Molecular Profiling of Merkel Cell Polyomavirus-Associated Merkel Cell Carcinoma and Cutaneous Melanoma.

Merkel cell carcinoma (MCC) is a rare, high-grade, aggressive cutaneous neuroendocrine malignancy most commonly associated with sun-exposed areas of older individuals. A relatively newly identified human virus, the Merkel cell polyomavirus (MCPyV) has been implicated in the pathogenesis of MCC. Our study aimed to examine nine MCC cases and randomly selected 60 melanoma cases to identify MCPyV status and to elucidate genetic differences between virus-positive and -negative cases. Altogether, seven MCPyV-positive MCC samples and four melanoma samples were analyzed. In MCPyV-positive MCC RB1, TP53, FBXW7, CTNNB1, and HNF1A pathogenic variants were identified, while in virus-negative cases only benign variants were found. In MCPyV-positive melanoma cases, besides BRAF mutations the following genes were also affected: PIK3CA, STK11, CDKN2A, SMAD4, and APC. In contrast to studies found in the literature, a higher tumor burden was detected in virus-associated MCC compared to MCPyV-negative cases. No association was identified between virus infection and tumor burden in melanoma samples. We concluded that analyzing the key morphologic and immunohistological features of MCC is critical to avoid confusion with other cutaneous malignancies. Molecular genetic investigations such as next-generation sequencing (NGS) enable molecular stratification, which may have future clinical impact.

5-Aminolevulinic acid photodynamic therapy with and without Er:YAG laser for actinic keratosis: Changes in immune infiltration.

INTRODUCTION: Ultraviolet light induced DNA damage, combined with immunosuppression and inflammation are involved in the pathogenesis of actinic keratosis. Photodynamic therapy not only destroys dysplastic cells via tissue destruction and vascular shutdown, but also induces an acute local inflammatory response and activates both the innate and adaptive immune system. In our current work we aimed to compare immunohistochemistry features of inflammatory infiltrate of actinic keratoses after 5-aminolevulinic acid photodynamic therapy with or without Er:YAG laser resurfacing. METHODS: Eleven patients with multiple actinic keratosis on the scalp, face, hands or forearms were treated by conventional and Er:YAG laser assisted 5-aminolevulinic acid PDT in split-site manner. Biopsies of AKs were taken before, 48 h and 3 months after the treatment. CD3, CD4, CD8, CD1a, Ki67 and p53 expressions were analyzed by immunohistochemical methods. RESULTS: The number of p53 and Ki67 positive cells decreased significantly 3 months after treatment, but the abnormal cells were not eliminated totally. The number of CD1a+ Langerhans cells significantly decreased 48 h after both treatments, while CD8+ T cell count was significantly lower 3 months after Er:YAG laser assisted photodynamic therapy. However, the number of CD3+ and CD4+ T cells were not changed significantly 48 h and 3 months later. CONCLUSIONS: One session of 5-aminolevulinic acid photodynamic therapy even with Er:YAG laser pretreatment could not terminate actinic damage totally. Photodynamic therapy induced immunological changes. However further investigations are needed to answer how the composition of actinic keratosis' immune infiltrate influence the effect of photodynamic therapy.

Mass spectrometric imaging of cysteine rich proteins in human skin.

Looking insight pathological processes, metallothioneins (MTs) are considered to be potential biomarkers for monitoring of a development of various types of diseases, such as cancer. The early identification of the MTs in biological tissues could be important tool for the estimation of appropriate clinical therapy. Therefore, here we investigated the application of matrix assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) together with immunohistochemical analyses (IHC) using MT-1/2 antibody for MT detection in formalin-fixed paraffin-embedded (FFPE) biopsy specimens of human skin. Principal component analyses revealed differences in the peptide/protein profiles separating healthy skin from the carcinoma specimens. Statistically significant ion peaks at m/z 6038, 6300, 6676, and 7026 were more frequently detected in squamous cell carcinoma (SCC), basal cell carcinoma (BCC) and melanoma. Using IHC, we found that MT-1/2 was significantly higher in SCC and melanoma compared to healthy skin. Surprisingly, significantly low levels of MT-1/2 were found in BCC. On one side, the results indicate important role of MTs in melanoma occurrence and progression, as on the second side, there are hidden processes associated with MTs based on differences of the occurrence of the MS peaks, which could be associated with cycling of MTs isoforms.

