Primary Irritation Research Articles

Exploring the Potent Combination of Quercetin-Boronic Acid, Epalrestat, and Urea Containing Nanoethosomal Keratolytic Gel for the Treatment of Diabetic Neuropathic Pain: In Vitro and In Vivo Studies.

Transdermal penetration of therapeutic moieties from topical dosage forms always remains a challenge due to the presence of permeation impeding keratin which should be addressed. The purpose of the study was to formulate quercetin and 4-formyl phenyl boronic acid (QB complex) used for the preparation of nanoethosomal keratolytic gel (EF3-G). The QB complex was confirmed by Fourier transform infrared spectroscopy while skin permeation, viscosity, and epalrestat entrapment efficiency were used for the optimization of nanoethosomal gel. The keratolytic effect of the proposed nanoethosomal gel with urea (QB + EPL + U) was calculated in rat and snake skin. The spherical shape of nanoethosomes was confirmed by scanning electron microscopy. According to the findings of stability studies, viscosity decreases as temperature increases, proving their thermal stability. The negative charge of optimized EF3 with 0.7 PDI proved narrow particle size distribution with homogeneity. Optimized EF3 showed two folds increase of epalrestat permeation in highly keratinized snake skin as compared to rats' skin after 24 h. Antioxidant behaviors of EF3 (QB) > QB complex > quercetin > ascorbic acid proved reduction of oxidative stress in DPPH reduction analysis. Interestingly, the hot plate and cold allodynia test in the diabetic neuropathic rat model reduced 3-fold pain as compared to the diabetic control group which was further confirmed by in vivo biochemical studies even after the eight week. Conclusively, ureal keratolysis, primary dermal irritation index reduction, and improved loading of epalrestat render the nanoethosomal gel (EF3-G) ideal for the treatment of diabetic neuropathic pain.

PHYSICAL STABILITY TEST HAIR TONIC COMBINATION Cinnamon (Cinnamomum burmanii) ESSENTIAL OIL AND VCO (VIRGIN COCONUT OIL) WITH CYCLING TEST METHOD AND IRRITATION TEST ON THE RABBIT

Cinnamon (Cinnamomum burmanii) and VCO (Virgin Coconut Oil) essential oils contain cinnamaldehyde and lauric acid which have potential for hair growth, they can affect the physical stability of hair tonic preparations. This study aims to determine the physical stability and determine the effect of the irritation effect on hair tonic preparations of a combination of cinnamon essential oil and VCO. Cinnamon essential oil hair tonic preparations were made with a concentration of 1% (F1), 3% (F2), and 5% (F3) and then tested for physical characteristics including organoleptic tests, homogeneity, pH, density test, and viscosity. Evaluation of physical stability was carried out using the cycling test method. Data were statistically analyzed using One Way Anova. The results of the irritation test were analyzed descriptively using the primary irritation index obtained from the appearance of erythema and edema on the rabbit's skin. Observation of the physical stability test on organoleptic and homogeneity showed that formula I had a cloudy white color, a characteristic cinnamon aroma, a slightly thick liquid, and was homogeneous; formula 2 and 3 has a clear color, characteristic of cinnamon aroma, slightly viscous and homogeneous liquid; stable pH in formula 2; stable density in formulas 1, 2 and 3; and stable viscosity in formulas 2 and 3. The results of the hair tonic irritation test showed that the primary irritation index for F1, F2, and F3 was 0.0; 0.3; 0.3 and it can be concluded that F2 and F3 can irritate with very mild categories.

Sericin: From Cocoons as a Raw Material to Cosmetics

Background: Sericin is a hydrophilic and adhesive protein that provides structural integrity to the silkworm cocoon. Objective: The study aimed to develop sericin isolated from Indian BV mulberry silk cocoons as a cosmetic raw material and study its sensory impact. Methods: Sericin was isolated using high-temperature and high-pressure techniques without the use of chemicals. Native and hydrolyzed sericin were studied for suitability in cosmetic formulations. Lastly, their safety was established using a primary irritation patch test in twenty-four healthy volunteers, and the impact of sericin on skin care formulation was studied. Results: Both native and hydrolyzed sericin were studied for their application in personal care products. Native sericin was difficult to process due to its gelling behavior. Hydrolyzed sericin was readily formulated in skin lotion and was found to be stable for 3 months under accelerated conditions. Sericin and its formulations did not cause any skin irritation and were found to be well-tolerated. Formulation with 0.01% w/w hydrolyzed sericin was best accepted among the panelists. Conclusion: The study establishes the use of sericin isolated from mulberry silkworms as a sensory enhancer for skin care applications.

