Methylprednisolone 100 mg tablet formulation with pea protein: experimental approaches over intestinal permeability and cytotoxicity.

OBJECTIVE: This study was carried out to transform the hydrolyzed pea protein into a pharmaceutical tablet form by masking methylprednisolone. SIGNIFICANCE: This study provides some crucial contributions in showing how functional excipients such as pea protein, which are generally used in food industries, can be used in pharmaceutical product formulations and their effects. METHODS: Methylprednisolone was formulated using spray drying technology. Design Expert Software (Version 13) was used for the statistical analysis. The in vitro cytotoxic effects for NIH/3T3 mouse fibroblast cells were investigated by XTT cell viability assay. HPLC was used to analyze the Caco-2 permeability studies and dissolution tests. RESULTS: The optimum formulation was evaluated against the reference product by performing cytotoxicity and cell permeability studies. According to our test results, Papp (apparent permeability) values of Methylprednisolone were measured around 3 × 10-6 cm/s and Fa (fraction absorbed) values around 30%. These data indicate that Methylprednisolone HCl has 'moderate permeability' and our study confirmed that it could have belonged to BCS Class II-IV since both low solubility and moderate permeability. CONCLUSION: The findings offer valuable information to guide and inform the use of pea protein in pharmaceutical formulations. Significant effects on methylprednisolone tablet formulation designed with the philosophy of quality by design (QbD) of pea protein have been demonstrated by both in vitro and cell studies.

Synthesis characterisation and neuroprotectivity of Silybum marianum extract loaded chitosan nanoparticles.

AIM: Silybum marianum extract (SME) possesses neuroprotective potency through its high antioxidant content. We attempted to increase the effectiveness of SME by encapsulating them in chitosan. Neuroprotective potency of SME and SME-loaded chitosan nanoparticles (SME-CNPs) were shown in SH-SY5Y cell line against H2O2-induced oxidative stress. METHODS: We produced CNPs and SME-CNPs by ionic gelation method and properly determined their physical characteristics. Encapsulation efficiency, loading capacity, and in vitro release tests were performed for SME-CNPs. The neurotoxicity and neuroprotective efficiency in SH-SY5Y cell line against H2O2 was also investigated. RESULTS: The size of SME-CNPs was 168.2 ± 11.12 nm with zeta potential 10.6 ± 1.0 mV. The encapsulation efficiency and loading capacity were successfully achieved at 96.6% and 1.89% respectively. SME and SME-CNPs improved cell viability higher than 80%, and SME-CNPs exhibited significant neuroprotective effects against H2O2 damage. CONCLUSIONS: It was concluded that SME and SME-CNPs highly prevent damage caused by H2O2 and reduce cell damage in vitro by their neuroprotective effects.

Development of piperine nanoemulsions: an alternative topical application for hypopigmentation.

This study, it was aimed to develop a topical piperine nanoemulsion (P-NE) using an ultrasonic emulsification process to find an alternative treatment option for some hypopigmentation disorders such as vitiligo. Results showed that 150 mg piperine loaded NE with 1:2 oil phase to Smix ratio and manufactured with 20 min ultrasonication duration with the pre-emulsification step was the most durable formulation with a mean globule size of 216.00 ± 2.65, a PdI value of 0.094 ± 0.02 and a zeta potential value of -27.50 ± 2.48 mV. After three months of storage, the selected P-NE (coded as F3P2) remained kinetically stable without visual changes. This formulation displayed a sustained release pattern with a release of 81.92% ± 3.04% piperine after 72 h. According to our in vitro activity experiments, it was determined that the P-NE had no toxic effect including the dose of 5 mg/mL, and the highest P-NE formulation dose of 5 mg/mL increased tyrosinase activity by 32.77% ± 9.09% and melanogenesis activity by 34.90% ± 0.73%. In conclusion, it was demonstrated that the P-NE formulation may serve as a promising therapy for the efficient treatment of vitiligo. Moreover, P-NE formulation may also help in preventing irregular pigmentation and skin cancer, associated with the conventional treatment methods.

