-fold faster (P< 0.0001) than placebo. In terms of risk, the group of patients treated with PTIC had a 63% lower risk of having a mean oxygen saturation < 92% vs placebo (P< 0.0001). Symptom duration in patients treated with PTIC was reduced by 6.1+/-3.2 days vs placebo. No differences in adverse effects were observed between the groups at 8, 15 and 97 days of follow-up. Conclusion. Treatment with PTIC down-regulated IP-10, IL-8, M-CSF and IL-Ra levels, which could explain the PTIC effect on the higher proportion of patients with mean SaO2 >=92% and a shorter duration of symptoms as compared with placebo. Serum cytokine and chemokine levels of SARS-CoV2-infected symptomatic outpatients at baseline and day 8 post-treatment with PTIC or placebo. Data are expressed as median with 95% confidence. (A) IP-10, IFN-gamma inducible protein-10;(B) IL-8, Interleukin-8;(C) M-CSF, Macrophage colony-stimulating factor;(D) IL-1Ra, IL-1 receptor antagonist;(E) TRAIL, TNF-related apoptosis inducing ligand;and (F) Forest plot (95% confidence intervals). (A) Probability of oxygen saturation 92% or greater while breathing ambient air. (B) Accelerated time failure model for oxygen saturation 92% or greater while breathing ambient air among polymerised type I collagen and placebo.

Emergent Medicine: Impact of Out-of-Hospital Red Lights and Sirens (RLS) on Time to Antibiotic Administration in Sepsis Alert Patients

Study Objectives: This study asks whether the use of red lights and sirens while transporting patients identified as out-of-hospital 'sepsis alerts' impacts the time to antibiotic administration. Emergency Medical Services (EMS) personnel use emergent transport-characterized by the use of red lights and sirens (RLS)-for those patients deemed 'critical' or in need of potentially life-saving interventions beyond the scope of the interventions available in the out-of-hospital scenario. Nationally, the use of RLS saves ~90 seconds in urban EMS settings, which may be the difference between life and death in a cardiac arrest, traumatic hemorrhage or airway emergency. However, the decision to transport with RLS is often at the discretion of the EMS personnel without formal guidelines or evidence-based recommendations. Emergent transport is associated with increased risk of motor vehicle collision (MVC), injuries to EMS personnel and/or patients and property damage. Additionally, RLS in transport may detrimentally impact the patient's physiological and psychologic response to EMS transport. Sepsis is a life-threatening systemic response to infection, leading to shock or death if not treated with antibiotics. After receiving initial resuscitation in the field, it is common for 'sepsis alert' patients to be transported to the hospital emergently with the use of red lights and sirens (RLS). Upon arrival to the hospital, antibiotics may not be given for prolonged periods of time. Out-of-hospital antibiotic administration has shown little to no difference in patient outcomes either. These findings question the value of emergent transport of out-of-hospital sepsis alert patients, particularly when patients transported with lights and sirens have ~2.4 times higher risk of MVC. Method(s): This is a prospective, non-randomized controlled trial evaluating the clinical impact of emergent transport for patients identified as out-of-hospital sepsis alerts and transported by the Sarasota County Fire Department (SCFD) using either RLS on even days or no RLS on odd days. The time to antibiotic administration was analyzed by the priority of transport. Absolute time to antibiotic administration was calculated in minutes and analyzed using two-tailed T-tests in Excel and SPSS. Patients who tested positive for COVID-19 were removed from the data analysis. Result(s): Data collection ran from May to October of 2021, with an total sample of 709 patients identified as out-of-hospital sepsis alerts. Preliminary data from June to July 2021 showed an average time to antibiotic administration of 112 minutes for emergent versus 125 minutes for non-emergent (n = 84;p-value = 0.448). The average difference in transport time between emergent and non-emergent transports was only 91 seconds. Conclusion(s): There was a marked but non-significant difference in time to antibiotic administration for patients identified as out-of-hospital sepsis alerts and transported with or without RLS in our preliminary analysis. This difference cannot be explained by the time saved using emergent transport. Regional prevalence of COVID-19 cases during the study may have increased the variability in time to antibiotic administration. The absolute time of transport cannot account for the observed difference in time to antibiotic administration and hints that other variables determine the expediency of care for out-of-hospital sepsis alerts seen in the emergency department. [Formula presented] No, authors do not have interests to disclose Copyright © 2022

