L-AP Alleviates Liver Injury in Septic Mice by Inhibiting Macrophage Activation via Suppressing NF-κB and NLRP3 Inflammasome/Caspase-1 Signal Pathways.

Liver injury and progressive liver failure are severe life-threatening complications in sepsis, further worsening the disease and leading to death. Macrophages and their mediated inflammatory cytokine storm are critical regulators in the occurrence and progression of liver injury in sepsis, for which effective treatments are still lacking. l-Ascorbic acid 6-palmitate (L-AP), a food additive, can inhibit neuroinflammation by modulating the phenotype of the microglia, but its pharmacological action in septic liver damage has not been fully explored. We aimed to investigate L-AP's antisepticemia action and the possible pharmacological mechanisms in attenuating septic liver damage by modulating macrophage function. We observed that L-AP treatment significantly increased survival in cecal ligation and puncture-induced WT mice and attenuated hepatic inflammatory injury, including the histopathology of the liver tissues, hepatocyte apoptosis, and the liver enzyme levels in plasma, which were comparable to NLRP3-deficiency in septic mice. L-AP supplementation significantly attenuated the excessive inflammatory response in hepatic tissues of septic mice in vivo and in cultured macrophages challenged by both LPS and ATP in vitro, by reducing the levels of NLRP3, pro-IL-1ß, and pro-IL-18 mRNA expression, as well as the levels of proteins for p-I-κB-α, p-NF-κB-p65, NLRP3, cleaved-caspase-1, IL-1ß, and IL-18. Additionally, it impaired the inflammasome ASC spot activation and reduced the inflammatory factor contents, including IL-1ß and IL-18 in plasma/cultured superannuants. It also prevented the infiltration/migration of macrophages and their M1-like inflammatory polarization while improving their M2-like polarization. Overall, our findings revealed that L-AP protected against sepsis by reducing macrophage activation and inflammatory cytokine production by suppressing their activation in NF-κB and NLRP3 inflammasome signal pathways in septic liver.

The Vitamin C Enantiomers Possess a Comparable Potency in the Induction of Oxidative Stress in Cancer Cells but Differ in Their Toxicity.

The use of vitamin C (VC) in high doses demonstrates a potent tumor suppressive effect by mediating a glucose-dependent oxidative stress in Kirsten rat sarcoma (KRAS) mutant cancer cells. VC with arsenic trioxide (ATO) is a promising drug combination that might lead to the development of effective cancer therapeutics. Considering that a tumor suppressive effect of VC requires its high-dose administration, it is of interest to examine the toxicity of two enantiomers of VC (enantiomer d-optical isomer D-VC and natural l-optical isomer L-VC) in vitro and in vivo. We show that the combinations of L-VC with ATO and D-VC with ATO induced a similar cytotoxic oxidative stress in KrasG12D-expressing mutant cancer cells as indicated by a substantial increase in reactive oxidative species (ROS) production and depolarization of mitochondria. To examine the L-VC and D-VC toxicity effects, we administered high doses of D-VC and L-VC to CD1 mice and carried out an evaluation of their toxic effects. The daily injections of L-VC at a dose of 9.2 g/kg for 18 days were lethal to mice, while 80% of mice remained alive following the similar high-dose administration of D-VC. Following the drug injection courses and histopathological studies, we determined that a natural form of VC (L-VC) is more harmful and toxic to mice when compared to the effects caused by the similar doses of D-VC. Thus, our study indicates that the two enantiomers of VC have a similar potency in the induction of oxidative stress in cancer cells, but D-VC has a distinctive lower toxicity in mice compared to L-VC. While the mechanism of a distinctive toxicity between D-VC and L-VC is yet to be defined, our finding marks D-VC as a more preferable option compared to its natural enantiomer L-VC in clinical settings.

Exogenous 2-keto-L-gulonic Acid Supplementation as a Novel Approach to Enhancing L-ascorbic Acid Biosynthesis in Zebrafish (Danio rerio).

