Antifungal Mechanism of Action of Lauryl Betaine Against Skin-Associated Fungus Malassezia restricta

Betaine derivatives are considered major ingredients of shampoos and are commonly used as antistatic and viscosity-increasing agents. Several studies have also suggested that betaine derivatives can be used as antimicrobial agents. However, the antifungal activity and mechanism of action of betaine derivatives have not yet been fully understood. In this study, we investigated the antifungal activity of six betaine derivatives against Malassezia restricta, which is the most frequently isolated fungus from the human skin and is implicated in the development of dandruff. We found that, among the six betaine derivatives, lauryl betaine showed the most potent antifungal activity. The mechanism of action of lauryl betaine was studied mainly using another phylogenetically close model fungal organism, Cryptococcus neoformans, because of a lack of available genetic manipulation and functional genomics tools for M. restricta. Our genome-wide reverse genetic screening method using the C. neoformans gene deletion mutant library showed that the mutants with mutations in genes for cell membrane synthesis and integrity, particularly ergosterol synthesis, are highly sensitive to lauryl betaine. Furthermore, transcriptome changes in both C. neoformans and M. restricta cells grown in the presence of lauryl betaine were analyzed and the results indicated that the compound mainly affected cell membrane synthesis, particularly ergosterol synthesis. Overall, our data demonstrated that lauryl betaine influences ergosterol synthesis in C. neoformans and that the compound exerts a similar mechanism of action on M. restricta.

Comparison of Single Piece of Dandruff DNA Extraction under Microscope and EZ-tape Method / 法医学杂志

OBJECTIVES@#To collect single piece of dandruff with microscopes to improve the regular EZ-tape method for DNA extraction and genotyping, increase the utilization of samples, reduce the miss rate as well as the proportion of genotyping results of mixed stains.@*METHODS@#The insides of the hats worn by two volunteers were stuck by EZ-tape and scotch tape respectively. DNA on EZ-tape was directly extracted using traditional method. Single piece of dandruff was collected from the scotch tapes under microscope. The two kinds of methods were both performed under continuous oscillation and standing digestion, respectively. DNA was extracted through Chelex-100 method, and STR genotypes were obtained after amplification and electrophoresis. The results of STR genotypes obtained by EZ-tape method and single piece of dandruff analytical method were compared.@*RESULTS@#Miss detections happened in 11 samples （45.8%） by EZ-tape method and only single-source typing results were obtained. Ten samples （41.7%） showed the genotype results of mixed stain and six of which showed allele insertions and deletions. The genotype results were obtained successfully using the single piece of dandruff analytical method, and two samples showed mixed stain genotype. The number of exact typing processed by oscillation was higher than that by standing digestion （ P<0.05）.@*CONCLUSIONS@#The oscillation during the DNA extraction process is in favour of the DNA releasing. Single piece of dandruff analytical method can be used to obtain single-source STR genotype with high successful ratio and low miss rate. This method can be a collection method of special samples such as dandruff in forensic practice.

Estrogenic activity of zinc pyrithione: an in vivo and in vitro study

Zinc pyrithione (ZP) is commonly used to prevent dandruff and seborrheic dermatitis. Many consumers are exposed daily to high doses of ZP, causing serious concerns about its toxicity. The reproductive and developmental toxicities were previously reported in pregnant rats. However, the estrogenic activity of ZP at varying degrees of exposure has been rarely studied. Thus, we performed an uterotrophic assay, E-screen assay, and gene expression profiling to assess the estrogenic activity of ZP. For the uterotrophic assay, ZP (2, 10, or 50 mg/kg/d) was subcutaneously administered to ovariectomized rats every day for three days. Uteri were extracted 24 hours after the last dose. Then, wet and blotted uterine weights were measured. For the E-screen essay, MCF-7 cells (a breast cancer cell line) were exposed to 10⁻⁹ to 10⁻⁶ M of ZP, and cell proliferation was then measured. For the gene expression analysis, changes of gene expression levels in uterine samples taken for the uterotrophic assay were analyzed. In the uterotrophic assay, the concentration of ZP had no significant effect on uterine weight. In the E-screen assay, ZP at any concentration showed no significant increase in MCF-7 cell proliferation, compared to the control group. However, 10⁻⁶ M of ZP significantly reduced cell viability. The changes in gene expression slightly differed between the ZP and control groups. The in vivo and in vitro assays, together with gene expression analysis, demonstrated that ZP showed no significant estrogenic activity.

