Generally Regarded As Safe Research Articles

Possible Use of Ganoderma Mushroom Extracts for the Treatment of Some Chronic Risk Factors Associated with Severe Covid-19

The novel coronavirus (SARS-CoV-2) emerged in Wuhan, China in December 2019 causing the coronavirus disease 2019 (COVID-19). The disease has caused a pandemic that had disrupted social and economic life globally. Intense research is ongoing to understand the nature of the virus and pathogenesis as well as the development of drugs and vaccines. Ample evidence suggests that the elderly people, those with pre-existing chronic medical conditions, such as lung disease, heart disease, diabetes, obesity, chronic kidney disease, compromised immunity and even men may be at higher risk of serious illness and intensive care. The virus tends to trigger the over-production of pro-inflammatory cytokines, which results in multiorgan damage, resulting in death. Following this review, we proposed the use of Ganoderma mushroom extracts, which have certain properties relevant to the management of COVID-19 complications including anti-inflammatory, antiviral, immune modulatory, and antioxidant, anti-aging and multiorgan protection. Most of this research has been done on rodent models with few clinical trials. Since, the mushroom is generally regarded as safe (GRAS) and have been used for centuries in Asian traditional medicine, we recommend the use of this mushroom in the management of the disease including as active ingredients in COVID-19 drugs, and alternatives to synthetic drugs, health supplements, adjuvant therapy or functional foods.

Salcaprozate sodium (SNAC) enhances permeability of octreotide across isolated rat and human intestinal epithelial mucosae in Ussing chambers

Octreotide is approved as a one-month injectable for treatment of acromegaly and neuroendocrine tumours. Oral delivery of the octapeptide is a challenge due mainly to low intestinal epithelial permeability. The intestinal permeation enhancer (PE) salcaprozate sodium (SNAC) has Generally Regarded As Safe (GRAS) status and is a component of an approved oral peptide formulation. The purpose of the study was to examine the capacity of salcaprozate sodium (SNAC), to increase its permeability across isolated rat intestinal mucosae from five regions and across human colonic mucosae mounted in Ussing chambers. Apical-side buffers were Kreb's-Henseleit (KH), fasted simulated intestinal fluid (FaSSIF-V2), rat simulated intestinal fluid (rSIF), and colonic simulated intestinal fluid (FaSSCoF). The basal apparent permeability coefficient (Papp) of [3H]-octreotide was equally low across rat intestinal regional mucosae in KH, rSIF, and FaSSIF-V2. Apical addition of 20 mM SNAC increased the Papp across rat tissue in KH: colon (by 3.2-fold) > ileum (3.4-fold) > upper jejunum (2.3-fold) > duodenum (1.4-fold) > stomach (1.4-fold). 20 mM and 40 mM SNAC also increased the Papp by 1.5-fold and 2.1-fold respectively across human colonic mucosae in KH. Transepithelial electrical resistance (TEER) values were reduced in the presence in SNAC especially in colonic regions. LC-MS/MS analysis of permeated unlabelled octreotide across human colonic mucosae in the presence of SNAC indicated that [3H]-octreotide remained intact. No gross damage was caused to rat or human mucosae by SNAC. Attenuation of the effects of SNAC was seen in rat jejunal mucosae incubated with FaSSIF-V2 and rSIF, and also to some extent in human colonic mucosae using FaSSCoF, suggesting interaction between SNAC with buffer components. In conclusion, SNAC showed potential as an intestinal permeation enhancer for octreotide, but in vivo efficacy may be attenuated by interactions with GI luminal fluid contents.

Post‐harvest quality of sunflower microgreens as influenced by organic acids and ethanol treatment

