CTC-5: A novel digital pathology approach to characterise circulating tumour cell biodiversity.

Metastatic progression and tumor evolution complicates the clinical management of cancer patients. Circulating tumor cell (CTC) characterization is a growing discipline that aims to elucidate tumor metastasis and evolution processes. CTCs offer the clinical potential to monitor cancer patients for therapy response, disease relapse, and screen 'at risk' groups for the onset of malignancy. However, such clinical utility is currently limited to breast, prostate, and colorectal cancer patients. Further understanding of the basic CTC biology of other malignancies is required to progress them towards clinical utility. Unfortunately, such basic clinical research is often limited by restrictive characterization methods and high-cost barrier to entry for CTC isolation and imaging infrastructure. As experimental clinical results on applications of CTC are accumulating, it is becoming clear that a two-tier system of CTC isolation and characterization is required. The first tier is to facilitate basic research into CTC characterization. This basic research then informs a second tier specialised in clinical prognostic and diagnostic testing. This study presented in this manuscript describes the development and application of a low-cost, CTC isolation and characterization pipeline; CTC-5. This approach uses an established 'isolation by size' approach (ScreenCell Cyto) and combines histochemical morphology stains and multiparametric immunofluorescence on the same isolated CTCs. This enables capture and characterization of CTCs independent of biomarker-based pre-selection and accommodates both single CTCs and clusters of CTCs. Additionally, the developed open-source software is provided to facilitate the synchronization of microscopy data from multiple sources (https://github.com/CTC5/). This enables high parameter histochemical and immunofluorescent analysis of CTCs with existing microscopy infrastructure without investment in CTC specific imaging hardware. Our approach confirmed by the number of successful tests represents a potential major advance towards highly accessible low-cost technology aiming at the basic research tier of CTC isolation and characterization. The biomarker independent approach facilitates closing the gap between malignancies with poorly, and well-defined CTC phenotypes. As is currently the case for some of the most commonly occurring breast, prostate and colorectal cancers, such advances will ultimately benefit the patient, as early detection of relapse or onset of malignancy strongly correlates with their prognosis.

Anatomical reconstruction of first ray instability hallux valgus with a medial anatomical TMTJ1 plate.

BACKGROUND: First tarsometatarsal joint (TMTJ1) arthrodesis is a powerful tool for hallux valgus correction. Past criticism of the TMTJ1 arthrodesis has focused on high non-union rates, and consequent need for delayed weightbearing as prevention. In this study we present a selection and treatment protocol to minimise non-union while allowing early weightbearing. METHODS: All TMTJ1 arthrodesis procedures for hallux valgus performed by the senior surgeon over the period June, 2016 to July, 2019 were included. An anatomically-designed, medial TMTJ1 plate and screw compression was utilised for TMTJ1 arthrodesis. The construct was augmented with synthetic intermetatarsal stabilisation. All patients were kept non-weightbearing for 2 weeks, followed by progressive weightbearing as tolerated for 4 weeks. Minimum follow-up was 1 year. RESULTS: 300 modified Lapidus procedures were performed for hallux valgus with mean IMA 17° (Range: 14-29). Mean age was 58 years, with 93% female. 284 (94%) had an Akin osteotomy, while 222 cases (74%) were associated with another forefoot procedure. Patients began progressive weight bearing as tolerated from 2 weeks. All were fully weight bearing by 8 weeks post-operatively. There was a 100% union rate in this group. Mean AOFAS Hallux MTP-IP scores rose from 59 pre-operatively to 97 post-operatively. One plate was removed due to tibialis anterior impingement. There were no recurrences at final follow-up. CONCLUSIONS: We describe a selection and treatment protocol for TMTJ1 arthrodesis for hallux valgus. This yields high union rates while allowing early weight bearing. LEVEL OF EVIDENCE: 4.

Transformed follicular lymphoma (tFL): consolidation therapy may improve survival.

