Identification of proteins associated with development of metastasis from cutaneous squamous cell carcinomas (cSCCs) via proteomic analysis of primary cSCCs.

BACKGROUND: Cutaneous squamous cell carcinoma (cSCC) is one of the most common cancers capable of metastasizing. Proteomic analysis of cSCCs can provide insight into the biological processes responsible for metastasis, as well as future therapeutic targets and prognostic biomarkers. OBJECTIVES: To identify proteins associated with development of metastasis in cSCC. METHODS: A proteomic-based approach was employed on 105 completely excised, primary cSCCs, comprising 52 that had metastasized (P-M) and 53 that had not metastasized at 5 years post-surgery (P-NM). Formalin-fixed, paraffin-embedded cSCCs were microdissected and subjected to proteomic profiling after one-dimensional (1D), and separately two-dimensional (2D), liquid chromatography fractionation. RESULTS: A discovery set of 24 P-Ms and 24 P-NMs showed 144 significantly differentially expressed proteins, including 33 proteins identified via both 1D and 2D separation, between P-Ms and P-NMs. Several differentially expressed proteins were also associated with survival in SCCs of other organs. The findings were verified by multiple reaction monitoring on six peptides from two proteins, annexin A5 (ANXA5) and dolichyl-diphosphooligosaccharide-protein glycosyltransferase noncatalytic subunit (DDOST), in the discovery group and validated on a separate cohort (n = 57). Increased expression of ANXA5 and DDOST was associated with reduced time to metastasis in cSCC and decreased survival in cervical and oropharyngeal cancer. A prediction model using ANXA5 and DDOST had an area under the curve of 0·93 (confidence interval 0·83-1·00), an accuracy of 91·2% and higher sensitivity and specificity than cSCC staging systems currently in clinical use. CONCLUSIONS: This study highlights that increased expression of two proteins, ANXA5 and DDOST, is significantly associated with poorer clinical outcomes in cSCC.

The histopathological mimics of inflammatory bowel disease: a critical appraisal.

The pathological diagnosis of inflammatory bowel disease (IBD) is often difficult because biopsy material may not contain pathognomonic features, making distinction between Crohn's disease, ulcerative colitis and other forms of colitides a truly challenging exercise. The problem is further complicated as several diseases frequently mimic the histological changes seen in IBD. Successful diagnosis is reliant on careful clinicopathological correlation and recognising potential pitfalls. This is best achieved in a multidisciplinary team setting when the full clinical history, endoscopic findings, radiology and relevant serology and microbiology are available. In this review, we present an up-to-date evaluation of the histopathological mimics of IBD.

A novel imaging approach to periocular basal cell carcinoma: in vivo optical coherence tomography and histological correlates.

PURPOSE: Optical coherence tomography (OCT) is a non-invasive imaging method widely used in ophthalmology. Recent developments have produced OCT devices for imaging the skin. The purpose of this study was to investigate Fourier Domain OCT morphological features of periocular basal cell carcinoma (BCC) in correlation with conventional histopathology. METHODS: Consecutive patients with periocular nodular BCC were prospectively examined with VivoSight OCT (Michelson Ltd) prior to surgical excision. OCT slice mode images were analysed using criteria defined for conventional and HD-OCT; the images were correlated to haematoxylin and eosin stained histology sections. RESULTS: A total of 15 patients with periocular BCC were recruited. Three categories of BCC morphological features were identified from slice mode OCT images: (1) Epidermal changes included epidermal thinning (15/15; 100%), ulceration and crusting (5/15, 33.3%) and decreased density of hair follicles (8/15; 53.3%); (2) Intralesional features included hyporeflective nodules (15/15; 100%), hyper-reflective edges (15/15; 100%) and hyporeflective central necrosis (3/15; 20%) (3) Perilesional features included hyporeflective borders (11/15; 73%), hypereflective margins (15/15; 100%) and dilated blood vessels (5/15; 33%). CONCLUSIONS: This study demonstrated that Fourier Domain OCT imaging offers additional information in the identification of morphological features of nodular BCC compared to conventional OCT diagnostic criteria. VivoSight produced fast, non-invasive imaging of skin lesions in the periocular region and high correlation with histology. Further studies are necessary to investigate OCT features of different histological subtypes of BCC.