Ultraviolet radiation-mediated development of cutaneous melanoma: An update.

Ultraviolet (UV) light is absorbed by nucleic acids, proteins or other endogenous chromophores, such as porphyrins, flavins and melanin, triggering biological processes in skin cells. Both UV-induced mutations in melanocytes and changes in the immune microenvironment are understood to play a role in the development of cutaneous melanoma. The degree of UV-induced stress and the protection against this stress are influenced by both intracellular and intercellular molecular interactions. The present review summarizes the known major molecular biological changes induced by UV light in the skin that play a role in melanoma initiation and promotion. Nevertheless, cutaneous melanoma is not a homogenous disease, and the interaction of variable environmental exposure and different genetic susceptibility and other host factors lead to the formation of melanomas with different biological behavior and clinical characteristics. This review highlights the challenges in the understanding of how UV radiation contributes to the formation of cutaneous melanoma, and reviews the new results of photobiology and their link to tumor genetics and tumor immunology with potential implications on melanoma prevention and therapeutic strategies. The information presented here is expected to add clarity to ongoing research efforts in this field to aid the development of novel strategies to prevent and treat melanoma.

Application of a low molecular weight antifungal protein from Penicillium chrysogenum (PAF) to treat pulmonary aspergillosis in mice.

PAF, a small antifungal protein from Penicillium chrysogenum, inhibits the growth of several pathogenic filamentous fungi, including members of the Aspergillus genus. PAF has been proven to have no toxic effects in vivo in mice by intranasal application. To test its efficacy against invasive pulmonary aspergillosis (IPA), experiments were carried out in mice suffering from IPA. Adult mice were immunosuppressed and then infected with Aspergillus fumigatus. After stable infection, the animals were inoculated with PAF intranasally at a concentration of 2.7 mg/kg twice per day. At this concentration-which is highly toxic in vitro to A. fumigatus-the mortality of the animals was slightly delayed but finally all animals died. Histological examinations revealed massive fungal infections in the lungs of both PAF-treated and untreated animal groups. Because intranasally administered PAF was unable to overcome IPA, modified and combined therapies were introduced. The intraperitoneal application of PAF in animals with IPA prolonged the survival of the animals only 1 day. Similar results were obtained with amphotericin B (AMB), with PAF and AMB being equally effective. Combined therapy with AMB and PAF-which are synergistic in vitro-was found to be more effective than either AMB or PAF treatment alone. As no toxic effects of PAF in mammals have been described thus far, and, moreover, there are so far no A. fumigatus strains with reported inherent or acquired PAF resistance, it is worth carrying out further studies to introduce PAF as a potential antifungal drug in human therapy.

Massive Localized Lymphedema Arising from Abdominal Wall: A Case Report and Review of the Literature.

Massive localized lymphedema (MLL) is a rare pseudosarcomatous lesion due to localized lymphatic obstruction from variable causes. It is most common on medial aspect of thigh and inguinal region. Abdominal localization is rare and may cause clinical diagnostic confusion with other malignant tumors due to its large size. We report a case of abdominal wall MLL of a 56-year-old male patient under clinical suspicion of well differentiated liposarcoma. The literature search and differential diagnosis will be addressed. In doubt cases, immunohistochemical stain or fluorescent in situ hybridization can help to separate this entity from the other mimickers.

Pleomorphic hyalinizing angiectatic tumor of soft parts: case report with unusual ganglion-like cells and review of the literature.

