Efficacy of LL-37 cream in enhancing healing of diabetic foot ulcer: a randomized double-blind controlled trial.

Wound healing in DFU (diabetic foot ulcer) has prolonged inflammation phase and defective granulation tissue formation. LL-37 has antimicrobial property, induces angiogenesis, and keratinocyte migration and proliferation. This study analyzes the efficacy of LL-37 cream in enhancing wound healing rate and decreasing the levels of IL-1α, TNF-α, and the number of aerobic bacteria colonization in DFU with mild infection. This study was conducted from January 2020 to June 2021 in Jakarta. Subjects were instructed to apply either LL-37 cream or placebo cream twice a week for 4 weeks. Wounds were measured on days 7, 14, 21, and 28 and processed with ImageJ. The levels of LL-37, IL-1α, and TNF-α from wound fluid were measured using ELISA. The number of aerobic bacteria colonization was counted from the isolate grown in culture. The levels of LL-37 in DFU at baseline were equally low in both groups which were 1.07 (0.37-4.96) ng/mg protein in the LL-37 group and 1.11 (0.24-2.09) ng/mg protein in the placebo group. The increase in granulation index was consistently greater in the LL-37 group on days 7, 14, 21, and 28 (p = 0.031, 0.009, 0.006, and 0.037, respectively). The levels of IL-1α and TNF-α increased in both groups on days 14 and 21 (p > 0.05). The decrease in the number of aerobic bacteria colonization was greater in the LL-37 group on days 7, 14 and 21, but greater in the placebo group on day 28 (p > 0.05). In conclusion, LL-37 cream enhanced the healing rate of DFU with mild infection, but did not decrease the levels of IL-1α and TNF-α and the number of aerobic bacteria colonization. This trial is registered at ClinicalTrials.gov, number NCT04098562.

Effect of adipose derived stromal vascular fraction on leprosy neuropathy: A Preliminary report.

BACKGROUND: Adipose derived stromal vascular fraction (SVF) contains a heterogeneous population of mononuclear cells, progenitor cells and about 1-10% are mesenchymal stromal cells. These cells are an ideal candidate for regenerative medicine for peripheral neuropathy. Leprosy is a disabling disorder with neuropathy, usually with consequences of permanent disability of the extremities. We conducted a preliminary study to evaluate the cell yield, its characteristics and clinical outcomes after SVF injections in four leprosy patients. METHODS: Four post leprosy patients were recruited and evaluated for sensory testing (warm detection, cold detection, vibration, pain and sensation) on the ulnar area of the hand. Liposuction was done and adipose tissue was processed into SVF with a closed system and injected to the ulnar area of the hand at the dorsal and palmar side. Evaluation of sensory testing was done after 3 days, 1 week, 1 month and 3 months following SVF injection. SVF was also characterized using flow cytometry, cell counting, sterility and presence of mycobacteria. RESULTS: The results showed that leprosy patients had a low count of mesenchymal cells and a high amount of CD34/CD45 positive cells. One patient was positive for mycobacteria from his adipose tissue and SVF. Sensory examination after SVF injection showed an improvement in temperature and pain sensation in the palmar and superficial branch. Meanwhile, touch sensation improved on the dorsal branch, and there was no improvement for vibration in all patients. CONCLUSIONS: The results showed that SVF had a potential to improve sensory loss in leprosy patients.

Clinico-laboratory findings of Malassezia folliculitis in Indonesia: A multicentre study.

