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Significance: Sentinel lymph node (SLN) biopsy is an important method for metastasis staging in, e.g., patients with malignant melanoma. Tools enabling prompt histopathological analysis are expected to facilitate diagnostics; optical technologies are explored for this purpose. Aim: The objective of this exploratory study was to investigate the potential of adopting multiphoton laser scanning microscopy (MPM) together with fluorescence lifetime analysis (FLIM) for the examination of lymph node (LN) tissue ex vivo. Approach: Five LN tissue samples (three metastasis positive and two negative) were acquired from a biobank comprising tissues from melanoma patients. Tissues were deparaffinized and subjected to MPM-FLIM using an experimental MPM set-up equipped with a time correlated single photon counting module enabling FLIM. Results: The data confirm that morphological features similar to conventional histology were observed. In addition, FLIM analysis revealed elevated morphological contrast, particularly for discriminating between metastatic cells, lymphocytes, and erythrocytes. Conclusions: Taken together, the results from this investigation show promise for adopting MPM-FLIM in the context of SLN diagnostics and encourage further translational studies on fresh tissue samples.

Objective To describe the radiographic and ultrasonographic characteristics of the spleen and abdominal lymph nodes in clinically healthy ferrets. Materials and Methods Fifty-five clinically healthy ferrets were prospectively recruited for this cross-sectional study. Three-view whole body radiographs and abdominal ultrasonography were performed on awake (23 out of 55) or sedated (32 out of 55) ferrets. On radiographs splenic and abdominal lymph node visibility was assessed. Splenic thickness and echogenicity and lymph node length, thickness, echogenicity, number and presence of cyst-like changes were recorded. Results The spleen was radiographically detectable in all ferrets. On ultrasound the spleen was hyperechoic to the liver (55 out of 55) and mildly hyperechoic (28 out of 55), isoechoic (15 out of 55) or mildly hypoechoic (12 out of 55) to the renal cortices. Mean splenic thickness was 11.80 ±0.34 mm. Lymph nodes were radiographically discernible in 28 out of 55 ferrets and included caudal mesenteric and sublumbar nodes. An average of 9 ±2 lymph nodes (mean± standard deviation; mode 10) were identified in each ferret using ultrasound. A single large jejunal lymph node was identified in all ferrets and had a mean thickness of 5.28 ± 1.66 mm. For other lymph nodes the mean thickness measurements plus one standard deviation were less than 4.4 mm (95% confidence interval: ≤ 3.72 mm). Clinical Significance The information provided in this study may act as a baseline for evaluation of the spleen and lymph nodes in ferrets.

Background We report our experience with mediastinoscopy at Auckland city hospital, a tertiary referral centre. We wished to examine correlations between clinical diagnosis and that made by histological sampling of enlarged mediastinal nodes particularly in patients with isolated mediastinal adenopathy. Methods We retrospectively reviewed clinical records of all patients who underwent mediastinoscopy in a five year period, mediastinoscopy was performed in the presence of enlarged lymph nodes (short axis > 1cm) found at CT. Mediastinoscopy was indicated for diagnostic staging of mediastinal adenopathy related to a parenchymal lung mass, diagnosis of isolated mediastinal adenopathy and diagnosis of mediastinal adenopathy with other CT findings. Data relating to indication, pre-test diagnosis, node stations sampled, histology, and operative complications were collected. Results Mediastinoscopy was performed in 137 consecutive patients. Seventy five patients had a lung mass, 47 had isolated mediastinal adenopathy and 15 had other CT findings. One operative complication occurred. In those patients with isolated adenopathy the following diagnoses were reached; sarcoidosis 23, TB 15, lymphoma 4, carcinoma 4, no diagnosis 1. Final diagnosis was significantly associated with patient's ethnicity. There was high sensitivity and specificity on comparison of clinical and histological diagnosis for both TB and sarcoidosis cases. Conclusions Mediastinoscopy proved to be safe and effective in nodal assessment of the mediastinum. In carefully selected cases procedural morbidity and mortality may be avoided by application of features related to patient's ethnicity and radiological findings.

Objective To determine the final histological and clinical diagnosis of patients with granulomatous lymphadenitis on fine needle aspiration cytology (FNAC). Method A retrospective cohort study was carried out over a five year period in a tertiary referral hospital. FNAC of 22 patients with granulomatous lymphadenitis was reviewed and correlated with the final histological diagnosis and clinical outcome. Results Fourteen cases (64%) underwent surgical biopsy for histological assessment. a definitive diagnosis on FNAC with ancillary investigations was achieved in 82% (18 out of 22) of the cases: four Hodgkin's lymphoma, two non-Hodgkin's lymphoma (NHL), five tuberculosis (TB), two toxoplasmosis, one sarcoidosis and four benign reactive changes. Conclusion A significant number of cases of FNAC diagnosed granulomatous lymphadenitis have an identifiable underlying cause. Patients with reactive cytological changes, who clinically appear benign, can avoid unnecessary surgery.