L-ascorbic acid (ASA) is a micronutrient that is essential for reproduction, growth, and immunity in animals. Due to the loss of enzyme L-gulono-1,4-lactone oxidase (GLO), most aquatic animals lack the capacity for ASA biosynthesis and therefore require supplementation with exogenous ASA. Recent studies have shown that 2-keto-L-gulonic acid (2KGA), a novel potential precursor of ASA, can enhance plant growth and improve stress resistance by promoting the synthesis and accumulation of ASA. Our hypothesis is that 2-keto-L-gulonic acid (2KGA) plays a similar role in aquatic animals. To investigate this, we conducted an in vivo trial to examine the effects of exogenous 2KGA supplementation on ASA metabolism and growth of zebrafish (Danio rerio). Zebrafish were categorized into groups based on their dietary intake, including a basal diet (CK group), a basal diet supplemented with 800 mg/kg ASA (ASA group), and 800 mg/kg 2KGA-Na (2KGA group) for a duration of three weeks. The results demonstrated a significant increase in ASA content in zebrafish treated with 2KGA (34.82% increase, p < 0.05) compared to the CK group, reaching a consistent level with the ASA group (39.61% increase, p < 0.05). Furthermore, the supplementation of 2KGA significantly improved growth parameters relevant to zebrafish (specific growth rate increased by 129.04%, p < 0.05) and enhanced feed utilization (feed intake increased by 15.65%, p < 0.05). Positive correlations were observed between growth parameters, feed utilization, whole-body chemical composition, and ASA content. Our findings suggest that supplementation with exogenous 2KGA can serve as a novel approach for elevating ASA synthesis in aquatic animals, and further investigation of its underlying mechanism is required.

Assessment of Phenolic Acid Content and Antioxidant Properties of the Pulp of Five Pumpkin Species Cultivated in Southeastern Poland.

Antioxidant properties and phenolic acid content in the pulp of five pumpkin species were evaluated. The following species cultivated in Poland were included: Cucurbita maxima 'Bambino', Cucurbita pepo 'Kamo Kamo', Cucurbita moschata 'Butternut', Cucurbita ficifolia 'Chilacayote Squash', and Cucurbita argyrosperma 'Chinese Alphabet'. The content of polyphenolic compounds was determined by ultra-high performance liquid chromatography coupled with HPLC, while the total content of phenols and flavonoids and antioxidant properties were determined by spectrophotometric methods. Ten phenolic compounds (protocatechuic acid, p-hydroxybenzoic acid, catechin, chlorogenic acid, caffeic acid, p-coumaric acid, syringic acid, ferulic acid, salicylic acid, kaempferol) were identified. Phenolic acids were the most abundant compounds; the amount of syringic acid was found to be the highest, ranging from 0.44 (C. ficifolia) to 6.61 mg∙100 g-1 FW (C. moschata). Moreover, two flavonoids were detected: catechin and kaempferol. They were found at their highest level of content in C. moschata pulp (catechins: 0.31 mg∙100 g-1 FW; kaempferol: 0.06 mg∙100 g-1 FW), with the lowest amount detected in C. ficifolia (catechins: 0.15 mg∙100 g-1 FW; kaempferol below the limit of detection). Analysis of antioxidant potential showed significant differences depending on the species and the test used. The DPPH radical scavenging activity of C. maxima was 1.03 times higher than C. ficiofilia pulp and 11.60 times higher than C. pepo. In the case of the FRAP assay, the multiplicity of FRAP radical activity in C. maxima pulp was 4.65 times higher than C. Pepo pulp and only 1.08 times higher compared to C. ficifolia pulp. The study findings show the high health-promoting value of pumpkin pulp; however, the content of phenolic acids and antioxidant properties are species dependent.

Membrane permeabilizers enhance biofortification of Brassica microgreens by interspecific transfer of metabolites from tea (Camellia sinensis).