PURPOSE: Retrospective study to evaluate the outcome of patients with transformed follicular lymphoma (tFL) treated with rituximab-containing chemotherapy and consolidation. PATIENTS AND METHODS: Patients diagnosed with tFL from 2003 to 2013 treated with consolidation therapy with last follow-up in December 2015 were identified from the institutional lymphoma database and included in this study. Data collected included age, gender, stage, interval to tFL diagnosis, R-IPI score, histological diagnosis and therapy. The treatment algorithm used was stratified for age, performance status (PS) and sibling donor availability using R-chemotherapy induction followed by consolidation with allogeneic stem cell transplant (SCT), autologous SCT, Zevalin or rituximab maintenance (RM). Patients with B-cell lymphoma with features intermediate between diffuse large B-cell lymphoma and Burkitt's lymphoma (BCL-U), with FISH-proven t(14;18) and t(8;14) and their variants were excluded. RESULTS: Four hundred patients were diagnosed with FL of whom 26 (7%) developed histologically proven tFL. The group was predominantly male (73%) with a median age at transformation of 53 (range 27-72) years and 85% presented with stage III/IV disease. Thirteen (50%) patients presented with de novo tFL and the remainder had previously diagnosed FL, with a median time to transformation of 5.7 (range 1-15) years. The median follow-up time from tFL diagnosis to December 2015 is 8 (range 4-14) years. All patients received immuno-chemotherapy achieving an overall response rate (ORR) of 100%. Fourteen (54%), patients were transplant eligible and based on donor availability, six had an auto-SCT only, five had an allo-SCT only and three had a matched unrelated allo-SCT for a post-auto-SCT relapse. The 12 patients (46%) who were not transplant eligible were consolidated with rituximab maintenance (RM) in nine (35%) and Zevalin in three (11%) cases. The overall survival (OS) and progression-free survival (PFS) for the series at 5 years were, 92 and 73%, respectively. CONCLUSION: This consecutively treated series of 26 patients with tFL have had a better outcome than expected which may be due to the use of rituximab-chemotherapy and a consolidation strategy based on age, PS and availability of a sibling donor.

A novel modification of the Stainsby procedure: surgical technique and clinical outcome.

BACKGROUND: The 'Stainsby procedure' is an effective salvage procedure for correction of fixed claw toe deformity. A novel approach is described involving the extensor tenotomy step of the procedure, which is easier, faster and safer to perform. METHODS: A retrospective single surgeon review was performed to assess this modification. 37 patients (92 lesser toes in 42 feet) underwent the modified Stainsby procedure. Mean follow-up was 17 months. Patients were interviewed and examined at a dedicated review clinic. American Orthopaedic Foot and Ankle Society (AOFAS) forefoot scores and Foot and Ankle Outcome (FAO) scores were calculated. RESULTS: Significant improvements in end-points including metatarsalgia, callosity and requirement for insoles or chiropody were noted. Median AOFAS and FAOS scores following the modified Stainsby procedure are reported and compare favourably to previously reported scores. Median FAOS scores were as follows: 92 for the category of pain, 84 for symptoms and stiffness, 96 for activities of daily living, 100 for sports and recreation and 81 for quality of life. Median AOFAS forefoot score was 80. There was a low rate of wound infection of 2.7% (n=1). CONCLUSION: We describe a novel technical modification to the Stainsby procedure and our results support the use of this modification, following assessment of patient outcome. However, limitations to this study were noted in terms of the length of follow-up, the absence of pre-operative clinical scores for comparative purposes, and the variety of surgical procedures performed concomitantly on the 1st ray.

Development of mapped stress-field boundary conditions based on a Hill-type muscle model.

Forces generated in the muscles and tendons actuate the movement of the skeleton. Accurate estimation and application of these musculotendon forces in a continuum model is not a trivial matter. Frequently, musculotendon attachments are approximated as point forces; however, accurate estimation of local mechanics requires a more realistic application of musculotendon forces. This paper describes the development of mapped Hill-type muscle models as boundary conditions for a finite volume model of the hip joint, where the calculated muscle fibres map continuously between attachment sites. The applied muscle forces are calculated using active Hill-type models, where input electromyography signals are determined from gait analysis. Realistic muscle attachment sites are determined directly from tomography images. The mapped muscle boundary conditions, implemented in a finite volume structural OpenFOAM (ESI-OpenCFD, Bracknell, UK) solver, are employed to simulate the mid-stance phase of gait using a patient-specific natural hip joint, and a comparison is performed with the standard point load muscle approach. It is concluded that physiological joint loading is not accurately represented by simplistic muscle point loading conditions; however, when contact pressures are of sole interest, simplifying assumptions with regard to muscular forces may be valid.