Dual implantation of a radio-telemeter and vascular access port allows repeated hemodynamic and pharmacological measures in conscious lean and obese rats.

Expansion of physiological knowledge increasingly requires examination of processes in the normal, conscious state. The current study describes a novel approach combining surgical implantation of radio-telemeters with vascular access ports (VAPs) to allow repeated hemodynamic and pharmacological measures in conscious rats. Dual implantation was conducted on 16-week-old male lean and obese Zucker rats. Continued viability one month after surgery was observed in 67% of lean and 44% of obese animals, giving an overall 54% completion rate. Over the five-week measurement period, reliable and reproducible basal mean arterial pressure and heart rate measures were observed. VAP patency and receptor-independent vascular reactivity were confirmed by consistent hemodynamic responses to sodium nitroprusside (6.25 µg/kg). Acutely, minimal hemodynamic responses to repeated bolus administration of 0.2 mL saline indicated no significant effect of increased blood volume or administration stress, making repeated acute measures viable. Similarly, repeated administration of the ß-adrenoceptor agonist dobutamine (30 µg/kg) at 10 min intervals resulted in reproducible hemodynamic changes in both lean and obese animals. Therefore, our study demonstrates that this new approach is viable for the acute and chronic assessment of hemodynamic and pharmacological responses in both lean and obese conscious rats. This technique reduces the demand for animal numbers and allows hemodynamic measures with minimal disruption to animals' welfare, while providing reliable and reproducible results over several weeks. In conclusion, dual implantation of a radio-telemeter and VAP introduces a valuable technique for undertaking comprehensive studies involving repeated pharmacological tests in conscious animals to address important physiological questions.

Effects of exenatide in poorly controlled type 2 diabetes.

AIM: The aim of this retrospective analysis was to assess the clinical effectiveness of exenatide in patients with type 2 diabetes in routine clinical practice. METHODS: Patients with type 2 diabetes mellitus and inadequate glycemic control were commenced on exenatide in an out-patient setting. Effects on Hba1c, weight and BMI at 3- and 6-month intervals were recorded by a retrospective review of medical records. RESULTS: We examined a cross-section of 61 patients. The mean weight at treatment initiation was 114 kg and baseline Hba1c was 9.8% (84 mmol/mol). Mean reduction in Hba1c at 3 months was 0.8% (10 mmol/mol, P < 0.01) and mean reduction at 6 months was 0.5% (6 mmol/mol, P < 0.05). Mean weight loss at 3 months was 4.2 kg (P < 0.0001) and at 6 months was 6.6 kg (P < 0.0001). Seventeen patients were prescribed exenatide in addition to insulin, against current guidelines. This cohort of patients showed a greater mean reduction in weight (7.4 vs 6.2 kg) as compared to the group on exenatide without insulin, but mean Hba1c increased at 6 months by 0.35% (4 mmol/mol). CONCLUSION: Adjunctive exenatide treatment in patients with suboptimally controlled type 2 diabetes on oral hypoglycaemic medications, achieved reductions in Hba1c and weight, in line with published studies. However, in patients already on insulin, favourable results can be achieved by the addition of exenatide by careful patient selection and follow-up.

Tumour angiogenesis: the gap between theory and experiments.

A common experimental technique for viewing in vivo angiogenesis utilises tumours implanted into a test animal cornea. The cornea is avascular but the tumour promotes vascularisation from the limbus and the new blood vessels can be readily observed through the transparent cornea. Many of the early mathematical models for tumour angiogenesis used this scenario as their experimental template and as such assumed that there is a large gap, of the order of 2mm, between the tumour and neighbouring vasculature at the onset of angiogenesis. In this work we consider whether the assumption that there is a significant gap between the tumour and neighbouring vasculature is unique to intra-cornea tumour implants, or whether this characterises avascular tumour growth more generally. To do this we utilise a simple scaling argument, derive a multi-compartment model for tumour growth, and consider in vivo images. This analysis demonstrates that the corneal implant experiments and the corresponding mathematical models cannot generally be applied to a clinical setting.

Primary peritoneal mesothelioma: case series and literature review.