Synthesis of biologically active copper oxide nanoparticles as promising novel antibacterial-antibiofilm agents.

In this study, we aimed to synthesize copper oxide nanoparticles (CuONPs) mediated by plant extract in an environmentally friendly way and to reveal their potential biological activities. Here we synthesized CuONPs by using different concentrations of aqueous leaf extract of Thymbra spicata at 80 °C to obtain Ts1CuONPs and Ts2CuONPs. Biosynthesized nanoparticles were characterized by using UV-Vis, AFM, FTIR, SEM-EDS, TEM, DLS and zeta potential analysis. The antibacterial activity of the nanoparticles was determined by calculation of the inhibition zone and minimum inhibitory concentration against selected bacterial strains. Moreover, the antioxidant activity of the as-synthesized nanoparticles was evaluated based on DPPH radical scavenging activity. The results indicate that the as-synthesized NPs have an average size of 26.8 and 21 nm for Ts1CuONPs and Ts2CuONPs, respectively. The formed CuONPs have more antibacterial action on gram-positive bacteria compared to gram-negative bacteria. In addition, CuONPs demonstrated good inhibition activity against biofilm formation of Staphylococcus aureus (S. aureus). Furthermore, the results showed that the smaller size of the CuONPs caused the higher cytotoxicity on L929 mouse fibroblast cells. The as-synthesized CuONPs exhibit antibacterial and antibiofilm potential against S. aureus, indicating that they may be attractive candidates to use in future therapeutic applications.

Papain Loaded Poly(ε-Caprolactone) Nanoparticles: In-silico and In-Vitro Studies.

Papain is a protease enzyme with therapeutic properties that are very valuable for medical applications. Poly(ε-caprolactone) (PCL) is an ideal polymeric carrier for controlled drug delivery systems due to its low biodegradability and its high biocompatibility. In this study, the three-dimensional structure and action mechanism of papain were investigated by in vitro and in silico experiments using molecular dynamics (MD) and molecular docking methods to elucidate biological functions. The results showed that the size of papain-loaded PCL nanoparticles (NPs) and the polydispersity index (PDI) of the NPs were 242.9 nm and 0.074, respectively. The encapsulation efficiency and loading efficiency were 80.4 and 27.2%, respectively. Human embryonic kidney cells (HEK-293) were used for determining the cytotoxicity of papain-loaded PCL and PCL nanoparticles. The in vitro cell culture showed that nanoparticles are not toxic at low concentrations, while toxicity slightly increases at high concentrations. In silico studies, which were carried out with MD simulations and ADME analysis showed that the strong hydrogen bonds between the ligand and the papain provide stability and indicate the regions in which the interactions occur.

Production and characterization of a conserved M2e peptide-based specific IgY antibody: evaluation of the diagnostic potential via conjugation with latex nanoparticles.

Antibodies play an important role in combating and controlling viral diseases such as influenza. Immunoglobulin Y (IgY) antibodies have several advantages such as a less invasive manufacturing process, ease of isolation, higher affinity compared with IgG antibodies, and cost-effectiveness. To date, although specific IgY production has been performed for different strains of influenza A, to the best of our knowledge, an IgY against the M2e peptide has not been produced. In the current study, IgY antibodies are produced, purified, and characterized using the M2e peptide sequence for the first time with the intent to apply them for the diagnosis of influenza A virus. Anti-M2e IgY antibodies are obtained from eggs using a two-step purification method. The activity and characterization of the antibodies are determined using an enzyme-linked immunosorbent assay, a nano-spectrophotometer, an SDS-Page assay, and a Western Blot analysis. Finally, anti-M2e IgY antibodies are conjugated to the latex nanoparticles, and the reaction between the influenza A virus and the nanoparticles is demonstrated using light microscopy, transmission electron microscopy, and energy dispersive X-ray spectroscopy. In conclusion, this study shows that anti-M2e IgY antibodies can contribute to the diagnosis, treatment, and prevention of the influenza A virus.

Production of anti-SAG1 IgY antibody against Toxoplasma gondii parasites and evaluation of antibody activity by ELISA method.