Sunflower microgreens are nutrient-dense food requiring minimal input but are highly perishable in nature. Low cost, generally regarded as safe (GRAS) chemical treatments (ethanol vapor [EV], citric acid [CA], ascorbic acid [AA], CA + ethanol [CA + E] and CA + AA, distilled water as a control) and packaging in polystyrene trays at 10 ± 1°C was used to extend their shelf life up to 16 days. Electrical conductivity decreased till 4th day and then increased from 4th to 16th day. Control followed by CA + AA had least electrical conductivity on 16th day. CA + AA followed by control and AA retained highest amount of total chlorophylls and carotenoids, respectively, whereas, a maximum decline was observed in EV and CA + E. AA. The treatments CA + AA, CA, and AA retained maximum amount of ascorbic acid, total phenols, and DPPH scavenging activity with least microbial load at the end of storage, respectively, whereas EV and CA + E exhibited a maximum decline. AA and CA + AA treatments were most effective in enhancing the shelf life of sunflower microgreens. Practical applications Microgreens are nutrient-dense functional crops that are consumed fresh. They have a short growing span, do not require much inputs for growth, and have high nutrient composition than their mature counterparts. They have a high seed to biomass ratio and are microbiologically safe when grown indoors under a controlled environment. The current food system revolves around the consumption of fresh fully developed vegetables from fields. Microgreens can transform this and shift the focus to a new product that can be grown in urban setups, even homes, with minimal input and maximum output. It is quick and delivers healthy produce that does not have to be transported to great distances to be consumed. It saves transportation costs, reduce wastage, and spoilage that might take place in the transport chain and brings nutrient-dense food to the masses. The actual price of the food will also be less due to reduced cost of labor, infrastructure, and transport.

Lutein Supplementation for Eye Diseases.

Lutein is one of the few xanthophyll carotenoids that is found in high concentration in the macula of human retina. As de novo synthesis of lutein within the human body is impossible, lutein can only be obtained from diet. It is a natural substance abundant in egg yolk and dark green leafy vegetables. Many basic and clinical studies have reported lutein’s anti-oxidative and anti-inflammatory properties in the eye, suggesting its beneficial effects on protection and alleviation of ocular diseases such as age-related macular degeneration, diabetic retinopathy, retinopathy of prematurity, myopia, and cataract. Most importantly, lutein is categorized as Generally Regarded as Safe (GRAS), posing minimal side-effects upon long term consumption. In this review, we will discuss the chemical structure and properties of lutein as well as its application and safety as a nutritional supplement. Finally, the effects of lutein consumption on the aforementioned eye diseases will be reviewed.

Functional production of human antibody by the filamentous fungus Aspergillus oryzae

BackgroundMonoclonal antibodies (mAbs) as biopharmaceuticals take a pivotal role in the current therapeutic applications. Generally mammalian cell lines, such as those derived from Chinese hamster ovaries (CHO), are used to produce the recombinant antibody. However, there are still concerns about the high cost and the risk of pathogenic contamination when using mammalian cells. Aspergillus oryzae, a filamentous fungus recognized as a GRAS (Generally Regarded As Safe) organism, has an ability to secrete a large amount of proteins into the culture supernatant, and thus the fungus has been used as one of the cost-effective microbial hosts for heterologous protein production. Pursuing this strategy the human anti-TNFα antibody adalimumab, one of the world’s best-selling antibodies for the treatment of immune-mediated inflammatory diseases including rheumatoid arthritis, was chosen to produce the full length of mAbs by A. oryzae. Generally, N-glycosylation of the antibody affects immune effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC) via binding to the Fc receptor (FcγR) on immune cells. The CRISPR/Cas9 system was used to first delete the Aooch1 gene encoding a key enzyme for the hyper-mannosylation process in fungi to investigate the binding ability of antibody with FcγRIIIa.ResultsAdalimumab was expressed in A. oryzae by the fusion protein system with α-amylase AmyB. The full-length adalimumab consisting of two heavy and two light chains was successfully produced in the culture supernatants. Among the producing strains, the highest amount of antibody was obtained from the ten-protease deletion strain (39.7 mg/L). Two-step purifications by Protein A and size-exclusion chromatography were applied to obtain the high purity sample for further analysis. The antigen-binding and TNFα neutralizing activities of the adalimumab produced by A. oryzae were comparable with those of a commercial product Humira®. No apparent binding with the FcγRIIIa was detected with the recombinant adalimumab even by altering the N-glycan structure using the Aooch1 deletion strain, which suggests only a little additional activity of immune effector functions.ConclusionThese results demonstrated an alternative low-cost platform for human antibody production by using A. oryzae, possibly offering a reasonable expenditure for patient’s welfare.

