BAP1 and PTEN mutations shape the immunological landscape of clear cell renal cell carcinoma and reveal the intertumoral heterogeneity of T cell suppression: a proof-of-concept study.

Clear cell renal cell carcinoma (ccRCC) is an immunologically vulnerable tumor entity, and immune checkpoint inhibitors are now widely used to treat patients with advanced disease. Whether and to what extent immune responses in ccRCC are shaped by genetic alterations, however, is only beginning to emerge. In this proof-of-concept study, we performed a detailed correlative analysis of the mutational and immunological landscapes in a series of 23 consecutive kidney cancer patients. We discovered that a high infiltration with CD8 + T cells was not dependent on the number of driver mutations but rather on the presence of specific mutational events, namely pathogenic mutations in PTEN or BAP1. This observation encouraged us to compare mechanisms of T cell suppression in the context of four different genetic patterns, i.e., the presence of multiple drivers, a PTEN or BAP1 mutation, or the absence of detectable driver mutations. We found that ccRCCs harboring a PTEN or BAP1 mutation showed the lowest level of Granzyme B positive tumor-infiltrating lymphocytes (TILs). A multiplex immunofluorescence analysis revealed a significant number of CD8 + TILs in the vicinity of CD68 + macrophages/monocytes in the context of a BAP1 mutation but not in the context of a PTEN mutation. In line with this finding, direct interactions between CD8 + TILs and CD163 + M2-polarized macrophages were found in BAP1-mutated ccRCC but not in tumors with other mutational patterns. While an absence of driver mutations was associated with more CD8 + TILs in the vicinity of FOXP3 + Tregs and CD68 + monocytes/macrophages, the presence of multiple driver mutations was, to our surprise, not found to be strongly associated with immunosuppressive mechanisms. Our results highlight the role of genetic alterations in shaping the immunological landscape of ccRCC. We discovered a remarkable heterogeneity of mechanisms that can lead to T cell suppression, which supports the need for personalized immune oncological approaches.

Kidney Cancer Models for Pre-Clinical Drug Discovery: Challenges and Opportunities.

Renal cell carcinoma (RCC) is among the most lethal urological malignancies once metastatic. The introduction of immune checkpoint inhibitors has revolutionized the therapeutic landscape of metastatic RCC, nevertheless, a significant proportion of patients will experience disease progression. Novel treatment options are therefore still needed and in vitro and in vivo model systems are crucial to ultimately improve disease control. At the same time, RCC is characterized by a number of molecular and functional peculiarities that have the potential to limit the utility of pre-clinical model systems. This includes not only the well-known genomic intratumoral heterogeneity (ITH) of RCC but also a remarkable functional ITH that can be shaped by influences of the tumor microenvironment. Importantly, RCC is among the tumor entities, in which a high number of intratumoral cytotoxic T cells is associated with a poor prognosis. In fact, many of these T cells are exhausted, which represents a major challenge for modeling tumor-immune cell interactions. Lastly, pre-clinical drug development commonly relies on using phenotypic screening of 2D or 3D RCC cell culture models, however, the problem of "reverse engineering" can prevent the identification of the precise mode of action of drug candidates thus impeding their translation to the clinic. In conclusion, a holistic approach to model the complex "ecosystem RCC" will likely require not only a combination of model systems but also an integration of concepts and methods using artificial intelligence to further improve pre-clinical drug discovery.

Targeting the Proteasome in Advanced Renal Cell Carcinoma: Complexity and Limitations of Patient-Individualized Preclinical Drug Discovery.

BACKGROUND: Systemic treatment options for metastatic renal cell carcinoma (RCC) have significantly expanded in recent years. However, patients refractory to tyrosine kinase and immune checkpoint inhibitors still have limited treatment options and patient-individualized approaches are largely missing. PATIENTS AND METHODS: In vitro drug screening of tumor-derived short-term cultures obtained from seven patients with clear cell RCC was performed. For one patient, a patient-derived xenograft (PDX) mouse model was established for in vivo validation experiments. Drug effects were further investigated in established RCC cell lines. RESULTS: The proteasome inhibitor carfilzomib was among the top hits identified in three of four patients in which an in vitro drug screening could be performed successfully. Carfilzomib also showed significant acute and long-term cytotoxicity in established RCC cell lines. The in vivo antitumoral activity of carfilzomib was confirmed in a same-patient PDX model. The cytotoxicity of carfilzomib was found to correlate with the level of accumulation of ubiquitinated proteins. CONCLUSIONS: In this proof-of-concept study, we show that patient-individualized in vitro drug screening and preclinical validation is feasible. However, the fact that carfilzomib failed to deliver a clinical benefit in RCC patients in a recent phase II trial unrelated to the present study underscores the complexities and limitations of this strategy.