Chicken egg yolk antibody, also known as immunoglobulin Y (IgY), is the predominant class of serum immunoglobulin in birds. IgY has many advantages over mammalian antibodies, such as enhanced immunogenicity conserved mammalian proteins exhibited in birds due to their phylogenetic distance, non-invasive rapid, and economical collection system. However, there are limited studies about IgY production against Toxoplasma, which is a worldwide veterinary and public health problem. In this study, the production of specific IgY antibodies against the surface antigen 1 (SAG1) protein of Toxoplasma gondii and the determination of antibody activity via the enzyme-linked immunosorbent assay (ELISA) method were conducted. According to ELISA, Western blot, and NanoDrop results, specific and higher amounts of IgY antibody against SAG1 were obtained with this study. Considering the advantages of IgY and importance of SAG1 for the diagnosis of toxoplasmosis, it is expected that anti-SAG1 IgY will play an increasing role and gain commercial value in research, diagnostics, and immunotherapy against toxoplasmosis in the future.

Ganglion cell-inner plexiform layer thickness in patients with Parkinson disease and association with disease severity and duration.

BACKGROUND: To evaluate the average, minimum, and 6-sectoral macular ganglion cell-inner plexiform layer (GC-IPL) thickness measured by spectral domain optical coherence tomography (SD-OCT) in patients with Parkinson disease (PD), as well as average and 4-sectoral retinal nerve fiber layer (RNFL) thickness and to determine whether thickness parameters are correlated to disease severity and duration. METHODS: Patients with PD (n = 54) and age-matched healthy controls (n = 54) were prospectively examined with SD-OCT. Randomly selected eyes of all subjects were included. The average, minimum, and 6-sectoral (superior, superotemporal, superonasal, inferonasal, inferior, and inferotemporal) GC-IPL thickness values were analyzed. Average and 4-sectoral (inferior, superior, temporal, and nasal) peripapillary RNFL thicknesses were also evaluated. Each parameter was compared between patients with PD and age-matched healthy controls. PD severity was quantified with the Hoehn and Yahr (HY) scale. A correlation analysis was performed to evaluate the association between SD-OCT measurements and the duration and severity of PD. RESULTS: The mean age of patients with PD and age-matched healthy controls was 66.62 ± 8.71 years and 66.68 ± 7.85 years, respectively. Disease duration ranged from 1 to 15 years with a mean of 5.12 years. The mean PD severity, according to the HY scale, was 2.26 (range, 1-5). SD-OCT measurements revealed significant differences in inferior and temporal peripapillary RNFL values between groups (P = 0.018 and P = 0.031, respectively). All GC-IPL thickness parameters were statistically lower in the patients with PD when compared with the healthy controls (P < 0.001). PD duration was not correlated to any of the RNFL thicknesses, but PD severity was correlated inversely only with inferior peripapillary RNFL thickness (P = 0.006). Average, inferior (P = 0.011), inferotemporal (P = 0.007), and superotemporal (P = 0.007) GC-IPL thicknesses were correlated inversely with both PD severity and duration. CONCLUSIONS: Retinal dopaminergic neurodegeneration in patients with PD can be detected with macular GC-IPL thickness measurements. Macular GC-IPL thickness was correlated with PD severity and duration. It may be used to follow disease progression and efficacy of the neuroprotective treatment in patients with PD.

Migraine-like headache in cerebral venous sinus thrombosis.

A 20-year-old female, university student presented with severe, throbbing, unilateral headache, nausea and vomiting that started 2 days ago. The pain was aggravated with physical activity and she had photophobia. She had been taking contraceptive pills due to polycystic ovary for 3 months. Cranial computed tomography was uninformative and she was considered to have the first attack of migraine. She did not benefit from triptan treatment and as the duration of pain exceeded 72 h further imaging was done. Cranial MRI and MR venography revealed a central filling defect and lack of flow in the left sigmoid sinus caused by venous sinus thrombosis. In search for precipitating factors besides the use of contraceptive pills, plasma protein C activity was found to be depressed (42%, normal 70-140%), homocystein was minimally elevated (12.7 µmol/L, normal 0-12 µmol/L) and anti-cardiolipin IgM antibody was close to the upper limit.

