Vladimir Vincek

Preservation of biomolecules in breast cancer tissue by a formalin-free histology system

BackgroundThe potential problems associated with the use of formalin in histology, such as health hazards, degradation of RNA and cross-linking of proteins are well recognized. We describe the utilization of a formalin-free fixation and processing system for tissue detection of two important biopredictors in breast cancer – estrogen receptor and HER2 – at the RNA and protein levels.MethodsParallel sections of 62 cases of breast cancer were fixed in an alcohol-based molecular fixative and in formalin. Molecular fixative samples were processed by a novel formalin-free microwave-assisted processing system that preserves DNA, RNA and proteins. Formalin-fixed samples were processed using the conventional method. Estrogen receptor was assessed by immunohistochemistry and real-time PCR. HER2 was assessed by immunohistochemistry, FISH, CISH and real-time PCR.ResultsThe immunohistochemical reaction for estrogen receptor was similar in molecular- and formalin-fixed samples (Spearman Rank R = 0.83, p < 0.05). Also HER2 result was similar to that of formalin-fixed counterparts after elimination of antigen retrieval step (Spearman Rank R = 0.84, p < 0.05). The result of HER2 amplification by FISH and CISH was identical in the molecular fixative and formalin-fixed samples; although a shorter digestion step was required when using the former fixative. Real-time PCR for both estrogen receptor and HER2 were successful in all of the molecular fixative specimens.ConclusionThe formalin-free tissue fixation and processing system is a practical platform for evaluation of biomolecular markers in breast cancer and it allows reliable DNA and RNA and protein studies.

Shave biopsy is a safe and accurate method for the initial evaluation of melanoma.

BACKGROUNDnShave biopsy of cutaneous lesions is simple, efficient, and commonly used clinically. However, this technique has been criticized for its potential to hamper accurate diagnosis and microstaging of melanoma, thereby complicating treatment decision-making.nnnSTUDY DESIGNnWe retrospectively analyzed a consecutive series of patients referred to the University of Florida Shands Cancer Center or to the Moffitt Cancer Center for treatment of primary cutaneous melanoma, initially diagnosed on shave biopsy to have Breslow depth < 2 mm, to determine the accuracy of shave biopsy in T-staging and the potential impact on definitive surgical treatment and outcomes.nnnRESULTSnSix hundred patients undergoing shave biopsy were diagnosed with melanoma from extremity (42%), trunk (37%), and head or neck (21%). Mean (± SEM) Breslow thickness was 0.73 ± 0.02 mm; 6.2% of lesions were ulcerated. At the time of wide excision, residual melanoma was found in 133 (22%), resulting in T-stage upstaging for 18 patients (3%). Recommendations for additional wide excision or sentinel lymph node biopsy changed in 12 of 600 (2%) and 8 of 600 patients (1.3%), respectively. Locoregional recurrence occurred in 10 (1.7%) patients and distant recurrence in 4 (0.7%) patients.nnnCONCLUSIONSnThese data challenge the surgical dogma that full-thickness excisional biopsy of suspicious cutaneous lesions is the only method that can lead to accurate diagnosis. Data obtained on shave biopsy of melanoma are reliable and accurate in the overwhelming majority of cases (97%). The use of shave biopsy does not complicate or compromise management of the overwhelming majority of patients with malignant melanoma.

Gene expression profiling reveals alteration of caspase 6 and 14 transcripts in normal skin of keloid-prone patients

Excessive scar formation in keloids points to altered tissue modeling and repair mechanisms. Dysregulation of cytokine and apoptotic cascades and their downstream signaling pathways might have a role in keloid development. Total RNA was isolated from biopsied keloidal tissue and adjacent normal skin of black patients, white patient’s scars, and normal skin of black and white patients, with normal wound healing. Apoptosis, cytokine and NFkB pathway microarrays were used to study and compare gene expression levels. Real-time PCR was used to verify microarray results in original samples and a separate, validation-set of samples. Significant differences were observed in the expression levels of members of caspase, cytokines and MAP kinase pathways, between the normal skin of keloid-prone and normal skin of keloid-resistant patients. Specifically, expression of caspase 6, and caspase 14 genes were different between normal skin of keloid-prone individuals and normal skin of keloid-resistant patients. Our results suggest that normal skin of keloid-prone individuals constitutively expresses a distinct gene profile which might contribute to their susceptibility to develop keloids.

Nonbullous neutrophilic dermatosis: Sweet’s syndrome, neonatal lupus erythematosus, or both?

