Re-evaluation of challenging telepathological diagnoses in support of a hospital in Tanzania.

Since 2007, a hospital in Tanzania has been supported with histopathological reports via telepathology (TP) by German pathologists. For this, the Internet-based platform iPath is used. The aim of this study was to analyse the rate of discrepancies in defined diagnostic groups. After shipment of paraffin-embedded tissue to Germany, specimens were processed according to recent diagnostic standards. All diagnoses were grouped into eight benign and 11 malignant main categories. The comparison comprised the following categories: 1, identical diagnosis; 2, mild discordance; 3, correct distinction between benign and malignant process, 4, false malignant; 5, false benign; and 6, no primary diagnosis possible. The cohort comprised 396 benign and 336 malignant diseases. Of the benign diseases, 62% were category 1, 23% category 2, 2% category 3, 6% category 4 and 7% category 6. Of the malignant diseases, 42% were category 1, 16% category 2, 12% category 3, 14% category 5 and 15% category 6. Exclusive support with static TP cannot meet all requirements of modern medical diagnostics. However, the project shows a approach for how pathologists in industrial countries can help low-income countries. In difficult cases, the opportunity for a final work-up using additional methods must be given for useful diagnostic purposes.

Prognostic Potential of the Expression of Podoplanin (D2-40) Within Cells of Squamous Cell Carcinoma of the Larynx and Hypopharynx.

BACKGROUND: Podoplanin (D2-20) stains immunohistochemically lymphatic vessels, regular mesothelium and tumor cells of different tumors, e.g. malignant mesothelioma or seminoma. In squamous cell carcinoma (SCC), the marker has been described as variously expressed. METHODS: This study has investigated the value of the immunohistochemical analysis for the prognostic relevance of the expression in 119 SCCs of the larynx and hypopharynx. The clinical data and documentation of follow-up for at least 5 years were available. RESULTS: The collective showed the expected distribution of patient age with accentuation of the male sex and a balanced spread of tumor stages including nodal status. The immunohistochemical stain intensity (negative, weak or strong) and the distribution (equal versus focal) were evaluated. In addition, the accentuation of the staining reaction was separately examined at the border of invasion. SCCs with a strong expression of podoplanin were associated with an unfavorable prognosis. A comparison of grouped cases showed a trend emerging with borderline results (negative to weakly positive, P = 0.51; negative to strongly positive, P = 0.054; weakly positive to strongly positive, P = 0.17). The staining at the border of invasion had no statistical effect on overall survival. Multivariate survival statistics however showed that lymphonodal metastasis and a reaction with podoplanin in tumor cells are associated with significant worse prognosis. CONCLUSION: In summary, regardless of the exact function of podoplanin in the process of cell migration and tumor progression, an immunohistochemical identification of expression in tumor cells of SCC of the larynx and hypopharynx can give additional information about the expectable prognosis.

Telepathological evaluation of paediatric histological specimens in support of a hospital in Tanzania.

BACKGROUND/OBJECTIVE: In a project of telepathology (TP) between German pathologists and a hospital in Tanzania, trained technical assistants have uploaded digital histological images onto the internet-based platform ipath. The diagnoses from 486 paediatric specimens were analysed. METHODS: The investigation included diagnoses, either primarily done via TP or secondarily after a further workup of the paraffin-embedded tissue, which was sent to Germany for cases which could not be solved via TP. In the latter, the initial TP-diagnoses were compared with the results after re-evaluation. RESULTS: The median age was 11 years. The cohort comprised 390 benign diseases (80.2%) and 96 malignant diseases (19.8%). For benign diseases, the most frequent anatomic sites were lymph nodes, skin, and soft tissue, breast, and head&-neck. Frequent diagnoses were non-specific inflammations and benign tumors. In malignant diseases, the most sites were lymph nodes, skin, soft tissue, head&neck, and ovary and the most frequent diseases sarcomas and lymphomas. The paraffin embedded tissue of 179 cases (36.3%) was shipped to Germany. With the concordance analysis, we could discover the mandatory necessity for the possibility of second opinion in difficult cases. CONCLUSION: An exclusively TP-support cannot meet all requirements of modern medical diagnostics. The education of local pathologists is imperative.

Diagnostic validity of static telepathology supporting hospitals without local pathologists in low-income countries.

