The second nationwide surveillance of antibacterial susceptibility patterns of pathogens isolated from skin and soft-tissue infections in dermatology departments in Japan.

The present study compared trends in antimicrobial resistance patterns in pathogens isolated from skin and soft-tissue infections (SSTIs) in Japan with those of a nationwide survey conducted in 2013. Three organisms that caused most of the SSTIs were collected from 12 dermatology departments in medical centers and 12 dermatology clinics across Japan between April 2019 and August 2020. A total of 390 strains, including 267 Staphylococcus aureus, 109 coagulase-negative staphylococci (CNS), and 14 Streptococcus pyogenes strains were submitted to a central laboratory for antimicrobial susceptibility testing. Patient demographic and clinical information was collated. Methicillin-resistant S. aureus (MRSA) was detected in 25.8% (69/267) of the S. aureus strains. The prevalence of MRSA between the present study and the 2013 survey did not differ significantly. Furthermore, there were no significant differences in MIC values and susceptibility patterns of the MRSA strains to other agents, regardless of a history of hospitalization within 1 year or invasive medical procedures. Methicillin-resistant CNS (MRCNS) was detected in 48.6% (53/109) of CNS isolates, higher than the 35.4% prevalence in the 2013 survey. This difference could be attributed to the heterogeneity in the members of the MRCNS, which comprises multiple staphylococci species, between the 2013 and 2019 surveys. However, it was noted that the susceptibility profiles of the MRCNS to each antibiotic were not significantly different from those identified in the 2013 survey. Most strains of S. pyogenes were susceptible to each antibiotic, similar to the 2013 survey. Continuous monitoring of trends in pathogen and susceptibility profiles is important to advise local public health efforts regarding the appropriate treatment of SSTIs.

Enhanced PD-L1 expression on tumor cells in primary cutaneous large T-cell lymphoma with CD30 expression as classic Hodgkin lymphoma mimics: A report of lymph node lesions of two cases.

Neoplastic PD-L1 (nPD-L1, clone SP142) expression remains unclear in cutaneous T-cell lymphoma (CTCL), although it is well-documented in classic Hodgkin lymphoma (CHL). Here, we report two cases of primary cutaneous large T-cell lymphoma (PCLTCL) with CD30 expression that developed secondary nodal lesions morphologically mimicking CHL, and describe their PD-L1 expression. Our two cases (52- and 60-year-old males) had long-standing clinical courses of CTCL. Their PCLTCL with CD30 expression developed nodal lesions, having a nodular growth pattern containing scattered CD30+ Hodgkin and Reed-Sternberg-like and/or lacunar cells that expressed CD15 but did not harbor Epstein-Barr virus. Their differential diagnosis from CHL was challenging. A diagnosis of PCLTCL with secondary nodal involvement featuring CHL mimicry was based on comparison of the primary and secondary lesions. In one case, shared expression of the same T-cell antigen was revealed by immunohistochemistry, and in the other, identical clonal TCR rearrangement was demonstrated by polymerase chain reaction (PCR). Interestingly, nPD-L1 was expressed on more than 50% of the tumor cells in the secondary nodal lesions, but on very few in the primary cutaneous lesions, in both cases. This is the first report of nPD-L1 expression greatly increasing with PCLTCL tumor progression to nodal involvement.

Comparison of the efficacy of ALA-PDT using an excimer-dye laser (630 nm) and a metal-halide lamp (600 to 740 nm) for treatment of Bowen's disease.