Interspecific metabolite transfer (ISMT) is a novel approach for plants biofortification. In this study, the effect of tea (Camellia sinensis; Cs), with or without membrane permeabilizers EDTA and Tween, as a donor plant on broccoli, cauliflower and kale sprouts was investigated. As a result, caffeine- and catechin-enriched broccoli, cauliflower and kale microgreens were produced. Kale sprouts were most permeable for catechins from Cs, while cauliflower was most permeable for caffeine. Cs + EDTA significantly increased vitamin C in broccoli and kale. Among the tested enzymes activity, pancreatic lipase was the most affected by the treatment with broccoli and cauliflower biofortified with Cs or Cs combined with permeabilizers. Broccoli sprouts biofortified with Cs most significantly inhibited α-amylase, while those biofortified with Cs combined with permeabilizers most significantly inhibited α-glucosidase. Results point to ISMT combined with membrane permeabilizers as a promising and eco-friendly biofortification strategy to improve the biopotential of Brassica microgreens.

Multitraits evaluation of a Solanum pennellii introgression tomato line challenged by combined abiotic stress.

Rising daily temperatures and water shortage are two of the major concerns in agriculture. In this work, we analysed the tolerance traits in a tomato line carrying a small region of the Solanum pennellii wild genome (IL12-4-SL) when grown under prolonged conditions of single and combined high temperature and water stress. When exposed to stress, IL12-4-SL showed higher heat tolerance than the cultivated line M82 at morphological, physiological, and biochemical levels. Moreover, under stress IL12-4-SL produced more flowers than M82, also characterized by higher pollen viability. In both lines, water stress negatively affected photosynthesis more than heat alone, whereas the combined stress did not further exacerbate the negative impacts of drought on this trait. Despite an observed decrease in carbon fixation, the quantum yield of PSII linear electron transport in IL12-4-SL was not affected by stress, thereby indicating that photochemical processes other than CO2 fixation acted to maintain the electron chain in oxidized state and prevent photodamage. The ability of IL12-4-SL to tolerate abiotic stress was also related to the intrinsic ability of this line to accumulate ascorbic acid. The data collected in this study clearly indicate improved tolerance to single and combined abiotic stress for IL12-4-SL, making this line a promising one for cultivation in a climate scenario characterized by frequent and long-lasting heatwaves and low rainfall.

Novel methylated flavoalkaloids from Echa 1 green tea inhibit fat accumulation and enhance stress resistance in Caenorhabditis elegans.

Methylation is a common structural modification of catechins in tea, which can improve the bioavailability of catechins. Flavoalkaloids are catechin derivatives with a nitrogen containing five-membered ring at the C-6 or C-8 position. Here we isolated three new methylated flavoalkaloids from Echa 1 green tea (Camellia sinensis cv. Echa 1) and synthesized another four new methylated flavoalkaloids. The structures of the new ester-type methylated catechins (etmc)-pyrrolidinone A-G (1-7) were elucidated by various spectroscopic techniques, including nuclear magnetic resonance (NMR), optical rotation, infrared, UV-vis, experimental and calculated circular dichroism (CD) spectra, and high-resolution mass. Among them, 6 and 7 showed the strongest α-glucosidase inhibitory activity and significantly lowered lipid content of Caenorhabditis elegans with 73.50 and 67.39% inhibition rate, respectively. Meanwhile, 6 and 7 also exhibited strong antioxidant activity in vitro and stress resistance to heat, oxidative stress, and UV irradiation in nematodes.

Influences of l-ascorbic acid on cytotoxic, biochemical, and genotoxic damages caused by copper II oxide nanoparticles in the rainbow trout gonad cells-2.

