Evaluating the Impact of Vitamin D3 on NF-κB and JAK/STAT Signaling Pathways in Drosophila melanogaster.

This study delved into the consequences of prolonged administration of vitamin D3 on innate immune systems, particularly NF-κB and JAK/STAT, in Drosophila melanogaster. The outcomes indicated that vitamin D3 treatment exhibited a notable capacity to improve the survival of adult flies with compromised immune functions, a condition induced by the loss of PGRP-LB, particularly when the flies were exposed to heat-killed Escherichia coli. The PGRP-LBΔ mutant line that was treated with heat-killed E. coli experienced reduced survival. Treatment of heat-killed E. coli-treated PGRP-LBΔ with vitamin D3 resulted in improved survival, and this phenotypic feature might be due to the downregulation of gene expression in the NF-κB and JAK/STAT pathways. However, a higher concentration of vitamin D3 was associated with decreased survival, potentially linked to intricate immunological responses. The research also underscored the influence of vitamin D3 on the expression of antioxidant genes, sod1 and sod2, indicating an augmented resistance to oxidative stress. Further, this study revealed the effect of vitamin D3 on the reproductive status of the autoinflammatory model, showing an increase in pupae and adult flies with a treatment of 10 mM vitamin D3, suggesting the potential benefits of vitamin D3 on the reproductive profile. Overall, this study provides preliminary insights into the complex interactions between vitamin D3, immune pathways, oxidative responses in the cell, and reproduction in Drosophila.

Validation of UV-Vis spectrophotometric and colorimetric methods to quantify methotrexate in plasma and rat skin tissue: Application to in vitro release, ex vivo and in vivo studies from dissolving microarray patch loaded pH-sensitive nanoparticle.

This research transformed MTX into smart nanoparticles that respond to the acidic conditions present in inflammation. These nanoparticles were then incorporated into a patch that dissolves over time, aiding their penetration. A method using UV-Vis spectrophotometry was validated to support the development of this new delivery system. This method was used to measure the quantity of MTX in the prepared patches in various scenarios: in laboratory solutions with pH 7.4 and pH 5.0, in skin tissue, and plasma. This validation was conducted in laboratory studies, tissue samples, and live subjects, adhering to established guidelines. The resulting calibration curve displayed a linear relationship (correlation coefficient 0.999) across these scenarios. The lowest quantity of MTX that could be accurately detected was 0.6 µg/mL in pH 7.4 solutions, 1.46 µg/mL in pH 5.0 solutions, 1.11 µg/mL in skin tissue, and 1.48 µg/mL in plasma. This validated method exhibited precision and accuracy and was not influenced by dilution effects. The method was effectively used to measure MTX levels in the developed patch in controlled lab settings and biological systems (in vitro, ex vivo, and in vivo). This showed consistent drug content in the patches, controlled release patterns over 24 h, and pharmacokinetic profiles spanning 48 h. However, additional analytical approaches were necessary for quantifying MTX in studies focused on the drug's effects on the body's functions.

Development of pH-Sensitive Nanoparticle Incorporated into Dissolving Microarray Patch for Selective Delivery of Methotrexate.