Investigation of metal-polyelectrolyte complex toxicity.

Water-soluble binary and ternary copper complexes of polyelectrolytes were synthesized, and the toxicity of these complexes was tested in mouse fibroblast cell line (L929) in vitro. Both the binary and ternary complexes were prepared at the ratio of 0.4 mole copper(II) ions per monomer of acrylic acid and 0.5 mole copper(II) ions per monomer of methyl vinyl ether maleic anhydride, furthermore at the ratio of 1 and 2 mole bovine serum albumin per mole of polyacrylic acid and poly(methyl vinyl ether-co-maleic anhydride), respectively. Compared to binary copper(II)-polyelectrolyte complexes, these ternary complexes have been determined to be of least toxicity.

Conjugation, characterization and toxicity of lipophosphoglycan-polyacrylic acid conjugate for vaccination against leishmaniasis.

Research on the conjugates of synthetic polyelectrolytes with antigenic molecules, such as proteins, peptides, or carbohydrates, is an attractive area due to their highly immunogenic character in comparison to classical adjuvants. For example, polyacrylic acid (PAA) is a weak polyelectrolyte and has been used in several biomedical applications such as immunological studies, drug delivery, and enzyme immobilization. However, to our knowledge, there are no studies that document immune-stimulant properties of PAA in Leishmania infection. Therefore, we aimed to develop a potential vaccine candidate against leishmaniasis by covalently conjugating PAA with an immunologically vital molecule of lipophosphoglycan (LPG) found in Leishmania parasites. In the study, LPG and PAA were conjugated by a multi-step procedure, and final products were analyzed with GPC and MALDI-TOF MS techniques. In cytotoxicity experiments, LPG-PAA conjugates did not indicate toxic effects on L929 and J774 murine macrophage cells. We assume that LPG-PAA conjugate can be a potential vaccine candidate, and will be immunologically characterized in further studies to prove its potential.

Development of an Antiviral Ion-Activated In Situ Gel Containing 18ß-Glycyrrhetinic Acid: A Promising Alternative against Respiratory Syncytial Virus.

The human respiratory syncytial virus (hRSV) is a major cause of serious lower respiratory infections and poses a considerable risk to public health globally. Only a few treatments are currently used to treat RSV infections, and there is no RSV vaccination. Therefore, the need for clinically applicable, affordable, and safe RSV prevention and treatment solutions is urgent. In this study, an ion-activated in situ gelling formulation containing the broad-spectrum antiviral 18ß-glycyrrhetinic acid (GA) was developed for its antiviral effect on RSV. In this context, pH, mechanical characteristics, ex vivo mucoadhesive strength, in vitro drug release pattern, sprayability, drug content, and stability were all examined. Rheological characteristics were also tested using in vitro gelation capacity and rheological synergism tests. Finally, the cytotoxic and antiviral activities of the optimized in situ gelling formulation on RSV cultured in the human laryngeal epidermoid carcinoma (HEp-2) cell line were evaluated. In conclusion, the optimized formulation prepared with a combination of 0.5% w/w gellan gum and 0.5% w/w sodium carboxymethylcellulose demonstrated good gelation capacity and sprayability (weight deviation between the first day of the experiment (T0) and the last day of the experiment (T14) was 0.34%), desired rheological synergism (mucoadhesive force (Fb): 9.53 Pa), mechanical characteristics (adhesiveness: 0.300 ± 0.05 mJ), ex vivo bioadhesion force (19.67 ± 1.90 g), drug content uniformity (RSD%: 0.494), and sustained drug release over a period of 6 h (24.56% ± 0.49). The optimized formulation demonstrated strong anti-hRSV activity (simultaneous half maximal effective concentration (EC50) = 0.05 µg/mL; selectivity index (SI) = 306; pre-infection EC50 = 0.154 µg/mL; SI = 100), which was significantly higher than that of ribavirin (EC50 = 4.189 µg/mL; SI = 28) used as a positive control against hRSV, according to the results of the antiviral activity test. In conclusion, this study showed that nasal in situ gelling spray can prevent viral infection and replication by directly inhibiting viral entry or modulating viral replication.