In parallel with the raising use of copper oxide nanoparticles (CuO NPs) in various industrial and commercial practices, scientific reports on their release to the environment and toxicity are increasing. The toxicity of CuO NPs is mostly based on their oxidative stress. Therefore, it is necessary to investigate the efficacy of well-known therapeutic agents as antioxidants against CuO NPs damage. This study aimed to investigate the mechanism of this damage and to display whether l-ascorbic acid could preserve against the cell toxicities induced by CuO NPs in the rainbow trout gonad cells-2 (RTG-2). While CuO NPs treatment significantly diminished cell viability, the l-ascorbic acid supplement reversed this. l-ascorbic acid treatment reversed the changes in expressions of sod1, sod2, gpx1a, and gpx4b genes while playing a supportive role in the changes in the expression of the cat gene induced by CuO NPs treatment. Moreover, CuO NPs treatment caused an upregulation in the expressions of growth-related genes (gh1, igf1, and igf2) and l-ascorbic acid treatment further increased these effects. CuO NPs treatment significantly up-regulated the expression of the gapdh gene (glycolytic enzyme gene) compared to the control group, and l-ascorbic acid treatment significantly down-regulated the expression of the gapdh gene compared to CuO NPs treatment. The genotoxicity test demonstrated that l-ascorbic acid treatment increased the genotoxic effect caused by CuO NPs by acting as a co-mutagen. Based on the findings, l-ascorbic acid has the potential to be sometimes inhibitory and sometimes supportive of cellular mechanisms caused by CuO NPs.

Pleiotropic Functions of Nitric Oxide Produced by Ascorbate for the Prevention and Mitigation of COVID-19: A Revaluation of Pauling's Vitamin C Therapy.

Linus Pauling, who was awarded the Nobel Prize in Chemistry, suggested that a high dose of vitamin C (l-ascorbic acid) might work as a prevention or treatment for the common cold. Vitamin C therapy was tested in clinical trials, but clear evidence was not found at that time. Although Pauling's proposal has been strongly criticized for a long time, vitamin C therapy has continued to be tested as a treatment for a variety of diseases, including coronavirus infectious disease 2019 (COVID-19). The pathogen of COVID-19, SARS-CoV-2, belongs to the ß-coronavirus lineage, which includes human coronavirus, severe acute respiratory syndrome (SARS), and Middle East respiratory syndrome (MERS). This review intends to shed new light on vitamin C antiviral activity that may prevent SARS-CoV-2 infection through the chemical production of nitric oxide (NO). NO is a gaseous free radical that is largely produced by the enzyme NO synthase (NOS) in cells. NO produced by upper epidermal cells contributes to the inactivation of viruses and bacteria contained in air or aerosols. In addition to enzymatic production, NO can be generated by the chemical reduction of inorganic nitrite (NO2-), an alternative mechanism for NO production in living organisms. Dietary vitamin C, largely contained in fruits and vegetables, can reduce the nitrite in saliva to produce NO in the oral cavity when chewing foods. In the stomach, salivary nitrite can also be reduced to NO by vitamin C secreted from the epidermal cells of the stomach. The strong acidic pH of gastric juice facilitates the chemical reduction of salivary nitrite to produce NO. Vitamin C contributes in multiple ways to the host innate immune system as a first-line defense mechanism against pathogens. Highlighting chemical NO production by vitamin C, we suggest that controversies on the therapeutic effects of vitamin C in previous clinical trials may partly be due to less appreciation of the pleiotropic functions of vitamin C as a universal bioreductant.

L-Ascorbic acid and phosphatidylcholine complex vesicles: formation and elucidation of their biological activities, and their molecular interactions.

The aim is to prepare, characterise, and evaluate the biological activities and key molecular interactions of L-ascorbic acid and phosphatidylcholine (PC-AA) complex vesicles. PC-AA complexes were prepared and characterised using DLS, TEM, FTIR, UV-Vis, in-vitro release, bioactivities, and cytotoxicity. The key interactions of the AA with the PC were studied with MD simulations. PC-AA complex provides improved stability towards the degradation of AA in aqueous solutions while also slowing its release profile. The PC-AA complexes with an optimal molar ratio of PC: AA = 2.5:1 was shown to have a hydrodynamic diameter of 368.67 ± 4.65 nm and an EE of 68.16 ± 0.23%. At low concentration, the PC-AA complexes have no toxicity towards human dermal fibroblast cells over 48 h. Importantly, MD suggests that AA only forms the PC-AA complex when in its neutral form which is the desired active form. PC-AA complex could be a potential active to use in medicinal and cosmeceutical applications.