We describe a 5-day-old infant who fulfilled the diagnostic criteria for Sweet’s syndrome, and the concurrent histologic and autoantibody features supporting the diagnosis of neonatal lupus. To our knowledge, this is the youngest case of Sweet’s syndrome reported in the literature. Importantly, our findings further support the hypothesis that lupus erythematosus should be considered in the differential diagnosis of a nonbullous neutrophilic dermatosis, as it may represent the initial manifestation of the disease.

Pyoderma gangrenosum masquerading as necrotizing fasciitis

Necrotizing fasciitis is a fulminant advancing soft tissue infection characterized by widespread fascial necrosis, which can result in significant morbidity and even death. This condition requires prompt diagnosis and aggressive management with extensive surgical debridement and appropriate antibiotic coverage. Pyoderma gangrenosum, in contrast, is a noninfectious inflammatory condition of the skin that typically does not require surgical management. Both conditions can present with extensive ulceration and tissue necrosis, and close clinical-pathologic correlation is required to make the appropriate diagnosis. We present a case in which the diagnosis of pyoderma gangrenosum was initially elusive and serves to illuminate the diagnostic pitfalls in dealing with these entities.

Comparative genome hybridization analysis of laser-capture microdissected in situ melanoma.

Background: The progression of melanoma occurs through discrete stages with known clinical and histologic features. Although many molecular events that occur during the progression of invasive and metastatic melanomas have been elucidated, there is limited knowledge of genetic changes that occur in the earliest stages of melanoma development. In this pilot study, we investigated genetic changes that happen in in situ melanoma so that we can better understand early melanoma development.

Signet ring cell melanoma, Brenner sign, and elevated vascular endothelial growth factor

few of the reported cases with drug-induced BP have been confirmed by a rechallenge or a positive patch test. We think that the interventions in our patient could be considered as a successful rechallenge with aspirin. Several mechanisms have been postulated for the pathogenesis of drug-induced BP. Sulfhydrylcontaining drugs may lead to direct dermoepidermal splitting with or without antibodies; however, aspirin does not contain this group. Some BP-inducing drugs, such as penicillamine, decrease suppressor T-cell activity, which stimulates the hyperproduction of autoantibodies against the BP antigens. By altering the antigenicity of the lamina lucida or attaching to a cell site, certain drugs may also act as a hapten and lead to the formation of autoantibodies. Aspirin may have acted as a hapten in our patient. Our case indicates a causal relationship between aspirin, probably the most widely used drug in the world, and BP.

Purpuric Eruption in a Patient Treated With Systemic Steroids—Quiz Case

A 58-year-old woman presented with Escherichia coli bacteremia, vancomycin-resistant Enterococcus faecium meningitis, and septic shock. Before this presentation, she had received systemic corticosteroids on 3 separate occasions at another hospital for exacerbations of chronic obstructive pulmonary disease as a result of 1-antitrypsin disease. Physical examination revealed an extensive purpuric rash on her abdomen (Figure1) and the anterior aspect of her thighs.Shediedshortlyafterward.Sectionsof skinobtained atautopsyshowedparasiticorganismsinthesuperficialdermis (Figure 2 and Figure 3) and subcutaneous fat. What is your diagnosis?

Clear cell hidradenocarcinoma with helpful immunohistochemistry: a case report

Clear cell hidradenocarcinomas are rare, malignant sweat gland neoplasms. This tumor usually arises de novo but can also arise from its benign and more common counterpart, nodular hidradenoma. These tumors have been described with many names, including nodular hidradenocarcinoma, malignant acrospiroma, malignant clear cell hidradenoma, solid-cystic adenocarcinoma, malignant clear cell myoepithelioma, and clear cell eccrine carcinoma. We would like to report a case of clear cell hidradenocarcinoma with a helpful immunohistochemical profile to aid in making the diagnosis.

Purely cutaneous rosai-dorfman disease with immunohistochemistry.

Background: The cutaneous form of Rosai-Dorfman disease (RDD) is a rare entity that manifests solely with skin papules or nodules and does not present with the usual myriad of symptoms of classical RDD. Aims: To analyze the most recent publications regarding cutaneous RDD to point out updated, relevant aspects regarding future directions for clinical recognition and management. To identify histopathologic and immunohistochemical findings in skin lesions that permit diagnosis. Materials and Methods: We present a case of a gentleman with a history of multiple lipomas with a new solitary nodule on physical exam; microscopic examination shows the typical findings of RDD with the associated diagnostic immunohistochemical profile, as well as the expected finding of histiocytes engulfing other intact inflammatory cells. Results: Our patient was managed with surgical excision of the entire lesion, one of the several available treatment options. Long-term follow-up 2 years later did not reveal any complications, recurrences, or new lesions. Conclusion: The diagnosis of cutaneous RDD is differentiated from other histiocytic conditions by the combination of clinical findings accompanied by histopathologic and immunohistochemical confirmation.