Primary peritoneal mesothelioma is a rare and aggressive tumour. We present six consecutive cases treated by our institution in the last three years. All were between 56-65 years old and only one gave a history of direct contact with asbestos. Four of the patients showed a thrombocytosis on presentation but other blood tests and evaluation of ascitic fluid were normal. In all cases, the diagnosis was made through investigation of mixed abdominal symptoms with CT scanning and laparoscopic biopsy. Despite the use of modern chemotherapy, response to treatment was unpredictable, with survival from ten weeks to three years.

Imaging of periocular basal cell carcinoma using en face optical coherence tomography: a pilot study.

AIM: To use en face optical coherence tomographic (OCT) imaging to identify features of tumour tissue and their correlation with histopathologic findings and to assess the effect of different wavelengths and resolutions of OCT in identifying tumour boundaries and features. METHODS: Excision specimens of consecutive biopsy-proven periocular basal cell carcinomas (BCCs) (n=8) were assessed by OCT, performing in vitro cross-section and en face scans of the tissues. Images were collected from three different machines: systems 1 and 2 had a wavelength of 1300 nm, and system 3 had a wavelength of 840 nm. System 2 used high numerical aperture interface optics that determines higher magnification and hence allows higher transversal resolution. All the eight specimens subsequently underwent routine histopathologic examination. RESULTS: Three common features of tumour tissue were observed in all the three systems: (1) lobular pattern of abnormal architecture, (2) dilated blood vessels and (3) high reflective margins. We compared the three systems based on their ability to pick up the three above-mentioned tumour features. In this respect, system 2 had the highest capability in picking up feature 1, followed by systems 1 and 3. In feature 2, similar results were obtained with all the three systems. System 3 was unable to pick up feature 3, whereas systems 1 and 2 performed equally. CONCLUSION: En face OCT imaging has the potential to identify tumour tissue from healthy tissue. It also showed correlation with corresponding histopathologic findings. Non-contact OCT imaging of the skin is a non-invasive and convenient method and can be useful for demarcating BCCs on the face and eyelids. Future larger studies on in vivo BCCs using en face ultra-high-resolution OCT should provide information on subtyping BCCs.

Combined adenocarcinoid and mucinous cystadenoma of the appendix: a case report.

INTRODUCTION: Adenocarcinoid of the appendix is a rare malignant tumour with features of both adenocarcinoma and carcinoid, showing both epithelial and endocrine differentiation. Mucinous cystadenoma is the commonest of the benign neoplasms of the appendix, with an incidence of 0.6% in appendicectomy specimens. We report a rare combination of these tumours and discuss the latest treatment options. To the best of our knowledge, only six cases have been reported in the literature to date. CASE PRESENTATION: A 71-year-old Caucasian man presented to our department with a right iliac fossa mass associated with pain. Laparoscopy revealed an adenocarcinoid of the appendix in combination with mucinous cystadenoma. He underwent a radical right hemicolectomy with clear margins and lymph nodes. CONCLUSION: Adenocarcinoids account for 2% of primary appendiceal malignancies. Most tumours are less than 2 cm in diameter and 20% of them metastasize to the ovaries. The mean age for presentation is 59 years and the 5-year survival rate ranges from 60% to 84%. Right hemicolectomy is generally advised if any of the following features are present: tumours greater than 2 cm, involvement of resection margins, greater than 2 mitoses/10 high-power fields on histology, extension of tumour beyond serosa. Chemotherapy mostly with 5-Fluorouracil and Leucovorin is advised for remnant disease after surgery. Cytoreductive surgery with intraperitoneal chemotherapy can offer improved survival for advanced peritoneal dissemination.

Rectosigmoid tumours: should we continue sitting on the fence?

Rectal cancers are currently defined as tumours below 15 cm from the anal verge on rigid sigmoidoscopy. Clinical trials have used this criterion to select patients for neoadjuvant chemoradiotherapy, but several authors have shown that the distance between the fully peritonealized sigmoid colon and the anal canal varies significantly between individuals. A fixed anatomical landmark would be a more reliable and reproducible method of demarcating the junction between the colon and the rectum. The distinction between rectal and sigmoid colon cancers is of particular importance as treatment protocols for rectal cancer management often involve neoadjuvant treatment in contrast to colonic cancers, so it is vital to get the anatomy right. As all rectal cancers are now assessed preoperatively by MRI, the use of a bony landmark is possible. We postulate that the fixed landmark to define the upper limit of the rectum should be the sacral promontory.