Background PET/CT scanning can determine suitability for curative therapy and inform decision making when considering radical therapy in patients with non-small cell lung cancer (NSCLC). Metastases to central mediastinal lymph nodes (N2) may alter such management decisions. We report a 2 year retrospective series assessing N2 lymph node staging accuracy with PET/CT compared to pathological analysis at surgery. Methods Patients with NSCLC attending our centre (excluding those who had induction chemotherapy) who had staging PET/CT scans and pathological nodal sampling between June 2006 and June 2008 were analysed. For each lymph node assessed pathologically, the corresponding PET/CT status was determined. 64 patients with 200 N2 lymph nodes were analysed. Results Sensitivity of PET/CT scans for indentifying involved N2 lymph nodes was 39%, specificity 96% and overall accuracy 90%. For individual lymph node analysis, logistic regression demonstrated a significant linear association between PET/CT sensitivity and time from scanning to surgery (p=0.031) but not for specificity and accuracy. Those scanned 9 weeks, 0% ≥9 weeks, p=0.013) and more accurate (94%

Aims—Human herpesvirus 8 (HHV-8) has been identified in multicentric Castleman's disease and in angioimmunoblastic lymphadenopathies. However, the presence of the virus does not necessarily indicate an aetiological role in these conditions. This study investigates the cell types infected by HHV-8 in Castleman's disease and examines the correlation between HHV-8 and Castleman's disease lymph node angiogenesis. Methods—Sixteen formalin fixed, paraffin wax embedded samples from patients with Castleman's disease (six multicentric, 10 solitary) were examined for the presence of HHV-8 using the polymerase chain reaction (PCR), non-isotopic in situ hybridisation, PCR in situ hybridisation (PCR-ISH), and real time quantitative TaqMan PCR to HHV-8 open reading frame 26 (ORF-26), and viral (v)-cyclin encoding regions. Vascularity was assessed using CD34, CD31, and factor VIII immunocytochemistry, and lymph nodes were scored as “low” or “high”. Results—Five multicentric Castleman's disease and two solitary Castleman's disease biopsies were positive for HHV-8. HHV-8 was identified in approximately 10% of intranodal B lymphocytes, in endothelial cells, and in subcapsular spindle cell proliferations. The copy number of HHV-8 was low at 10–50 copies/1000 cells. The highest copy number was in subcapsular spindle cells. There was no correlation between vascularity score and HHV-8 status. Conclusion—The preferential localisation of HHV-8 in subcapsular spindle cell proliferations (where early intranodal Kaposi's sarcoma initiates) and endothelial cells in Castleman's disease might finally explain the link between intranodal Kaposi's sarcoma and Castleman's disease.

The karyotypic abnormality t(14;18)(q32;q21) is reported to occur in 75% of follicular lymphomas. This translocation results in the rearrangement of a putative oncogene bcl-2, which resides at chromosome 18 band q21 (the 18q21 gene). Using two human genomic DNA fragments cloned from the chromosome 18 band q21 as probes, we analyzed 65 uncultured human lymphoma samples by the Southern blot technique. The 18q21 gene was rearranged in 18 of 26 (69%) follicular lymphomas, 3 of 5 (60%) follicular lymphomas transformed to large cell lymphomas, 8 of 20 (40%) diffuse large cell lymphomas (DLCLs), and 2 of 7 (29%) small noncleaved cell lymphomas (SNCs). Our analysis detected rearrangement of the 18q21 gene in 10 of 13 (77%) cases in which the t(14;18)(q32;q21) translocation was found by cytogenetic techniques. Our analysis also proved helpful in difficult karyotyping situations: (a) identifying the donor chromosome fragment as chromosome 18 band q21 in 4 of 9 (44%) cases that cytogenetically displayed a 14q+ chromosome of unknown origin, and (b) identifying a rearrangement of chromosome 18 band q21 in 12 of 18 (67%) cases that cytogenetically yielded no cells in metaphase. We also demonstrated three cases of submicroscopic rearrangement of the 18q21 gene. In our studies, patients with DLCLs and rearrangement of the 18q21 gene had a significantly higher incidence of extranodal involvement when compared with patients with DLCLs and no 18q21 gene rearrangement (P = 0.03).

The monoclonal antibodies HMFG-1 and HMFG-2 are directed against different epitopes on a large molecule in the human milk fat globule and are also expressed on breast tumors. Here we examine the relative intensity of staining of the two antibodies in formalin fixed paraffin embedded tissue sections from malignant and benign mammary lesions. Most ductal carcinomas and their lymph node metastases stained more strongly with HMFG-2 whereas primary lobular carcinomas are more variable but the majority of their lymph node metastases stained more strongly with HMFG-1. Western blots of gel separated extracts of formalin fixed, paraffin embedded tumor tissue, stained with the antibodies in an ELISA assay, show that the molecular weight of components expressing the antigenic determinants varies. Smaller molecular weight components recognised by HMFG-2 and present in metastatic tumors may be lost during tissue processing. To assess the importance of the molecular weight parameter in tumor classification and prognosis it will be necessary to use frozen tissue.

Serum from a patient suspected of having AIDS showed positive ELISA tests but the diagnostic p 41 band was absent and the p 24 band was barely discernible on a Western blot. Before another Western blot was performed lentivirions were detected between dendritic reticulum cells in a lymph node biopsy. It is suggested that samples of biopsies of lymph nodes from patients with or at risk for AIDS should be embedded in resin for future ultrastructural study if indicated.

During the development of B and T cells, a number of genes undergo rearrangement. In this paper we have studied the structure of the T cell gamma-chain gene in primary lymphomas and cell lines derived from patients with lymphoma. Samples derived from patients with angioimmunoblastic lymphadenopathy (AIL), Lennert's lymphoma, and Hodgkin's disease were also examined. TcR gamma was rearranged in all T cell lymphomas and in some B cell lymphomas and B cell lymphoma cell lines. Rearrangement of the gamma-chain gene was also found in AIL, Lennert's lymphoma, and four of eight cases of Hodgkin's disease. These studies indicate that rearrangement of TcR gamma is a useful clonal marker but does not aid in the identification of cell lineage.