Development of Low-Contact-Impedance Dry Electrodes for Electroencephalogram Signal Acquisition.

Dry electroencephalogram (EEG) systems have a short set-up time and require limited skin preparation. However, they tend to require strong electrode-to-skin contact. In this study, dry EEG electrodes with low contact impedance (<150 kΩ) were fabricated by partially embedding a polyimide flexible printed circuit board (FPCB) in polydimethylsiloxane and then casting them in a sensor mold with six symmetrical legs or bumps. Silver-silver chloride paste was used at the exposed tip of each leg or bump that must touch the skin. The use of an FPCB enabled the fabricated electrodes to maintain steady impedance. Two types of dry electrodes were fabricated: flat-disk electrodes for skin with limited hair and multilegged electrodes for common use and for areas with thick hair. Impedance testing was conducted with and without a custom head cap according to the standard 10-20 electrode arrangement. The experimental results indicated that the fabricated electrodes exhibited impedance values between 65 and 120 kΩ. The brain wave patterns acquired with these electrodes were comparable to those acquired using conventional wet electrodes. The fabricated EEG electrodes passed the primary skin irritation tests based on the ISO 10993-10:2010 protocol and the cytotoxicity tests based on the ISO 10993-5:2009 protocol.

Design of an Ante-enhancer with an Azone-Mimic Structure using Ionic Liquid.

Laurocapram (Azone) was broadly examined as a representative enhancer of skin penetration in the 1980s. However, it was not approved for treatment because it caused skin irritation following its penetration into the epidermis through the stratum corneum. In the present study, a so-called ante-enhancer with an Azone-mimic structure was designed based on an ante-drug with negligible systemic toxic effects following its permeation through the skin. The ante-enhancer was designed using ionic liquid technology: an ionic liquid-type ante-enhancer (IL-Azone) with an Azone-mimic structure was prepared from ε-caprolactam and myristic acid as cationic and anionic substances, respectively. The enhancing effects of IL-Azone on the permeation by the following model drugs through pig skin were examined: isosorbide 5-mononitrate (ISMN), antipyrine (ANP), and fluorescein isothiocyanate dextran (FD-4). Skin irritation by IL-Azone was assessed using the Draize method. The primary irritation index (P.I.I.) of IL-Azone by the Draize method was markedly lower than that of Azone (6.9). Although the ability of IL-Azone to enhance skin penetration was not as high as Azone, IL-Azone moderately increased skin permeation by the model compounds tested (ISMN: 4.7 fold, ANP: 4.5 fold, FD-4: 4.0 fold). These results suggest the usefulness of designing a skin penetration enhancer using ionic liquid technology. Further trials on the ionic liquid design with an Azone-mimic structure using other cations and anions may lead to the development of better ante-enhancers.

Acenocoumarol, an Anticoagulant Drug, Prevents Melanogenesis in B16F10 Melanoma Cells.