PURPOSE: Rheumatoid arthritis (RA) is a systemic autoimmune disease that attacks human joints. Methotrexate (MTX), as one the most effective medications to treat RA, has limitations when administered either orally or by injection. To overcome this limitation, we formulated MTX through a smart nanoparticle (SNP) combined with dissolving microarray patch (DMAP) to achieve selective-targeted delivery of RA. METHODS: SNP was made using the combination of polyethylene glycol (PEG) and polycaprolactone (PCL) polymers, while DMAP was made using the combination of hyaluronic acid and polyvinylpyrrolidone K-30. SNP-DMAP was then evaluated for its mechanical and chemical characteristics, ex vivo permeation test, in vivo pharmacokinetic study, hemolysis, and hen's egg test-chorioallantoic membrane (HET-CAM) test. RESULT: The results showed that the characteristics of the SNP-DMAP-MTX formulas meet the requirements for transdermal delivery, with the particle size of 189.09 ±12.30 nm and absorption efficiency of 65.40 ± 5.0%. The hemolysis and HET-CAM testing indicate that this formula was non-toxic and non-irritating. Ex vivo permeation shows a concentration of 51.50 ± 3.20 µg/mL of SNP-DMAP-MTX in PBS pH 5.0. The pharmacokinetic profile of SNP-DMAP-MTX showed selectivity and sustained release compared with oral and DMAP-MTX with values of t1/2 (4.88 ± 0 h), Tmax (8 ± 0 h), Cmax (0.50 ± 0.04 µg/mL), AUC (3.15 ± 0.54 µg/mL.h), and mean residence time (MRT) (9.13 ± 0 h). CONCLUSION: The developed SNP-DMAP-MTX has been proven to deliver MTX transdermal and selectively at the RA site, potentially avoiding conventional MTX side effects and enhancing the effectiveness of RA therapy.

Vitamin D was Superior to Omega-3 as a Simvastatin Adjuvant in Improving Blood Lipids and Atherogenic Index in Type-I Dyslipidemic Rats.

Objectives: Adjuvant therapy is often used to optimize the antihyperlipidemic effect of simvastatin. Omega-3 and vitamin D supplementation are recommended as adjuvant therapies to low-intensity statins. This study aimed to compare the effects of vitamin D and omega-3 as adjuvant therapy to simvastatin to improve the lipid profiles and atherogenic index of plasma (AIP) in type-I dyslipidemic rats. Materials and Methods: Thirty-six male rats were randomized and divided into six groups: healthy control, dyslipidemic rats with no treatment, and dyslipidemic rats treated with either low-dose simvastatin only or omega-3 or vitamin D at low and high doses. Dyslipidemia was induced with high-fat diets for four weeks, followed by treatment for the next two weeks. Blood samples were withdrawn before and after simvastatin treatment. In addition, aspartate transaminase (AST) and alanine transaminase (ALT) levels were analyzed to assess liver function. Results: Administration of a high-fat diet-induced type 1 dyslipidemia and increased ALT levels (p < 0.05). Treatment with low-dose simvastatin did not significantly improve triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDLc) or non-HDLc levels. When combined with a high-dose vitamin D, simvastatin significantly reduced TG and increased HDLc levels (p < 0.05), thereby improving AIP levels. This improvement was not observed in rats treated with omega-3 or vitamin D at a lower dose. Conclusion: We concluded that high-dose vitamin D as an adjuvant to simvastatin therapy was superior to omega-3 in improving TG, HDL, and AIP levels. High-dose vitamin D also improved ALT levels in type-I dyslipidemic rats. This result may be translated in clinics to reduce the risk of coronary syndrome in patients with type-I dyslipidemia.

Piper retrofractum ameliorates imiquimod-induced skin inflammation via modulation of TLR4 axis and suppression of NF-κB activity.

Chronic inflammation is a significant concern due to its association with various pathological conditions. As a result, extensive research has been conducted to identify new natural products that can effectively treat acute inflammation, which has the potential to inhibit the chronic inflammation. In our study, we aimed to identify Indonesian medicinal plants with the ability to inhibit proinflammatory agents, specifically targeting NF-κB, a crucial regulator of gene transcription involved in the production of proinflammatory proteins/cytokines. Through a series of identification processes, we found that Piper retrofractum (Javanese chili) extract demonstrated promising inhibitory effects on NF-κB and proinflammatory molecules. Further investigation was conducted using a variety of assays, including reporter assay, viability test, ELISA, and Western blotting. The results revealed that the extract significantly reduced LPS, NO, COX-2, IL-6, IL-1, and NF-κB through the TLR4 axis. Notably, Piper retrofractum extract was found to enhance the survival of human keratinocytes by protecting them from cell death induced by TRAIL, a member of the TNF superfamily. Moreover, immunohistochemistry analysis in an Imiquimod-induced skin inflammation mice model showed downregulation of COX-2 and IL-1ß expression upon treatment with the extract. In conclusion, our findings suggest that Piper retrofractum extract possesses anti-inflammatory properties by reducing proinflammatory cytokine production through inhibition of NF-κB signaling pathway. These promising results highlight the potential of Piper retrofractum extract as a candidate for future drug development in the clinical treatment of inflammation-related conditions, offering hope for the advancement of therapeutic interventions.