Effects of the In Ovo and Dietary Supplementation of L-Ascorbic Acid on the Growth Performance, Inflammatory Response, and Eye L-Ascorbic Acid Concentrations in Ross 708 Broiler Chickens.

Effects of the dietary and in ovo administration of L-ascorbic acid (L-AA) on the performance, plasma nitric oxide, and eye L-AA concentrations of Ross 708 broilers were investigated. At 17 days of incubation, live embryonated hatching eggs were randomly assigned to a non-injected or sham-injected (100 µL of saline) control group, or a group injected with either 12 or 25 mg of L-AA suspended in 100 µL of saline. Chicks received a commercial diet with or without 200 mg/kg of supplemental L-AA and were randomly assigned to each of 6 replicate floor pens in each in ovo injection-dietary treatment combination. Weekly live performance variables through 14 days of post hatch age (doa) and the eye weights in both sexes at 0, 7, and 14 doa were determined. At 0 and 14 doa, plasma nitric oxide levels and eye L-AA concentrations of one bird of each sex in each pen were determined. Dietary supplemental L-AA decreased feed intake and growth between 0 and 7 doa, but from 8 to 14 doa; all birds fed supplemental L-AA had a lower feed conversion ratio. At 14 doa, male chicks had higher eye L-AA concentrations and lower plasma nitric oxide levels when treated in ovo with 12 mg of L-AA. In conclusion, dietary L-AA may be used to improve feed conversion in the second week of broiler post hatch growth. However, the in ovo administration of 12 mg of L-AA can increase male eye L-AA concentrations and is effective in reducing their general inflammatory response.

Biogenic Nanocopper: Eco-Friendly Synthesis, Characterization, and Their Anti-Trichomonas vaginalis, Anticancer, and Antimicrobial Effects.

Biogenic nanocopper (BNC) agents exhibit strong anticancer, antimicrobial, and antiparasitic effects. Their fewer side effects to normal cells cause them to be preferred to treat various diseases. Metal nanoparticles, particularly copper nanoparticles, are attracting more significant interest as therapeutic agents with the improvement of green synthesis methods. Studies to reduce the side effects of copper nanoparticles to exhibit strong pharmacological properties are progressing intensively. Here, BNCs with reduced side effects were synthesized using L-ascorbic acid as the reducing agent and various concentrations of copper (II) chloride. BNCs exhibited significant pharmacological activity on cancer, bacteria, and Trichomonas vaginalis cells. The newly synthesized BNCs were characterized by scanning electron microscopy, UV-Vis spectrophotometry, Fourier transform infrared spectroscope, and Differential/Thermal Gravimetric Analysis. The pharmacological activity of BNCs was evaluated by obtaining their inhibitor concentration and minimum inhibitory concentrations against some cancer, bacteria, and T. vaginalis cells. Newly synthesized BNCs have various shapes such as cubic, spherical, or rod and particle size distribution between 70 and 100 nm. According to experiment results, the newly synthesized BNCs were a significantly antiproliferative, antibacterial, and anti-T. vaginalis effect on cells compared to the control drugs. These findings confirm newly synthesized BNCs and their in vitro pharmacological potential. Further research should be targeted on the preclinical study of absorption, distribution, metabolism, excretion/toxicity (ADME/Tox) and in vivo effects on cancer and microbial pathogens.

Effects of the In Ovo Injection of L-Ascorbic Acid on Broiler Hatching Performance.