Hyperpigmentation can occur in abnormal skin conditions such as melanomas, as well as in conditions including melasma, freckles, age spots, seborrheic keratosis, and café-au-lait spots (flat brown spots). Thus, there is an increasing need for the development of depigmenting agents. We aimed to repurpose an anticoagulant drug as an effective ingredient against hyperpigmentation and apply cosmeceutical agents. In the present study, the anti-melanogenic effects of two anticoagulant drugs, acenocoumarol and warfarin, were investigated. The results showed that both acenocoumarol and warfarin did not cause any cytotoxicity and resulted in a significant reduction in intracellular tyrosinase activity and melanin content in B16F10 melanoma cells. Additionally, acenocoumarol inhibits the expression of melanogenic enzymes such as tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2, suppressing melanin synthesis through a cAMP-dependent, protein kinase (PKA)-dependent downregulation of microphthalmia-associated transcription factor (MITF), a master transcription factor in melanogenesis. Furthermore, anti-melanogenic effects were exerted by acenocoumarol through downregulation of the p38 and JNK signaling pathway and upregulation of extracellular signal-regulated kinase (ERK) and phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt)/glycogen synthesis kinase-3β (GSK-3β) cascades. In addition, the β-catenin content in the cell cytoplasm and nucleus was increased by acenocoumarol through a reduction in the phosphorylated β-catenin (p-β-catenin content). Finally, we tested the potential of acenocoumarol for topical applications by conducting primary human skin irritation tests. Acenocoumarol did not induce any adverse reactions during these tests. Based on the results, it can be concluded that acenocoumarol regulates melanogenesis through various signaling pathways such as PKA, MAPKs, PI3K/Akt/GSK-3β, and β-catenin. These findings suggest that acenocoumarol has the potential to be repurposed as a drug for treating hyperpigmentation symptoms and could provide new insights into the development of therapeutic approaches for hyperpigmentation disorders.

Aspirin microcrystals deposited on high-density microneedle tips for the preparation of soluble polymer microneedles.

To reduce mucosal damage in the gastrointestinal tract caused by aspirin, aspirin microcrystals were loaded in soluble polymeric microneedle (MN) tips. Aspirin was prepared into aspirin microcrystals by jet milling. Aspirin microcrystals with particle sizes of 0.5-5μm were loaded on MN tips with a height of 250µm or 300µm. The aspirin microcrystals suspended in a polymer solution were concentrated in the MN tips under negative pressure. The aspirin microcrystals had high stability in the MNs since they were not dissolved in solution during the fabrication process. The MN patch packaged in an aluminum-plastic bag containing silica gel desiccant can be stored at 4°C. The MN tips implanted in the skin of Institute of Cancer Research (ICR) mice dissolved within 30min. Isolated porcine ear skin was punctured by MNs with heights of 300μm and 250μm to depths of 130μm and 90μm, respectively. The fluorescent red (FR) release from MNs reached 98.59% within 24h. The MNs delivered aspirin microcrystals to the epidermis and dermis, providing a smooth plasma concentration in rats. The MNs loaded with aspirin microcrystals did not evoke primary irritation on the dorsal skin of Japanese white rabbits. In summary, MNs loaded with aspirin microcrystals provide a new approach to improve the stability of aspirin in MN patches.

A 7-Hydroxy 4-Methylcoumarin Enhances Melanogenesis in B16-F10 Melanoma Cells.

The objectives of this study were to investigate the melanogenetic potentials of the naturally occurring 7-hydroxy coumarin derivatives 7-hydroxy 5,6-dimethoxycoumarin (7H-5,6DM), 7-hydroxy 6,8-dimethoxycoumarin (7H-6,8DM), 7-hydroxy 6-methoxycoumarin (7H-6M), and 7-hydroxy 4-methylcoumarin (7H-4M) in the melanogenic cells model for murine B16F10 melanoma cells. The initial results indicated that melanin production and intracellular tyrosinase activity were significantly stimulated by 7H-4M but not by 7H-5,6DM, 7H-6,8DM, or 7H-6M. Therefore, our present study further investigated the melanogenic effects of 7H-4M in B16-F10 cells, as well as its mechanisms of action. In a concentration-dependent manner, 7H-4M increased intracellular tyrosinase activity, leading to the accumulation of melanin without affecting the viability of B16-F10 cells. Our study further investigated the effects of 7H-4M on melanogenesis, including its ability to promote tyrosinase activity, increase melanin content, and activate molecular signaling pathways. The results indicate that 7H-4M effectively stimulated tyrosinase activity and significantly increased the expression of melanin synthesis-associated proteins, such as microphthalmia-associated transcription factor (MITF), tyrosinase, tyrosinase-related protein-1 (TRP1), and TRP2. Based on our findings, we can conclude that 7H-4M has the ability to activate the melanogenesis process through the upregulation of cAMP-dependent protein kinase (PKA) and the cAMP response element-binding protein (CREB). Additionally, our study showed that 7H-4M induced melanogenic effects by downregulating the extracellular signal-regulated kinase (ERK) and the phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt)/glycogen synthesis kinase-3β (GSK-3β) cascades, while upregulating the JNK and p38 signaling pathways. Finally, the potential of using 7H-4M in topical applications was tested through primary human skin irritation tests. During these tests, no adverse reactions were induced by 7H-4M. In summary, our results indicate that 7H-4M regulates melanogenesis through various signaling pathways such as GSK3β/β-catenin, AKT, PKA/CREB, and MAPK. These findings suggest that 7H-4M has the potential to prevent the development of pigmentation diseases.