Maturation of dendritic cells derived from type 1 diabetes-susceptible NOD mice is differentially regulated by two strains of generally regarded as safe (GRAS) Lactobacillus

Abstract Although commensal bacteria have been implicated in type 1 diabetes (T1D), specific mechanistic insight is limited. We hypothesized that specific strains of commensal bacteria can modulate the diabetogenic peripheral T cell effector repertoire (PTER) through differential dendritic cell (DC) maturation. To test this, we assessed the presence of an IL17+ PTER and compared the microbial profiles present within female non-obese diabetic (NOD) mice, which spontaneously develop TID, and non-obese resistant (NOR) mice, which are resistant to T1D. We also assessed changes in DC maturation mediated by generally regarded as safe (GRAS) Lactobacillus johnsonii N6.2 (LjN6.2) versus Lactobacillus reuteri TD1 (LrTD1), which were previously shown to inhibit and accelerate T1D, respectively, in bio-breeding diabetes-prone (BBDP) rats. We found that NOR mice possessed a distinct Th17 bias and higher CD11c+MHCII+ DCs frequencies within lymph nodes and pancreas when compared to NOD mice. In vitro, LjN6.2 induced production of IL-6, IL-10, and TGF-b, previously shown to prime protective IL17+ T lymphocytes. Additionally, LjN6.2 promoted dendritic cell expression of the tolerogenic signaling molecules SOCS1, B7-H4, and PD-L1. In contrast, LrTD1 promoted production of proinflammatory cytokines, IL-12 and IL-23, which share a common p40 subunit. Together these data suggest that certain bacterial strains may enhance tolerance mechanisms and, thereby, contribute toward an environment that might protect against T1D, whereas bacterial strains such as LrTD1 may act as an “inducing” environmental factor by enhancing IL-12 production. Hence, these results emphasize a need to elucidate immunological mechanisms mediated by GRAS bacteria.

Systems metabolic engineering of Bacillus subtilis for efficient biosynthesis of 5-methyltetrahydrofolate.

5-Methyltetrahydrofolate (5-MTHF) is the major form of folate in human plasma and is the only folate form that can penetrate the blood-brain barrier. It has been widely used for the prevention and treatment of various diseases. It is mainly produced by chemical synthesis. However, the low production rate cannot meet the increasing demand. In addition, chemical synthesis is potentially detrimental to the environment. Despite various microorganisms synthetizing 5-MTHF, an efficient 5-MTHF bioproduction approach is lacking because of the tight regulation of the 5-MTHF pathway and limited metabolic flux toward the folic acid pathway. In this study, the 5-MTHF synthetic pathway in Bacillus subtilis was systematically engineered to realize 5-MTHF accumulation and further improve 5-MTHF production. Specifically, the 5-MTHF synthesis pathway with dihydrofolate (DHF) as the precursor was strengthened to shift the metabolic flux to 5-MTHF biosynthesis by replacing the native yitJ gene with Escherichia coli metF, knockout of purU, and overexpressing dfrA. The intracellular level of 5-MTHF increased 26.4-fold, reaching 271.64 µg/L. Next, the 5-MTHF precursor supply pathway was strengthened by co-overexpression of folC, pabB, folE, and yciA. This resulted in a 93.2-fold improvement of the 5-MTHF titer, which reached 960.27 µg/L. Finally, the clustered regularly interspaced short palindromic repeats interference system was used to identify key genes in the competitive and catabolic pathways for repression to further shift the metabolic flux toward 5-MTHF biosynthesis. The repression of genes thyA (existing in the purine metabolic pathway), pheA (existing in the competitive metabolic pathway), trpE (existing in the competitive metabolic pathway), and panB (existing in the pantoate synthesis pathway) significantly increased the titer of 5-MTHF. By repressing the pheA gene, the 5-MTHF titer reached 1.58 mg/L, which was 153.8-fold that of the wild-type strain of B. subtilis 168. Through medium optimization, the 5-MTHF titer reached 1.78 mg/L, which was currently the highest titer of 5-MTHF in B. subtilis. Apart from the highest titer of 5-MTHF, the highest titer of total folates including 5-MTHF, 5-FTHF, folic acid, and THF could reach 3.31 mg/L, which was 8.5-fold that in B. subtilis. To the best of our knowledge, the 5-MTHF and total folate titers reported here are the highest using a Generally regarded as safe (GRAS) bacterium as the production host. Overall, this study provides a good starting point for further metabolic engineering to achieve efficient biosynthesis of 5-MTHF by GRAS bacteria.

