马拉色菌属的研究进展

马拉色菌是一种寄生于人和动物正常皮肤表面的真菌,可导致机会感染而引起各种马拉色菌属相关疾病。马拉色菌的致病机制主要是分解脂质,导致角质形成细胞形态学改变和细胞凋亡。马拉色菌可引起花斑癣、脂溢性皮炎、马拉色菌毛囊炎等,在特应性皮炎中也起到变应原的作用。对于这些疾病的治疗可使用抗真菌药物。     

Distribution of Malassezia Species on the Scalp in Korean Seborrheic Dermatitis Patients

Background

Malassezia species play an important role in the pathogenesis of seborrheic dermatitis. In particular, M. restricta and M. globosa are considered to be the predominant organisms in seborrheic dermatitis of Western countries. However, species distribution of Malassezia in seborrheic dermatitis has not been clearly determined yet in Asia.

Objective

To identify the distribution of Malassezia species on the scalp of seborrheic dermatitis patients in Korea using 26S rDNA PCR-RFLP analysis.

Methods

A total of 40 seborrheic dermatitis patients and 100 normal healthy volunteers were included in this study. For the identification of Malassezia species, the scalp scales of the subjects were analyzed by 26S rDNA PCR-RFLP analysis.

Results

The most commonly identified Malassezia species were M. restricta in the seborrheic dermatitis patients, and M. globosa in the normal controls. In the seborrheic dermatitis group, M. restricta was identified in 47.5%, M. globosa in 27.5%, M. furfur in 7.5%, and M. sympodialis in 2.5% of patients. In the healthy control group, M. globosa was identified in 32.0%, M. restricta in 25.0%, M. furfur in 8.0%, M. obtusa in 6.0%, M. slooffiae in 6.0%, and M. sympodialis in 4.0% of subjects.

Conclusion

M. restricta is considered to be the most important Malassezia species in Korean seborrheic dermatitis patients.     

Epidemiologic Study of Malassezia Yeasts in Patients with Malassezia Folliculitis by 26S rDNA PCR-RFLP Analysis

Background

So far, studies on the inter-relationship between Malassezia and Malassezia folliculitis have been rather scarce.

Objective

We sought to analyze the differences in body sites, gender and age groups, and to determine whether there is a relationship between certain types of Malassezia species and Malassezia folliculitis.

Methods

Specimens were taken from the forehead, cheek and chest of 60 patients with Malassezia folliculitis and from the normal skin of 60 age- and gender-matched healthy controls by 26S rDNA PCR-RFLP.

Results

M. restricta was dominant in the patients with Malassezia folliculitis (20.6%), while M. globosa was the most common species (26.7%) in the controls. The rate of identification was the highest in the teens for the patient group, whereas it was the highest in the thirties for the control group. M. globosa was the most predominant species on the chest with 13 cases (21.7%), and M. restricta was the most commonly identified species, with 17 (28.3%) and 12 (20%) cases on the forehead and cheek, respectively, for the patient group.

Conclusion

Statistically significant differences were observed between the patient and control groups for the people in their teens and twenties, and in terms of the body site, on the forehead only.     

Epidemiologic Study of Malassezia Yeasts in Seborrheic Dermatitis Patients by the Analysis of 26S rDNA PCR-RFLP

Background

This case-control study concerns a molecular biological method based on the data gathered from a group of Korean subjects to examine the distribution of Malassezia yeasts in seborrheic dermatitis (SD) patients. Cultures for Malassezia yeasts were taken from the foreheads, cheeks and chests of 60 patients with SD and in 60 healthy controls of equivalent age.

Objective

The purpose of this study is to identify the relationship between certain species of Malassezia and SD. This was done by analyzing the differences in the distribution of Malassezia species in terms of age and body parts of the host with healthy controls.

Methods

26S rDNA PCR-RFLP, a fast and accurate molecular biological method, was used to overcome the limits of morphological and biochemical methods.

Results

The positive Malassezia culture rate was 51.7% in patients with SD, which was lower than that of healthy adults (63.9%). M. restricta was dominant in patients with SD (19.5%). Likewise, M. restricta was identified as a common species (20.5%) in healthy controls. In the ages 31~40, M. restricta was found to be the most common species (31.6%) among SD patients.

Conclusion

According to the results of the study, the most frequently isolated species was M. restricta (19.5%) in patients with SD. There was no statistically significant difference in the distribution of Malassezia species between the SD patients and healthy control groups.     