Moisturized and non-irritating hand gel based on sappan wood (Caesalpinia sappan l.) and limau citrus peel (Citrus amblycarpa (hassk.) ochse) extracts.

Hand gel is a preparation used to protect hand skin from dryness. The antioxidant compounds in sappan wood and limau citrus peel can moisturize hands by preventing the oxidation of oils and fats on the surface of the (sebaceous glands). This study aims to formulate hand gel from ethanol extract of sappan wood with a combination of limau citrus peel extract that can moisturize hands and not to cause irritation. Sappan wood and limau citrus peel simplicia were macerated using 96% ethanol. The ethanolic extracts were then formulated into hand gel by varying the concentration of the ethanol extracts. The evaluation of the hand gel involved an organoleptic test, homogeneity test, pH test, spreadability test, viscosity test, and skin moisture test. A primary irritation test was carried out to ensure the hand gel's safety. The results shows that the three hand gel formulas produce an orange-red color and soft scent of roses and citrus, with a pH of 7.8 and with good homogeneity. The formulas also have a spreading level of FI (5 cm), FII (5.2 cm), and FIII (5, 1 cm), and viscosity values were FI 12000 mPas, F II 12500 mPas, and F III 12400 mPas. The humidity test results on the three formulas shows different moisture values of 45.16%, 46.17%, and 45.09%, respectively (category of normal or moisture). The hand gel formulas have an irritation index of 0 (no irritation category). The hand gel formulas meet the quality requirements of hand gel, have moisturizing agent, and are not irritating.

Formulasi Masker Peel-Off Ekstrak Biji Buah Pepaya (Carica papaya L.) Sebagai Antijerawat dengan Variasi Konsentrasi Polivinil Alkohol

Papaya seeds have the ability to an antibacterial because they contain triterpenoid compounds, papain enzymes, and fatty acids such as oleic acid, palmitic acid, and linoleic acid which can moisturize the skin and shed dead skin cells so that they are suitable for making peel-off masks. The study aimed to obtain a peel-off mask formulation of papaya seed extract (Carica papaya L.) as a stable anti-acne and has good pharmaceutical properties. The research design used was laboratory experimental research. Papaya fruit seed extract peel-off masks are made in 2 formulas with variations in PVA concentrations, namely 10% and 12%. The peel-off masks obtained were tested for stability including organoleptic testing, homogeneity, viscosity, dispersion ability test, pH test, drying speed test, and primary irritation test. The results showed that papaya fruit seed extract can be made into a peel-off mask formula that meets the pharmaceutical stability test, but formula 2 shows the most optimal stability and good pharmaceutical properties compared to formula 1. In addition, the two formulas do not irritate the skin so they are safe to use when applied.

Uji Aktivitas Krim Anti-Aging Ekstrak Etanol Daun Bayam Merah (Amaranthus tricolor L.) pada Kulit Punggung Kelinci New Zealand yang dipapar Sinar UV-A

The strongest cause of premature aging is sunlight (photoaging), especially UV A rays. Anti-aging creams containing antioxidant compounds can be used in the treatment of aging on the skin of the face. Red spinach leaves (Amaranthus tricolor L.) are a natural source of antioxidants that have very strong potential as an anti-aging. Objectives: This study aims to determine the activity of red spinach leaf extract cream as an anti-aging using New Zealand rabbits exposed to UV-A rays. The skin of the rabbit's back is smeared with cream F1, F2, F3, F4, F5 for 30 days. Parameter observation using a skin analyzer is carried out before, after induction, and after cream smearing. Parameters include percent collagen, elasticity, and moisture. The results of the study were analyzed statistically using one way anova. All test creams pass the physical quality test requirements, but the creams that pass the stability test are F1, F2, F4. The primary irritation test F1, F2, F4 is not irritating whereas F3 is very little irritating. Test the ocular irritation of all creams is not irritating. The test results of anti-aging activity show that F2 provides the most effective anti-aging effect because the percent increase in parameters is the greatest.