The importance of accurate pathological assessment of lymph node involvement in colorectal cancer.

This review presents an up-to-date analysis of the importance of accurate pathological lymph node staging in colorectal cancer. Lymph node staging is reliant on the technique of the surgeon and the pathologist as well as methods employed in the histopathology laboratory, and is vital for determining appropriate therapy. The significance of micrometastatic nodal disease is evaluated and new techniques for pathological evaluation are discussed. Recommendations for evaluation of lymph node status in colorectal cancer are provided based on current scientific evidence, and standardization of pathological dissection and laboratory handling is advocated.

Phthalocyanine-mediated photodynamic therapy induces cell death and a G0/G1 cell cycle arrest in cervical cancer cells.

We have developed a series of novel photosensitizers which have potential for anticancer photodynamic therapy (PDT). Photosensitizers include zinc phthalocyanine tetra-sulphonic acid and a family of derivatives with amino acid substituents of varying alkyl chain length and degree of branching. Subcellular localization of these photosensitizers at the phototoxic IC(50) concentration in human cervical carcinoma cells (SiHa Cells) was similar to that of the lysosomal dye Lucifer Yellow. Subsequent nuclear relocalization was observed following irradiation with 665nm laser light. The PDT response was characterized using the Sulforhodamine B cytotoxicity assay. Flow cytometry was used for both DNA cell cycle and dual Annexin V-FITC/propidium iodide analysis. Phototoxicity of the derivatives was of the same order of magnitude as for tetrasulphonated phthalocyanine but with an overall trend of increased phototoxicity with increasing amino acid chain length. Our results demonstrate cell death, inhibition of cell growth, and G(0)/G(1) cell cycle arrest during the phthalocyanine PDT-mediated response.

Audit of local recurrence rates following 'ultra'-conservative surgery for invasive breast cancer--a boost to the breast?

BACKGROUND: Conservative breast surgery with postoperative radiotherapy and appropriate systemic therapy is associated with similar outcomes when compared with mastectomy. The reported 5 year local recurrence rate varies between 3% and 15%. We prefer a more conservative 'complete' local excision rather than 'wide' local excision combined with post-operative radical radiotherapy and tumour bed boost with the aim of achieving optimal cosmesis. AIMS: Our review was undertaken to assess whether or not this 'ultra' conservative approach was compromising long-term local control. METHODS: Case notes and pathology reports of patients who underwent conservative surgery for breast cancer from January 1983 to February 2001 were accessed for this audit. Patient demographic data and tumour characteristics were noted. The primary outcome data were the number of local recurrences following invasive breast cancer at 5 and 10 years and the distance from the tumour to the closest margin of excision. RESULTS: At 5 and 10 years there were 16/451 and 5/124 local recurrences, with a local recurrence rate of 3.5% (95% CI, 1.7-4.7%) and 4.1% (95% CI, 0.47-6.5%), respectively. Complete data with regards to the closest histological margin of excision were available in 423 patients. One hundred and sixty-five patients (39%) had their tumours excised with a distance of less than 1 mm to the closest margin. Nearly, all tumours (97.8%) were excised with the distance to the closest margin less than 1 cm and 81% with 5 mm or less. CONCLUSION: It is possible to achieve low local recurrence rates after very conservative surgery for breast cancer when this is combined with radical radiotherapy and an additional tumour bed boost.

Monitoring patients on methotrexate: hepatic fibrosis not seen in patients with normal serum assays of aminoterminal peptide of type III procollagen.