INTRODUCTION: Static telepathology (TP) was used to support a hospital in Tanzania that cannot employ a resident pathologist but has a basic laboratory. Histological slides were prepared by the local technical staff and digital images were uploaded into an Internet-based system; consultant pathologists in Germany could give their opinion. The aim of the study was to examine the diagnostic validity of this project without local pathologists. METHODS: The set-up period for special training of local technical assistants was 10 weeks. Diagnoses of the first 545 cases that were processed via TP were compared with the results of a second opinion on the basis of routine slides created from the corresponding paraffin blocks, which were sent to Germany. RESULTS: Of all cases, 384 (70%) TP diagnoses were completely confirmed by the second opinion. Minor deviations (e.g. divergent subtypes of tumours or other aetiology of non-specific reactive processes) were documented in 76 cases (14%), so that overall, 84% of diagnoses were useful in the setting of the available therapeutic possibilities. The results were better in some subgroups of diseases (90-100% useful diagnoses) and suboptimal (minimum 63%) in a few subgroups with rare diseases. Thirty (5%) malignant diseases were primarily misinterpreted as being benign and 12 (2%) benign diseases as malignant. Forty-three (8%) cases were insufficient for diagnosis using TP and could not be provided with a primary assessment. DISCUSSION: Static TP can help support medical services in low-income countries in the absence of local pathologists with a potentially high diagnostic validity, especially for selected groups of diseases. The procedure can significantly improve the diagnostic procedures before commencement of therapy - a substantial contribution within a globalised world.

Levels of uPA and PAI-1 in breast cancer and its correlation to Ki67-index and results of a 21-multigene-array.

BACKGROUND: Conventional parameters including Ki67, hormone receptor and Her2/neu status are used for risk stratification for breast cancer. The serine protease urokinase plasminogen activator (uPA) and the plasminogen activator inhibitor type-1 (PAI-1) play an important role in tumour invasion and metastasis. Increased concentrations in tumour tissue are associated with more aggressive potential of the disease. Multigene tests provide detailed insights into tumour biology by simultaneously testing several prognostically relevant genes. With OncotypeDX®, a panel of 21 genes is tested by means of quantitative real-time polymerase chain reaction. The purpose of this pilot study was to analyse whether a combination of Ki67 and uPA/PAI-1 supplies indications of the result of the multigene test. METHODS: The results of Ki67, uPA/PAI-1 and OncotypeDX® were analysed in 25 breast carcinomas (luminal type, pT1/2, max pN1a, G2). A statistical and descriptive analysis was performed. RESULTS: With a proliferation index Ki67 of < 14%, the recurrence score (RS) from the multigene test was on average in the low risk range, with an intermediate RS usually resulting if Ki67 was > 14%. Not elevated values of uPA and PAI-1 showed a lower rate of proliferation (average 8.5%) than carcinomas with an increase of uPA and/or PAI-1 (average 13.9%); p = 0.054, Student's t-test. When Ki67 was > 14% and uPA and/or PAI-1 was raised, an intermediate RS resulted. These differences were significant when compared to cases with Ki67 < 14% with non-raised uPA/PAI-1 (p < 0.03, Student's t-test). Without taking into account the proliferative activity, an intermediate RS was also verifiable if both uPA and PAI-1 showed raised values. CONCLUSION: A combination of the values Ki67 and uPA/PAI-1 tended to depict the RS to be expected. From this it can be deduced that an appropriate analysis of this parameter combination may be undertaken before the multigene test in routine clinical practice. The increasing cost pressure makes it necessary to base the implementation of a multigene test on ancillary variables and to potentially leave it out if not required in the event of a certain constellation of results (Ki67 raised, uPA and PAI-1 raised).

Practical Consequences Resulting from the Analysis of a 21-Multigene Array in the Interdisciplinary Conference of a Breast Cancer Center.

During the multidisciplinary planning of postoperative therapy after breast cancer, borderline cases can arise with no clear rationale for or against adjuvant chemotherapy. In 50 hormone- receptor-positive, Her2neu-negative carcinomas of the breast with no or only minimal lymph node involvement (max. pT1a) we initiated an Oncotype DX® multigene assay in addition to the evaluation of usual parameters. In the oncology conference a vote for or against chemotherapy was taken on the basis of the conventional criteria for decision-making before the test results were available. The final recommendation was made after the multigene test. In 32 breast carcinomas (64%) a low recurrence score could be documented, while 26 (32%) showed an intermediate RS and 3 (6%) showed a high RS. In most cases the result of the test could validate the choice of therapy established using conventional criteria. In 5 cases the initial recommendation for adjuvant therapy was revised, and in 3 cases chemotherapy was secondarily recommended after evaluation of the test results. Conversely, in some cases a low or intermediate risk constellation did not argue against a recommendation for adjuvant chemotherapy. Altogether, the results of our study do not indicate that a multigene assay should be used as a routine diagnostic tool. Instead a thorough compilation and careful analysis of conventional parameters for therapeutic decision-making should take precedence, with special emphasis on histopathological and immunohistochemical results. In selected cases, however, a multigene assay can be a useful tool in the deliberation for or against a therapeutic pathway.