Antimicrobial ability and free-irritation effect of neem-based lotion cosmeceutical for skin care

Neem (Azadirachta indica) is well-known as a medical plant to treat some diseases. The antibacterial activity and primary skin irritation test were evaluated by using ethanolic neem extract. The main objective of the study is to determine the effectiveness of the neem-based lotions against Staphylococcus aureus sp and Pseudomonas aeruginosa sp. A series of lotions were prepared by adding different formulations of pure neem extract, coconut oil (CO) and aloe vera. The comparison was made with base lotion and commercial neem lotion. The lotion with diluted neem in CO and aloe vera had the best result against Staphylococcus aureus sp. and Pseudomonas aeruginosa sp. with the inhibition diameter zone of 19.3 mm and 9.6 mm, respectively. The commercial neem lotion with aloe vera displayed an inhibition diameter zone of 11.1 mm and 14.1 mm against Staphylococcus aureus sp. and Pseudomonas aeruginosa sp, respectively. Furthermore, this lotion was examined in a primary skin irritation test to ensure the possible effect on the skin. Erythema and oedema analyses exhibited a 0 irritation score means no skin irritation was observed on the both right and left back of the rabbit after 72 hrs. Hence, the primary irritation index of the lotion with diluted neem oil lotion and aloe vera was 0 and falls in the standard category of ‘negligible’. The study confirmed that neem lotion has the potential an antimicrobial agent for skin disease treatment and does not cause irritation effect to the skin.

ANTIPSORIATIC ACTIVITY OF HYDROGEL CONTAINING NANOSTRUCTURED LIPID CARRIER (NLC) ENTRAPPED WITH TRIAMCINOLONE ACETONIDE

Objective: The aim of present study was to prepare triamcinolone acetonide (TA) loaded NLCs hydrogel for antipsoriatic activity. Methods: A Nanostructured lipid carrier (NLCs) was prepared by using solvent diffusion and high pressure homogenization methods. NLCs dispersion was characterized by particle size, zeta potential, scanning electron microscopy (SEM), differential scanning calorimetry, and an in vitro release study. Optimized NLC incorporated into the hydrogel and characterized for rheological properties, drug content, in vitro drug release, stability study, skin irritation and antipsoriatic activity for optimized batch of hydrogel. Results: Optimized NLCs loaded with TA were exhibited spherical shape with particle size 286±0.07 nm, polydispersity index 0.317, zeta potential-21.91±0.05mV and entrapment efficiency 86.19±0.06% respectively. The drug release of optimized batch was 8.34 % and 88.84±0.08% at 1h and 8h respectively. The release kinetics of the optimized NLCs best fitted the peppas-korsmeyer model. The results of NLC hydrogel formulations were spreadability 27.4±0.06-11.76±0.07 g. cm2/sec, drug content 65.60±0.05%-74.50±0.02%, in vitro drug release 87.52±0.04%, primary irritation index was 0.0752, it indicates barely perceptible irritation. Histopathological studies showed that, in psoriasis-induced animal treated with TA loaded NLC hydrogel, marked reduction in thickness of epidermis, as compared to conventional gel formulation. It shows the increase % orthokeratosis 88.69% and % drug activity 54.23% than the marketed formulation. Conclusion: The present results demonstrated that hydrogel based NLC shows the better and effective drug delivery for the management of psoriasis.

Novel Siprulina platensis Bilosomes for Combating UVB Induced Skin Damage.