Lavandula stoechas extract incorporated polylactic acid nanofibrous mats as an antibacterial and cytocompatible wound dressing.

In recent years, great efforts have been devoted to the design and production of bioactive wound dressings that promote skin regeneration and prevent infection. Many plant extracts and essential oils have been widely accepted in traditional medicine for a wide variety of medicinal purposes, especially wound healing. Over the past decade, many studies have focused on manufacturing and designing wound dressings containing plant compounds and extracts. In this study, Lavandula stoechas extract (LSE) (0.25 %, 0.5 %, and 1%wt) incorporated-polylactic acid (PLA) nanofibrous mats were successfully produced and characterized. Microstructural analysis by SEM revealed that the fiber diameter changed with the increase in the amount of LSE. Also, the nanofibrous mats were evaluated for their in vitro antibacterial, cytotoxicity, and wound healing properties for their use as a wound dressing material. According to the results of the disc diffusion test, PLA nanofibrous mats containing LSE %1 showed 9.65 ± 0.46 and 7.37 ± 0.03 inhibition zone (mm) against E. coli and S. aureus, respectively. According to the results of the in vitro wound healing assay, mats containing 0.5 % LSE showed better-wound closure activity compared to the control. Our results show that LSE-incorporated nanofibrous dressings can be an effective alternative with good antimicrobial activity.

Immunogenic evaluation of multi-epitope peptide-loaded PCPP microparticles as a vaccine candidate against Toxoplasma Gondii.

Toxoplasmosis is a major health problem and socioeconomic burden, affecting around 30-50% of the global population. Poly(dicarboxylatophenoxy)phosphazene (PCPP) polymer was chosen as adjuvant for the immunogenic peptide antigen. Peptide-loaded PCPP microparticles were synthesized via the coacervation method and the characterization studies of microparticles were conducted to determine their size, charge, morphology, encapsulation efficacy, and loading capacity. To evaluate in vivo efficacy of the vaccine candidate, Balb/c mice were immunized with the formulations. Brain and spleen tissues were isolated from animals to investigate cytokine levels, lymphocyte proliferation, and brain cyst formation. As a result, antibody and cytokine responses in groups immunized with peptide-loaded PCPP microparticles were found to be significantly higher when compared to the control group. In conclusion, our novel multi-epitope peptide-loaded PCPP microparticle-based vaccine formulation demonstrated considerable humoral and cellular immune responses against T. gondii and protected mice against T. gondii infection during Toxoplasmosis.

Safety and immunogenicity of an inactivated whole virion SARS-CoV-2 vaccine, TURKOVAC, in healthy adults: Interim results from randomised, double-blind, placebo-controlled phase 1 and 2 trials.