Antibody Responses to COVID-19 Infection are Blunted in Hematology/Oncology Patients

Introduction: Coronavirus disease (COVID-19) causes critical illnesses in a large proportion of patients, leading to significant morbidity & mortality. In this regard, the poor prognosis and high mortality rates due to COVID-19 in those with immune dysfunction has been noted. The reason for this has been attributed to poor host immune response in this cohort. However, serological responses have not been well studied in these settings. Aims & Objectives: To describe the antibody response to COVID19 infections among Immunocompromised patients. Material(s) and Method(s): In this prospective observational study, antibody responses to SARS CoV-2 in patients with hematological/ oncologic conditions requiring chemotherapy, immuno-suppression or post transplantation was studied. Patients included were diagnosed positive between July 2020 to February 2022. Patients were recruited in the study on a follow-up visit where in addition to capturing. Demographics, clinical details and a blood sample was collected for antibody testing against nucleoprotein (anti-N) and spike receptor binding domain (anti-S) (Roche Elecsys ECLIA) platform. Serological response was studied using chi-square tests, comparing anti-N and anti-S antibody titres across groups. These were compared against serological response in a parallellytested set of healthy control individuals. Result(s): A total of 134 immunocompromised patients were included 99(73%) patients on chemotherapy, 19(14%) post- stem cell transplantation and 17(13%) on other immunosuppression. The median age was 24 years with a male predominance (84/134). Of these 97%(130/134) had been vaccinated at time of testing serological response. Serological response with positive anti -S antibody titres was noted in 88% (118/134) of these patients while only 71% (95/134) showed positive anti -N antibody response. In comparison, among 235 healthy control, positive anti-S antibody responses were found in 97% (p<0> Among the immune-compromised, vaccination(mainly Covishield) also appeared to confer better anti-S antibody responses in comparison to those un-vaccinated prior to COVID-19 infections, although the difference was not statistically significant (94.4 vs 86.6, p = 0.348). Conclusion(s): This is the first study that has prospectively documented serological response to COVID-19 infection in patients with underlying hematology/oncology disorders. Overall a pattern of inferior antibody response to Sars Cov2 was observed in this immunologically dysfunctional cohort, as compared to a healthy control set. (Table Presented).

Bringing health care closer to patients - Two years of the home care ALS programme in Slovenia

Ljubljana ALS Center takes care of the majority of patients with ALS in Slovenia. Due to limited access to health care during COVID-19 pandemic, we have started a home care ALS programme to improve care of patients with ALS. We developed eligibility criteria to decide which patients could benefit from home visits. The criteria included problematic transport due to patient immobility, mechanical ventilation use, gastrostomy-related problems and need for advanced directives discussion. During March 2020 and March 2022, we performed 190 home visits in 67 patients. This represents 20% of all ALS out-patient visits and 30% of the patients treated at our center during this period. The patients were visited on average every 3.3 months (range between visits 3 days -15 months). Only 4% of the visits were done due to a sudden unexpected clinical deterioration. The neurologist that performed home visits was usually accompanied by one or two members of our multidisciplinary team (mostly by respiratory therapist and sometimes by nurse, social worker or team coordinator). 58% of the patients visited were using gastrostomy, 72% were using noninvasive ventilation (NIV) and 9% were using invasive ventilation (IV). The main procedures/tasks performed at home visits were: arterial blood gasses analysis (in 72% of all visits), assessment of NIV (in 60%), adjustment of symptomatic therapy (in 41%), advance directive regarding mechanical ventilation (in 28%), prescription of existing therapy (in 13%), discussion on possible gastrostomy (in 13%), gastrostomy assessment (in 10%), gastrostomy care (in 10%), replacement of gastric tube (in 5%), assessment of IV (in 5%), botulinum toxin application (in 5%), decision to withhold treatment (in 4%), introduction of NIV (in 2%), introduction of cough assist (in 2%), discussion on treatment withdrawal (in 1%). The home care ALS programme provides an improved health care for patients with ALS, especially for those in advance stages of the disease. Many of these patients would probably not be able to attend regular out-patient visits at the hospital. The multidisciplinary programme integrates different aspects of ALS care, the most important being home ventilation and palliative care. Based on our experience, it can be cost and time effective with appropriate planning. The programme has recently received long-term funding by the Health Insurance Institute of Slovenia as a part of mobile palliative teams initiative.