The recent interest in bioactive compounds from natural sources has led to the evolution of the skin care industry. Efforts to develop biologically active ingredients from natural sources have resulted in the emergence of enhanced skin care products. Spirulina (SPR), a nutritionally enriched cyanobacteria-type microalga, is rich in nutrients and phytochemicals. SPR possesses antioxidant, immunomodulatory, and anti-inflammatory activities. Spirulina-loaded bilosomes (SPR-BS), a novel antiaging drug delivery system, were designed for the first time by incorporation in a lecithin−bile salt-integrated system for bypassing skin delivery obstacles. The optimized BS had good entrapment efficiency, small particle size, optimal zeta potential, and sustained drug release pattern. Blank and SPR-loaded BS formulations were safe, with a primary irritancy index of <2 based on the Draize test. In vivo tests were conducted, and photoprotective antiaging effects were evaluated visually and biochemically by analyzing antioxidant, anti-inflammatory, and anti-wrinkling markers following ultraviolet (UV) B irradiation. Results of biochemical marker analysis and histopathological examination confirmed the superior antiaging effect of SPR-BS compared with SPR. Thus, SPR-loaded BS is a promising nanoplatform for SPR delivery, can be used for treating UV-induced skin damage, and offers maximum therapeutic outcomes.

Miglitol, an Oral Antidiabetic Drug, Downregulates Melanogenesis in B16F10 Melanoma Cells through the PKA, MAPK, and GSK3β/β-Catenin Signaling Pathways

Hyperpigmentation is a common condition that causes darker spots or patches on the skin, which often look brown, black, gray, red, or pink. This results in unresolved psychological impact due to high anxiety, depression, and somatoform disorder. We aimed to repurpose an antidiabetic drug, miglitol, as an effective compound against hyperpigmentation when applied as a cosmeceutical agent. The present study investigated the antimelanogenic effects of miglitol and the trehalase inhibitor validamycin A. Miglitol in isolation exhibited no cytotoxicity and significantly reduced the melanin production and intracellular tyrosinase activity in B16F10 melanoma cells. The Western blotting results showed that miglitol reduces the expression of melanogenic regulatory factors, including tyrosinase, tyrosinase-related protein (TRP)-1, TRP-2, and microphthalmia-associated transcription factor (MITF). Mechanistically, miglitol appears to suppress melanin synthesis through cAMP-dependent protein kinase (PKA)-dependent downregulation of MITF, a master transcription factor in melanogenesis. The antimelanogenic effects of miglitol was mediated by downregulation of the p38 signaling pathway and upregulation of extracellular signal-regulated kinase (ERK). Moreover, miglitol decreases P-GSK3β and β-catenin levels compared to those in the untreated group. However, miglitol activated P-β-catenin expression compared to that in the untreated group. Finally, we tested the potential of miglitol in topical application through primary human skin irritation tests on the normal skin (upper back) of 33 volunteers. In these assays, miglitol (125 and 250 μM) did not induce any adverse reactions. Taken together, these findings suggest that the regulation of melanogenesis by miglitol may be mediated by the PKA, MAPK, and GSK3β/β-Catenin signaling pathways and that miglitol might provide new insights into drug repurposing for the treatment of hyperpigmentation symptoms.

Irritation Test of Bawang Dayak (Eleutherine Bulbosa (Mill.) Urb.) Loose Powder for Acne Vulgaris

Several studies revealed that the ethanolic extract, cream, and loose powder of Bawang Dayak bulbs (Eleutherine bulbosa (Mill.) Urb.) are effective in inhibiting the growth of different acne-causing bacteria, including S. aureus, S. epidermidis, and P. acne. Several product series have also been developed using the plant for the treatment of acne-prone skin. Therefore, this study aims to carry out a primary irritation test of E. bulbosa loose powder using the patch test method on rabbits and humans to determine its safety before it is marketed. The results showed that the primary irritation index of E.bulbosa loose powder on rabbits was 0.125, which was classified in the negligible category, and there were no signs of erythema or edema in humans. This indicates that it does not cause irritation and has the potential to be developed into anti-acne products.

DYNAMICS OF MORPHOLOGICAL CHANGES IN CELLS OF THE CEREBELLUM IN ACUTE POISONING WITH CLOZAPINE IN COMBINATION WITH ALCOHOL