BACKGROUND: Development of safe and effective vaccine options is crucial to the success of fight against COVID-19 pandemic. Herein, we report interim safety and immunogenicity findings of the phase 1&2 trials of ERUCoV-VAC, an inactivated whole virion SARS-CoV-2 vaccine. METHODS: Double-blind, randomised, single centre, phase 1 and 2 trials included SARS-CoV-2 seronegative healthy adults aged 18-55 years (18-64 in phase 2). All participants, except the first 4 in phase 1 who received ERUCoV-VAC 3 µg or 6 µg unblinded and monitored for 7 days for safety purposes, were assigned to receive two intramuscular doses of ERUCoV-VAC 3 µg or 6 µg (an inactivated vaccine containing alhydrogel as adjuvant) or placebo 21 days apart (28 days in phase 2) according to computer-generated randomisation schemes. Both trials are registered at ClinicalTrials.gov (phase 1, NCT04691947 and phase 2, NCT04824391). RESULTS: Forty-four participants (3 µg [n:17], 6 µg [n:17], placebo [n:10]) in phase 1 and 250 (3 µg [n:100], 6 µg [n:100], placebo [n:50]) in phase 2 received ≥1 dose. In phase 1 trial, 25 adverse events AEs (80 % mild) occured in 15 participants (34.1 %) until day 43. There was no dose-response relationship noted in safety events in ERUCoV-VAC recipients (p = 0.4905). Pain at injection site was the most common AE (9/44;20.5 %). Both doses of ERUCoV-VAC 3 µg and 6 µg groups were comparable in inducing SARS-CoV-2 wild-type neutralising antibody (MNT50): GMTs (95 %CI) were 8.3 (6.4-10.3) vs. 8.6 (7.0-10.2) at day 43 (p = 0.7357) and 9.7 (6.0-13.4) vs. 10.8 (8.8-12.8) at day 60 (p = 0.8644), respectively. FRNT50 confirmed MNT50 results: SARS-CoV-2 wild-type neutralising antibody GMTs (95 %CI) were 8.4 (6.3-10.5) vs. 9.0 (7.2-10.8) at day 43 (p = 0.5393) and 11.0 (7.0-14.9) vs. 12.3 (10.3-14.5) at day 60 (p = 0.8578). Neutralising antibody seroconversion rates (95 %CI) were 86.7 % (59.5-98.3) vs 94.1 % (71.3-99.8) at day 43 (p = 0.8727) and 92.8 % (66.1-99.8) vs. 100 % (79.4-100.0) at day 60 (p = 0.8873), in ERUCoV-VAC 3 µg and 6 µg groups, respectively. In phase 2 trial, 268 AEs, (67.2 % moderate in severity) occured in 153 (61.2 %) participants. The most common local and systemic AEs were pain at injection site (23 events in 21 [8.4 %] subjects) and headache (56 events in 47 [18.8 %] subjects), respectively. Pain at injection site was the only AE with a significantly higher frequency in the ERUCoV-VAC groups than in the placebo arm in the phase 2 study (p = 0.0322). ERUCoV-VAC groups were comparable in frequency of AEs (p = 0.4587). ERUCoV-VAC 3 µg and 6 µg groups were comparable neutralising antibody (MNT50): GMTs (95 %CI) were 30.0 (37.9-22.0) vs. 34.9 (47.6-22.1) at day 43 (p = 0.0666) and 34.2 (23.8-44.5) and 39.6 (22.7-58.0) at day 60, (p = 0.2166), respectively. FRNT50 confirmed MNT50 results: SARS-CoV-2 wildtype neutralising antibody GMTs were 28.9 (20.0-37.7) and 30.1 (18.5-41.6) at day 43 (p = 0.3366) and 34.2 (23.8-44.5) and 39.6 (22.7-58.0) at day 60 (p = 0.8777). Neutralising antibody seroconversion rates (95 %CI) were 95.7 % (91.4-99.8) vs. 98.9 % (96.9-100.0) at day 43 (p = 0.8710) and 96.6 % (92.8-100.0) vs 98.9 % (96.7-100.0) at day 60 (p = 0.9129) in ERUCoV-VAC 3 µg and 6 µg groups, respectively. CONCLUSIONS: Two-dose regimens of ERUCoV-VAC 3 µg and 6 µg 28 days both had an acceptable safety and tolerability profile and elicited comparable neutralising antibody responses and seroconversion rates exceeding 95 % at day 43 and 60 after the first vaccination. Data availability Data will be made available on request.

The Effect of Smoking on Inactivated and mRNA Vaccine Responses Applied to Prevent COVID-19 in Multiple Sclerosis.