BACKGROUND/PURPOSE: Topical 5-aminolevulinic acid-based photodynamic therapy (ALA-PDT) is an effective treatment for Bowen's disease (BD). In order to compare the efficacy of two different light sources, using either an excimer-dye laser (EDL) (630 nm) or a metal-halide lamp (MHL) (600 to 740 nm) a protocol for topical ALA-PDT for treatment of BD of the extremities was established, and responses during 12 months follow-up were assessed. METHODS: From 25 patients a total of 26 lesions that had been histopathologically diagnosed as BD from 2005 to 2010 in the Department of Dermatology at the Aichi Medical University Hospital were randomly selected. The light source used for the topical ALA-PDT was EDL in 17 lesions and MHL in 9 lesions. The photosensitizing protoporphyrin IX that is produced within BD lesions 4 h after application of 20% ALA cream was mostly consumed after exposure to 100 J/cm(2) irradiation using 630 nm EDL. Each lesion was irradiated once a week for 3 weeks, for a total dosage of 300 J/cm(2) (100 mW/cm(2)). Patients were followed up clinically every 3 months for 12 months, and at 1 month after the final treatment lesions were evaluated histopathologically. RESULTS: Histologically, the complete response (CR) rate at 1-month follow-up was 82% (14/17 lesions) in the EDL treatment group and 100% (9/9 lesions) in the MHL treatment group (P > 0.05). The recurrence rate at 12 months after PDT was 46% (6/13 lesions, one patient lost to follow-up) in the EDL group and 0% in the MHL group (P < 0.05) (χ(2) test with Fisher's exact test). The average period before recurrence after EDL treatment was 6.5 months. CONCLUSION: A novel protocol for topical ALA-PDT in Japanese in Asian patients with BD was developed and implemented. The protocol improved the CR rate compared with previous studies. Moreover, the present results indicate that the efficacy of topical ALA-PDT using MHL was superior to that using EDL for BD patients.

Clinical utility of botulinum toxin type A local injection therapy for head and forehead hyperhidrosis.

Head and forehead hyperhidrosis (HFH) is a disease that causes a large amount of sweating from the head region, and it significantly reduces patients' quality of life. Only a few reports have shown the effectiveness of botulinum toxin type A (BTX-A) local injection therapy (BTX-A therapy) for HFH. To clarify the benefits of BTX-A for HFH, BTX-A therapy was performed in 15 patients, and its efficacy was evaluated. The amount of sweating was measured by the ventilation capsule method and Minor's iodine-starch test. Evaluation was also performed using the Hyperhidrosis Disease Severity Scale (HDSS) and the Dermatology Life Quality Index (DLQI). In most cases, a remarkable antiperspirant effect was observed from 2 weeks after the injection, and the effect lasted for approximately 30 weeks. HDSS and DLQI improved along with the decrease in sweating. Two patients (13.3%) complained of transient mild ptosis. There were no serious side-effects. This study showed that BTX-A therapy is a safe and effective treatment for HFH.

Cost-of-illness study for axillary hyperhidrosis in Japan.

The prevalence of primary axillary hyperhidrosis in Japan is 5.75% (males, 6.60%; females, 4.72%) in the population aged 5-64 years. No study on comprehensively evaluated direct medical costs, hygiene product costs, and productivity loss in axillary hyperhidrosis patients has been published in Japan. The aim of this study was to estimate the cost of illness for axillary hyperhidrosis in Japan by conducting a nationwide insurance claims database analysis and a cross-sectional Web-based survey. Among patients diagnosed with primary axillary hyperhidrosis at least once between November 2012 and October 2019, health insurance receipt data of 1447 patients were analyzed. A cross-sectional Web-based survey was conducted on 321 patients aged 16-59 years with axillary hyperhidrosis to calculate hygiene product costs and productivity loss using a Work Productivity and Activity Impairment questionnaire. Furthermore, nationwide estimation was performed for the hygiene product costs and productivity loss based on the number of patients estimated from the prevalence. The annual direct medical costs per axillary hyperhidrosis patient were ¥91 491 in 2016, ¥93 155 in 2017, and ¥75 036 in 2018. In all of these years, botulinum toxin type A injection accounted for approximately 90% of the total costs. The annual total cost of hygiene products per axillary hyperhidrosis patient was ¥9325. The overall work impairment (%) of working patients with axillary hyperhidrosis was 30.52%, and its monthly productivity loss was ¥120 593/patient. The activity impairment (%) of full-time housewives with axillary hyperhidrosis was 49.05% and its monthly productivity loss was ¥176 368/patient. The annual hygiene product cost based on the nationwide estimation was ¥24.5 billion and the monthly productivity loss was ¥312 billion. The significant cost associated with axillary hyperhidrosis was clarified. If out-of-pocket expenses for treatments not covered by health insurance are included in the estimation, the cost will further increase.