Development of Solid Lipid Nanoparticle-Loaded Polymeric Hydrogels Containing Antioxidant and Photoprotective Bioactive Compounds of Safflower (Carthamus tinctorius L.) for Improved Skin Delivery.

Safflower (Carthamus tinctorius L.) is a potent natural antioxidant because of active compounds such as quercetin (QU) and luteolin (LU). These components prevent damage to the skin caused by free radicals from UV rays. However, due to the poor solubility and transdermal permeation, the effectiveness of the compounds in showing their activity was limited. In this study, we develop solid lipid nanoparticle (SLN)-based hydrogel formulations to enhance the solubility and penetration of two bioactive compounds found in safflower petals extract (SPE). The hot emulsification-ultrasonication method was used to produce SLNs, and to obtain high antioxidant activity, 100% v/v ethanol was used in the extraction procedure. The results showed that this approach could encapsulate >80% of both QU and LU. Moreover, Fourier transform infrared (FTIR), differential scanning calorimetry (DSC), and powder X-ray diffraction (PXRD) spectra indicated that most of the QU and LU were trapped in a lipid matrix and dispersed homogeneously at the molecular level, increasing the solubility. Additionally, SLN-hydrogel composites are able to release two lipophilic bioactive compounds for 24 h, which also demonstrated increased skin retention and penetrability of the QU and LU up to 19-fold. In vitro blood biocompatibility showed that no hemolytic toxicity was observed below 500 µg/mL. Accordingly, the formulation was considered safe for use. Sun protective factor (SPF) test shows a value above 15, showing an excellent promising application as the photoprotective agent to prevent symptoms associated with photoinduced skin aging.

A Combination of Virgin Coconut Oil and Extra Virgin Olive Oil Elicits Superior Protection Against Doxorubicin Cardiotoxicity in Rats

Objectives: The use of the chemotherapy agent doxorubicin (DOX) is associated with free radical formation that may lead to cardiotoxicity. Virgin coconut oil (VCO) and extra virgin olive oil (EVOO) are plant-based oil that is rich in antioxidants. This study examined the protective effects of VCO and EVOO combination to reduce DOX acute cardiotoxicity in rats. Materials and Methods: Twenty-five male rats (180-200 g) were divided into the following groups: Group I as a control, group II was given DOX i.p. injection of 25 mg/kg body weight (b.w.), group III to V received peroral administration of either VCO, EVOO or VCO-EVOO (1:1) combination at a dose of 10 mL/kg b.w. for 6 days before receiving DOX i.p. injection. After 24 hours from DOX injection, blood samples and organs were collected. Cardiac biomarkers, such as serum glutamic-oxaloacetic transaminase (SGOT), lactate dehydrogenase (LDH), and creatine kinase-MB (CKMB) were analyzed followed by histopathological examination. Results: The administration of EVOO alone was found to reduce the marked elevation of SGOT, LDH, and CKMB levels in DOX-treated rats (p<0.05), while VCO administration only significantly reduced LDH and CKMB levels. However, when both oils were used in combination, the protective effect was shown to be more powerful since all cardiac biomarker levels were maintained at near-normal levels (p<0.05). Histopathological analysis showed a significant improvement in the myocardial tissue structures after pre-treatment with VCO-EVOO combination. Conclusion: The administration of VCO and EVOO in combination was superior to elicit protection against DOX-induced cardiotoxicity compared to their individual application in rats.

Increased aldehyde dehydrogenase 1 (ALDH1) levels are associated with chemo-responsiveness in breast cancer patients treated with taxane-adriamycin-cyclophosphamide regimen.