Introduction: Coronavirus disease 2019 (COVID-19) is the biggest health challenge of recent times. Studies so far reveal that vaccination is the only way to prevent this pandemic. There may be factors that decrease or increase vaccine effectiveness. In multiple sclerosis (MS), some of these factors may cause changes in the effectiveness of the vaccine, depending on the nature of the disease and disease-modifying treatments (DMT). In this study, we aimed to investigate the relationship between antibody titer and smoking in non-treated and DMT-treated MS patients who received inactivated vaccine (Sinovac) and messenger RNA BNT162b2 (BioNTech) mRNA vaccines. Method: Vaccine antibody responses were measured between 4-12 weeks after two doses of inactivated vaccine and mRNA vaccines. Patients were separated into 6 groups as: patients with MS without treatment PwMS w/o T, ocrelizumab, fingolimod, interferons (interferon beta-1a and interferon beta-1b), dimethyl fumarate, and teriflunomide. Antibody titers of smokers and non-smokers were compared for both vaccines and for each group. Results: The study included 798 patients. In the mRNA vaccine group, smokers (n=148; 2982±326 AU/mL) had lower antibody titers compared to the non-smokers (n=244; 5903±545 AU/mL) in total (p=0.020). In the inactivated vaccine group, no significant difference was detected between smokers (n=136; 383±51 AU/mL) and non-smokers (n=270; 388±49 AU/mL) in total (p=0.149). In both vaccine groups, patients receiving ocrelizumab and fingolimod had lower antibody titers than those receiving other DMTs or PwMS w/o T. In untreated MS patients, antibody levels in smokers were lower than in non-smokers in the mRNA vaccine group. No difference was found between antibody levels of smokers and non-smokers in any of the inactivated vaccine groups. Conclusion: Ocrelizumab and fingolimod have lower antibody levels than PwMS w/o T or other DMTs in both mRNA and inactivated vaccine groups. Smoking decreases antibody levels in the mRNA vaccine group, while it has no effect in the inactivated vaccine group.

mRNA versus inactivated virus COVID-19 vaccines in multiple sclerosis: Humoral responses and protectivity-Does it matter?

BACKGROUND: COVID-19 vaccines are recommended for people with multiple sclerosis (pwMS). Adequate humoral responses are obtained in pwMS receiving disease-modifying therapies (DMTs) after vaccination, with the exception of those receiving B-cell-depleting therapies and non-selective S1P modulators. However, most of the reported studies on the immunity of COVID-19 vaccinations have included mRNA vaccines, and information on inactivated virus vaccine responses, long-term protectivity, and comparative studies with mRNA vaccines are very limited. Here, we aimed to investigate the association between humoral vaccine responses and COVID-19 infection outcomes following mRNA and inactivated virus vaccines in a large national cohort of pwMS receiving DMTs. METHODS: This is a cross-sectional and prospective multicenter study on COVID-19-vaccinated pwMS. Blood samples of pwMS with or without DMTs and healthy controls were collected after two doses of inactivated virus (Sinovac) or mRNA (Pfizer-BioNTech) vaccines. PwMS were sub-grouped according to the mode of action of the DMTs that they were receiving. SARS-CoV-2 IgG titers were evaluated by chemiluminescent microparticle immunoassay. A representative sample of this study cohort was followed up for a year. COVID-19 infection status and clinical outcomes were compared between the mRNA and inactivated virus groups as well as among pwMS subgroups. RESULTS: A total of 1484 pwMS (1387 treated, 97 untreated) and 185 healthy controls were included in the analyses (male/female: 544/1125). Of those, 852 (51.05%) received BioNTech, and 817 (48.95%) received Sinovac. mRNA and inactivated virus vaccines result in similar seropositivity; however, the BioNTech vaccination group had significantly higher antibody titers (7.175±10.074) compared with the Sinovac vaccination group (823±1.774) (p<0.001). PwMS under ocrelizumab, fingolimod, and cladribine treatments had lower humoral responses compared with the healthy controls in both vaccine types. After a mean of 327±16 days, 246/704 (34.9%) of pwMS who were contacted had COVID-19 infection, among whom 83% had asymptomatic or mild disease. There was no significant difference in infection rates of COVID-19 between participants vaccinated with BioNTech or Sinovac vaccines. Furthermore, regression analyses show that no association was found regarding age, sex, Expanded Disability Status Scale score (EDSS), the number of vaccination, DMT type, or humoral antibody responses with COVID-19 infection rate and disease severity, except BMI Body mass index (BMI). CONCLUSION: mRNA and inactivated virus vaccines had similar seropositivity; however, mRNA vaccines appeared to be more effective in producing SARS-CoV-2 IgG antibodies. B-cell-depleting therapies fingolimod and cladribine were associated with attenuated antibody titer. mRNA and inactive virus vaccines had equal long-term protectivity against COVID-19 infection regardless of the antibody status.