Pharmacologic treatment of seborrheic dermatitis

Seborrheic dermatitis is a chronic, recurrent papulosquamous skin disorder characterized by flaking of the skin, erythema, pruritus, and oily skin. It is a common dermatosis, with a prevalence of approximately 1% to 3% of the general population in the United States. The incidence of this skin disease has a bimodal distribution, with peaks noted in newborns and in adults between 30 and 60 years of age. In adults, it predominantly occurs over areas of the body with a high concentration of sebaceous glands. The etiology of this disease is thought to be an inflammatory response of the body to Malassezia yeasts. It is also related to abnormal sebum secretion, as well as a heightened immune response of the host. Based on the characteristic features of this inflammatory skin disease, the mainstays of treatment for seborrheic dermatitis are antifungals, corticosteroids, and immunomodulatory agents. The primary goal of therapy for seborrheic dermatitis is to manage acute symptoms, including pruritus, erythema, and scaling. Maintenance treatment is directed toward preventing the recurrence of acute exacerbations. Patients should be informed that seborrheic dermatitis is a chronic condition, with a tendency to recur despite proper treatment. Clinicians should offer suggestions about lifestyle modifications to avoid triggers and choose proper treatment options that ensure the patient's safety.

Estrogenic activity of zinc pyrithione: an in vivo and in vitro study

Zinc pyrithione (ZP) is commonly used to prevent dandruff and seborrheic dermatitis. Many consumers are exposed daily to high doses of ZP, causing serious concerns about its toxicity. The reproductive and developmental toxicities were previously reported in pregnant rats. However, the estrogenic activity of ZP at varying degrees of exposure has been rarely studied. Thus, we performed an uterotrophic assay, E-screen assay, and gene expression profiling to assess the estrogenic activity of ZP. For the uterotrophic assay, ZP (2, 10, or 50 mg/kg/d) was subcutaneously administered to ovariectomized rats every day for three days. Uteri were extracted 24 hours after the last dose. Then, wet and blotted uterine weights were measured. For the E-screen essay, MCF-7 cells (a breast cancer cell line) were exposed to 10⁻⁹ to 10⁻⁶ M of ZP, and cell proliferation was then measured. For the gene expression analysis, changes of gene expression levels in uterine samples taken for the uterotrophic assay were analyzed. In the uterotrophic assay, the concentration of ZP had no significant effect on uterine weight. In the E-screen assay, ZP at any concentration showed no significant increase in MCF-7 cell proliferation, compared to the control group. However, 10⁻⁶ M of ZP significantly reduced cell viability. The changes in gene expression slightly differed between the ZP and control groups. The in vivo and in vitro assays, together with gene expression analysis, demonstrated that ZP showed no significant estrogenic activity.

Efficacy of Cistanche Tubulosa and Laminaria Japonica Extracts (MK-R7) Supplement in Preventing Patterned Hair Loss and Promoting Scalp Health