FOXO3a loss is a frequent early event in high-grade pelvic serous carcinogenesis.

Serous ovarian carcinoma is the most lethal gynecological malignancy in Western countries. The molecular events that underlie the development of the disease have been elusive for many years. The recent identification of the fallopian tube secretory epithelial cells (FTSECs) as the cell-of-origin for most cases of this disease has led to studies aimed at elucidating new candidate therapeutic pathways through profiling of normal FTSECs and serous carcinomas. Here we describe the results of transcriptional profiles that identify the loss of the tumor suppressive transcription factor FOXO3a in a vast majority of high-grade serous ovarian carcinomas. We show that FOXO3a loss is a hallmark of the earliest stages of serous carcinogenesis and occurs both at the DNA, RNA and protein levels. We describe several mechanisms responsible for FOXO3a inactivity, including chromosomal deletion (chromosome 6q21), upregulation of miRNA-182 and destabilization by activated PI3K and MEK. The identification of pathways involved in the pathogenesis of ovarian cancer can advance the management of this disease from being dependant on surgery and cytotoxic chemotherapy alone to the era of targeted therapy. Our data strongly suggest FOXO3a as a possible target for clinical intervention.

Lateral thigh swelling: an unusual presentation of psoas abscess in a 61-year-old male.

INTRODUCTION: Psoas abscess is a rare pathology with significant morbidity and mortality, if not diagnosed early and treated appropriately. METHODS: We report a case of 61-year-old male who presented with only lateral thigh swelling. A brief discussion regarding the case follows. CONCLUSION: Psoas abscess is a rare entity and can present in atypical fashion. Clinicians need to be aware of these variation so to not miss this diagnosis.

A finite-element analysis study of the metatarsophalangeal joint of the hallux rigidus.

Hallux rigidus was first described in 1887. Many aetiological factors have been postulated, but none has been supported by scientific evidence. We have examined the static and dynamic imbalances in the first metatarsophalangeal joint which we postulated could be the cause of this condition. We performed a finite-element analysis study on a male subject and calculated a mathematical model of the joint when subjected to both normal and abnormal physiological loads. The results gave statistically significant evidence for an increase in tension of the plantar fascia as the cause of abnormal stress on the articular cartilage rather than mismatch of the articular surfaces or subclinical muscle contractures. Our study indicated a clinical potential cause of hallux rigidus and challenged the many aetiological theories. It could influence the choice of surgical procedure for the treatment of early grades of hallux rigidus.

A clinical test to avoid sural nerve injuries in percutaneous Achilles tendon repairs.

Percutaneous repairs of Achilles tendon ruptures has gained popularity due the reduced incidence of wound complications, however its use is still limited by the high incidence of sural nerve injuries associated with these repairs. The only technique described to avoid this adverse event is to surgically expose the nerve peri-operatively. In our study, we describe and validate a clinical technique to identify the sural nerve. The sural nerve was mapped using this technique both clinically and by Ultrasound (US) in a cohort of male subjects with intact Achilles tendons. We demonstrated an excellent correlation between the clinical and US mapping. This study demonstrates an accurate and repeatable clinical technique for mapping the sural nerve in conjunction with percutaneous Achilles tendon repairs.

Expression microarray analysis of papillary thyroid carcinoma and benign thyroid tissue: emphasis on the follicular variant and potential markers of malignancy.

The most common sub-variant of papillary thyroid carcinoma (PTC) is the so-called follicular variant (FVPTC), which is a particularly problematic lesion and can be challenging from a diagnostic viewpoint even in resected lesions. Although fine needle aspiration cytology is very useful in the diagnosis of PTC, its accuracy and utility would be greatly facilitated by the development of specific markers for PTC and its common variants. We used the recently developed Applied Biosystems 1700 microarray system to interrogate a series of 11 benign thyroid lesions and conditions and 14 samples of PTC (six with classic morphology and eight with follicular variant morphology). TaqMan(R) reverse transcriptase-polymerase chain reaction was used to validate the expression portfolios of 50 selected transcripts. Our data corroborates potential biomarkers previously identified in the literature, such as LGALS3, S100A11, LYN, BAX, and cluster of differentiation 44 (CD44). However, we have also identified numerous transcripts never previously implicated in thyroid carcinogenesis, and many of which are not represented on other microarray platforms. Diminished expression of metallothioneins featured strongly among these and suggests a possible role for this family as tumour suppressors in PTC. Fifteen transcripts were significantly associated with FVPTC morphology. Surprisingly, these genes were associated with an extremely narrow repertoire of functions, including the major histocompatibility complex and cathepsin families.