BACKGROUND: Malassezia folliculitis (MF) is a humid-favoured fungal skin disease caused by Malassezia species. Inaccurate treatments, changes in skin flora and disease exacerbation are often occurred due to oversights in the diagnosis. Several diagnostic methods are established for MF. OBJECTIVE: To identify clinico-laboratory findings of Malassezia folliculitis in Indonesia. METHODS: The study was conducted from January 2014 to December 2018 in seven referral teaching hospitals. Medical records of MF-diagnosed patients were obtained and analysed using the binomial test, chi-square test and Cohen's Kappa coefficient in SPSS 26.0. RESULTS: A total of 353 cases of MF were identified in seven referral teaching hospitals in Indonesia, 66.3% of which were males and 33.7% were females, dominated by the 17-25 years old group (44.5%). Itchy sensation (83.9%) was a major subjective complaint. Lesions were majorly found on the trunk-chest, back and shoulder (68.3%), while the clinical manifestation are mostly follicular papule-pustular lesions (62.1%). Patients were 87.4% positive by KOH examination (modified Jacinto Jamora's criteria) and 69.1% positive by Wood's lamp. Generally, sex, age, subjective complaint, lesion location, clinical manifestation and both examinations were statistically significant (p < .001). A significant relationship between all the clinical criteria of the patients in the KOH especially the clinical manifestation was significantly related to Wood's lamp. The Cohen's Kappa assessment suggested that there was an agreement between KOH and Wood's lamp (κ = -0.272, p < .001). CONCLUSION: The clinical symptoms of Malassezia folliculitis are dominated by pruritus, papulopustular follicular lesions on the trunk and the presence of spore load.

Serum Interleukin 6 Level and Nutrition Status as Potential Predictors of Clinical Leprosy Development Among Household Contacts in Endemic Areas.

BACKGROUND: Leprosy is a chronic infectious disease that can lead to severe lifelong disabilities. Close contacts of patients with leprosy have a higher risk of acquiring the disease. Nevertheless, there is a lack of reliable markers to predict Mycobacterium leprae infection. We aimed to identify new potential markers for developing clinical leprosy among contacts. METHODS: Serum levels of interleukin (IL) 6, IL-8, IL-10, hemoglobin, ferritin, and transferrin saturation were measured in 67 patients with multibacillary leprosy (MB), 65 household contacts (HHCs) of MB patients, and 127 endemic controls (ECs). By means of multivariate logistic regression and receiver operating characteristic (ROC) analyses, we analyzed baseline variables and laboratory parameters that showed significant differences between MB in the HHC and EC groups and obtained the respective areas under the curve (AUC). Optimal cutoff values of the associated cytokines were also determined. RESULTS: Elevated IL-6 level was observed in MB patients compared to HHCs and ECs (P = .022 and .0041, respectively). Anemia and iron deficiency were also higher in the MB group compared to HHCs or ECs (P < .001). Likewise, we observed an increased risk of having MB leprosy in underweight HHCs (odds ratio [OR], 2.599 [95% confidence interval {CI}, .991-6.820]) and underweight ECs (OR, 2.176 [95% CI, 1.010-4.692]). Further ROC analysis showed that high serum IL-6 level, underweight, anemia, and iron deficiency can discriminate leprosy from their HHCs (AUC, 0.843 [95% CI, .771-.914]; P = .000; optimal cutoff value of IL-6 = 9.14 pg/mL). CONCLUSIONS: Our results suggest that serum IL-6 and nutrition status could serve as potential prognostic markers for the development of clinical leprosy in infected individuals.

Annular infectious dermatoses.

Annular configuration is conspicuous in the clinical manifestation of many skin diseases and can be helpful for the diagnosis and differential diagnosis. Variations may include arciform, ring-form, annular, circinate, serpiginous, gyrated, polycyclic, targeted or figurate forms, in different colors, sizes, and numbers, with various textures and surfaces. In infectious dermatoses, the annular reactions can be specific or nonspecific, while the underlying mechanisms remain largely unknown. In the specific reactions caused by direct invasion of the pathogens, the contest between the centrifugal outspread of the infectious agents and the centripetal impedance of the host immune response is supposed to determine the final conformation. Examples include erythema infectiosum, orf, erythema multiforme, and pityriasis rosea of viral origin. Bacterial infections that may display annular lesions include erythrasma, erythema (chronicum) migrans of Lyme borreliosis, secondary syphilis, cutaneous tuberculosis, and leprosy. Superficial mycosis, such as dermatophytosis, candida intertrigo, tinea imbricata, and subcutaneous mycosis, such as chromoblastomycosis, and algae infection protothecosis, are characterized by annular progression of the skin lesions. The creeping serpiginous extension is an alarming sign for the diagnosis of cutaneous larva migrans. A better understanding of the virulence and pathogenicity of the pathogens and the way and type of immune response will help to clarify the pathogenesis.