Post-COVID-19 vaccine SARS-CoV-2 antibody investigation in healthcare professionals

Objectives: Main purpose of this study was evaluating inactive severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) vaccine subsequent anti-S1 IgG feedback and the aspects involved in such reactions for professionals in healthcare (HCP) as the dominant risk group. Method(s): Thirty-six HCPs with previous COVID-19 infection and 164 with no priors, 200 in total, who was working in the Ankara Public Health Molecular Diagnosis Laboratory were included. Main tool of identifying humoral immune response quantifably in serum samples which were obtained 28 days after administering each of two doses of vaccine was Roche Elecsys SARS-CoV-2 kit. Result(s): Average antibody levels of 164 negative HCPs were 15.82 +/- 8.59 IU/mL and 26.042 +/- 10.73 IU/mL while 36 positive HCPs demonstrated antibody responses as 66.083 +/- 33.927 IU/mL and 90 +/- 27.012 IU/mL 28 days after each of two doses of vaccine for both individual groups respectively. A statistically meaningful difference was found in antibody levels after two vaccine doses in both groups (p < 0.0001). The authors observed statistically higher average antibody levels after initial vaccine dosage in HCPs with infection than the antibody levels of naive individuals after second dose (p < 0.0001). Age, gender and vaccination feedback did not have a statistically meaningful disparity (p > 0.05). Conclusion(s): It was concluded that the average antibody level achieved after inital dose n HCPs with COVID-19 infection was surpassing the average antibody level obtained after the second dose in naive HCPs. The authors recommend further clinical researches on antibody levels and the extent of protection to prohibit COVID-19. Copyright © 2022 by Prusa Medical Publishing.

The key role of plasminogen activator inhibitor-1 (PAI-1) in the thrombogenesis difference in COVID-19- associated pneumonia compared to bacterial pneumonia

Background: PAI-1 plays a key role in a wide range of physiological and pathological processes, including coagulation, fibrinolysis, inflammation. Aim(s): The aim was to estimate the diagnostic role of PAI-1 on admission in hospitalized patients with COVID-19 pneumonia, comparing with bacterial pneumonia. Method(s): We observed 3 groups: Main (1) -85 patients (mean age -59 (52;65) years, men -45 (52.9%)), hospitalized with pneumonia on the background of laboratory-confirmed (PCR) COVID-19, divided into 3 subgroups: Subgroup 1 -40 patients with moderate COVID-19, subgroup 2 -25 patients with severe COVID-19, subgroup 3 -20 patients with critical COVID-19;Comparative (2) -55 patients (mean age -48.9 (34;62) years, men -30 (54.5%)), hospitalized with community-acquired pneumonia of bacterial etiology (CABP) without COVID-19;Control (3) -25 healthy volunteers (average age -50.0 (35;65) years, men -13 (52.0%)). General analysis, plasma level of PAI-1 (Human PAI-1 ELISA Kit, Elabscience) performed at admission before starting of anticoagulant treatment, statistical analysis. Result(s): At admission the highest level of PAI-1 (6.1 [0.15;18] ng/ ml) was in COVID-19 patients (Main group) and exceeds the Control group (0.1 [0.09;0.11] ng/ml), p1-3 = 0.000) in more than 60 times. Whereas the level of PAI-1 in patients with CABP didn't differ from Control group (0.1 [0.09;0.11] ng/ml), p2-3 = 0.29) (Figure 1).Among COVID-19 group the levels of PAI-1 correlated with the disease severity (Figure 2). Conclusion(s): 1) significantly increase of plasma levels of PAI-1 in COVID-19 pneumonia demonstrates the cornerstone in thrombogenesis of this disease -problems in fibrinolysis system, which is the main difference between CABP;2) in hospitalized patients with COVID-19 pneumonia the level of PAI-1 is associated with the disease severity and could be the crucial marker for patients' distribution. (Figure Presented).