Cistanche tubulosa and Laminaria japonica have been reported to have anti-oxidative, anticoagulant, anti-cancer and anti-inflammatory properties. They are expected to be a promising candidates for promoting hair growth and treating dandruff and scalp inflammation as a consequence. In this double-blinded, placebo-controlled clinical trial, we investigated the efficacy of Cistanche tubulosa extract and Laminaria japonica extract complex (MK-R7) in promoting hair health in patients with mild to moderate patterned hair loss. Using phototrichogram (Folliscope 4.0, LeadM, Seoul, Korea), we compared the density and diameter of hairs in patients receiving a placebo or Cistanche tubulosa extract and Laminaria japonica extract complex (MK-R7) at baseline, 8 and 16 weeks of the study. In order to determine the efficacy of treatment on dandruff and scalp inflammation, investigator's assessment score and patient's subjective score were also performed. We found a statistically significant increase in the hair density of the test group (n = 45, MK-R7 400 mg) after 16 weeks of consuming the MK-R7 (test group: 23.29 n/cm2 +/- 24.26, control: 10.35 n/cm2 +/- 20.08, p < 0.05). In addition, we found a statistically significant increase in hair diameter in the test group compared to control group at week 16 (test group: 0.018 mm +/- 0.015, control: 0.003 mm +/- 0.013, p < 0.05). There were also significant outcomes regarding the investigator's visual assessment and patient's subjective score of dandruff and scalp inflammation in the test group compared to those in control group. Based on the results of this clinical study, we conclude that Cistanche tubulosa extract and Laminaria japonica extract complex (MK-R7) are promising substances for promoting health of the scalp and hair.

Effectiveness of Home-use Light Emitting Diode Device for Seborrheic Dermatitis of the Scalp: a Preliminary Study / 대한의진균학회지

BACKGROUND: Seborrheic dermatitis (SD) is a chronic, relapsing inflammatory disease, mainly affecting the scalp and face. The pathogenesis of SD has been not fully understood yet, but may be related to the skin colonization of Malassezia species, lipophilic yeasts. Phototherapy with light emitting diode (LED) device has been become a new therapeutic modality for some skin diseases such as acne. OBJECTIVE: To evaluate effectiveness and safety of phototherapy with home-use LED in the treatment of patients with SD of the scalp. METHODS: Eight patients with mild-to-moderate SD of the scalp participated. The patients used a home-use LED device combined 395 nm blue light with 660 nm red light for 6~7 min twice daily for 8 weeks. Patients' assessments were made by clinical findings including erythema (0~15), scales (0~15), itching (0~10), and lesional extent (0~15). And clinical photographs were taken at 0, 2, 4 and 8 weeks, respectively. RESULTS: At 8 weeks after LED treatment, erythema (7.75 to 5.13, p=0.018), scales (7.38 to 4.13, p=0.017), itching (6 to 3, p=0.011) and lesional extent (8.25 to 5.25, p=0.017) were significantly decreased, respectively. Satisfactory scores were also relatively high (mean 8.13 of 10). No severe adverse reaction was reported, excepting hair dryness (n=1) and brief stinging sense (n=1). CONCLUSION: Home-use LED was effective and safe in the treatment of SD of the scalp. These results suggest the LED device might be an adjuvant therapeutic tool in the treatment of Malassezia species associated diseases such as dandruff and SD. Further long-term and large-scale studies are required to assess the efficacy and safety of home-use LED.

[Microscopic examination in quantifying of Malassezia yeast in scalp and rapid diagnosis of fungi invasive condition: a brief report]

Malassezia Species are often commensal of the human skin and scalp that opportunistically in exist of particular predisposing factors, their proliferation increases; as, in dandruff and seborrheic dermatitis which both togather affect more than 50% of humans, the excess proliferation of yeast in scalp, leads to scalp-flaking and causes physical and mental disorder in peaple, spacially in youth that their health and hiar hygiene and beauty is more important for them. Thus, this survey has been done for rapid, easy and inexpensive method to diagnosis of abnormal proliferation and invasive condition of Malassezia yeast and can be more benefical for proper treatment. Sampling with scalpel scraping from scalp of volunteer persons that had not bathed at least two day ago were done and preparation of direct microscopic slides and staining with methylene blue were accomplished. Then, survey of morpholgic characteristics, yeast quantification and mycelium detection were done by direct microscopic examination. From 140 scalp samples of adult persons of both gender [male and female] with different age groups, observation of malassezia yeast in 93.5% [131] were positive and 6.5% [9] were negative in direct microscopic examination. Results of yeast quantification in positive cases were: mild or normal flora 25.2%, intermediate 24.5%, severe 50.3%. Detection of mycelium in positive cases were 22.9% [30] [P=0.007 df=2]. Application of an accessible, easy and inexpensive method and a determinated pattern [yeast quantification with direct microscopic examination] to distinguish normal flora from abnormal condition [excess proliferation and mycelium production] in cases of Malassezia yeasts can be more useful to rapid diagnosis of abnormal proliferation and invasive condition in order to initiate a proper antifungal treatment.

