Iota-Carrageenan Inhibits Replication of SARS-CoV-2 and the Respective Variants of Concern Alpha, Beta, Gamma and Delta.

The COVID-19 pandemic continues to spread around the world and remains a major public health threat. Vaccine inefficiency, vaccination breakthroughs and lack of supply, especially in developing countries, as well as the fact that a non-negligible part of the population either refuse vaccination or cannot be vaccinated due to age, pre-existing illness or non-response to existing vaccines intensify this issue. This might also contribute to the emergence of new variants, being more efficiently transmitted, more virulent and more capable of escaping naturally acquired and vaccine-induced immunity. Hence, the need of effective and viable prevention options to reduce viral transmission is of outmost importance. In this study, we investigated the antiviral effect of iota-, lambda- and kappa-carrageenan, sulfated polysaccharides extracted from red seaweed, on SARS-CoV-2 Wuhan type and the spreading variants of concern (VOCs) Alpha, Beta, Gamma and Delta. Carrageenans as part of broadly used nasal and mouth sprays as well as lozenges have the potential of first line defense to inhibit the infection and transmission of SARS-CoV-2. Here, we demonstrate by using a SARS-CoV-2 spike pseudotyped lentivirus particles (SSPL) system and patient-isolated SARS-CoV-2 VOCs to infect transgenic A549ACE2/TMPRSS2 and Calu-3 human lung cells that all three carrageenan types exert antiviral activity. Iota-carrageenan exhibits antiviral activity with comparable IC50 values against the SARS-CoV-2 Wuhan type and the VOCs. Altogether, these results indicate that iota-carrageenan might be effective for prophylaxis and treatment of SARS-CoV-2 infections independent of the present and potentially future variants.

Carrageenan-Based Acyclovir Mucoadhesive Vaginal Tablets for Prevention of Genital Herpes.

Women are the most affected by genital herpes, which is one of the most common sexually transmitted infections, affecting more than 400 million people worldwide. The application of vaginal microbicides could provide a safe method of protection. Acyclovir is a safe and effective medication for vaginal administration, and numerous benefits have been observed in the treatment of primary or recurrent lesions due to genital herpes. Vaginal tablets based on a combination of the polymers iota-carrageenan and hydroxypropyl methylcellulose were developed for the controlled release of acyclovir. Swelling, mucoadhesion and drug release studies were carried out in simulated vaginal fluid. The tablets, containing a combination of iota-carrageenan and hydroxypropyl methylcellulose, have an adequate uptake of the medium that allows them to develop the precise consistency and volume of gel for the controlled release of acyclovir. Its high mucoadhesive capacity also allows the formulation to remain in the vaginal area long enough to ensure the complete release of acyclovir. These promising formulations for the prevention of genital herpes deserve further evaluation.

Matrix tablets based on a carrageenan with the modified-release of sodium riboflavin 5'-phosphate.

The focus of this work was to produce modified-release monolithic matrix tablets containing sodium riboflavin 5'-phosphate (vitamin B2) as active pharmaceutical ingredient (API). Riboflavin 5'-phosphate is absorbed from the upper gastrointestinal tract by a specific transport mechanism. The aim of this work was the development of modified-release tablets from which most or the entire API can dissolve within 5 h. The dissolution was started in medium pH 1.2 (gastric juice) and finished in medium pH 4.5. The matrix former was iota-carrageenan combined with microcrystalline cellulose (MCC) and lactose in different ratios. Factorial design was used in this work so as to study the effects of the MCC/lactose ratio on the parameters of the tablets, and especially on the dissolution process. The dissolution data were subjected to statistical analysis, and the release profiles were fitted with different models. It was found that the MCC/lactose ratio influenced the quality of the tablets to a high degree. The Korsmeyer-Peppas model proved to characterize the total dissolution profile best, but fitting of the separate sections was also possible with a linear model.

Exploring the amyloid degradation potential of nanoformulated carrageenan-bridging in vitro and in vivo perspectives.

Amyloids, with their ß-sheet-rich structure, contribute to diabetes, neurodegenerative diseases, and amyloidosis by aggregating within diverse anatomical compartments. Insulin amyloid (IA), sharing structural resemblances with amyloids linked to neurological disorders, acts as a prototype, while compounds capable of degrading these fibrils hold promise as therapeutic agents for amyloidosis intervention. In this research, liposomal nanoformulated iota carrageenan (nCG) was formulated to disrupt insulin amyloids, demonstrating about a 17-20 % higher degradation efficacy compared to conventional carrageenan through thioflavin T fluorescence, dynamic light scattering analysis, and turbidity quantification. The biocompatibility of the nCG and nCG-treated insulin amyloids was evaluated through MTT assay, live-dead cell assay on V79 cells, and hemolysis testing on human blood samples to establish their safety for use in vitro. Zebrafish embryos were utilized to assess in vivo biocompatibility, while adult zebrafish were employed to monitor the degradation capacity of IA post subcutaneous injection, with fluorescence emitted by the fish captured via IVIS. This demonstrated that the formulated nCG exhibited superior anti-amyloid efficacy compared to carrageenan alone, while both materials demonstrated biocompatibility. Furthermore, through docking simulations, an exploration was conducted into the molecular mechanisms governing the inhibition of the target protein pancreatic insulin by carrageenan.