According to most researchers, the central mechanism of clozapine poisoning in combination with alcohol is their cholinolytic effect. A life-threatening condition with clozapine occurs as a result of an unintentional overdose of the prescribed drug, suicidal behaviour, or criminal behaviour. Despite the urgency of the problem of clozapine poisoning in combination with alcohol, the pathogenesis of these conditions and the resulting morphological changes in the brain have not been sufficiently investigated. In this connection, the purpose of our experimental research is the study of the dynamics of changes in Purkinje cells of the cerebellum in the early period of acute poisoning with clozapine in combination with alcohol. In sections of the cerebellum of the control group, reversible changes in Purkinje cells (15–20%) predominated in the form of primary irritation and acute swelling. Irreversible damage to Purkinje cells (2-5%). 3 hours after poisoning with clozapine in combination with ethanol, Purkinje cells showed irreversible damage to cerebellar cells with signs of severe changes, neuronophagia, and pericellular edema, with the following ratio: reversible 25-30% and irreversible 60-70%. 24 hours after the combined poisoning with clozapine in combination with alcohol, in percentage terms, the ratio of the changes was the following: reversible 25-30% and irreversible 60-70%. The study revealed the dynamics of reversible and irreversible changes in Purkinje cells with a predominance of irreversible damage was established, especially after 24 hours from the start of the experiment.

Topical garlic treatment for verruca plana triggers Koebner phenomenon: A case report.

Verruca plana is a benign proliferation of the skin caused by human papilloma virus (HPV) infection. Fresh garlic can serve as an alternative therapy, and it has shown considerable effectiveness as a topical treatment for verruca plana. However, topical garlic treatment for verruca plana triggered Koebner phenomenon (KP), which has not been previously reported. The aim of our report is to explore the possible causes of this adverse reaction. We here describe a 20-year-old female patient who developed a beaded rash after garlic treatment for facial wart plana, known as autoinoculation or KP. Garlic may have caused damage to the surrounding normal skin through primary irritation or allergic reactions. Then, the HPV virus on the primary verruca plana took the opportunity to spread to the surrounding skin injured by garlic stimulation, triggering the KP. When using garlic to treat verruca plana, the operator needs to precisely apply the mashed garlic to the warts, and this treatment is strictly prohibited for patients who are allergic to garlic. Avoid such adverse reactions.

Evaluation of skin-irritancy potential of a skin ointment containing the essential oil of ocimum sanctum l (Basil/Thulasi) leaves in Wistar albino rats

Skin irritant potential of a skin ointment containing 4% essential oil extracted from the Ocimum sanctum L. (Basil/Thulassi) leaves was evaluated on the hair clipped skin of female Wistar albino rats. The calculated Primary Irritation Index was ‘0” and it falls under the category of “negligible irritant.

Imperatorin Positively Regulates Melanogenesis through Signaling Pathways Involving PKA/CREB, ERK, AKT, and GSK3β/β-Catenin.

The present study investigated the melanogenic effects of imperatorin and isoimperatorin and the underlying mechanisms of imperatorin using a mouse melanoma B16F10 model. Interestingly, treatment with 25 μM of either imperatorin or isoimperatorin, despite their structural differences, did not produce differences in melanin content and intracellular tyrosinase activity. Imperatorin also activated the expression of melanogenic enzymes, such as tyrosinase (TYR) and tyrosinase-related proteins TYRP-1 and TYRP-2. Mechanistically, imperatorin increases melanin synthesis through the cyclic adenosine monophosphate (cAMP)-dependent protein kinase (PKA)/cAMP-responsive element-binding protein (CREB)-dependent upregulation of microphthalmia-associated transcription factor (MITF), which is a key transcription factor in melanogenesis. Furthermore, imperatorin exerted melanogenic effects by downregulating extracellular signal-regulated kinase (ERK) and upregulating phosphatidylinositol 3 kinase (PI3K)/protein kinase B (AKT)/glycogen synthesis kinase-3β (GSK-3β). Moreover, imperatorin increased the content of β-catenin in the cell cytoplasm and nucleus by reducing the content of phosphorylated β-catenin (p-β-catenin). Finally, we tested the potential of imperatorin in topical application through primary human skin irritation tests. These tests were performed on the normal skin (upper back) of 31 volunteers to determine whether 25 or 50 µM of imperatorin had irritation or sensitization potential. During these tests, imperatorin did not induce any adverse reactions. Taken together, these findings suggest that the regulation of melanogenesis by imperatorin can be mediated by signaling pathways involving PKA/CREB, ERK, AKT, and GSK3β/β-catenin and that imperatorin could prevent the pathogenesis of pigmentation diseases when used as a topical agent.