Mapping the teaching of honeybee veterinary medicine in the European Union and European Free Trade Area.

BACKGROUND: Honey bee (Apis mellifera) is a very important species for human beings, animals, environmental biodiversity, crop production and economic sustainability in Europe and worldwide. This study investigates whether future veterinarians are trained to deal with the particular needs of the only traditional food-producing insect in Europe. METHODS: This study analyses data collected from 77 European veterinary education establishments in EU and the European Free Trade Area. RESULTS: The results show that 75 per cent of those establishments (58 out of 77) teach honeybee veterinary medicine. There is a clear geographical differentiation. In north-western countries only about half of the establishments include honeybee health, production and product inspection in their undergraduate curricula, while in eastern, central and southern countries, which are also important beekeeping countries, the great majority of the establishments incorporate honeybee veterinary medicine in their undergraduate curriculum. Eighty-six per cent of all the establishments teaching honeybee veterinary medicine (50 of the 58) incorporate it in their core curriculum either as separate subject or as part of other subjects. Twenty-five per cent of all the establishments (19 out of 77) organise postgraduate training courses in this field. CONCLUSIONS: Veterinarians have an important role in ensuring the health, sustainability and productivity of managed honeybee colonies as they do for other animal species. It seems however that teaching of honeybee veterinary medicine receives less attention in undergraduate veterinary curricula in EU compared with other fields of veterinary medicine. Seeing the increasing importance of honey bees for crop protection, environmental protection and economic sustainability, it would be beneficial to further strengthen the education of honeybee veterinary medicine in the future. Establishments should encourage and prepare veterinarians for practising science-based veterinary medicine in honey bees by incorporating such teaching in undergraduate curricula and by providing postgraduate opportunities to qualified veterinarians wishing to enhance their basic skills in this field.

Mapping the teaching of aquatic animal veterinary medicine in the European Union and European Free Trade Area.

Aquatic animal production is the fastest growing food sector globally. Aquaculture and fisheries are very dynamic sectors in the EU and the number of ornamental aquarium pets is increasing. Veterinarians have a fundamental role to play by ensuring health and welfare of aquatic species, productivity and profitability of fish farming, public health and ecosystem conservation. This study investigates how the undergraduate curriculum prepares future veterinarians for such roles by analysing data from the 77 European veterinary education establishments based in EU and the European Free Trade Area. Over 95 per cent of these establishments incorporate teaching in aquatic animal veterinary medicine in their curriculum, while the great majority do so within the core curriculum. Almost half of the establishments provide teaching in aquatic animal veterinary medicine as separate subjects. Many establishments (>40 per cent) provide such training as elective option in their undergraduate curricula or as postgraduate opportunities to enhance Day One Competences. The veterinary education establishments integrating adequately aquatic animal veterinary medicine in their curriculum are evenly distributed in all regions of Europe. Veterinarians are trained and empowered by legislation to assess health of aquatic animals, to diagnose, to prescribe medicines, to notify for diseases and to ensure safe food for the consumers. Veterinary education establishments should encourage training of veterinarians to follow a career in aquatic animal veterinary medicine.

Indoor air quality survey of nail salons in Boston.

Employees in nail salons, largely Vietnamese immigrant women in Boston, are exposed to a range of volatile organic chemicals from the products used in salons, including solvents, glues and polishes. Some of these chemicals have the potential to cause short and long-term adverse health effects. Only limited research has been performed on assessing occupational exposures. This project aimed to characterize total volatile organic compound (TVOC) and PM2.5 concentrations in nail salons as a function of ventilation, building characteristics, customer and employee occupancy, and type of services being performed. Students conducted sampling in 21 salons in Boston, MA from September to December, 2011. Study visits included: indoor environmental quality measurements (TVOCs, PM2.5 and carbon dioxide), site observations, and an interview. CO2 levels in 15 of 21 salons exceeded 800 ppm, suggesting that these salons may have insufficient ventilation. Higher TVOC and PM2.5 levels were found in salons with less ventilation (as estimated using CO2 concentrations). Contrary to our a priori hypothesis, average levels of TVOCs, CO2 and PM2.5 were consistent throughout salons, indicating that exposures may not be restricted to areas in the salon where work is being performed (e.g., at the manicure table). Higher TVOC concentrations were observed when tasks were being performed, yet were not dependent upon the number of tasks being performed. Improving ventilation conditions in salons to meet minimum outdoor air delivery requirements can reduce exposures to TVOCs.