Comparison between Malassezia Folliculitis and Non-Malassezia Folliculitis

Background

Among the various types of folliculitis, differentiation of Malassezia folliculitis (MF) from other forms of folliculitis is important because it is usually treated with antifungal agents.

Objective

We attempted to find a method to enhance the detection rate of MF, and examined the differences in the clinical manifestation between MF and non-MF (NMF).

Methods

We performed a retrospective study involving patients with folliculitis who were previously diagnosed with MF or NMF on the basis of serial tissue sectioning and diastase-Periodic acid-Schiff (d-PAS) staining findings. The clinical features of MF and NMF were compared.

Results

Among a total of 100 folliculitis patients, 20 were diagnosed with MF and 80 with NMF. Tissues from the 80 patients with NMF were sectioned serially into 10 slices and stained with hematoxylin and eosin stain; among these, 10 had many round-to-oval yeast organisms in the hair follicles that confirmed MF. Finally, d-PAS staining was used to detect the presence of yeast in the NMF slides. Notably, among the 70 d-PAS-stained samples, yeast organisms were found in 6 samples, confirming MF. As a result, the diagnosis of 16 patients changed from NMF to MF. Compared with NMF, MF showed major involvement of the trunk and low involvement of the face and legs as well as male predilection.

Conclusion

Physicians should consider serial sectioning and/or d-PAS staining of folliculitis lesions, particularly of those on the trunk of male patients, even if no yeast organisms are detected initially.     

Molecular Analysis of Malassezia Microflora on the Skin of the Patients with Atopic Dermatitis

Background

The yeasts of the genus Malassezia are members of the normal flora on human skin and they are found in 75~80% of healthy adults. Since its association with various skin disorders have been known, there have been a growing number of reports that have implicated Malassezia yeast in atopic dermatitis (AD).

Objective

The aim of the present study is to isolate the various Malassezia species from AD patients by using 26S rDNA (ribosomal Deoxyribonucleic acid) PCR-RFLP and to investigate the relationship between a positive Malassezia culture and the severity of AD.

Methods

Cultures for Malassezia yeasts were taken from the scalp, cheek, chest, arm and thigh of 60 patients with atopic dermatitis. We used a rapid and accurate molecular biological method 26S rDNA PCR-RFLP, and this method can overcome the limits of the morphological and biochemical methods.

Results

Positive Malassezia growth was noted on 51.7% of the patients with atopic dermatitis by 26S rDNA PCR-RFLP analysis. The overall dominant species was M. sympodialis (16.3%). M. restricta was the most common species on the scalp (30.0%) and cheek (16.7%). M. sympodialis (28.3%) was the most common species on the chest. The positive culture rate was the highest for the 11~20 age group (59.0%) and the scalp showed the highest rate at 66.7%. There was no significant relationship between the Malassezia species and SCORing for Atopic Dermatitis (SCORAD).

Conclusion

The fact that the cultured species was different for the atopic dermatitis lesion skin from that of the normal skin may be due to the disrupted skin barrier function and sensitization of the organism induced by scratching in the AD lesion-skin. But there was no relationship between the Malassezia type and the severity score. The severity score is thought to depend not on the type, but also on the quantity of the yeast.     

Evaluation of Expression of Lipases and Phospholipases of Malassezia restricta in Patients with Seborrheic Dermatitis

Background

Malassezia species (spp.) are cutaneous opportunistic pathogens and associated with various dermatological diseases including seborrheic dermatitis, dandruff and atopic dermatitis. Almost all Malassezia spp. are obligatorily lipid-dependent, which might be caused by lack of the myristic acid synthesis. Recent genome analysis of M. restricta and M. globosa suggested that the absence of a gene encoding fatty acid synthesis might be compensated by abundant genes encoding hydrolases, which produce fatty acids, and that lipases and phospholipases may play a role in virulence of the fungus.

Objective

The current study aimed to investigate the contribution of lipases and phospholipases in virulence of the M. restricta as being the most frequently isolated Malassezia spp. from the human skin.

Methods

Swap samples of two different body sites of at least 18 patients with seborrheic dermatitis were obtained and in vivo expression of lipases and phospholipases of M. restricta was analyzed by the gene specific two-step nested RT-PCR.

Results

The results of the current study suggest that majority of the patients display expression of lipase RES_0242.

Conclusion

These data imply a possible role of lipase in the host environment to produce free fatty acids for the fungus.     