Leishmania tropica: the effect of darkness and light on biological activities in vitro.

Leishmania parasites can be exposed to effects of light in their vectors and hosts, at various periods. However, there is no information about the effects of light on Leishmania parasites. The aim of this study is to investigate the effects of light on various cell parameters of Leishmania tropica, in vitro. All experiments were conducted on L. tropica promastigotes and amastigote-macrophage cultures, using flow cytometric analysis, MTT and phenol-sulfuric acid assay, DAPI and Giemsa. The results showed that the morphology of parasites has changed; the cell cycle has been affected and this caused parasites to remain at G0/G1 phase. Furthermore the proliferation, infectivity, glucose consumption and mitochondrial dehydrogenase activities of parasites were decreased. Thus, for the first time, in this study, the effects of light on biological activities of Leishmania parasites were shown. These new information about parasites' biology, would be very important to investigate the effects of light on the parasites in infected vectors and hosts.

Comparison of the anticancer effect of microalgal oils and microalgal oil-loaded electrosprayed nanoparticles against PC-3, SHSY-5Y and AGS cell lines.

Many of the bioactive substances used in pharmaceutical industry are easily affected by temperature, light and oxygen, and are easily degraded during storage and processing, and exhibit poor adsorption properties during digestion, which limits their direct use. Microalgae are rich in oils which have antimicrobial properties and antioxidants that attract attention in both food and pharmaceutical sectors in recent years. Studies to encapsulate bioactive compound-rich microalgae oils with nanotechnological approaches to improve the physical and chemical stability are relatively new, and it is promising to apply these approaches for pharmaceutical purposes. In this study, cytotoxic effects of oil extracts of Botryococcus braunii and Microcystis aeruginosa and their oil-loaded nanoparticles on L929 cell line, PC-3 prostate cell line, SHSY-5Y neuroblastoma cell line and AGS gastric adenocarcinoma cell line were investigated. The obtained extracts were found to have no cytotoxic effect on L929 cells. However, they showed cytotoxic effect on cancer cells. As for the nanoparticles; a gradual release was determined and the stability of the nanoparticle structure was shown. In the light of obtained findings, it was considered that nanoparticles produced with oil extracts of microalgae which have bioactive substances, have potential to be evaluated especially in pharmaceutical and cosmetic fields.

Preparation of chitosan nanoparticles as Ginkgo Biloba extract carrier: In vitro neuroprotective effect on oxidative stress-induced human neuroblastoma cells (SH-SY5Y).

Ginkgo biloba (Gb) is an ancient Chinese tree cultivated for its health-promoting properties. Moreover, Gb extract has a therapeutic effect, especially on neurodegenerative diseases. In this study, Gb extract-loaded chitosan nanoparticles (Gb-CsNPs) were synthesized by ionic gelation method. Size and zeta potential of the nanoparticles were analyzed and Scanning Electron Microscopy (SEM) and Fourier Transform Spectroscopy (FT-IR) were performed. Besides, encapsulation efficacy and loading capacity were calculated, and in vitro release, and cellular uptake studies were carried out. The biocompatibility of Gb-CsNPs was demonstrated and their neuroprotective activity was investigated on oxidative stress-induced SH-SY5Y cells. Apoptotic cells were monitored by DAPI, and cell migration was examined by in vitro scratch assay. Results showed that Gb-CsNPs had an average size of 104.4 nm, their zeta potential and polydispersity index (PDI) values were 29.3 mV, and 0.09 respectively. Encapsulation efficacy and loading capacity were found as 97.4% and 40%, respectively. It has been revealed that Gb-CsNPs were biocompatible and showed neuroprotective activity by increasing cell viability from 60% to 92.3%. Consequently, neuroprotective effect of the Gb extract was increased by chitosan encapsulation. This formulation is a candidate to be used as a food supplement after being supported by future in vivo studies.