Effects of the in ovo injection of various concentrations of L-ascorbic acid (L-AA) on the hatchability and retention levels of L-AA in the serum of broiler embryos were investigated. A total of 960 Ross 708 broilers hatching eggs were randomly divided into four treatment groups: non-injected control, saline-injected control, and saline containing either 12 or 25 mg of L-AA. At 18 days of incubation (doi), injected eggs received a 100 µL volume of sterile saline (0.85%) alone or containing one of the two L-AA levels. Percentage egg weight loss was also determined from 0 to 12 and 12 to 18 doi. Hatch residue analysis was conducted after candling to determine the staging of embryo mortality. At approximately 21 doi, hatchability of live embryonated eggs (HI) and hatchling body weight (BW) were determined. Blood samples were taken at 6 and 24 h after L-AA in ovo injection to determine serum L-AA concentrations. Serum L-AA concentrations, HI, and hatchling BW did not differ among all treatment groups. However, chicks in the non-injected group had a higher (p = 0.05) embryonic mortality at hatch in comparison to those in the 12 mg of L-AA in saline and saline alone treatment groups. These results suggest that the in ovo injection of high levels of L-AA (12 and 25 mg) does not negatively affect HI or serum concentrations of L-AA but has the potential to promote embryonic livability. Further research is needed to determine the retention time of L-AA in the other tissues of broilers, including the cornea of the eye, in response to different levels of supplemental L-AA.

Spermatozoa Survival in Egg Yolk-Based and Soybean-Based Extenders at Ambient and Chilling Temperature in Domestic Turkeys (Meleagris gallopavo).

Populations of many galliform species have declined mainly due to habitat loss and over-hunting, notably the Congo peacock, which has been classified as a vulnerable species by the International Union for Conservation of Nature (IUCN). The domestic turkey, being a species of least concern, which has been reported to be closely related to peacocks, could serve as a model for the optimization of assisted reproductive technologies for the Congo peacock. This study was aimed at developing a suitable turkey semen extender for artificial insemination in field conditions. Semen was collected using the dorso-abdominal massage technique from seven turkey toms and analyzed. Ejaculates with >70% motility and >80% live spermatozoa were pooled and divided into four aliquots (four treatments). Each of the four treatments was extended in a soybean-based extender or an egg yolk-based extender, with or without L-ascorbic acid. Two liquid preservation protocols (ambient temperature (35 °C) and chilled (4 °C)) were employed, and quality parameters including motility, viability and morphology were evaluated. The results show that the two extenders were similar with regard to semen quality parameters, and L-ascorbic acid supplementation of the turkey semen extenders improved semen quality during liquid storage.

Bioconversion of cyanobacteria by black soldier fly larvae (Hermetia illucens L.): Enhancement by antioxidants.

Cyanobacterial blooms have been a global environmental problem for decades. Bioconversion by black soldier fly larvae (BSFL) has been widely reported to be a clean and efficient method to remove organic pollutants. In this study, BSFL bioconversion was used to treat cyanobacterial blooms. Antioxidants (a mixture of l-ascorbic acid [180 mg/kg fresh feed] and α-tocopherol [360 mg/kg fresh feed]) were added to compare bioconversion performance against a non-supplemented group. With increasing proportions of cyanobacteria (0%-25% dry mass), the bioconversion efficiency of the antioxidant group improved significantly compared to the control group, and the survival rate of larvae rose from 96.50-45.50% to 98.00-55.83% with antioxidant addition. The toxic effects of exogenous anti-nutrients could be reduced by the antioxidants through inactivation of trypsin inhibitor and enhancement of the microcystin-LR degradation rate. Overall, the BSFL bioremediation capacity was improved with addition of exogenous antioxidants, verifying both the effects and mechanism of antioxidant addition in promoting the bioconversion of cyanobacteria by BSFL and providing a basis for future application and study.

Zinc and L-Ascorbic Acid Dietary Supplementation Affected Antioxidant Status, Heat Shock Protein 70, and Some Immunoglobulins in Japanese Quail Under Heat Stress Conditions.

