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

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.

Alternaria alternata as a Seed-Transmitted Pathogen of Sida hermaphrodita (Malvaceae) and Its Suppression by Aureobasidium pullulans

Background: Sida hermaphrodita (Virginia fanpetals) was introduced to Poland nearly 70 years ago as a potential fodder plant, and it is gaining importance as an energy crop. Alternaria alternata transmitted by seeds may exert a negative effect on the health of Virginia fanpetals plants. Methods: The virulence of the A. alternata pathogen, isolated from Virginia fanpetals seeds, was tested on detached leaves of Virginia fanpetals plants. The isolates were identified as A. alternata based on partial sequence analysis of Alta1, TEF1a and gdp genes and the ITS 1–5.8SrDNA–ITS 2 region. Pathogen transmission from seeds to seedlings and the influence of seed dressing with a suspension of Aureobasidium pullulans on seedling health were analyzed in a greenhouse experiment. Results: Three of the nine analyzed A. alternata isolates were highly pathogenic for S. hermaphrodita. The initial symptoms of leaf infection were small, round dark brown or black spots which grew into larger dark brown spots surrounded by a chlorotic halo. Alternaria alternata was re-isolated from inoculated plants and was identified as the causal agent of Alternaria leaf spot disease. In the greenhouse experiment, S. hermaphrodita seeds dressed with a suspension of A. pullulans and inoculated with A. alternata produced a higher number of seedlings with a higher health status than non-dressed seeds. Conclusions: The study demonstrated that A. alternata is transmitted from infected S. hermaphrodita seeds to developing plants and biological control limits this phenomenon.

Disease-modifying immune-modulatory effects of the N-163 strain of Aureobasidium pullulans-produced 1,3-1,6 Beta glucans in young boys with Duchenne muscular dystrophy: Results of an open-label, prospective, randomized, comparative clinical study

Background: Duchenne muscular dystrophy (DMD) is an inherited neuromuscular disorder causing progressive muscle weakness and premature death. Steroids remain the mainstream approach for supportive care but have side effects; other targeted therapies and gene therapies are also being developed. As there is limited evidence on the use of disease modifying nutritional supplement adjuncts in DMD, this pilot trial is to evaluate the effects of supplementation of Aureobasidium pullulans-derived 1,3 1,6 beta glucan from the N163 strain in young patients with DMD. Methods: Twenty-seven patients with Duchenne muscular dystrophy (DMD), nine in the control arm (undergoing conventional therapies) participated. The patients were divided into groups: those not administered steroids (Steroid negative) (n = 5), those administered steroids (Steroid positive) (n = 4), and 18 in the treatment arm (N163 beta glucan supplement along with conventional therapies; N-163 Steroid negative and N-163 Steroid positive); they participated in the study for 45 days. Assessments of muscle function, disease status, and levels of IL6, IL13, TGF beta;, creatinine kinase (CK), titin, TNF Alpha;, haptoglobin, and dystrophin in the blood and myoglobin in the urine were performed at baseline and at the end of the study. Results: IL6 showed a significant decrease in the N163 Steroid negative group, from a baseline value of 7.2 pg/ml to 2.7 pg/ml. IL13 decreased in both treatment groups, from 157.76 pg/ml to 114.08 pg/ml (N-163 Steroid negative) and from 289.56 pg/ml to 255.56 pg/ml (N-163 Steroid positive). TGF beta levels showed a significant decrease in the N163 Steroid negative group, from a baseline value of 3302 ng/ml to 1325.66 ng/ml post intervention. Dystrophin levels increased by up to 32% in both Steroid positive and negative groups. Medical research council (MRC) grading showed muscle strength improvement in 12 out of 18 patients (67%) in the treatment group and four out of nine (44%) subjects in the control group. Conclusion: Supplementation with the N163 beta glucan food supplement produced disease-modifying beneficial effects: a significant decrease in inflammation and fibrosis markers, increase in dystrophin and improvement in muscle strength in DMD subjects over 45 days, thus making this a potential adjunct treatment for DMD after validation. A longer duration of follow up and further research on the mechanism of action and commonalities with other diseases provoked by hyperactive inflammation and/or fibrosis may pave the way for their extended applications in other dystrophinopathies and neuroinflammatory diseases.

Modified Rhodopsins From Aureobasidium pullulans Excel With Very High Proton-Transport Rates

Aureobasidium pullulans is a black fungus that can adapt to various stressful conditions like hypersaline, acidic, and alkaline environments. The genome of A. pullulans exhibits three genes coding for putative opsins ApOps1, ApOps2, and ApOps3. We heterologously expressed these genes in mammalian cells and Xenopus oocytes. Localization in the plasma membrane was greatly improved by introducing additional membrane trafficking signals at the N-terminus and the C-terminus. In patch-clamp and two-electrode-voltage clamp experiments, all three proteins showed proton pump activity with maximal activity in green light. Among them, ApOps2 exhibited the most pronounced proton pump activity with current amplitudes occasionally extending 10 pA/pF at 0 mV. Proton pump activity was further supported in the presence of extracellular weak organic acids. Furthermore, we used site-directed mutagenesis to reshape protein functions and thereby implemented light-gated proton channels. We discuss the difference to other well-known proton pumps and the potential of these rhodopsins for optogenetic applications.