Primary cutaneous T-cell lymphoma of unspecified type with cytotoxic phenotype: clinicopathological analysis of 27 patients.

The objective of our study was to investigate the clinicopathological features of the currently ill-defined subtype of primary cutaneous T-cell lymphoma of unspecified type (CTCLU) with a cytotoxic phenotype and no Epstein-Barr virus (EBV) association. A series of 27 patients with CTCLU (median age 49 years; range 25-87 years; 18 men) was reviewed. Performance status scores above 1 (7%), clinical stages above 2 (15%), B symptoms (26%), extracutaneous involvement (30%), and a fatal course within 1 year of diagnosis (19%) were observed infrequently. The International Prognostic Index was high or high to intermediate in 11%, and the Prognostic Index for Peripheral T-cell Lymphoma unspecified was above group 2 in 22%. Notably, the rates of spontaneous regression and T-cell receptor gene rearrangements by polymerase chain reaction analysis were seen in 26 and 17% of our cases, respectively. Histologically, 22 patients had subcutaneous involvement of whom eight showed a lethal clinical course, and five patients without subcutaneous involvement were all survivors. Immunophenotypical and morphological features allowed us to subclassify our cases according to the following four categories: (1) epidermotropic CD8+ T-cell lymphoma (n=5); (2) cutaneous gamma/delta T-cell lymphoma (n=8); (3) cutaneous alpha/beta pleomorphic T-cell lymphoma (n=8); and (4) cutaneous medium/large pleomorphic T-cell lymphoma, not otherwise specified (n=6). All four of these groups of lymphomas exhibited a relatively favorable clinical course compared to previous reports. However, epidermotropic CD8+ T-cell lymphoma appeared to be unique with a higher ratio (80%) of spontaneous regression, a lower ratio (40%) of subcutaneous involvement, and a more favorable clinical course than the other three subcategories.

Reduction in QSART and vasoactive intestinal polypeptide expression in the skin of Parkinson's disease patients and its relation to dyshidrosis.

BACKGROUND: With regards to dyshidrosis in Parkinson's disease (PD), there is no established and consistent view on the occurrence sites, frequency and etiology, although there have been several reports on hypohidrosis of the limbs and sudoresis on the face/cervical region. METHODS: Hydrosis in the forearms of PD patients and healthy individuals were compared by quantitative sudomotor axon reflex test (QSART). The expression of various neuropeptides and alpha-synuclein was examined with immunohistochemical staining. RESULTS: There was a significant reduction in QSART of PD patients but not of healthy controls. Reduced expression of vasoactive intestinal polypeptide (VIP) was also detected in the sweat glands of PD patients. CONCLUSION: Reduction in QSART and VIP expression in the sweat glands might be involved in the dyshidrosis of PD patients.

Photodynamic therapy using direct-current pulsed iontophoresis for 5-aminolevulinic acid application.

In photodynamic therapy (PDT) for skin cancer, 5-aminolevulinic acid (ALA) is applied topically to the affected area to be absorbed percutaneously through passive diffusion, and typically requires 4-6 h before performing PDT. In this study, we attempted to reduce the absorption period in PDT by ionizing ALA using direct-current pulsed iontophoresis to treat actinic keratosis (AK). Twenty percent ALA solution was applied to AK lesions of five patients using direct-current pulsed iontophoresis. ALA-PDT was repeated three times with a total irradiation of 150 J/cm(2) (50 J/cm(2) per irradiation, weekly). One week after the last PDT, therapeutic results were assessed by skin biopsy. In all subjects, protoporphyrin IX (PpIX) production was confirmed after iontophoresis, and its production levels were comparable to the conventional occlusive dressing technique (ODT). Skin biopsies from the treated lesion showed the disappearance of tumour cells. These results indicated that direct-current pulsed iontophoresis for applying ALA before PDT is useful to treat AK.

Oncolytic virotherapy for malignant melanoma with herpes simplex virus type 1 mutant HF10.