Mucocutaneous mycoses in people living with human immunodeficiency virus in Indonesia.

BACKGROUND: To date, integrated care for people living with human immunodeficiency virus (PLHIV) has improved. However, although the management of mucocutaneous mycosis cases has improved, disease progression might be different in immunocompromised patients, which leads to variable clinical manifestations. OBJECTIVES: To describe the characteristics of mucocutaneous mycosis cases in the PLHIV population and its associated factors in Indonesia. METHODS: This retrospective study was conducted from January 2014 to December 2018 in four academic hospitals. Data were acquired from medical records with the inclusion of mucocutaneous mycosis patients with concurrent HIV infection. Analysis with the chi-squared test was performed using Statistical Package for the Social Sciences (SPSS) version 20.0. RESULTS: A total of 1,796 cases of mucocutaneous mycoses were identified in 1782 PLHIV. The most common types of infection were candidiasis (63%), followed by dermatophytosis (35.1%), and malasseziosis (1.9%), which were significantly higher in PLHIV with CD4 level <200 cells/mm3 . The proportions of male gender (78.6% vs. 56.3%, p < .001), high level of education (48.0% vs. 64.1%, p < .001), office workers (73.8% vs. 64.1%, p < .001), combination of topical and systemic antifungal agents (59.1% vs. 48.5%, p = .006) and not receiving antiretroviral therapy (63.2% vs. 7.8%, p < .001) were significantly higher in PLHIV with a CD4 level <200 cells/mm3 . CONCLUSION: In Indonesia, the most common fungal infection in PLHIV is candidiasis. This study also addressed the important matters regarding mucocutaneous mycoses in PLHIV. Education is an important measure to prevent the incidence of cutaneous mycoses in PLHIV, especially in high-risk groups.

The mycological efficacy and safety of selenium sulfide 1.8% versus ketoconazole 2% shampoo in pityriasis versicolor: A double-blind randomized controlled trial.

Introduction: Although it has long been used as a topical treatment of pityriasis versicolor (PV), the efficacy of topical selenium disulfide as compared to topical ketoconazole in the treatment of PV remains unclear. This study aims to assess the mycological efficacy and safety of SeS2 1.8% shampoo and ketoconazole 2% shampoo for the treatment of PV. Methods: A double-blind randomized controlled trial was performed in patients with PV during September-December 2018, based on block randomization. Physical examinations, scale provocation test, Wood's lamp, and potassium hydroxide examination were conducted. Intention to treat analysis was performed to evaluate the mycological efficacy. Results: The mycological efficacy and side effect were not significantly different between the ketoconazole group and the selenium group; 94% vs 86% (RR=2.3(95%CI 0.6-8.5), p=0.182), and 22% versus 8%. Conclusions: SeS2 1.8% has the same mycological efficacy and side effect as compared to ketoconazole 2% in the treatment of PV.

Type 2 Diabetes Mellitus with Early Dry Skin Disorder: A Comparison Study Between Primary and Tertiary Care in Indonesia.

