The gut microbiome has attracted significant attention as a disease modifier of melanoma. A recent study has shown that the microbial diversity in the skin microbiome was lower in patients with melanoma than in patients with nevi. 1 However, no study has reported the association, if any, between the skin microbiome and clinical findings in melanoma. Therefore, we investigated the clinical significance of the skin microbiome in patients with acral melanoma. This study was performed in accordance with the Declaration of Helsinki principles and was approved by the institutional review board of Kumamoto University Hospital. Written informed consent was obtained from all patients included in this study. We retrospectively investigated 27 patients with acral melanoma who presented to Kumamoto University Hospital between April 2013 and April 2020. Cancer staging was performed based on the 8th edition of the American Joint Committee on Cancer/Union for International Cancer Control melanoma staging system. Immunohistochemistry was performed using a primary antibody against interleukin (IL)-17 (Abcam, Cambridge, UK) as described previously.

Background Extramammary Paget's disease (EMPD) is a rare malignant skin cancer. One of the hallmarks of cancers, including EMPD, is an enhancement of aerobic glycolysis, which is also known as the Warburg effect. In the last step of glycolysis, the enzyme lactate dehydrogenase A (LDHA) catalyzes the conversion of pyruvate to lactic acid, the accumulation of which contributes to the creation of an acidic tumour microenvironment. This in turn results in immunosuppression in various types of cancers. However, the contribution of these pathways has not been well-studied in EMPD. Objective To investigate the significance of the Warburg effect and its contribution to the tumour immune microenvironment in EMPD. Methods The mRNA expression levels of molecules involved in glycolysis and immune-related cytokines were examined by ddPCR. The number of immune cells was assessed by immunohistochemistry (IHC). Results The levels of two glycolytic enzymes, HK2 and LDHA, in tumour tissues were significantly increased compared to those in paired-normal tissues. IHC analyses revealed increased numbers of PD-L1 + , PD-1 + , CD163 + M2 macrophages, Iba1 + macrophages and Foxp3 + Tregs that were associated with high LDHA levels in EMPD. ddPCR demonstrated that multiple cytokines including IL-4, IL-6, IL-10, TGF-b and CCL-2 were upregulated and associated with high LDHA levels in EMPD. Statistical analyses showed that IL-6 mRNA expression correlated with the number of CD163 + , Iba-1 + and Foxp3 + cells. Conclusion The Warburg effect contributes to immunomodulation in the tumour microenvironment and further elucidation may lead to better understanding of the pathogenesis of EMPD.

Background: Angiosarcoma is a rare malignant tumor with a poor prognosis. It is known that hypoxic condition activates tumor progression in several cancers. Additionally, hypoxic tumor microenvironment accelerates immune escape. However, the presence and significance of hypoxia in angiosarcoma has not been adequately investigated. Objective: To study the role of hypoxia in the progression of angiosarcoma. Methods: The protein level of hypoxia inducible factor-1α (HIF-1α) in angiosarcoma was examined using immunohistochemistry and immunoblotting. To study the effect of hypoxia on tumor progression, cell proliferation, migration, invasion, and tube formation assays were performed in angiosarcoma cells. The influence of tumor cell supernatant in hypoxia from angiosarcoma cells on immune escape and angiogenesis was analysed to investigate the modulatory effect of hypoxia on tumor microenvironment of angiosarcoma. The molecular mechanism related to these results was investigated using immunoblotting and real time RT-PCR. Results: HIF-1α protein was over-expressed in angiosarcoma tissues and cell lines under hypoxic conditions, and there was heterogeneity of oxygen supply in angiosarcoma. Hypoxia enhanced the proliferation, migration, and invasion abilities and inhibited tube formation in angiosarcoma cells. Tumor cell supernatant in hypoxia from angiosarcoma cells activated the monocyte invasion ability, facilitated its differentiation into M2-like macrophages, and suppressed cell-adhesion. These in vitro results were compatible to the pathological findings of angiosarcoma patients. Conclusion: Hypoxia plays a major role in progression of angiosarcoma cells by enhancing cell proliferation, migration, and invasion and by modulating the tumor microenvironment.

A 65-year-old woman had undergone a thymectomy for thymoma and 1 year after surgery she developed scattered erosive erythema with scaling and crusting. Examination findings exhibited the elevation of anti-dsDNA antibody, anti-desmoglein 1 antibody, anti-acetylcholine receptor antibody and anti-thyroglobulin antibody. A skin biopsy showed intraepidermal blisters containing neutrophils and acantholytic keratinocytes and direct immunofluorescence revealed the deposition of immunoglobulin G in the epidermis and on the basement membrane. These findings indicated the presence of systemic lupus erythematosus (SLE), myasthenia gravis, pemphigus foliaceus and chronic thyroiditis. Only 1% of SLE patients have three other autoimmune diseases according to previous publications. Our case is rare because she suffered four autoimmune diseases after the thymectomy.

Three adult dogs with chronic lymphocytic leukaemia (CLL) were successfully treated with melphalan and prednisolone. Based on the immunophenotypic analysis of leukaemic cells, two dogs were diagnosed with B cell CLL and one dog was tentatively diagnosed as having T cell CLL. One dog with B cell CLL had IgM monoclonal gammopathy. The clinical signs and haematological abnormalities associated with CLL in the three dogs improved with the administration of cytoreductive melphalan (3 to 5 mg/m2/day) and prednisolone (4.3 to 30 mg/m2/day) for eight to 210 days. There were no severe adverse effects except a mild increase in plasma alkaline phosphatase activity. Melphalan and prednisolone therapy may achieve remission with few side effects in dogs with CLL.