A randomized controlled trial comparing the efficacy of 10% guava extract shampoo versus 2% ketoconazole shampoo and clobetasol 0.05% shampoo in the treatment of scalp seborrheic dermatitis

BACKGROUND: Seborrheic dermatitis is a common chronic, relapsing inflammatory skin condition occurring most often on the face, scalp and chest.OBJECTIVE: To compare the efficacy and safety of 10% guava extract shampoo versus 2% ketoconazole shampoo and clobetasol 0.05% shampoo in the treatment of scalp seborrheic dermatitis.METHODS: Forty-five patients clinically diagnosed with scalp seborrheic dermatitis were randomized to one of the three treatment groups: 10% guava extract shampoo, 2 % ketoconazole shampoo and 0.05% clobetasol shampoo. Patients were followed-up weekly for two weeks to assess the Seborrheic Area Severity Index (SASI) scores, pruritus score, patient's global assessment of improvement and adverse events.RESULTS: After two weeks of treatment, there was no significant difference in the three groups in terms of the mean change in SASI scores (p= 0.46, ANOVA), pruritus scores and in the patient's global assessment of improvement, and frequency of adverse events.CONCLUSION: 10% guava extract shampoo is a safe and effective alternative treatment for scalp seborrheic dermatitis.

Clinical Characteristics of Adolescence and Adult Atopic Dermatitis in Korea / 대한피부과학회지

BACKGROUND: Atopic dermatitis (AD) is a very common disease with peak prevalence among infants and children. Until now, adolescence and adult AD (AAD) patients have been overlooked because they were relatively small population and AAD were considered merely consecutive of infant and childhood AD. OBJECTIVE: The aim of this study is to investigate clinical characteristics of AAD in Korea and different characteristics of childhood AD. METHODS: We evaluated the clinical features of AD in 48 patients. We obtained informations about the age of onset, associated allergic diseases, aggravating factors and seasonal aggravation from the history using a standardized questionnaire. And clinical features of all subjects were evaluated by one investigator and recorded according to anatomical region. RESULTS: 23 patients (48%) reported onset of AD was after 12 years old and 25 patients (52%) reported onset before 12 years old. Only nine patients have suffered AD from infancy. 25 patients (52%) were associated with allergic asthma and/or rhinitis. Most important aggravating factor was climate with seasonal variation. Other common aggravating factors were stress, sweating, dryness and house dust. And numerous patients who experienced seasonal aggravation replied that symptoms had flared from July to August and December. The face, trunk, extremities, scalp and neck were common sites of dermatitis and most of the patients (93%) showed the facial involvement. Red face (68%) and dandruff (45%) were the most common features of the face and the scalp, respectively. About one third of the patients showed the involvement of the neck, presenting as dirty neck or dermatitis. Dryness of the trunk and eczema of flexural areas of the extremities were also common. And 13 patients had hand-foot dermatitis. Keratosis pilaris, hyperlinear palm and ichthyosis vulgaris were detected in 21 (43%), 14 (29%) and 9 patients (18%). CONCLUSION: This study shows AAD is so different from childhood AD, suggesting that AAD may not be merely consecutive of infant and childhood AD.

Pityrosporum(Malassezia) Related Diseases Especially Seborrheic Dermatitis / 대한의진균학회지