BACKGROUND: Increased plasma aldehyde dehydrogenase 1 (ALDH1) levels have been proposed to predict cancer chemoresistance. However, studies have reported inconsistent results, depending on the type of cancer cells used. OBJECTIVE: This study aimed to investigate the correlation between plasma levels of ALDH1 and chemotherapy responses to the taxane-adriamycin-cyclophosphamide (TAC) regimen in breast cancer patients. METHODS: Thirty breast cancer patients who underwent chemotherapy using the TAC regimen were included in this study. Blood sampling was performed before chemotherapy was initiated and after the first and third cycles of chemotherapy administration. After 3 cycles of chemotherapy, patients were categorized as non-responsive if the tumor size was reduced <30%, if the tumor size remained the same or increased, or if any new tumors were discovered. Patients were defined as responsive after 3 cycles of chemotherapy if the tumor mass disappeared, if the tumor size was reduced by at least 30% of the initial size and if no new tumors were found. RESULTS: Among the 30 patients, only five were responsive to the TAC regimen. The clinical response to TAC was not correlated with the patient's age, cancer grading, or tumor stage. A change in the ALDH1 levels was observed after the third cycle of TAC administration, with significantly higher ALDH1 levels observed in responsive compared with non-responsive patients (p < 0.05). CONCLUSION: The results of this study may indicate a role for ALDH1 in chemoresponsiveness, rather than chemoresistance, for the TAC regimen in breast cancer patients. Further research remains necessary to confirm this result.

Potential Roles of Kleinhovia hospita L. Leaf Extract in Reducing Doxorubicin Acute Hepatic, Cardiac and Renal Toxicities in Rats.

BACKGROUND: Doxorubicin (DOX) is a potent chemotherapy agent; however, its use may lead to cardiac, hepatic, and renal dysfunction. Kleinhovia hospita L extract contains antioxidant compounds that have been shown to reduce chemical-induced hepatotoxicity. OBJECTIVES: This study aimed to examine the protective effects of Kleinhovia sp. extract to reduce DOX acute toxicities. MATERIALS AND METHODS: Thirty male rats were assigned to the following groups: Group I as controls, Group II was given DOX i.p. injection (25 mg/kg); Groups III, IV, and V were treated with Kleinhovia sp. extract 100, 250, and 500 mg/kg orally for 5 days, respectively, prior to DOX i.p. injection. After 24 h, blood and organs were analyzed for biomarker levels and histopathological changes. RESULTS: DOX treatment in Group II significantly increased creatine kinase-MB (CK-MB), aspartate transaminase (AST), alanine transaminase (ALT), and urea levels compared to controls. Kleinhovia sp. extract at any given dose significantly improved ALT and AST; yet, CK-MB levels only reduced with 250 mg/kg dose (Group IV). Urea and creatinine levels in Kleinhovia sp. groups were also lower compared to DOX-treated rats, but it was not significant. Histopathological analysis showed improved liver, heart, and renal tissue structures in Kleinhovia sp-treated rats, especially at higher doses. CONCLUSION: Kleinhovia sp. extract at any dose given protected the rats from liver toxicity, but only at dose 250 mg/kg reduced cardiac toxicity. Although renal biomarkers were insignificantly lower, renal architecture was improved with Kleinhovia sp. treatment. SUMMARY: Doxorubicin (25 mg/kg) i.p injection led to elevated ALT, AST, CK-MB and urea levels in rats.At the given dose, doxorubicin induced pathological changes in cardiac, liver and renal tissues.Pretreatment with Kleinhovia sp. extract prior to doxorubicin injection significantly reduced the elevation of ALT, AST and CK-MB, especially at the dose of 250 mg/kg.Improvement in histological structures of cardiac, liver and renal tissues was shown in Kleinhovia sp. (250 mg/kg) treated rats, indicating a protective effect of the extract on doxorubicin acute toxicity. Abbreviations Used: DOX: Doxorubicin; CK-MB: creatine kinase-MB, AST: Aspartate transaminase; ALT: Alanine transaminase.