The development of peptide‐coated nanoparticles for targeting and drug delivery to cancer cells

Ovarian Cancer (OC) is a most lethal female malignancy worldwide, mostly due to the late‐stage diagnosis and high recurrence rate. Survival statistics remain stuck at ~30% for the past four decades highlighting the necessity to improve the early stage detection and therapeutic outcomes. Nanotherapy in OC may provide significant and unique benefits for drug efficacy as well as for reducing the therapeutically limiting adverse effects associated with current treatment regimens. Herein, we report a unique nanoparticle (NP)‐based platform of peptide‐stabilized poly (lactic‐co‐glycolic acid) (PLGA). The ultimate goal is NP‐mediated targeted delivery of anticancer therapy and contrast agents. Morphology and small size (~100 nm) of NPs provide them an ideal candidate for mechanistic studies. We present detailed imaging studies of NPs uptake by OC cells as well as NPs distribution within cellular organelles obtained with high resolution live cell imaging. The results show a preferential accumulation of NPs in mitochondria. In addition, the in vitro studies include the NP‐based delivery of anticancer agents, such as Pt(II) and mitochondria‐targeting lonidamine. This project demonstrates potential of peptide‐stabilized nanocarrier as an effective nanomedicine against cancer. The PLGA is FDA approved and the amino acids of the peptides have Generally Regarded as Safe (GRAS) status. The NP platform is completely biodegradable, and as we demonstrate biologically active.

Eukaryotic Probiotic Saccharomyces boulardii Application in Clinical Trails: A Review

Probiotics are live microbes that assist in restoring the symbiotic intestinal gut flora balance, and thus, bestow health benefits to the host. The most commonly used human probiotics are members of the Lactobacillus and Bifidobacterium species. Besides these bacteria, Saccharomyces boulardii, a yeast strain, is also widely used as a probiotic to treat a variety of conditions, including antibiotics-associated diarrhea and recurrent Clostridium difficile infection. A primary advantage of using Saccharomyces boulardii as a probiotic is that it can be used by patients undergoing antibiotic regimen due to its natural resistance to antibiotics. The genetic transfer of antibiotic resistance genes, a frequent event between pathogenic and gastrointestinal tract (GIT) bacteria, is not as frequent between yeast and bacteria. Furthermore, Saccharomyces boulardii is also tolerant to various local stresses such as the presence of gastrointestinal (GI) enzymes, bile salts, organic acids, etc. and can withstand considerable variations in pH and temperature while transiting through the human GIT. Saccharomyces boulardii is known as generally regarded as safe (GRAS)with probiotic activity against a wide range of pathogens. This yeast is often marketed in a lyophilized form, “S. boulardii lyo”, complete sequence of the genome was performed, and a comparative analysis of the genome was performed. From clinical studies, the results of randomized controlled trials in patients repeatedly confirmed the significant positive effect in the treatment of acute and chronic diseases.

The Fabrication and in vitro Evaluation of Retinoic Acid-Loaded Electrospun Composite Biomaterials for Tracheal Tissue Regeneration.

Although relatively rare, major trauma to the tracheal region of the airways poses a significant clinical challenge with few effective treatments. Bioengineering and regenerative medicine strategies have the potential to create biocompatible, implantable biomaterial scaffolds, with the capacity to restore lost tissue with functional neo-trachea. The main goal of this study was to develop a nanofibrous polycaprolactone-chitosan (PCL-Chitosan) scaffold loaded with a signaling molecule, all-trans retinoic acid (atRA), as a novel biomaterial approach for tracheal tissue engineering. Using the Spraybase® electrospinning platform, polymer concentration, solvent selection, and instrument parameters were optimized to yield a co-polymer with nanofibers of 181–197 nm in diameter that mimicked tracheobronchial tissue architecture. Thereafter, scaffolds were assessed for their biocompatibility and capacity to induce mucociliary functionalization using the Calu-3 cell line. PCL-Chitosan scaffolds were found to be biocompatible in nature and support Calu-3 cell viability over a 14 day time period. Additionally, the inclusion of atRA did not compromise Calu-3 cell viability, while still achieving an efficient encapsulation of the signaling molecule over a range of atRA concentrations. atRA release from scaffolds led to an increase in mucociliary gene expression at high scaffold loading doses, with augmented MUC5AC and FOXJ1 detected by RT-PCR. Overall, this scaffold integrates a synthetic polymer that has been used in human tracheal stents, a natural polymer generally regarded as safe (GRAS), and a drug with decades of use in patients. Coupled with the scalable nature of electrospinning as a fabrication method, all of these characteristics make the biomaterial outlined in this study amenable as an implantable device for an unmet clinical need in tracheal replacement.