Pleomorphic hyalinizing angiectatic tumor (PHAT) is a recently described, non-metastasizing tumor of uncertain lineage. This tumor distributes equally between the genders and has a predilection for the subcutaneous soft tissue, particularly in lower extremity, other locations are rare. Based on the recent literature, PHAT is suspected to encompass the morphological spectrum with other tumors such as myxoinflammatory fibroblastic sarcoma (MIFS) and hemosiderotic fibrolipomatous tumor (HFLT), although cytogenetic data remain inconsistent. We report a case of PHAT that arose in the upper arm with unusual morphology which showed ganglion-like cells similar to Reed-Sternberg-like cells found in MIFS. The tumor had strong immunohistochemical expression of CD34, CD99, and was negative for S-100. The ganglion-like cells were positive for both CD34 and CD68 but negative for CD30. The translocation between chromosome 1 and 10, a frequent finding of MIFS and HFLT, was not identified by FISH excluding the possibility of hybrid PHAT and MIFS. We conclude FISH can be a potential useful tool to separate PHAT with atypical morphology from hybrid tumor in doubted cases. Due to the rarity of PHAT and lack of consistent pathogenetic signatures, more cases and further studies will be needed to elucidate the pathogenesis and nature of this tumor.

In vivo application of a small molecular weight antifungal protein of Penicillium chrysogenum (PAF).

The antifungal protein of Penicillium chrysogenum (PAF) inhibits the growth of important pathogenic filamentous fungi, including members of the Aspergillus family and some dermatophytes. Furthermore, PAF was proven to have no toxic effects on mammalian cells in vitro. To prove that PAF could be safely used in therapy, experiments were carried out to investigate its in vivo effects. Adult mice were inoculated with PAF intranasally in different concentrations, up to 2700 µg·kg⁻¹ daily, for 2 weeks. Even at the highest concentration--a concentration highly toxic in vitro for all affected molds used, animals neither died due to the treatment nor were any side effects observed. Histological examinations did not find pathological reactions in the liver, in the kidney, and in the lungs. Mass spectrometry confirmed that a measurable amount of PAF was accumulated in the lungs after the treatment. Lung tissue extracts from PAF treated mice exerted significant antifungal activity. Small-animal positron emission tomography revealed that neither the application of physiological saline nor that of PAF induced any inflammation while the positive control lipopolysaccharide did. The effect of the drug on the skin was examined in an irritative dermatitis model where the change in the thickness of the ears following PAF application was found to be the same as in control and significantly less than when treated with phorbol-12-myristate-13-acetate used as positive control. Since no toxic effects of PAF were found in intranasal application, our result is the first step for introducing PAF as potential antifungal drug in therapy.

[Intrathoracic migration of Kirschner wires]. / Kirschner-drótok intrathoracalis vándorlása.

Orthopedic surgeons apply metallic pins to stabilize the clavicule and humerus on a daily basis. Migration of these pins into the thoracic cavity is rare. We present the case of an elderly female patient, whose right humeroscapular joint was fixed with Kirschner wires due to recurrent luxation. Six weeks later, a follow-up X-ray revealed that the pins have migrated into the right thoracic cavity, confirmed by a CT chest. Videothoracoscopic removal of the metallic pins was not possible because of dense adhesions. Right anterolateral thoracotomy was carried out, and after pneumolysis one pin was taken out from the 2nd lung segment. The other one, which was running along the cupola and entering the spinal cord, was also removed. There was no postoperative surgical complication. The authors review the literature of this rare complication and point out that pins migrating into the thoracic cavity should be removed to avoid life threatening complications.

[Anaesthesia of the myasthenic patients during thymectomy]. / Myastheniás betegek altatása thymectomia során.

Myasthenia is a rare autoimmune disease characterized by fluctuating muscle weakness and fatigability due to a reduction in available acetylcholine receptors at the neuromuscular junction. Data of 186 patients suffering from myasthenia were collected retrospectively. All patients underwent thymectomy over a 23 years period from 1981 to 2006 without surgical mortality. Postoperative ventilation was required for more than 24 hours in seven patients and one patient needed postoperative ventilatory support more than seven days. Thymectomy for myasthenia was performed using promethazine and atropine in general anaesthesia. Introduction could be facilitated with propofol, etomidate or thiopental and sevoflurane, avoiding use of any muscle relaxants. Non-depolarizing muscle relaxants were not used during the procedures. Adequate surgical conditions were provided by short-acting inhaled anaesthetics (sevoflurane) and small doses of opiates. 95% of the narcotized patients were immediately extubated after the procedure in the operating room. Length of stay in intensive care unit could have been reduced without any postoperative ventilatory support. Nonsteroid analgesics and nalbuphine were used for pain relief. Anaesthesia of thymectomy is based on volatile gases. Airway complications can be prevented with use of small amounts of anticholinergic drugs, perhaps steroids.

