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Isolation of Haycocknema perplexum genomic DNA via laser capture microdissection of individual nematodes
ABSTRACTS
proposed to predict malignant behaviour and recurrence including cellularity, mitoses, nuclear pleomorphism, necrosis, size and margin status. However, radiotherapy effects can make interpretation of the significance of some of these features problematic and hence complicate assessment of malignancy. Conclusion: We report a case of SFT in the prostate treated with neo-adjuvant radiotherapy and resection, where assessment of malignancy is potentially confounded by neo-adjuvant treatment effect. LIQUID BASED CYTOLOGY IN THYROID FNAs… CONTROVERSIAL!? Wendy McBurnie Canterbury Health Laboratories, Christchurch Hospital, New Zealand The RCPA/ASC Structured Reporting Protocol for Thyroid Cytology, 2014, states that: ‘Liquid based cytology (LBC) preparations are appropriate for assessing needle wash material, but their use as a replacement for direct smears is to be strongly discouraged. There is no consistent evidence supporting use of liquid based preparation methods for thyroid cytology samples, either as an adjunct or a replacement for direct smears, in terms of adequacy and accuracy rates. Direct smears are essential for on-site adequacy assessment. There are subtle differences in the appearance of the cytological material following LBC preparation which requires experience for reliable interpretation.’ Canterbury Health laboratories have been solely using LBC, and have not been attending radiology for rapid onsite assessment of thyroid FNA samples since 2010. We present our local data and discussions. ISOLATION OF HAYCOCKNEMA PERPLEXUM GENOMIC DNA VIA LASER CAPTURE MICRODISSECTION OF INDIVIDUAL NEMATODES Lewis Melville1,2, Ryan Farid2, James McCarthy2 1 Pathology Queensland, and 2QIMR Berghofer Medical Research Institute, Herston, Qld, Australia Aims: Haycocknema perplexum is an emerging pathogen in Australia. It exhibits behaviour which is unique among human parasitic nematodes. Adult nematodes penetrate skeletal muscle fibres of their host, where they reproduce, perpetuating the infection and provoking an inflammatory response. The result is a progressive, debilitating and sometimes fatal polymyositis. Muscle biopsy plays an essential role in making the diagnosis. The life cycle, natural reservoirs and modes of transmission of this Australian parasite are unknown. The current study aims to isolate H. perplexum genomic DNA from paraffin sections for Next Generation DNA sequence analysis to enable phylogenetic analysis that may inform understanding of its biology. Methods: Laser capture microdissection is used to extract H. perplexum nematodes from formalin fixed, paraffin embedded muscle biopsy tissue. DNA is isolated, and quantified and tested by PCR amplification using universal nematode 18S ribosomal DNA primers.
S77
Results: H. perplexum genomic DNA is successfully isolated from formalin fixed tissue. The morphology and tissue diagnosis of the disease is reviewed. Discussion: A successful protocol for the identification, microdissection and DNA extraction of this intramuscular parasite has been outlined. Further genomic analysis will shed light on the relatedness of H. perplexum to other human and animal parasites and its likely natural reservoir and life cycle. ENCRUSTED CYSTITIS PRESENTING AS AN URACHAL CARCINOMA, A CASE REPORT Laura Mogie, Georgina England Royal Adelaide Hospital and Queen Elizabeth Hospital, SA Pathology, SA, Australia We report a case of a 67-year-old male who presented with a bladder mass, suspicious for urachal carcinoma. The sections comprised ulcerated mucosa with granulation tissue and overlying calcified necrotic debris with associated fibrinous exudate. No mucinous material or malignant epithelium was present. A diagnosis of encrusted cystitis (EC) was made. Encrusted cystitis is a rare form of chronic cystitis.1 Risk factors include catheterisation, immunocompromise, urological diseases and urinary tract infections in patients with chronic debilitating conditions.2 The aetiology is not well understood and a variety of hypotheses exist.3 It is largely believed to occur secondary to urea-splitting bacteria, most commonly Corynebacterium urealyticum.4 Histologically, it comprises mucosal fibrin-covered ulcers with calcified necrotic debris and inflammatory cells with calcium deposits. The surrounding tissue displays inflammation in early disease and fibrosis in the later stages.5 The management of EC differs significantly from its mimics and it is important for the anatomical pathologist to be aware of this rare entity to prevent unnecessary aggressive surgical management. This case highlights a rare complication of urological instrumentation, a mimic of malignancy and emphasises the importance of clinicopathological correlation. References 1. Namsupak J, Headley T, Morabito RA, et al. Encrusted cystitis managed with multimodal therapy. Can J Urol 2008; 15; 3917–9. 2. Soriano F, Tauch A. Microbiological and clinical features of Corynebacterium urealyticum: urinary tract stones and genomics as the Rosetta stone. Clin Microbiol Infect 2008; 14: 632–43. 3. Jelic T, Roque R, Yasar U, et al. Calcifying nanoparticles associated encrusted urinary bladder cystitis. Int J Nanomedicine 2008; 3: 385–90. 4. Johnson MH, Strope SA. Encrusted cystitis. Urology 2012; 79: e31–2. 5. Zhou M, Magi-Galluzzi C. Genitourinary Pathology. 2nd edition. Philadelphia: Elsevier Saunders, 2015; 158–9.