BACKGROUND: Many viruses have been engineered and evaluated for their potential as therapeutic agents in the treatment of malignant neoplasm, including malignant melanoma. OBJECTIVE: In this study, we investigated the efficacy of HF10, an attenuated, replication-competent HSV, in immunocompetent animal models with malignant melanoma. METHODS: For in vitro study, viral cytotoxicity assays and replication assays were performed both in human and mouse melanoma cells. For the study in vivo, intraperitoneally disseminated or subcutaneous melanoma models were prepared in DBA/2 mice using clone M3 cells, then HF10 was inoculated intraperitoneally or intratumorally. Therapeutic efficacy of HF10 was assessed by survival, tumor growth, and histopathological analysis. RESULTS: HF10 infection produced cytolytic effects in melanoma cells at various multiplicities of infection (MOI). In the intraperitoneal melanoma model, all mice survived when given intraperitoneal injections of HF10 compared with 100% fatality in the control mice. In the subcutaneous tumor model, intratumoral inoculation of HF10 significantly reduced tumor growth. Histology and immunohistochemistry showed tumor lysis and inflammatory cell infiltration after intratumoral HF10 inoculation. Viral antigen was retained at the inoculation site until 7 days post-infection. HF10-treated intraperitoneal tumor mice were also protected against tumor rechallenge. HF10 also affected the non-inoculated contralateral tumor when injected into the ipsilateral tumor of mice, suggesting that HF10 can induce systemic antitumor immune responses in mice. CONCLUSION: Oncolytic viral therapy using HF10 was effective in melanoma mouse models, and intratumoral injection of HF10 induced systemic antitumor responses. These results suggest that HF10 is a promising agent for the treatment of advanced melanoma.

Thioredoxin upregulation by 5-aminolaevulinic acid-based photodynamic therapy in human skin squamous cell carcinoma cell line.

BACKGROUND/PURPOSE: 5-aminolaevulinic acid-based photodynamic therapy (ALA-PDT) is widely performed in the clinical setting for superficial skin cancers, giving favorable results, but residual tumor and recurrence occur occasionally. Thioredoxin is a common antioxidant that suppresses apoptosis and facilitates cell growth. We investigated the expression of thioredoxin following ALA-PDT in human skin squamous cell carcinoma cell line, HSC-5. METHODS: ALA-PDT was performed in HSC-5 cells using low-dose (5 J/cm(2), 100 mW/cm(2)) or high-dose (30 J/cm(2), 100 mW/cm(2)) irradiation, and the expression of thioredoxin was measured by Western blotting. An MTT assay was used to assess cell growth following a low dose of multiple irradiations. Cell death was examined by Western blotting for caspase-3 and PARP. Immunofluorescence double staining using annexin V and propidium iodine was also performed. RESULTS: Expression of thioredoxin was only observed following low-dose exposure ALA-PDT. Multiple low-dose exposure ALA-PDT significantly proliferated cell growth. With high-dose exposure ALA-PDT, caspase-3 and PARP expression were seen, and cell death due to apoptosis and/or necrosis was observed, but thioredoxin was barely detected. CONCLUSION: Low-dose exposure ALA-PDT increased the expression of thioredoxin and facilitated the growth of HSC-5 cells.

Local injection of botulinum toxin A for palmar hyperhidrosis: usefulness and efficacy in relation to severity.

Botulinum toxin A is widely used in Europe and the USA for the treatment of localized hyperhidrosis, and its efficacy has been recognized. In this study, botulinum toxin A (Botox) was locally injected at 30 sites (2 U/injection) on the right palm in 27 patients with palmar hyperhidrosis (14 severe patients, 13 mild patients), and the results confirmed the efficacy of injection. The amount of sweat was then quantified for the left and right hands every month after local injection. The quantity of sweat on the treated hand was approximately one-fifth that on the untreated hand. In addition, the quantity of sweat on the untreated hand decreased slightly. Over time, the quantity of sweat on the treated hand increased slightly, but the quantity of sweat on the treated hand at 6 months after injection was less than half that before injection, and there were significant differences before and after injection. In the present study, severe sweating was defined as 1 mg/cm2/min or more and mild sweating as less than 1 mg/cm2/min, and the therapeutic effects of botulinum toxin A were analyzed in relation to severity. When compared to the mild cases, the quantity of sweat remained higher in the severe cases after botulinum toxin A therapy. Therefore, to achieve satisfactory effects in severe cases, it would be necessary to increase the number of injection sites, as well as injection dose.