Testicular Inflammatory Myofibroblastic Tumor: A Known Entity at a Very Rare Site.

Inflammatory myofibroblastic tumors (IMT) are distinctive lesions of unknown etiology, composed of myofibroblastic spindle cells with an associated inflammatory background. They can occur in a wide age range and at all anatomic sites, but most frequently they can be observed in the lung (especially in pediatric cases), abdomen, or retroperitoneum. The urinary bladder is one of the most common sites in urological cases. We present a very rare case of IMT of the testis. Clinically, a 40-year-old patient showed a palpable painless lesion of the right testis. Ultrasound examination showed two solid intratesticular foci. During surgical intervention, the intraoperative frozen section revealed mesenchymal tumors admixed with an uncommon inflammatory infiltrate, consistent with a reorganized abscess. Despite the benign result, orchiectomy was performed due to the multifocal presentation and the large size of 3 cm. The final diagnosis was IMT without ALK-rearrangement. Incomplete resection increases the risk of local relapses to 30%. In this case, a complete resection could be achieved and the patient is free of tumor 15 months later.

LASP-1, a novel urinary marker for detection of bladder cancer.

OBJECTIVES: The LIM and SH3 (LASP)-1 protein is a focal adhesion protein that has been linked to oncogenesis. The aim of this study was to evaluate the diagnostic use of the detection of LASP-1 in tumor specimens and in urine for noninvasive detection of transitional cell carcinoma (TCC). MATERIALS AND METHODS: Immunohistochemical staining for LASP-1 was performed on 72 archived bladder tumor specimens, and LASP-1 content was measured in 132 spontaneous urine sample sediments by Western blot analysis. RESULTS: In the histologic specimens, immunohistochemical staining for LASP-1 showed abundant expression throughout the urothelium of the bladder and ureter. However, modest overexpression of LASP-1 was observed in the TCC specimens. Measurement of the LASP-1 protein concentrations in urinary cell pellets from healthy donors and bladder cancer patients was highly sensitive for the presence of TCC. The cut-off value was determined by receiver operating characteristic (ROC) analysis. When a cut-off value of 1 ng LASP-1/500 µl of urine was used, the sensitivity, specificity, and positive and negative predictive values of the assay were 83.1%, 85.3%, 83.1%, and 80.6%, respectively.The increased urinary LASP-1 content in TCC patients was attributable in part to a specific increase in cell shedding presumably caused by changes in cell adhesion, as confirmed by LASP-1 knockdown. Contamination with erythrocytes above 250 cells/µl and urinary infection gave false positive results and are therefore sample exclusion criteria. CONCLUSIONS: In the absence of urinary infection or gross hematuria, urinary LASP-1 level is a promising marker for transitional cell carcinoma.

Leiomyolipoma of the orbit.

Orbital leiomyolipoma is a rare benign tumor characterized by a mixture of mature smooth muscle cells and adipocytes. The authors present a case which, to the best of their knowledge, is the second case of leiomyolipoma of the orbit reported in the ophthalmic literature. A 65-year-old patient presented with a painful and swollen left eye, the symptoms having been present for 1 week. Imaging (CT, MRI follow up) confirmed the presence of a nearly 2-cm lesion medial to the lacrimal gland. A biopsy was planned, and 3 specimens were taken for histologic examination. Histology showed diffusely intermingling smooth muscle cells and mature adipocytes; immunohistochemistry demonstrated positive staining for smooth muscle actin and negativity for HMB45, MART-1, and Cytokeratin AE1/3. Although it is very rare, ophthalmologists should consider leiomyolipoma in the differential diagnosis of orbital tumors. The long-term prognosis for patients affected by this benign tumor is good.

Biomaterial-induced sarcomagenesis is not associated with microsatellite instability.

Sarcomagenesis, in contrast to carcinogenesis, is poorly understood. Microsatellite instability has been implicated in the development of many cancers, in particular those associated with chronic inflammatory conditions. In an experimental animal model, rats developed not only a peri-implantational chronic inflammatory reaction, but also malignant mesenchymal tumors in response to different biomaterials. Therefore, it was the aim of our study to test if the development of biomaterial-induced sarcomas is characterized by a mutator phenotype. A multiplex-PCR approach was designed to screen biomaterial-induced sarcomas for the presence of microsatellite instability. Seven different microsatellite loci were tested in ten tumors for microsatellite instability using a fluorochrome-labelled multiplex-PCR and subsequent fragment analysis. All tumors provided a microsatellite-stable phenotype at all loci tested. Our data suggest that microsatellite instability is rarely or not at all a feature of malignant transformation of biomaterial-induced soft tissue tumors. Thus, there is no evidence that a mutator phenotype is a hallmark of biomaterial-induced sarcomagenesis.