Pitfalls and recommendations in cases of laser removal of decorative tattoos with pigmented lesions: case report and review of the literature.

IMPORTANCE: Pigmented lesions in decorative tattoos cause diagnostic difficulties at a clinical and dermoscopic level. In cases of laser removal of tattoos, hidden suspicious nevi may be revealed gradually. OBSERVATIONS: We describe the first case of a malignant melanoma that developed on a preexisting nevus within a tattoo during and between the phases of laser removal. The patient refused to undergo excision of the nevus until we made excision conditional for continued laser treatment. CONCLUSIONS AND RELEVANCE: The English literature reports 16 cases of malignant melanoma developing in tattoos. Correlation between the placement of tattoos and the development of malignant melanoma remains unclear. Our case emphasizes the diagnostic problems of pigmented lesions within tattoos. For safety reasons, tattoos should never be placed on pigmented lesions; if they are, the tattoos should not undergo laser treatment. We suggest an excision before starting laser tattoo removal. Dermoscopic assessments on a regular basis during the period of tattoo removal are recommended.

Treatment errors resulting from use of lasers and IPL by medical laypersons: results of a nationwide survey.

BACKGROUND: The demand for hair and tattoo removal with laser and IPL technology (intense pulsed light technology) is continually increasing. Nowadays these treatments are often carried out by medical laypersons without medical supervision in franchise companies, wellness facilities, cosmetic institutes and hair or tattoo studios. This is the first survey is to document and discuss this issue and its effects on public health. PATIENTS AND METHODS: Fifty patients affected by treatment errors caused by medical laypersons with laser and IPL applications were evaluated in this retrospective study. We used a standardized questionnaire with accompanying photographic documentation. Among the reports there were some missing or no longer traceable parameters, which is why 7 cases could not be evaluated. RESULTS: The following complications occurred, with possible multiple answers: 81.4% pigmentation changes, 25.6% scars, 14% textural changes and 4.6% incorrect information. The sources of error (multiple answers possible) were the following: 62.8% excessively high energy, 39.5% wrong device for the indication, 20.9% treatment of patients with darker skin or marked tanning, 7% no cooling, and 4.6% incorrect information. CONCLUSIONS: The causes of malpractice suggest insufficient training, inadequate diagnostic abilities, and promising unrealistic results. Direct supervision by a medical specialist, comprehensive experience in laser therapy, and compliance with quality guidelines are prerequisites for safe laser and IPL treatments. Legal measures to make such changes mandatory are urgently needed.

Pyogenic granuloma: treatment with the 1,064-nm long-pulsed neodymium-doped yttrium aluminum garnet laser in 20 patients.

BACKGROUND: There are various therapeutic options for the treatment of pyogenic granuloma (PyG), but the results are frequently unsatisfactory, especially at difficult sites and with extensive lesions. OBJECTIVE: To evaluate the success of treatment of PyG using the 1,064-nm neodymium-doped yttrium aluminum garnet (Nd:YAG) laser and to compare it with state-of-the-art treatment methods. MATERIALS AND METHODS: Twenty patients with PyG were treated using the long-pulsed 1,064-nm Nd:YAG laser with fluences of 60 to 180 J/cm(2) , a spot size of 7 mm, and a pulse duration of 40 ms. One to four treatment sessions were necessary for complete removal. RESULTS: Recurrence-free healing occurred in 19 of 20 patients (follow-up ≥ 6 months, maximum 22 months). Because of heavy bleeding, one nonresponder was successfully treated using a carbon dioxide laser. The cosmetic results were good; textural changes of the skin were slight, if present at all. CONCLUSION: When used with the right strategy and patient cohort, the long-pulse 1,064-nm Nd:YAG laser is an effective, low-risk, minimally invasive method of treating PyG. This type of laser is a good therapeutic option that achieves good cosmetic results, particularly in PyG with a large diameter that are not suitable for treatment using the pulsed dye laser.