Purification, biochemical and secondary structural characterisation of β-mannanase from Lactobacillus casei HDS-01 and juice clarification potential

A 36.7 kDa extracellular mannanase was originally purified from a generally regarded as safe (GRAS) lactic acid bacteria (LAB) strain Lactobacillus casei HDS-01. The LC-GC/GC determined that HDS-01 mannanase contained 338 amino acid residues. This protein belonged to glycoside hydrolase (GH) 26 Family and shared high similarity (94%) but low coverage (54%) with mannanase from Bacillus licheniformis (GQ859489.1). Circular dichroism (CD) illustrated that the proportion of α + β structural elements closely related to enzyme activity and were both negatively influenced by temperature. Fourier-transform infrared (FT-IR) showed the existence of typical protein bonds such as CO, CC, NH, CCH and CC. The CD and FT-IR collectively suggested HDS-01 mannanase a β-conformation rich (60–70%) protein. The optimal reaction condition of HDS-01 mannanase was 40 °C and pH 5.0. More than 50% activity was maintained after 2 h incubation under the condition of pH value 5.0–7.0 and 30–60 °C. This enzyme was strongly activated by Mn2+ and exhibited a high affinity for locust bean gum (Km = 2.68 ± 0.02 mg mL−1, Vmax = 400.03 ± 1.22 μmol mL−1·min−1). The HDS-01 mannanase clarified fruit juices more efficiently than commercial mannanase. These results promised HDS-01 mannanase a bio-safe addictive used in various especially food grade industrial fields.

Metabolic Engineering of Bacillus licheniformis for Production of Acetoin.

Acetoin is a potential platform compound for a variety of chemicals. Bacillus licheniformis MW3, a thermophilic and generally regarded as safe (GRAS) microorganism, can produce 2,3-butanediol with a high concentration, yield, and productivity. In this study, B. licheniformis MW3 was metabolic engineered for acetoin production. After deleting two 2,3-butanediol dehydrogenases encoding genes budC and gdh, an engineered strain B. licheniformis MW3 (ΔbudCΔgdh) was constructed. Using fed-batch fermentation of B. licheniformis MW3 (ΔbudCΔgdh), 64.2 g/L acetoin was produced at a productivity of 2.378 g/[L h] and a yield of 0.412 g/g from 156 g/L glucose in 27 h. The fermentation process exhibited rather high productivity and yield of acetoin, indicating that B. licheniformis MW3 (ΔbudCΔgdh) might be a promising acetoin producer.

Evaluation of oil-in-water (O/W) emulsifying properties of galactan exopolysaccharide from Weissella confusa KR780676.

Galactan exopolysaccharide (EPS) was extracted from Weissella confusa KR780676 isolated from idli batter. The present study reports the effect of galactan EPS concentration, pH, ionic strength, temperature, salinity, monovalent salts on the emulsion formed with vegetable oils (virgincoconut oil, groundnut oil, olive oil and mustard oil). Emulsion determination test revealed the oil-in-water type of emulsion. Microscopic observation showed the phase of oil droplet distribution of emulsions. Groundnut and olive oil showed significant emulsifying activity (50-60%) and stability (90%) irrespective of the EPS concentration. One percent of EPS also showed good emulsifying activity (50-70%) and stability (100%) with groundnut and olive oil in wide range of temperature (- 20, 4, 30, 40, 50 and 60°C), pH (3-8), salinity (1, 5, 10, 20 and 30% of NaCl) and monovalent salt (1, 5, 10, 20 and 30% of KCl). Galactan could be used as a prominent emulsifier for applications in the food industry as it possesses significant activity with most of the influential factors in wide range and also obtained from lactic acid bacteria that are regarded as Generally Regarded as Safe (GRAS).

Pharmaceutical cocrystals of nebivolol hydrochloride with enhanced solubility

Abstract Nebivolol hydrochloride is an antihypertensive BCS class II drug with low solubility and high permeability prescribed as antihypertensive. Crystal engineering was applied to prepare novel cocrystal forms of nebivolol hydrochloride with general GRAS (Generally regarded as safe) molecules. Pharmaceutical cocrystals of nebivolol hydrochloride were prepared with GRAS molecules, such as 4-hydroxybenzoic acid and nicotinamide (NA), with different ratios like 1:1, 1:2, 1:3. Liquid assisted grinding and solvent evaporation method was used to prepare nebivolol hydrochloride cocrystals. All prepared crystalline forms were characterized by X-ray diffraction, differential scanning calorimetry, and FT-IR to confirm their purity and formation of cocrystals. X-ray crystal data provided details of hydrogen bonding and interactions between drug and co-formers and differential scanning calorimetry provided changes in thermal behaviour and formation of cocrystals. Physicochemical properties such as solubility and dissolution rate were measured and found that all the co-crystals dissolved faster and their equilibrium solubility is higher than the parent drug. Nebivolol hydrochloride - 4 - hydroxy benzoic acid cocrystals showed highest solubility among all crystalline forms.