Adsorption and antibacterial studies of a novel hydrogel adsorbent based on ternary eco-polymers doped with sulfonated graphene oxide developed from upcycled plastic waste.

A novel ternary blended polymer composed of cost-effective and readily available polymers was synthesized using poly (vinyl alcohol) (PVA), iota carrageenan (IC), and poly (vinyl pyrrolidone) (PVP). Sulfonated graphene oxide (SGO), prepared from recycled drinking water bottles, was utilized as a doping agent. Varying amounts (1-3 wt%) were combined into the polymer matrix. The produced hydrogel film was examined as a potential adsorbent hydrogel film for the removal of methylene blue (MB) and Gentamicin sulfate (GMS) antibiotic from an aqueous solution. The experimental results demonstrate that the presence of SGO significantly increased the adsorption efficiency of PVA/IC/PVP hydrogel film. The antimicrobial tests revealed that the PVA/IC/PVP-3% SGO hydrogel film exhibited the most potent activity against all the tested pathogenic bacteria. However, the adsorption results for MB and GMS showed that the addition of 3 wt% SGO resulted in a removal percentage that was a two fold increase in the removal percentage compared with the undoped PVA/IC/PVP hydrogel film. Furthermore, the response surface methodology (RSM) model was utilized to examine and optimize several operating parameters, including time, pH of the solution, and initial pollutant concentration. The adsorption kinetics were better characterized by the pseudo-second-order kinetics model. The composite film containing 3 wt% SGO had a maximum adsorption capacity of 606 mg g-1 for MB and 654 mg g-1 for GMS, respectively. The generated nanocomposite hydrogel film demonstrated promising potential for application in water purification systems.

Use of high-intensity ultrasound as a pre-treatment for complex coacervation from whey protein isolate and iota-carrageenan.

The aim of this work was to evaluate the influence of high intensity ultrasound (HIUS) treatment on the molecular conformation of whey protein isolated (WPI) as a previous step for complex coacervation with iota carrageenan (IC) and its effect on the surface functional properties of complex coacervates (CC). Both biopolymers were hydrated (1% w/w) separately. A WPI suspension was treated with an ultrasonic bath (40 kHz, 600 W, 30 and 60 min, 100% amplitude). A non-sonicated protein was used as a control. Coacervation was achieved by mixing WPI and IC dispersions (10 min). FTIR-ATR analysis (400-4000 cm-1) detected changes after sonication on WPI secondary structure (1600-1700 cm-1), electrostatic interaction between WPI and IC by electronegative IC charged groups like sulfate (1200-1260 cm-1), anhydrous oxygen of the 3.6 anhydro-D-galactose (940-1066 cm-1) and the electropositive regions of WPI. Rheology results showed pseudoplastic behavior of both IC and WPI-IC with a significant change in viscosity level. Further, HIUS treatment had a positive effect on the emulsifying properties of the WPI-IC coacervates, increasing the time foaming (30 min) and emulsion stability (1 month) percentage. HIUS and complex coacervation proved to be an efficient tool to improve the surface functional properties of WPI.

Layer-by-layer vaginal films for acyclovir controlled release to prevent genital herpes.

Genital herpes is one of the most common sexually transmitted infections worldwide. It mainly affects women, as the rate of sexual transmission from male-to-female is higher than from female-to-male. The application of vaginal antivirals drugs could reduce the prevalence of genital herpes and prevent future infections. Layer-by-layer vaginal films were prepared by the solvent evaporation method using iota-carrageenan, hydroxypropyl methylcellulose and the polymethacrylates Eudragit® RS PO and Eudragit® S100, for the controlled release of acyclovir. The films were characterized by texture analysis and Raman spectroscopy. Swelling, mucoadhesion, and drug release studies were conducted in simulated vaginal fluid. The results show that Layer-by-Layer films exhibited adequate mechanical properties. The structuring of the layer-by-layer films allowed the controlled release of acyclovir and produced a prolonged mucoadhesion residence time of up to 192 h. The films formed in layer 2 by the combination of Eudragit® RS PO and S100 showed a controlled release of acyclovir for eight days, and adequate mechanical properties. These promising formulations for the prevention of genital herpes deserve further evaluation.

Antimicrobial and cytocompatible chitosan, N,N,N-trimethyl chitosan, and tanfloc-based polyelectrolyte multilayers on gellan gum films.