Skin Characteristics in Patients with Pityriasis Versicolor Using Non-Invasive Method, MPA5

Background

Skin pigmentary changes of pityriasis versicolor may occur as either hyperpigmented or hypopigmented lesions, depending on the outcome of interactions between Malassezia yeasts and the skin, such as lipoperoxidation process, stimulus of inflammatory cell to melanocytes, and increased thickness of keratin layer.

Objective

To investigate skin characteristic factors that enhance the susceptibility to Malassezia yeasts and provoke different color changes of pityriasis versicolor patients.

Methods

To clarify these factors, we investigated the skin characteristics of pityriasis versicolor patients, using a non-invasive method known as MPA 5® (Courage and Khazaka, Germany). A total of 90 normal healthy subjects and 30 pityriasis versicolor patients were included in this study.

Results

Both hyperpigmented and hypopigmented pityriasis versicolor skin lesions showed higher humidity, increased sebum excretion rate and increased transepidermal water loss (TEWL) values than normal healthy subjects. But no significant difference of specific Malassezia yeasts species between hyperpigmented and hypopigmented skin lesions was evident.

Conclusion

These results indicate that higher humidity and increased sebum level provide a better growing environment of Malassezia yeasts in the skin, leading to the assumption that interaction between Malassezia yeasts and skin barrier materials makes disruption of skin barrier causing increased TEWL.     

花斑糠疹和马拉色菌毛囊炎菌种分布特点分析

目的探讨花斑糠疹和马拉色菌毛囊炎的菌种分布特点。方法从临床诊断为花斑糠疹和马拉色菌毛囊炎的患者分离培养菌种,通过形态学和生理生化学方法鉴定菌种,并比较两组患者菌种分布情况。结果共收集花斑糠疹病例161例,培养阳性121株,其中合轴马拉色菌46株,糠秕马拉色菌13株,球形马拉色菌29株,钝性马拉色菌33株;马拉色菌毛囊炎135例,培养阳性114株,其中合轴马拉色菌52株,糠秕马拉色菌43株,球形马拉色菌13株,钝性马拉色菌6株。两种疾病菌种分布差异有显著性(P<0.005)。结论花斑糠疹和马拉色菌毛囊炎菌种分布存在差异,可能为两种疾病不同临床表现的原因之一,但结果尚需慎重解释,需要分子水平的进一步研究。     

Molecular Biological Identification of Malassezia Yeasts Using Pyrosequencing

Background

A Pyrosequencing assay has been used in identification of fungal species such as Candida or Aspergillus and diagnosis of pathogenic bacteria such as Helicobacter pylori but there has been no report on successful isolation and identification of Malassezia yeasts using the pyrosequencing method.

Objective

Examine the applicability and plausibility of the pyrosequencing method in identification of the Malassezia species.

Methods

At internal transcribed spacer (ITS) sites 1 and 2, three primers were developed using Pyrosequencing Assay Design Software (Biotage AB). Pyrosequencing was performed on 11 standard strains and 83 genomic DNA samples obtained from 66 healthy controls aged from 1 to 80.

Results

The eleven Malassezia standard species and 83 genomic DNA samples were successfully identified using the pyrosequencing assay.

Conclusion

The pyrosequencing method is a new tool for analysis of Malassezia yeasts, and its precision and rapidity suggests its clinical applicability.     

Antibiotic Susceptibility of Staphylococcus aureus in Atopic Dermatitis: Current Prevalence of Methicillin-Resistant Staphylococcus aureus in Korea and Treatment Strategies

Background

Staphylococcus aureus is a well-known microbe that colonizes or infects the skin in atopic dermatitis (AD). The prevalence of methicillin-resistant S. aureus (MRSA) in AD has recently been increasing.

Objective

This study aimed to determine the antimicrobial susceptibility patterns in AD skin lesions and evaluate the prevalence of MRSA in Korea. We also recommend proper first-line topical antibiotics for Korean patients with AD.

Methods

We studied S. aureus-positive skin swabs (n=583) from the lesional skin of infants, children, and adults who presented to our outpatient clinic with AD from July 2009 to April 2012.

Results

S. aureus exhibited high susceptibility against most antimicrobial agents. However, it exhibited less susceptibility to benzylpenicillin, erythromycin, clindamycin, and fusidic acid. The prevalence of MRSA was 12.9% among 583 S. aureus isolates, and the susceptibility to oxacillin was significantly lower in infants in both acute and chronic AD lesions.