This study assessed the impact of the dietary inclusion of L-ascorbic acid and organic zinc (Availa-Zn) on heat-stressed Japanese quails. Growth performance, antioxidant status, immune status, heat shock protein 70 (HSP70), and some blood biochemical parameters were assessed. One-day-old, unsexed Japanese quail chicks (n = 240) were randomly allocated into 4 dietary treatments (6 replicates per treatment; 10 birds per replicate). Birds were fed a basal corn-soybean meal diet (control treatment) with different supplemental levels of L-ascorbic acid and/or Availa-Zn (200 mg L-ascorbic acid/kg diet, 62 mg Availa-Zn/kg diet, and 200 mg L-ascorbic acid + 62 mg Availa-Zn/kg diet) from July to August 2020 for 35 days. The average minimum and maximum ambient temperatures varied from 85.4 to 98.8 °F, and the relative humidity was between 69 and 74%. Supplemented L-ascorbic acid and Availa-Zn, either as separate supplements or as combined supplements, increased bird growth performance, blood hemoglobin, thyroid hormones, total protein, globulin, total antioxidant capacity, HSP70, catalase, superoxide dismutase enzyme activity, and immunoglobulin A and G (P < 0.05), while heterophil/lymphocytes decreased (P < 0.01) during the entire rearing period (1-35 days). Most of the assessed parameters showed stronger responses when L-ascorbic acid and Availa-Zn were added together, which may suggest a synergistic effect. In conclusion, the combined supplementation of L-ascorbic acid and Availa-Zn at 200 and 62 mg/kg, respectively, could be considered an efficient dietary supplement to enhance Japanese quail growth performance, antioxidant capacity, immune status, and general health under heat stress conditions.

L-Ascorbic Acid Shapes Bovine Pasteurella multocida Serogroup A Infection.

Bovine Pasteurella multocida serogroup A (bovine PmA) is one of the most important pathogens causing fatal pneumonia in cattle. However, it is largely unknown how nutrition shapes bovine PmA infection. Here, we discovered that the infected lung held the highest bacterial density than other tissues during infection. By screening the different metabolites between high (lung)- and low (liver)-bacterial density tissues, the present work revealed that L-ascorbic acid and L-aspartic acid directly influenced bovine P. multocida growth. Interestingly, L-ascorbic acid, which is expressed at higher levels in the infected livers, inhibited bovine PmA growth as well as virulence factor expression and promoted macrophage bactericidal activity in vitro. In addition, ascorbic acid synthesis was repressed upon bovine PmA infection, and supplementation with exogenous L-ascorbic acid significantly reduced the bacterial burden of the infected lungs and mouse mortality. Collectively, our study has profiled the metabolite difference of the murine lung and liver during bovine PmA infection. The screened L-ascorbic acid showed repression of bovine PmA growth and virulence expression in vitro and supplementation could significantly increase the survival rate of mice and reduce the bacterial load in vivo, which implied that L-ascorbic acid could serve as a potential protective agent for bovine PmA infection in clinic.

Ascorbic acid analogue 6-Deoxy-6-[18F] fluoro-L-ascorbic acid as a tracer for identifying human colorectal cancer with SVCT2 overexpression.

L-ascorbic acid (AA) was reported to have an anti-cancer effect over 40 years. In recent years, several ongoing clinical trials are exploring the safety and efficacy of intravenous high-dose AA for cancer treatment. The lack of appropriate imaging modality limits the identification of potentially suitable patients for AA treatment. This study focuses on identifying AA-sensitive tumor cells using molecular imaging. 6-Deoxy-6-[18F] fluoro-L-ascorbic Acid (18F-DFA), a structural analog of AA, was synthesized and labeled to visualize the metabolism of AA in vivo. Colorectal cancer (CRC) cell lines with high and low expression of sodium-dependent vitamin C transporters 2 (SVCT2) were used for a series of cellular uptake tests. PET imaging was performed on xenograft tumor-bearing mice. More AA uptake was observed in CRC cells with high SVCT2 expression than in cells with low SVCT2 expression. The substrate (unlabeled AA) can competitively inhibit the 18F-DFA tracer uptake by CRC cells. The biodistribution of 18F-DFA in mice showed high radioactivity was seen in organs such as adrenal glands, kidneys, and liver that were known to have high concentrations of AA. Both PET imaging and tissue distribution showed that cancer cells with high SVCT2 expression enhanced the accumulation of 18F-DFA in mice after tumor formation. Immunohistochemistry was used to verify the corresponding results. As a radiotracer, 18F-DFA can provide powerful imaging information to identify tumor with high affinity of AA, and SVCT2 can be a potential biomarker in this process.