BACKGROUND: The prevalence of Type 2 Diabetes Mellitus (T2DM) in Indonesia has continued to increase over the years. Management of T2DM is challenging across clinical settings, including primary and tertiary care. Uncontrolled T2DM puts patients at risk of the development of T2DM complications, especially early-stage dry skin that is neglected by most of the patients. This study aimed to investigate the comparison between the T2DM management profile and dry skin clinical profile of T2DM patients in primary care and tertiary care settings. METHODS: The study was conducted as a cross-sectional epidemiological study by comparing T2DM patient profiles in primary and tertiary care. The data collected included sociodemographic, clinical, and laboratory data that were correlated with T2DM and early dry skin related-T2DM. This study included early dry skin within the SRRC score of 3-11 and excluded infection, ulcer, and severe erythema. RESULTS: The patients in primary and tertiary care presented poorly controlled T2DM with median HbA1c levels of 7.8% and 7.6%. The patients in primary care also presented with high triglyceride, 179 mg/dl. Furthermore, several significant differences were found in the duration of T2DM, duration of dry skin, and DM treatment (OAD and insulin). CONCLUSION: Significant differences in the duration of T2DM, duration of dry skin, and DM treatment (OAD and insulin) might be affected by the parameter of T2DM glycemic control (Blood pressure (BP), body mass index (BMI), HbA1c, random blood glucose (RBG), and triglyceride).

Oral and Topical Centella asiatica in Type 2 Diabetes Mellitus Patients with Dry Skin: A Three-Arm Prospective Randomized Double-Blind Controlled Trial.

INTRODUCTION: Uncontrolled diabetes mellitus (DM) is related to skin disorders, particularly dry skin. Pathogenesis of dry skin in type 2 diabetes mellitus (T2DM) rises from the chronic hyperglycemia causing an increase in advanced glycation end-products (AGEs), proinflammatory cytokines, and oxidative stress. Combination of oral and topical Centella asiatica (CA) is expected to treat dry skin in T2DM patients more effectively through decreasing N(6)-carboxymethyl-lysine (CML) and interleukin-1α (IL-1α) and increasing superoxide dismutase (SOD) activity. METHODS: A three-arm prospective, double-blind, randomized, controlled study was performed to evaluate the efficacy of the oral and topical CA extract in 159 T2DM patients with dry skin. The subjects were divided into the CA oral (CAo) 2 × 1.100 mg + CA topical (CAt) 1% ointment group, oral placebo (Plo) + CAt group, and Plo and topical placebo (Plt) group. Dry skin assessment was performed on day 1, 15, and 29, while evaluation of CML, IL-1α, and SOD activity was on day 1 and 29. RESULT: Effectivity of CAo + CAt combination was assessed based on HbA1c and random blood glucose (RBG). In well-controlled blood glucose, on day 29, the percentage of SRRC decrement was greater in the CAo + CAt group compared to the control group (p = 0.04). SCap value in the CAo + CAt group was greater than that in the control group (p = 0.01). In the partially controlled blood glucose, increment of SOD activity in the CAo + CAt group was greater than that in the control group (p = 0.01). There were medium-to-strong correlation between CML with SOD (r = 0.58, p < 0.05) and IL-1α with SOD (r = 0.70, p < 0.05) in well-controlled blood glucose. Systemic and topical adverse events were not significantly different between groups. CONCLUSION: CAo and CAt combination can be used to significantly improve dry skin condition through increasing SOD activity in T2DM patients with controlled blood glucose.

Prognostic factors influencing the treatment outcome of onychomycosis Candida.

Onychomycosis contributes as many as half of all nail disorder cases. In 2017, the incidence of onychomycosis was 15% of all dermatomycosis cases at our hospital, a tertiary hospital in Indonesia, with only 25% of the patients achieving mycological cure. This study aims to identify the prognostic factors influencing the treatment outcome of onychomycosis Candida. This is a retrospective study, using data obtained from outpatient registry at our hospital. Fifty-four onychomycosis patients were included in this study. Potential prognostic factors were analysed by STATA15.0. Retrospective analysis with cox proportional-hazard was used to measure the contribution of each variable to the treatment's outcome. Onset of disease, history of nail disorder, and site of infection were not associated with mycological cure (P > .05). Based on retrospective analysis, age[odds ratio (OR)1.46; 95% confidence interval (CI)1.07-2.03], onset of disease (OR 1.14; 95%CI 1.11-1.17), comorbidities (OR 1.07; 95%CI 1.03-1.11), type of onychomycosis (OR 1.08; 95%CI 1.05-1.16), site of infection (OR 1.12; 95%CI 1.04-1.22) and number of infected nails (OR 1.50; 95%CI 1.25-1.68) were significantly associated with poor treatment outcome, while type of treatment and type of systemic agents showed no significant association with the outcome. Kaplan-Meier curves showed that subjects elderly age and more than 3 infected nails had the lowest median survival. Elderly, longer onset, presence of comorbidities, multiple sites of infection, and high number of infected nails can affect the mycological cure negatively. Unstandardised treatment was associated with the mycological cure despite not affecting the prognosis. Therefore, the management's goal is to identify these specific prognostic features.