Impact of Processing Parameters on the Quality of Pharmaceutical Solid Dosage Forms Produced by Fused Deposition Modeling (FDM).

Fused deposition modeling (FDM) three-dimensional (3D) printing is being increasingly explored as a direct manufacturing method to product pharmaceutical solid dosage forms. Despite its many advantages as a pharmaceutical formulation tool, it remains restricted to proof-of-concept formulations. The optimization of the printing process in order to achieve adequate precision and printing quality remains to be investigated. Demonstrating a thorough understanding of the process parameters of FDM and their impact on the quality of printed dosage forms is undoubtedly necessary should FDM advance from a proof-of-concept stage to an adapted pharmaceutical manufacturing tool. This article describes the findings of an investigation into a number of critical process parameters of FDM and their impact on quantifiable, pharmaceutically-relevant measures of quality. Polycaprolactone, one of the few polymers which is both suitable for FDM and is a GRAS (generally regarded as safe) material, was used to print internally-exposed grids, allowing examination of both their macroscopic and microstructural reproducibility of FDM. Of the measured quality parameters, dimensional authenticity of the grids was found to poorly match the target dimensions. Weights of the grids were found to significantly vary upon altering printing speed. Printing temperature showed little effect on weight. Weight uniformity per batch was found to lie within acceptable pharmaceutical quality limits. Furthermore, we report observing a microstructural distortion relating to printing temperature which we dub The First Layer Effect (FLE). Principal Component Analysis (PCA) was used to study factor interactions and revealed, among others, the existence of an interaction between weight/dosing accuracy and dimensional authenticity dictating a compromise between the two quality parameters. The Summed Standard Deviation (SSD) is proposed as a method to extract the optimum printing parameters given all the perceived quality parameters and the necessary compromises among them.

Experimental Cannabinoid 2 Receptor Activation by Phyto-Derived and Synthetic Cannabinoid Ligands in LPS-Induced Interstitial Cystitis in Mice.

Interstitial cystitis (IC) is a chronic bladder disorder with unclear etiology. The endocannabinoid system has been identified as a key regulator of immune function, with experimental evidence for the involvement of cannabinoid receptors in bladder inflammation. This study used intravital microscopy (IVM) and behavioral testing in lipopolysaccharide-induced IC, to investigate the anti-inflammatory analgesic effects of a natural dietary sesquiterpenoid, beta-caryophyllene (BCP), which is present in cannabis among other plants, and has reported agonist actions at the cannabinoid 2 receptor (CB2R). BCP’s anti-inflammatory actions were compared to the synthetic CB2R-selective cannabinoid, HU308, and to an FDA-approved clinical treatment (dimethyl sulfoxide: DMSO). IVM data revealed that intravesical instillation of BCP and/or HU308 significantly reduces the number of adhering leukocytes in submucosal bladder venules and improves bladder capillary perfusion. The effects of BCP were found to be comparable to that of the selective CB2R synthetic cannabinoid, HU308, and superior to intravesical DMSO treatment. Oral treatment with BCP was also able to reduce bladder inflammation and significantly reduced mechanical allodynia in experimental IC. Based on our findings, we believe that CB2R activation may represent a viable therapeutic target for IC, and that drugs that activate CB2R, such as the generally regarded as safe (GRAS) dietary sesquiterpenoid, BCP, may serve as an adjunct and/or alternative treatment option for alleviating symptoms of inflammation and pain in the management of IC.

L-serine: a neglected amino acid with a potential therapeutic role in diabetes.

L‐serine is classified as a non‐essential amino acid; however, L‐serine is indispensable having a central role in a broad range of cellular processes. Growing evidence suggests a role for L‐serine in the development of diabetes mellitus and its related complications, with L‐serine being positively correlated to insulin secretion and sensitivity. L‐serine metabolism is altered in type 1, type 2, and gestational diabetes, and L‐serine supplementations improve glucose homeostasis and mitochondrial function, and reduce neuronal death. Additionally, L‐serine lowers the incidence of autoimmune diabetes in NOD mice. Dietary supplementations of L‐serine are generally regarded as safe (GRAS) by the FDA. Therefore, we believe that L‐serine should be considered as an emerging therapeutic option in diabetes, although work remains in order to fully understand the role of L‐serine in diabetes.