ATYPICAL PRESENTATION OF INVASIVE COMPLETE HYDATIDIFORM MOLE WITH METASTASES TO CERVIX, VULVA AND LUNGS Mudiwa Catherine Muronda, Patricia Guzman Douglass Hanly Moir, Anatomical Pathology, NSW, Australia Gestational trophoblastic disease encompasses a spectrum of pregnancy related proliferative disorders derived from the
proposed to predict malignant behaviour and recurrence including cellularity, mitoses, nuclear pleomorphism, necrosis, size and margin status. However, radiotherapy effects can make interpretation of the significance of some of these features problematic and hence complicate assessment of malignancy. Conclusion: We report a case of SFT in the prostate treated with neo-adjuvant radiotherapy and resection, where assessment of malignancy is potentially confounded by neo-adjuvant treatment effect. LIQUID BASED CYTOLOGY IN THYROID FNAs… CONTROVERSIAL!? Wendy McBurnie Canterbury Health Laboratories, Christchurch Hospital, New Zealand The RCPA/ASC Structured Reporting Protocol for Thyroid Cytology, 2014, states that: ‘Liquid based cytology (LBC) preparations are appropriate for assessing needle wash material, but their use as a replacement for direct smears is to be strongly discouraged. There is no consistent evidence supporting use of liquid based preparation methods for thyroid cytology samples, either as an adjunct or a replacement for direct smears, in terms of adequacy and accuracy rates. Direct smears are essential for on-site adequacy assessment. There are subtle differences in the appearance of the cytological material following LBC preparation which requires experience for reliable interpretation.’ Canterbury Health laboratories have been solely using LBC, and have not been attending radiology for rapid onsite assessment of thyroid FNA samples since 2010. We present our local data and discussions. ISOLATION OF HAYCOCKNEMA PERPLEXUM GENOMIC DNA VIA LASER CAPTURE MICRODISSECTION OF INDIVIDUAL NEMATODES Lewis Melville1,2, Ryan Farid2, James McCarthy2 1 Pathology Queensland, and 2QIMR Berghofer Medical Research Institute, Herston, Qld, Australia Aims: Haycocknema perplexum is an emerging pathogen in Australia. It exhibits behaviour which is unique among human parasitic nematodes. Adult nematodes penetrate skeletal muscle fibres of their host, where they reproduce, perpetuating the infection and provoking an inflammatory response. The result is a progressive, debilitating and sometimes fatal polymyositis. Muscle biopsy plays an essential role in making the diagnosis. The life cycle, natural reservoirs and modes of transmission of this Australian parasite are unknown. The current study aims to isolate H. perplexum genomic DNA from paraffin sections for Next Generation DNA sequence analysis to enable phylogenetic analysis that may inform understanding of its biology. Methods: Laser capture microdissection is used to extract H. perplexum nematodes from formalin fixed, paraffin embedded muscle biopsy tissue. DNA is isolated, and quantified and tested by PCR amplification using universal nematode 18S ribosomal DNA primers.
S77
Results: H. perplexum genomic DNA is successfully isolated from formalin fixed tissue. The morphology and tissue diagnosis of the disease is reviewed. Discussion: A successful protocol for the identification, microdissection and DNA extraction of this intramuscular parasite has been outlined. Further genomic analysis will shed light on the relatedness of H. perplexum to other human and animal parasites and its likely natural reservoir and life cycle. ENCRUSTED CYSTITIS PRESENTING AS AN URACHAL CARCINOMA, A CASE REPORT Laura Mogie, Georgina England Royal Adelaide Hospital and Queen Elizabeth Hospital, SA Pathology, SA, Australia We report a case of a 67-year-old male who presented with a bladder mass, suspicious for urachal carcinoma. The sections comprised ulcerated mucosa with granulation tissue and overlying calcified necrotic debris with associated fibrinous exudate. No mucinous material or malignant epithelium was present. A diagnosis of encrusted cystitis (EC) was made. Encrusted cystitis is a rare form of chronic cystitis.1 Risk factors include catheterisation, immunocompromise, urological diseases and urinary tract infections in patients with chronic debilitating conditions.2 The aetiology is not well understood and a variety of hypotheses exist.3 It is largely believed to occur secondary to urea-splitting bacteria, most commonly Corynebacterium urealyticum.4 Histologically, it comprises mucosal fibrin-covered ulcers with calcified necrotic debris and inflammatory cells with calcium deposits. The surrounding tissue displays inflammation in early disease and fibrosis in the later stages.5 The management of EC differs significantly from its mimics and it is important for the anatomical pathologist to be aware of this rare entity to prevent unnecessary aggressive surgical management. This case highlights a rare complication of urological instrumentation, a mimic of malignancy and emphasises the importance of clinicopathological correlation. References 1. Namsupak J, Headley T, Morabito RA, et al. Encrusted cystitis managed with multimodal therapy. Can J Urol 2008; 15; 3917–9. 2. Soriano F, Tauch A. Microbiological and clinical features of Corynebacterium urealyticum: urinary tract stones and genomics as the Rosetta stone. Clin Microbiol Infect 2008; 14: 632–43. 3. Jelic T, Roque R, Yasar U, et al. Calcifying nanoparticles associated encrusted urinary bladder cystitis. Int J Nanomedicine 2008; 3: 385–90. 4. Johnson MH, Strope SA. Encrusted cystitis. Urology 2012; 79: e31–2. 5. Zhou M, Magi-Galluzzi C. Genitourinary Pathology. 2nd edition. Philadelphia: Elsevier Saunders, 2015; 158–9.
ATYPICAL PRESENTATION OF INVASIVE COMPLETE HYDATIDIFORM MOLE WITH METASTASES TO CERVIX, VULVA AND LUNGS Mudiwa Catherine Muronda, Patricia Guzman Douglass Hanly Moir, Anatomical Pathology, NSW, Australia Gestational trophoblastic disease encompasses a spectrum of pregnancy related proliferative disorders derived from the