Beneficial Effects of novel Aureobasidium Pullulans strains produced Beta-1,3-1,6 Glucans on Interleukin-6 and D-Dimer levels in COVID-19 patients; results of a randomized multiple-arm pilot clinical study (preprint)

Objective: Cytokine storm and Coagulopathy have been implicated as major causes of morbidity and mortality in COVID-19 patients. A black yeast Aureobasidium pullulans AFO-202 strain produced beta 1,3-1,6 glucan has been reported to offer potential immune enhancement and metabolism balancing, as well as mitigation of coagulopathy risks. The N-163 strain produced beta glucan is an efficient anti-inflammatory immune modulator. In this pilot clinical study, we report the beneficial effects of these two beta glucans on the biomarkers for cytokine storm and coagulopathy in COVID-19 patients. Methods: : A total of 24 RT-PCR positive COVID-19 patients were recruited (Age range: 18~62; 17 males and 7 females). Patients were randomly divided into three groups (Gr): Gr. 1 control (n=8); Gr. 2: AFO-202 beta glucan (n=8); and Gr. 3, a combination of AFO-202 and N-163 beta glucans (n=8). All three groups received the standard care while groups 2 and 3 received additional supplementation of beta glucans for 30 days. In addition to basic clinical parameters, we periodically evaluated D-Dimer, IL-6, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), the neutrophil to lymphocyte ratio (NLR), the lymphocyte to CRP ratio (LCR) and the leukocyte-CRP ratio (LeCR). Results: : The duration of hospital stay for all three groups was nearly equivalent. There was no mortality of the subjects in any of the groups. Intermittent oxygen was administered from day of admission for up to four to five days with mask (two to four Lpm) to two subjects in Gr. 2 and one subject in Gr. 3. None of the subjects required ventilation. The D-Dimer values in Gr. 1, which was on average 751 ng/ml at baseline, decreased to 143.89 ng/ml on day 15, but increased to 202.5 ng/ml on day 30, which in groups 2 and 3 decreased on day 15 and continued to remain at normal levels until day 30. IL-6 levels decreased on day 15 from an average of 7.395 pg/ml to 3.16 pg/ml in the control, 26.18 pg/ml to 6.94 pg/ml in Gr. 2 and 6.25 pg/ml to 5.22 pg/ml in Gr. 3. However, when measured on day 30, in Gr. 1, the IL-6 increased to 55.37 pg/ml while there was only slight marginal increase in Gr. 2 but within normal range, and the levels further decreased to less than 0.5 pg/ml in Gr. 3. The same trend was observed with ESR. LCR and LeCR increased significantly in Gr. 3. NLR decreased significantly in groups 2 and 3. There was no difference in CRP within the groups. Conclusion: In this exploratory study, consumption of Aureobasidium pullulans produced beta glucans for thirty days, results in a significant control of IL6, D-Dimer and NLR, a significant increase in LCR, LeCR and marginal control of ESR in COVID-19 patients. As these beta glucans are well known food supplements with decades of a track record for safety, based on these results, we recommend larger multi-centric clinical studies to validate their use as an adjunct in the management of COVID-19 and the ensuing long COVID-19 syndrome.

Efficient control of IL-6, CRP and Ferritin in Covid-19 patients with two variants of Beta-1,3-1,6 glucans in combination, within 15 days in an open-label prospective clinical trial (preprint)

Objective: Cytokine storm and coagulopathy being the major causes of morbidity in Covid 19 patients, based on an earlier pilot study yielding control of IL 6 and D-Dimer, when AFO 202 and N 163 strains of Aureobasidium Pullulans produced beta glucans, were studied separately and in combination for 30 days. In this study, we evaluated the efficiency of the two variants combinations in comparison with the control arm, which underwent a conventional regimen of treatment alone, for a shorter duration. Methods: A total of 40 RT-PCR positive Covid-19 patients divided into two groups (Gr): Gr. 1 control (n=22). Standard treatment; Gr. 2 (n =18) Standard treatment + combination of AFO 202 and N 163 beta glucans for 15 days. Biomarkers of relevance to cytokine storm and coagulopathy were evaluated at baseline on Day 7 and Day 15. Results: The C-reactive protein (CRP), which declined from 33.95 ng/ml to 5.07 ng/ml in control and from 33.95 ng/ml to 5.64 ng/ml in the treatment arm on Day 7, increased to 14.6 ng/ml in the former while it continued to be under control in the treatment arm at 5.68 ng/ml on Day 15. The same trend was observed in Ferritin, whose values were 560.58 pg/ml at baseline to 261.44 pg/ml (day 7) and 182.40 pg/ml (day 15) in the treatment group, while it was 535.24 pg/ml at baseline, 116.66 pg/ml on day 7 and 291.95 pg/ml on day 15 in the control group. IL-6 showed a higher decrease in treatment group compared to the control group. The difference between day 7 and day 15 values were statically significant. Conclusion: A statistically significant control of IL-6, CRP and Ferritin in Covid 19 patients who orally consumed AFO 202 and N 163 strains of Aureobasidium Pullulans produced Beta glucans together in 15 days make us recommend this safe food supplement be consumed by Covid 19 patients along with conventional treatments, especially to the vulnerable population, as a prophylaxis amidst the prolonged pandemic with evolution of mutated strains of SARS COV2.