In this work free-standing gels formed from gellan gum (GG) by solvent evaporation are coated with polysaccharide-based polyelectrolyte multilayers, using the layer-by-layer approach. We show that PEMs composed of iota-carrageenan (CAR) and three different natural polycationic polymers have composition-dependent antimicrobial properties, and support mammalian cell growth. Cationic polymers (chitosan (CHT), N,N,N-trimethyl chitosan (TMC), and an amino-functionalized tannin derivative (TN)) are individually assembled with the anionic iota-carrageenan (CAR) at pH 5.0. PEMs (15-layers) are alternately deposited on the GG film. The GG film and coated GG films with PEMs are characterized by infrared spectroscopy with attenuated total reflectance (FTIR-ATR), X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), and water contact angle (WCA) measurements. The TN/CAR coating provides a hydrophobic (WCA = 127°) and rough surface (Rq = 243 ± 48 nm), and the TMC/CAR coating provides a hydrophilic surface (WCA = 78°) with the lowest roughness (Rq = 97 ± 12 nm). Polymer coatings promote stability and durability of the GG film, and introduce antimicrobial properties against Gram-negative (Salmonella enteritidis) and Gram-positive (Staphylococcus aureus) bacteria. The films are also cytocompatible. Therefore, they have properties that can be further developed as wound dressings and food packaging.

Review of the use of nasal and oral antiseptics during a global pandemic.

A review of nasal sprays and gargles with antiviral properties suggests that a number of commonly used antiseptics including povidone-iodine, Listerine®, iota-carrageenan and chlorhexidine should be studied in clinical trials to mitigate both the progression and transmission of SARS-CoV-2. Several of these antiseptics have demonstrated the ability to cut the viral load of SARS-CoV-2 by 3-4 log10 in 15-30 s in vitro. In addition, hypertonic saline targets viral replication by increasing hypochlorous acid inside the cell. A number of clinical trials are in process to study these interventions both for prevention of transmission, prophylaxis after exposure, and to diminish progression by reduction of viral load in the early stages of infection.

Optimized acid hydrolysis of the polysaccharides from the seaweed Solieria filiformis (Kützing) P.W. Gabrielson for bioethanol production / Hidrólise ácida otimizada dos polissacarídeos da macroalga Solieria filiformis (Kützing) P.W. Gabrielson para produção de bioetanol

The seaweeds are bio-resource rich in sulfated and neutral polysaccharides. The tropical seaweed species used in this study (Solieria filiformis), after dried, shows 65.8% (w/w) carbohydrate, 9.6% (w/w) protein, 1.7% (w/w) lipid, 7.0% (w/w) moisture and 15.9% (w/w) ash. The dried seaweed was easily hydrolyzed under mild conditions (0.5 M sulfuric acid, 20 min.), generating fermentable monosaccharides with a maximum hydrolysis efficiency of 63.21%. Galactose and glucose present in the hydrolyzed were simultaneously fermented by Saccharomyces cerevisiae when the yeast was acclimated to galactose and cultivated in broth containing only galactose. The kinetic parameters of the fermentation of the seaweed hydrolyzed were Y(P/S) = 0.48 ± 0.02 g.g−1, PP = 0.27 ± 0.04 g.L−1.h−1, η = 94.1%, representing a 41% increase in bioethanol productivity. Therefore, S. filiformis was a promising renewable resource of polysaccharides easily hydrolyzed, generating a broth rich in fermentable monosaccharides for ethanol production.

As algas marinhas são recursos naturais ricos em polissacarídeos sulfatados e neutros. A espécie de macroalga tropical utilizada neste estudo (Solieria filiformis) apresentou teores de carboidratos de 65,8% (m/m), proteínas de 9,6% (m/m), lipídios de 1,7% (m/m), umidade de 7,0% (m/m) e 15,9 % (m/m) de cinzas. A macroalga seca foi facilmente hidrolisada em condições brandas, na presença de ácido sulfúrico 0,5 M, por 20 min, produzindo monossacarídeos fermentáveis com uma eficiência de hidrólise máxima de 63,21%. A galactose e a glicose presentes no hidrolisado foram fermentadas simultaneamente por Saccharomyces cerevisiae, após aclimatação da levedura cultivada em meio contendo apenas galactose como fonte de carbono. Os parâmetros cinéticos da fermentação do hidrolisado algáceo pela levedura aclimatada a galactose foram Y(P/S) = 0,48 ± 0,02 g.g-1, PP = 0,27 ± 0,04 g.L- 1.h-1, η = 94,1%. Portanto, a macroalga S. filiformis se mostrou um recurso renovável promissor como fonte de polissacarídeos facilmente hidrolisados, gerando um meio nutritivo rico em glucose e galactose para a produção de etanol.