Conclusion

S. aureus from AD has a high prevalence of MRSA and multidrug resistance, especially in infants. In addition, the rate of fusidic acid resistance is high among all age groups, and mupirocin resistance increases with age group regardless of lesional status. This is the first study comparing the antimicrobial susceptibility rates of S. aureus isolates from AD cases with respect to age and lesion status in Korea.     

Epidemiologic Study of Malassezia Yeasts in Acne Patients by Analysis of 26S rDNA PCR-RFLP

Background

Although acne is a common follicular inflammatory dermatosis, studies of the relationship between Malassezia yeasts and acne have rarely been conducted.

Objective

We sought to identify Malassezia yeasts from acne patients and establish a relationship between specific types of species of Malassezia and acne.

Methods

Sixty acne patients were enrolled. Each strain obtained was identified as one of eleven species by 26S rDNA PCR-RFLP. We then compared these data with those of age- and sex-matched healthy subjects.

Results

Growth of Malassezia was evident in fewer patients with acne (50%) in comparison to controls (70.6%). M. restricta was dominant in patients with acne (23.9%), whereas M. globosa was most common (26.7%) in healthy controls. In the patients group, the rate was the highest (71.7%) in the twenties and, in terms of body site, the rate was the highest (60%) in the chest. In the control group, the rate was the highest (75.0%) in the thirties and in the forehead (85.0%).

Conclusion

The detection rate of Malassezia yeasts was conspicuously low in the acne patients group. Statistically significant differences were observed between the patient and the control groups in the twenties and thirties, and in terms of body site, in the forehead and chest.     

The Investigation on the Distribution of Malassezia Yeasts on the Normal Korean Skin by 26S rDNA PCR-RFLP

Background

Malassezia yeasts are normal flora of the skin that are discovered in 75~98% of health subjects, but since its association with various skin disorders have been known, many studies have been conducted in the distribution of the yeasts.

Objective

To isolate, identify, and classify Malassezia yeasts from the normal human skin of Koreans by using the rapid and accurate molecular biology method (26S rDNA PCR-RFLP) which overcome the limits of morphological and biochemical methods, and to gather a basic database that will show its relation to various skin diseases.

Methods

Malassezia yeasts were cultured from clinically healthy human skin using scrub-wash technique at five sites (forehead, cheek, chest, upper arm, and thigh) and swabbing technique at scalp in 160 participants comprised of 80 males and 80 females aged from 0 to 80. Identification of obtained strains were placed into the one of the 11 species by 26S rDNA PCR-RFLP.

Results

An overall positive culture rate was 62.4% (599/960). As shown in the experiment groups by their age, the positive culture rate was the highest (74.2%) in the age 21~30 and 31~40 (89/120). In the experiment groups by different body areas, the scalp showed the highest positive culture rate of 90% (144/160). On analysis of 26S rDNA PCR-RFLP, M. globosa was the most predominant species in the age 0~10 (32.8%), 11~20 (28.9%), 21~30 (32.3%). M. restricta was identified as predominant species in the age 41~50 (27.9%), 61~70 (31.5%) and 71~80 (24.0%). In the age 31~40 years, M. sympodialis was found to be the most common species (24.6%). According to body site, M. restricta was more frequently recovered in the scalp (56.8%), forehead (39.8%) and cheek (24.0%) and while M. globosa was more frequently recovered in the chest (36.8%). Higher positive culture rates of Malassezia yeasts were shown in male subjects than female counterparts in all body areas except scalp (p<0.05). Especially in this study, M. dermatis, newly isolated Malassezia species from atopic dermatitis patient in Japan, was isolated and identified in 19 cases (1.9%) in healthy subjects.

Conclusion

The key is to recognize the existence of a difference in the type of Malassezia species in different ages as well as body areas, which reflects differing skin lipid levels in various ages and different body areas. Moreover, 26S rDNA PCR-RFLP analysis which was opted in this study could provide a sensitive and rapid identification system for Malassezia species, which may be applied to epidemiological surveys and clinical practice.     

糠秕马拉色菌所致荷兰男性脂溢性皮炎一例

糠秕马拉色菌是人体皮肤表面的常驻嗜脂性酵母,可引起皮肤浅部真菌感染如花斑糠疹和马拉色菌毛囊炎.脂溢性皮炎为发生在头皮、面部和躯干上半部的慢性红斑,常伴油性鳞屑,本病的病因尚不清楚,但近年研究提示,其发病机制与马拉色菌酵母有关.     