Heterologous Hyaluronic Acid Production in Kluyveromyces lactis.

Hyaluronic Acid (HA) is a biopolymer composed by the monomers Glucuronic Acid (GlcUA) and N-Acetyl Glucosamine (GlcNAc). It has a broad range of applications in the field of medicine, being marketed between USD 1000–5000/kg. Its primary sources include extraction of animal tissue and fermentation using pathogenic bacteria. However, in both cases, extensive purification protocols are required to prevent toxin contamination. In this study, aiming at creating a safe HA producing microorganism, the generally regarded as safe (GRAS) yeast Kluyveroymyces lactis is utilized. Initially, the hasB (UDP-Glucose dehydrogenase) gene from Xenopus laevis (xlhasB) is inserted. After that, four strains are constructed harboring different hasA (HA Synthase) genes, three of humans (hshasA1, hshasA2, and hshasA3) and one with the bacteria Pasteurella multocida (pmhasA). Transcript values analysis confirms the presence of hasA genes only in three strains. HA production is verified by scanning electron microscopy in the strain containing the pmHAS isoform. The pmHAS strain is grown in a 1.3 l bioreactor operating in a batch mode, the maximum HA levels are 1.89 g/L with a molecular weight of 2.097 MDa. This is the first study that reports HA production in K. lactis and it has the highest HA titers reported among yeast.

Toxic and in vitro anti-acetylcholinesterase and anti-carboxylesterase effects of various plant extracts on Aphis gossypii Glover, 1877 (Hemiptera: Aphididae)

The significance of discovering new active substances that are environment friendly when compared to pesticides, agriculturally sustainable, plant-based, and in the status of GRAS (generally regarded as safe) has been increasing every day. For this purpose, leaves of Daphne odora L., Dieffenbachia amoena L., Eucalyptus camaldulensis L. , Ficus carica L. , Lantana camara L. , Matricaria chamomilla L., Mentha pulegium L. and Nerium oleander L. were collected from Adana in 2018. Toxic and in vitro anti-acetylcholinesterase (AChE) and anti-carboxylesterase (CE) activities of aqueous leaf extracts of these species on the important polyphagous pest, Aphis gossypii Glover, 1877 (Hemiptera: Aphididae), were determined after 24 and 72 h. The fastest and greatest toxic effect was obtained with 20% F. carica extract giving 75.6% mortality. This was followed by N. oleander with 71.6% and D. odora with 62.1% mortality. Aphis gossypii in vitro anti-AcHe and anti-CE activities were highest at 10% concentration of the plant extracts and inhibition levels were 51.8-82.5% with F. carica extract, 40.9-54.9% with D. odora extract and 40.2-82.5% with E. camaldulensis extract. In conclusion, D. odora, E. camaldulensis, F. carica and N. oleander extracts gave promising results for future studies on the discovery of potential xenobiotics against A. gossypii and for pest control.

The in vivo toxicological profile of cationic solid lipid nanoparticles.

Cationic solid lipid nanoparticles (cSLNs) are considered as one of the most effective lipid nanocarriers for delivery of low water-solubility compounds and genetic materials. As the excipients used in the cSLN production are generally regarded as safe (GRAS), the formulations are granted as non-toxic. However, the toxicological profile of new SLN-based formulations should always be performed to confirm that the delivery systems themselves may not impose risks to the human health. Therefore, in this study, we delineate the toxicological profile of the cSLN formulation at 24 and 72h after single intravenous injection to male Wistar rats. Hematological, biochemical, and histopathological evaluations of the spleen, lungs, liver, and kidneys indicated short-lived alterations including neutrophilia. We found increases in the population of macrophages in the lungs, liver, and spleen and also migration of circulating neutrophils into inflamed tissue and a decrease in blood urea nitrogen. We also observed the presence of cSLNs within the brain parenchyma without any sign of damage to the blood-brain barrier. These side effects appeared to be mild and transitory (< 72h). These findings reinforce the importance of investigating the toxicity of SLN-based formulations before the incorporation of drugs/genetic material to the formulation and its translation to the clinic.