Rapid detection of human herpesvirus 8 DNA using loop-mediated isothermal amplification.

The reliability of a loop-mediated isothermal amplification (LAMP) method for the detection of human herpesvirus 8 (HHV-8) DNA was evaluated. Although LAMP products were produced with the DNA sample extracted from BCP-1 cells, LAMP products were not produced with the DNAs from seven other human herpesviruses. The detection limit of the HHV-8 LAMP method was 100 copies of target sequence/tube. To determine whether the HHV-8 LAMP method could be used to quantify viral DNA, threshold times, which are defined as the time (in s) it takes to reach the threshold turbidity level (0.1), were measured for the amplification of serial dilutions of a DNA plasmid containing the target sequence. The standard curve possessed a correlation coefficient of 0.9428 with a slope of -84.079 and y-intercept value of 1936.2. Additionally, an attempt was made to detect viral DNA in 17 specimens collected from Kaposi's sarcomas and two cell lines obtained from primary effusion lymphomas. HHV-8 DNA was detected in 14 of the 17 Kaposi's sarcoma tissue samples and both of the primary effusion lymphoma cell lines. Viral DNA was not detected in HHV-8 LAMP-negative samples using the real-time PCR method.

Construction of novel in vitro epithelioid cell granuloma model from mouse macrophage cell line.

There have been several attempts to make granuloma model to clarify the mechanism of granulomatous diseases like sarcoidosis. However, a unique in vitro model that generates multinucleated giant cell (MGC) through epithelioid cells resembled to human granuloma, has not yet been clearly established. In this study, the generation of granuloma model that forms MGC via epithelioid cells from the mouse macrophage cell line was investigated. A RAW 246.7 mouse macrophage cell line was cultured with lipopolysaccharide (LPS) and concanavalin A (Con A) in various concentrations either alone or both. We found that separate treatment of LPS and Con A induced around 35 and 20% MGC respectively whereas cotreatment of these chemicals drastically accelerated granuloma formation rate and it was around 80%. The highest fusion index (MGC formation rate) was observed at days 7. A gradual increase of tumor necrosis factor alpha (TNF-alpha) production in the culture supernatant was analyzed by enzyme-linked immunosorbent assay (ELISA). And the neutralization of the elevated level of TNF-alpha production by its monoclonal antibody leads to significant decrease of MGC formation. Interestingly, we found that the RAW cells were changed into spindle cells, which morphologically resembled to epithelioid cells and eventually MGC was formed from these spindle cells. Our in vitro granuloma model appeared to be similar with in vivo epithelioid cell granulomas like sarcoidosis. Thus, our model would be useful as in vitro epithelioid granuloma model for analyzing the mechanisms and screening the effective drugs of granulomatous diseases in future.

Epstein-Barr virus-associated post-transplant lymphoproliferative disorder presenting with skin involvement after CD34-selected autologous peripheral blood stem cell transplantation.

We report a case of Epstein-Barr virus (EBV)-associated post-transplant lymphoproliferative disorder (PTLD) after CD34-selected autologous peripheral blood stem cell transplantation (PBSCT). A 54-year-old woman with multiple myeloma underwent CD34-selected autologous PBSCT. The patient's post-transplantation course was complicated by fever, pancytopenia and CMV antigenemia. On day 128 post PBSCT, a skin biopsy from an erythematous nodule on the right anterior chest revealed a deep dermal infiltrate of atypical CD20 and CD79a-positive B-cells with centroblastic large cell morphology. EBV reactivation was confirmed by immnohistochemistry, in situ hybridization and Southern blot analysis. These findings represent monomorphic PTLD having pathological features of a large cell-type B-cell lymphoma. Bone marrow aspiration also demonstrated hemophagocytic syndrome (HPS), accompanied with infiltration of EBV-positive B-cells. Despite treatment with rituximab and hydroxyurea, the patient died 155 days after transplantation.