Effect of Emollients Containing Vegetable-Derived Lactobacillus in the Treatment of Atopic Dermatitis Symptoms: Split-Body Clinical Trial

Background

Atopic dermatitis (AD) patients suffer from xerosis. Proper skin care, including the use of emollients, may help improve xerosis and minimize disease exacerbation. Lactobacillus sakei probio 65, isolated from the Korean vegetable-based product kimchi, can decrease interleukin 4 and immunoglobulin E levels and inhibit Staphylococcus aureus. Moreover, it has reportedly shown positive dermatological effects in both animal and clinical studies.

Objective

To compare the effects of an emollient that contains Lactobacillus (treated) with a normal emollient (control) on AD.

Methods

This double-blind, randomized, split-body clinical trial involved 28 patients with AD. The patients applied the Lactobacillus-containing emollient on one side of their body and the control emollient on the other side twice daily for 4 weeks. Trans-epidermal water loss (TEWL) and skin capacitance were evaluated and investigator global assessment and the visual analogue scale (VAS) were administered on weeks 0, 1, 2, and 4.

Results

The treated sides had significantly lower TEWL and VAS values and significantly higher skin capacitance values over time than the control sides.

Conclusion

Topical application of Lactobacillus-containing emollients may improve the skin permeability of patients with AD.     

Comparison of Nested PCR and RFLP for Identification and Classification of Malassezia Yeasts from Healthy Human Skin

Background

Malassezia yeasts are normal flora of the skin found in 75~98% of healthy subjects. The accurate identification of the Malassezia species is important for determining the pathogenesis of the Malassezia yeasts with regard to various skin diseases such as Malassezia folliculitis, seborrheic dermatitis, and atopic dermatitis.

Objective

This research was conducted to determine a more accurate and rapid molecular test for the identification and classification of Malassezia yeasts.

Methods

We compared the accuracy and efficacy of restriction fragment length polymorphism (RFLP) and the nested polymerase chain reaction (PCR) for the identification of Malassezia yeasts.

Results

Although both methods demonstrated rapid and reliable results with regard to identification, the nested PCR method was faster. However, 7 different Malassezia species (1.2%) were identified by the nested PCR compared to the RFLP method.

Conclusion

Our results show that RFLP method was relatively more accurate and reliable for the detection of various Malassezia species compared to the nested PCR. But, in the aspect of simplicity and time saving, the latter method has its own advantages. In addition, the 26S rDNA, which was targeted in this study, contains highly conserved base sequences and enough sequence variation for inter-species identification of Malassezia yeasts.     

Analysis of Colonization and Genotyping of the Exotoxins of Staphylococcus aureus in Patients with Atopic Dermatitis

Background

The skin of atopic dermatitis (AD) patients has a high susceptibility to Staphylococcus aureus colonization, and the toxins produced by S. aureus may aggravate AD by acting as superantigens.

Objective

The purpose of this study was to evaluate the relationship of the skin barrier function, colonization of S. aureus, and the clinical severity of AD. We also examined the predominant toxin genes produced in Korean AD patients.

Methods

Thirty-nine patients with AD were evaluated for clinical severity and skin barrier function by using Severity Scoring of Atopic Dermatitis (SCORAD) index and transepidermal water loss (TEWL). S. aureus was isolated from the forearm, popliteal fossa, and anterior nares of AD patients (n=39) and age-matched controls (n=40); the toxin genes were analyzed by performing multiplex polymerase chain reaction.

Results

TEWL showed a statistically significant correlation with clinical severity in patients with AD (p<0.05). TEWL was correlated with the number of S. aureus colonization sites and the presence of nasal colonization, but these results were not statistically significant. S. aureus strains were isolated in 64.1% of the 39 AD patients. The SCORAD index and AD severity were strongly correlated with the number of colonization sites. The predominant toxin gene found in AD patients was staphylococcal enterotoxin a (sea) only, which was produced in 52.6% of patients. The toxin genes sea and toxic shock syndrome toxin-1 (tsst-1) were found together in 42.1%, while tsst-1 only was found in 5.3% of the patients.

Conclusion

S. aureus strains were isolated in 64.1% of the 39 AD patients. Skin barrier function, as measured by TEWL, revealed a statistically significant correlation with clinical severity in AD patients. The SCORAD index and severity of AD was strongly correlated with the number of colonization. The most common toxin gene was sea in the Korean AD patients and this gene might have an important role in the pathogenesis of AD.