Apoptosis of Adipose-Derived Stem Cells Induced by Liposomal Soybean Phosphatidylcholine Extract.

BACKGROUND: Recently, Phosphatidylcholine (PC) has been used as an off-label treatment for lipolysis injection, which is associated with inflammatory reaction due to sodium deoxycholate, an emulsifier, so that inflammation as side effect occurs in those patients. Liposome formulation from soybean lipid was thought to be a better and safer alternative. This study aimed to analyze the mechanism of Liposomal Soybean Phosphatidylcholine (LSPC) extract from Indonesian soybeans (containing 26% PC) to induce Adipose-derived Stem Cells (ASCs) death in vitro. METHODS: Liposomes were prepared using thin film hydration method followed by a stepwise extrusion process to produce a small amount of 41.0-71.3 nm. Liposomal soybean phosphatidylcholine extract (LSPCE), liposomal purified PC (LPCC), and solution of PC+SD were used for comparison. Annexin V fluorescein Isothiocyanate/Propidium Iodide (FITC/PI) double staining by flow cytometry and also measurement of caspase-3 activity using ELISA were used to quantify the rate of apoptosis. ASCs viability was measured using MTT assay after induction with liposomes. Morphological changes were shown using a phase-contrast, inverted microscope and Transmission-Electron Microscope (TEM). RESULTS: The flow cytometry results showed that cells treated with both LSPCE and LPCC showed increase in early apoptosis beginning at 6 hr after incubation, which was confirmed by caspase 3 measurement. MTT assay showed that both LSPCE and LPCC could decrease viability of cells. Cells treated with LSPCE and LPCC showed some rounded cells, which was an early sign of cell death. Cells treated with SD showed extensive membrane damage with necrosis features using TEM. CONCLUSION: The results above demonstrated that LSPCE induced apoptosis of ASCs.

Correlation of CD4 counts with clinical and histopathological findings in disseminated histoplasmosis: a 10-year retrospective study.

BACKGROUND: Disseminated cutaneous histoplasmosis (DCH) is one of the manifestations of systemic histoplasmosis infection in HIV-positive patients. Interaction between host immune status and histoplasmosis infection is still poorly understood. It is thought that immune status, represented by CD4 counts, may be correlated with clinical and histopathological findings. OBJECTIVE: To correlate the CD4 counts with the different clinical and histopathological findings in cutaneous histoplasmosis. METHODS: This was a serial case report of seven HAART-treated HIV positive patients with disseminated histoplasmosis observed within the period of January 2004 through December 2014 from the Dermatology and Venereology Department, Cipto Mangunkusumo Hospital, Jakarta, Indonesia. The patients came with different complaints and clinical findings. CD4 counts were recorded prior to lesion biopsy. RESULTS: The CD4 counts were independent from clinical morphology and distribution of lesions. Lower CD4 counts were associated with the presence of intraepidermal yeast-like cells, whereas there is the ability of forming granulomas at higher CD4 counts. CONCLUSION: CD4 count correlates to histopathological findings of cutaneous histoplasmosis.

Identification of Malassezia species from pityriasis versicolor in Indonesia and its relationship with clinical characteristics.