Topical delivery of l-ascorbic acid spanlastics for stability enhancement and treatment of UVB induced damaged skin.

l-Ascorbic acid (LAA) is considered a powerful antioxidant that protects skin from premature aging. Maintaining the stability of vitamin C remains the biggest challenge in cosmeceuticals. Our main aim is the entrapment of high dose of vitamin C in spanlastic vesicles to provide maximum stability and efficacy. LAA-loaded spanlastics were prepared by ethanol injection method and were characterized for entrapment efficiency (EE%), particles size (PS), polydispersity index (PDI), zeta potential, deformability index (DI) and in vivo skin permeation. Selected spanlastics formula composed of span 60 and tween 60 (5:1) showed highest EE% of 89.77 ± 3.61% (w/w), high deformability of 11.13 ± 1.145 as well as good physical and chemical stability for 6 months. Improved drug penetration into stratum corneum (SC) was obtained from spanlastics compared to topical LAA solution. Quantitative real time PCR revealed that MMP2 and MMP9 levels were significantly suppressed in response to LAA spanlastics treated rats by 30.4% and 65.3%, respectively, when compared to the control group after exposure to UV irradiation. Results were confirmed by western blot analysis. Histopathological study of rat skin after UV irradiation revealed that application of LAA-loaded spanlastics provided the highest skin protection compared to UVB and LAA solution treated group which was evident by the normal thick epidermal morphology and the densely arranged dermal collagen fibers. LAA-loaded spanlastics successfully improved LAA stability, skin permeation and antioxidant protection against skin photodamage.

The Effect of L-Ascorbic Acid and Serum Reduction on Tenogenic Differentiation of Human Mesenchymal Stromal Cells.

BACKGROUND AND OBJECTIVES: Despite significant improvement in the treatment of tendon injuries, the full tissue recovery is often not possible because of its limited ability to auto-repair. The transplantation of mesenchymal stromal cells (MSCs) is considered as a novel approach in the treatment of tendinopathies. The question about the optimal culture conditions remains open. In this study we aimed to investigate if serum reduction, L-ascorbic acid supplementation or a combination of both factors can induce tenogenic differentiation of human adipose-derived MSCs (ASCs). METHODS AND RESULTS: Human ASCs from 3 healthy donors were used in the study. The tested conditions were: 0.5 mM of ascorbic acid 2-phosphate (AA-2P), reduced serum content (2% FBS) or combination of these two factors. The combination of AA-2P and 2% FBS was the only experimental condition that caused a significant increase of the expression of all analyzed genes related to tenogenesis (SCLERAXIS, MOHAWK, COLLAGEN_1, COLLAGEN_3, DECORIN) in comparison to the untreated control (evaluated by RT-PCR, 5th day of experiment). Moreover, this treatment significantly increased the synthesis of SCLERAXIS, MOHAWK, COLLAGEN_1, COLLAGEN_3 proteins at the same time point (evaluated by Western blot method). Double immunocytochemical staining revealed that AA-2P significantly increased the extracellular deposition of both types of collagens. Semi-quantitative Electron Spin Resonance analysis of ascorbyl free radical revealed that AA-2P do not induce harmful transition metals-driven redox reactions in cell culture media. CONCLUSIONS: Obtained results justify the use of reduced content of serum with the addition of 0.5 mM of AA-2P in tenogenic inducing media.