Mycobacterium tuberculosis at autopsy--exposure and protection: an old adversary revisited.

BACKGROUND: The risk of encountering tuberculosis (TB) has reduced with the decreased incidence of the disease; however, it still can be found at autopsy. AIM: To assess the magnitude of exposure to Mycobacterium tuberculosis at autopsy in a large general hospital setting, in a country with low incidence. METHODS: Retrospective search of the autopsy records from 1991 to 2004. Patients' records and histological slides were reviewed, and medical personnel interviewed. RESULTS: 15 cases of active TB were identified in the 14-year period, during which 4930 autopsies were performed (1 case per 329 autopsies); of these, 10 cases were unsuspected (67%). Five of these cases contained abundant acid-fast bacilli. Patients tended to be middle aged and males with complex clinical histories; two were HIV positive. Two patients were brought in dead to hospital, with no clinical indication of TB. Of 15 autopsy staff, 1 required chemoprophylaxis but none contracted TB. CONCLUSION: The risk of unexpectedly encountering TB at autopsy continues even in a low-risk European setting. It has implications for the health of autopsy room staff, autopsy room design and ventilation, choice of protective equipment and for the public health service. Protective strategies include assessment of the risk of a case being infected, early recognition of gross lesions, use of methods for reducing the production of infected aerosols and protection against any aerosols created.

Cannonball metastases with favourable prognosis.

BACKGROUND: Secondary 'cannonball' metastases to the lung are frequent and usually associated with disseminated malignancy and poor prognosis. AIM: To report the case of a patient with metastatic pulmonary endometrial stromal sarcoma who had a previous hysterectomy for benign uterine fibroids and no past history of malignancy. RESULT: A 70-year-old female presented with cannonball metastases in her lung. Four years previously she had a hysterectomy for 'fibroids'. Review of the original histology revealed endometrial stromal sarcoma, similar to the lung metastasis. She currently has a good prognosis. CONCLUSION: A patient with 'cannonball' metastases can have a favourable prognosis. A female patient with a previous hysterectomy for uterine fibroids, should be considered to have metastatic sarcoma until proven otherwise.

ret/PTC and BRAF act as distinct molecular, time-dependant triggers in a sporadic Irish cohort of papillary thyroid carcinoma.

The purpose of this study was to assess BRAF mutation rates in various thyroid tissues and to investigate if concomitant mutations with ret/PTC activation occurred in inflammatory and neoplastic lesions. To this end, we developed a novel Taqman based screening assay for the common T1799A BRAF mutation. Heterozygous T1799A mutations were detected in 13 of 34 (44%) papillary thyroid carcinomas (PTCs) tested. No such mutations were detected in the other tissue types tested. Concomitant presence of both oncogenes was reported in 5 of the 34 PTCs. A significant temporal trend was observed, with ret/PTC chimera detected for the most part before 1997 and BRAF mutations being more prevalent after 1997. The results suggest that some environmental/etiological agent(s) may have influenced the pathobiology of thyroid tumor development, among the population examined, over time.

Chondro-osseous respiratory epithelial adenomatoid hamartoma of the nasal cavity: a case report.

Chondro-osseous respiratory epithelial adenomatoid hamartoma (COREAH) of the nasal cavity is an exceedingly rare benign lesion of the nasal cavity. We report a case arising in an 11-year-old male patient, who presented with a 6-month history of right-sided nasal obstruction. Examination revealed a large fleshy, tender polyp in the right nasal cavity. Computed tomography (CT) and magnetic resonance imaging (MRI) confirmed the large mass in the anterior nasal cavity extending superiorly to the cribriform plate on the right hand side. The polyp was resected endoscopically and confirmed histologically as a COREAH. The clinical differential diagnosis includes cartilaginous tumours and other hamartomatous lesions. Local resection should be curative.