Pityriasis versicolor (PV) is a chronic superficial fungal disease caused by Malassezia spp. The incidence is as high as 30-40% in tropical climates. Previous studies indicate that the geographic factor has influence on the main species isolated from PV. Our aim was to identify Malassezia spp. from PV patients in Indonesia and their correlation to clinical characteristics. Isolates of Malassezia were collected from 98 PV patients (62 males and 36 females). Identification was based on morphological observation and biochemical evaluation. Leeming Notman agar was used as isolation culture medium. The biochemical evaluation consisted of specimen culture onto Sabouraud dextrose agar, Cremophor EL, Esculin, Dixon's agar at 37 degrees C and catalase reaction. The isolates found were Malasseziafurfur (42.9%), M. sympodialis (27.5%), M. globosa (13.3%), M. slooffiae (7.7%), M. obtusa (7.7%) and M. restricta (2.2%), and 7.14% specimens were unidentified. There was no statistically significant association between Malassezia spp. and demographic characteristics and clinical characteristics of the patients. Unlike reports from temperate climate countries, this study in Indonesia found M. furfur as the most frequently isolated Malassezia spp. in PV patients.

Comparison of proteinase, lipase and alpha-glucosidase activities from the clinical isolates of Candida species.

Proteinase, lipase and alpha-glucosidase activity in 81 clinically isolated Candida strains grown in nutrient-restricted media were comparatively measured, and the correlation between inducible enzyme activity and fungal growth was analyzed. Enzyme activity was assayed by colorimetric methods. Extracellular proteinase activity was significantly higher in Candida albicans, followed by C. parapsilosis and C. tropicalis. Extracellular lipase activity was observed in all 6 Candida spp. in the descending order of C. albicans, C. tropicalis and C. parapsilosis. Cell surface alpha-glucosidase activity was significantly higher in C. tropicalis, C. albicans, and C. parapsilosis, but was not detected in the other three species. A relatively strong correlation was observed between proteinase activity and fungal growth (correlation coefficient: 0.72); a mild correlation was observed between lipase activity and fungal growth (0.55); and a slight correlation was observed between alpha-glucosidase activity and fungal growth (0.32). There was no correlation among the strains in the activity levels of the three enzymes. These results suggest that pathogenic fungi produce larger amounts of inducible hydrolytic enzymes, and that proteinase is most likely to be related to fungal growth in nutrient-restricted conditions.

Epidemiology of onychomycosis in Indonesia: data obtained from three individual studies.

In terms of both diagnosis and treatment, onychomycosis remains a problem in Indonesia. To examine this situation, we performed a retrospective study of the incidence of onychomycosis involving 10 state university hospitals across the country. We collected data from medical records of patients treated at these hospitals in 1997-1998 (study 1) and from a 1998-1999 multi-center Indonesian study on pulse-dose itraconazole therapy (study 2), both of which were part of onychomycosis campaigns with the objective of increasing the awareness of physicians of onychomycotic problems. Further, we analyzed data from year 2003 medical records of patients from 4 hospitals in Java (study 3). The average incidence of onychomycosis among fungal diseases increased to 4.7% in large cities that, from an average incidence within Indonesia of 3.5% in 1997-1998. At only 0.5%, the incidence of onychomycosis among all skin diseases was low in both study 1 and 3. Female patients outnumbered male patients with a ratio of 1.5:1 to 2:1. In study 2, the fingernail was the site most frequently affected. KOH examinations and fungal culture in study 1 (n=557) showed causative organisms to be Candida (50.1%), dermatophytes (26.2%), moulds (3.1%) and mixed infections (1.8%), with the remainder (18.7%) unidentified. Similar frequency of causative organisms was reported in study 2 (n=113) and in study 3 (n=183). People's low awareness of nail disease and consequent lack of concern as well as the limited diagnostic ability of KOH examination and fungal culture might influence the low incidence of the disease and the frequency of detection of the causative organisms.