The cause of seborrheic dermatitis is unknown, although many attempts have been made to relate it to infection with bacteria or with ,Malassezia furfur. Recently, there have been many studies concerning the efficacy of anti-fungal agents against seborrheic dermatitis. In this occasion, I introduce the result of studies concerning the efficacy of antimycotic drugs against dandruff and seborrheic dermatitis in Japan and in our outpatient clinic of dermatology. I also report the results of our laboratory studies concerning the mechanism of seborrheic dermatitis. We produced the animal models of seborrheic dermatitis. Lesions in immunized guinea pigs developed faster than those of non immunized guinea pigs. The anti-Malassezia IgY seems to delay the development of the lesion. Malassezia spores were detected from lesional skin by direct light microscopy method in our outpatient clinic. Approximately 85% of the facial sites and 75% of the scalp sites were positive in total. One of our clinical studies was a double-blind, comparative study conducted at 8 institutions on the usefulness of shampoo containing 0.75% miconazole nitrate (MZS) for the treatment of dandruff. Shampoo vehicle (BSS) without the reagent served as the control. MZS was evaluated as useful in 34 of the 58, and BSS in 19 of the 50 subjects, thus MZS was significantly more useful than BSS (p=0.020). The other clinical study concerned on the efficacy of ketoconazole cream used in 168 patients for the treatment of seborrheic dermatitis. This study was done as an open trial by multiple institutes. Approximately 80% evaluated as effective, in total. In 23 valuable cases, fungus disappeared in 16 cases (69.6%). It is concluded that anti-fungal agents have potential value as a new therapeutic agent -an alterative to steroids- for the treatment of seborrheic dermatitis.

The study of incidence and distribution of pityrospoum on clinically normal skin / 대한피부과학회지

BACKGROUND: Previous studies have suggested that pityrosporurr yeasts, normal resident flora of skin, may be important in the pathogenesis of dandruff seborrheic dermatitis and confluent and reticulated papillomatosis. OBJECTIVE: We investigated the incidence and distribution of pityiosorum yeasts on clinically normal skin as a more reasonable method for providing the basic data or proving the relationship of pityrosporum and pathogenesi, of the diseases. MEHTODS: A total of 350 subjects were studied. Skin scrapings were stained with 20% Parker ink-KOH(P-KOH) solution. The numbers of pityrosporum spores and hyphe per high power field( x 400) were counted and designated according to a new grading method using bacterial index(BI) of lepra bacilli. RESULTS: The incidence rates of pityrosporum orbiculare(p. orbiculare) and pityrosporum ovale(p. ovale) were 60-70% and 20-50% of subjects respectively on seborrheic area., The incidence of p. orbicilare was highter than that of p. ovale on all examined sites except for the iterior scalp. And pityrosporal hyphae was present on anteriand posterior scalp and earwax in 24-25% of subjects respectively. The incidence of pityrosporum was higher in summer than other seasons The incidence rate of p. orbiculare and hyphae in the group aged below 9 (except neonates) and above 60, were lower than that of other age groups. The inciderice rates and distribution of grades of pityrosporum of neonates were lower than those of older subjects. CONCLUSION: The results of this study showed that the incidence and distribution of pityrosporum spores were similar to previoureports and hyphal forms were found 25% approximately on clinically normal skin of the scalp and carwax. The significance of the presenet of pityosporal hyphae and the relationship between normal skin and pityrosporal hyphae are to be firther elucidated.

Statistical and mycologic studies on dandruff / 대한피부과학회지

BACKGROUND: Previous studies suggested that Pityrosporum may play an important role in the pathogenesis of dandruff. OBJECTIVE: Our purpose was to study the statistical and mycobgic findings of dandruff in normal subjects for better understanding of this condition. MEHTODS: Scales frorn the scalp of 384 students were studied stastistically by answer sheets and clinically by a dermatologist. Mycologic studies of Pityrosporum were also performed by direct smear, culture and inoculation to guinea pigs. RESULTS: There were 76.0% subjects with dandruff. Clinically grading the subjects by Kligrnan's scale, grade III and IV(mild caling) were the most. Dandruff developed commonly at the age of 14 to 16, and no seasonal change was observed. Spores of Pityrosporum were observed in all subjects by KOH microscopic examination with Cohens method, and the grading of the dandruff was not related with the number of the spores. In most subjects, P. orbiculare and P. ovae were found together. Positive culture colonies were observed in 93.7%. Inoculation of the cultured colonies to guinea pigs resulted in an experimental dermatitis. CONCLUSION: Our study shows that Pityrosporum may involve in the pathogenesis of dandruff; however, it is still unclear whether Pityrosporum is a main causative agent of dandruff.