[Changes in surgical therapy of pulmonary hydatid cysts]. / Változások a tüdo echinococcus ciszta sebészi kezelésében.

The lung is the second most common site of hydatid cysts after the liver. The authors analyse retrospectively the results of patients treated with pulmonary hydatid cysts in the past 18 years, considering video-thoracoscopy. Twenty eight patients were treated during this period in 31 cases. Hydatid disease affected only the lung in case of 22 patients, while in 6 cases it was present in the liver and lung simultaneously. Pulmonary hydatid disease affected one side in 24 and both sides in 4 cases. For surgical treatment pericystectomy in one, atypical segment resection in 18, anatomical segmentectomy in three, lobectomy in 7 and video-thoracoscopy in 3 cases were performed without surgical complications. The mean hospital stay was 10.5 days in case of thoracotomies and 8.5 days in case of video-thoracoscopy. There was one recurrence in conventional surgery and reoperation was necessary. After video-thoracoscopy no recurrence was detected. Mean follow-up was 120 months, after video-thoracoscopy it was 20 months. Three patients have uncertain chest pain after thoracotomy, but none has any complaints after video-thoracoscopy. Fifteen patients took mebendazole permanently after the final histological result. According to the authors' practice the indication of lung resections for pulmonary hydatid cysts is limited, in selective cases video-thoracoscopic cystectomy can be a successful treatment of choice.

[Video-assisted thoracoscopy (VATS) during the last ten years (1992-2001)]. / Video-asszisztált torakoszkópia (VATS) klinikánk 10 éves anyagában (1992-2001).

We summarize and analyse the video-assisted thoracoscopies (VATS) performed in our department during the last ten years. In this period 296 patients underwent VATS for diagnosis or therapy. We describe indications, advantages and disadvantages, we also analyse the complications. Video-assisted thoracoscopy is less demanding to the patients than thoracotomy, it reduces the length and cost of inpatient treatment. We recommend extensive use of VATS in suitable patients.

[Surgical aspects in the treatment of myasthenia gravis]. / A myasthenia gravis gyógykezelésének sebészeti vonatkozásai.

We treated 81 patients suffering from myasthenia with thymectomy in a 10-year period (1991-2000). We think, that thymectomy must be carried out in the treatment of myasthenia gravis, unless contraindications are present. The operation is not urgent and in the preoperative period patients must reach optimal condition with the help of standard medical treatment. The result of the operation is influenced by the length of time between the beginning of the complaints and the operation. The best results can be expected when the operation is performed in less than 2 years time, however an operation performed later may also be successful. We operated on 63% (n = 51) of the patients between 4-12 months, on 18% (n = 15) between 13-24 months and on 18.6% (n = 15) more than 25 months after the beginning of the complaints. Complete thymectomy was performed in all patients through median sternotomy. We had no operative mortality. Our postoperative results were evaluated with Jaretzki classification: 75% of our patients are in remission and asymptomatic (n = 61).

[Surgical treatment of pleural metastases in breast cancer]. / Az emlórák pleurametasztázisainak sebészi kezelése.

Pleural metastases of breast cancer is a severe progression of the disease and the treatment is difficult. Distant metastases are mainly treated by chemo-, radio- or hormone therapy, but in recent years surgical intervention is increasingly important. Between the 1st of January 1992 and 31st of December 2001 in the 2nd Department of Surgery University of Debrecen Medical and Health Science Center Medical School of Medicine 43 patients with breast cancer were operated on because of pleural metastases. In these patients biopsy and pleurodesis with talcum insufflation were performed. The surgical treatment was followed by chemotherapy. Pleurodesis was successful at 76.7 percent. The 6, 12 and 24 months survival was 58.1, 39.5 and 16.3 percent. Based on literature data and own experience we conclude that surgical intervention with adjuvant therapy extends life expectancy for patients with pleural metastases. Pleurodesis can prevent or delay hydrothorax, it improves vital functions and the quality of life for patients with pleural metastases.