Beneficial Effects of novel Aureobasidium Pullulans strains produced Beta-1,3-1,6 Glucans on Interleukin-6 and D-Dimer levels in COVID-19 patients; results of a randomized multiple-arm pilot clinical study (preprint)

ObjectiveCytokine storm and Coagulopathy have been implicated as major causes of morbidity and mortality in COVID-19 patients. A black yeast Aureobasidium pullulans AFO-202 strain produced beta 1,3-1,6 glucan has been reported to offer potential immune enhancement and metabolism balancing, as well as mitigation of coagulopathy risks. The N-163 strain produced beta glucan is an efficient anti-inflammatory immune modulator. In this pilot clinical study, we report the beneficial effects of these two beta glucans on the biomarkers for cytokine storm and coagulopathy in COVID-19 patients. MethodsA total of 24 RT-PCR positive COVID-19 patients were recruited (Age range: 18[~]62; 17 males and 7 females). Patients were randomly divided into three groups (Gr): Gr. 1 control (n=8); Gr. 2: AFO-202 beta glucan (n=8); and Gr. 3, a combination of AFO-202 and N-163 beta glucans (n=8). All three groups received the standard care while groups 2 and 3 received additional supplementation of beta glucans for 30 days. In addition to basic clinical parameters, we periodically evaluated D-Dimer, IL-6, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), the neutrophil to lymphocyte ratio (NLR), the lymphocyte to CRP ratio (LCR) and the leukocyte-CRP ratio (LeCR). ResultsThe duration of hospital stay for all three groups was nearly equivalent. There was no mortality of the subjects in any of the groups. Intermittent oxygen was administered from day of admission for up to four to five days with mask (two to four Lpm) to two subjects in Gr. 2 and one subject in Gr. 3. None of the subjects required ventilation. The D-Dimer values in Gr. 1, which was on average 751 ng/ml at baseline, decreased to 143.89 ng/ml on day 15, but increased to 202.5 ng/ml on day 30, which in groups 2 and 3 decreased on day 15 and continued to remain at normal levels until day 30. IL-6 levels decreased on day 15 from an average of 7.395 pg/ml to 3.16 pg/ml in the control, 26.18 pg/ml to 6.94 pg/ml in Gr. 2 and 6.25 pg/ml to 5.22 pg/ml in Gr. 3. However, when measured on day 30, in Gr. 1, the IL-6 increased to 55.37 pg/ml while there was only slight marginal increase in Gr. 2 but within normal range, and the levels further decreased to less than 0.5 pg/ml in Gr. 3. The same trend was observed with ESR. LCR and LeCR increased significantly in Gr. 3. NLR decreased significantly in groups 2 and 3. There was no difference in CRP within the groups. ConclusionIn this exploratory study, consumption of Aureobasidium pullulans produced beta glucans for thirty days, results in a significant control of IL6, D-Dimer and NLR, a significant increase in LCR, LeCR and marginal control of ESR in COVID-19 patients. As these beta glucans are well known food supplements with decades of a track record for safety, based on these results, we recommend larger multi-centric clinical studies to validate their use as an adjunct in the management of COVID-19 and the ensuing long COVID-19 syndrome.

Beneficial immune-regulatory effects of novel strains of Aureobasidium pullulans AFO-202 and N-163 produced beta glucans in Sprague Dawley rats (preprint)

Background: Immune system dysregulation plays a significant role in the pathogenesis of COVID-19. A balanced immune response is essential to mounting anti-viral defences, and biomarkers such as the white blood cell (WBC) count, the neutrophil-to-lymphocyte ratio (NLR) and the lymphocyte-to-C-reactive-protein (LCR) ratio have been reported as potential predictors of immune status. The beneficial immunomodulation effects of a biological response modifier glucan (BRMG) produced by two strains of Aureobasidium pullulans , AFO-202 and N-163, were reported in earlier in vitro studies. In this study, we compared their efficacy on immune-inflammatory parameters in Sprague Dawley (SD) rats. Methods: : This study was performed on four groups of healthy SD rats, with six subjects in each group: Group 1, which was euthanised on Day 0 to obtain baseline values; Group 2, which served as the control (drinking water); Group 3, which received AFO-202 beta glucan at a dose of 200 mg/kg/day; and Group 4, which received N-163 beta glucan at a dose of 300 mg/kg/day. Test solutions were administered to the animals in Groups 2–4 by gavage once daily for 28 consecutive days. Biochemical analyses were conducted on haematological, immunological and inflammatory biomarkers on Days 15 and 29. Results: : The NLR decreased, whereas the LCR and leukocyte-to-C-reactive protein ratio (LeCR) increased in Group 3 (AFO-202) at 15 days, but the values were within the normal physiological range only. At 29 days, this difference among the groups was not observed. There were no significant differences between the groups in the other parameters, such as red blood cell (RBC) count, WBC count, CRP, IgA, IL-6, IFN-γ and sFAS. Conclusion: AFO-202 beta glucan helps marginally decrease NLR and increase LCR and LeCR in healthy SD rats within 15 days. This might be beneficial to tackling infections such as COVID-19 that involve immune system dysregulation. These results warrant further investigations in larger numbers of healthy and diseased models to develop appropriate strategies for balancing immune system dysregulation using these beta glucan food supplements with proven safety.