BACKGROUND: The aminoterminal peptide of type III procollagen (PIIINP) is formed during the synthesis of type III collagen and can be measured in the serum. It has been used as a marker for hepatic fibrosis in patients on long-term methotrexate and it has been suggested that serial assay of PIIINP could reduce or eliminate the need for liver biopsies in these patients. OBJECTIVES: To determine whether routine use of the PIIINP assay in a cohort of patients on methotrexate would reliably identify those who were developing hepatic fibrosis and exclude those who were not, thereby reducing or eliminating the need for liver biopsies in this latter group. METHODS: Data were available from a clinical series of 38 patients on methotrexate, who had undergone a total of 70 liver biopsies and 306 PIIINP assays. Liver biopsies were graded using the Roenigk classification. RESULTS: In 34 patients, the findings on 46 liver biopsies could be compared with the results of contemporaneous PIIINP assays. Apart from two biopsies from two patients where fibrosis was no longer detected on a subsequent biopsy, all four biopsies showing fibrosis had abnormal results on over half of the associated PIIINP assays. There were no biopsies showing fibrosis where all associated PIIINP assays were normal. However, 50% of biopsies without fibrosis had at least one abnormal associated assay. In 23 patients, the results of serial PIIINP assays performed between two sequential liver biopsies were correlated with changes in the biopsy in terms of fibrosis. Data were available for 32 pairs of liver biopsies. Apart from a biopsy pair in one patient where fibrosis on the second biopsy was not detected on a third biopsy, all four biopsy pairs defined as showing deterioration had abnormal results on over half of the intervening PIIINP assays. There were no biopsy pairs showing deterioration where all intervening assay results were normal. However, 63% of stable biopsy pairs had at least one abnormal intervening assay. Two patients with nonalcoholic steatohepatitis, which manifests a pattern of fibrosis not scored under the Roenigk classification, had persistently and substantially elevated PIIINP levels. CONCLUSIONS: The data presented support the view that follow-up liver biopsies, as recommended by published guidelines, for patients on long-term low-dose methotrexate can be avoided if PIIINP levels are consistently normal. This approach would have reduced the number of patients requiring biopsy in our series by 45%. The PIIINP assay will also be helpful in the management of patients on methotrexate in whom liver biopsy is contraindicated, and in patients with nonalcoholic steatohepatitis.

Lack of pharmacokinetic interactions between cilomilast and theophylline or smoking in healthy volunteers.

The pharmacokinetic profile of cilomilast (Ariflo), a selective phosphodiesterase 4 (PDE4) inhibitor, was investigated in three separate studies. Two of these studies explored the drug interaction potential of cilomilast with the nonselective PDE inhibitor, theophylline, and a third study compared the pharmacokinetic profile of cilomilast in smokers and nonsmokers. Repeated administration of cilomilast had no effect on the steady-state pharmacokinetics of theophylline in either a pilot dose-ranging or definitive therapeutic study. At therapeutic doses, the point estimate and 90% confidence interval for theophylline AUC(0-12) and C(max) were completely contained within the range (0.8, 1.25). Similarly, repeated administration of theophylline had little clinically relevant effect on the steady-state pharmacokinetics of cilomilast when compared to placebo, as only slight average increases in cilomilast AUC(0-12) and C(max) (6% and 3%, respectively) were observed. In addition, mean cilomilast exposure (AUC(0- infinity )) was found to be similar in both smokers and nonsmokers (8.47 +/- 2.20 microg*h/mL and 7.70 +/- 2.25 microg*h/mL, respectively). Throughout all three studies, cilomilast was well tolerated, and concomitant use of these selective and nonselective inhibitors, although unlikely in the clinic, is hypothetically feasible. Taken together, these studies clearly differentiate cilomilast from theophylline for drug-drug liability issues in a smoker and nonsmoker population, as well as highlight the potential to switch from one drug to another without undue clinical concern.

Efficacy of incisional vs punch biopsy in the histological diagnosis of periocular skin tumours.

AIMS: The aim of this study was to compare the accuracy of incisional and punch biopsy techniques in obtaining correct histological diagnosis of periorbital eyelid tumours. The technique of punch biopsy is presented and described in detail. METHODS: A retrospective analysis was made of 20 consecutive incisional biopsies and 20 consecutive punch biopsies. In each case, the histology obtained at biopsy was compared with that identified at the time of tumour excision. RESULTS: A total of 40 consecutive biopsies on 38 patients were analysed. The first 20 were incisional; the second 20 were punch biopsies. Of the 20 incisional biopsy specimens, 19 were confirmed accurate at the time of excision of the lesion. Of the 20 punch biopsies, 17 were confirmed accurate at the time of excision. These correspond to accuracy rates of 95 and 85%, respectively. CONCLUSIONS: Both incisional and punch biopsy techniques have relatively high accuracy rates and there is a high concordance between tissue diagnoses made by each of these techniques. Incisional techniques should preferably be performed on any atypical lesion. Punch biopsy is a quick and simple procedure. It is easy to perform in an outpatient environment and requires a minimum of surgical equipment and no specific surgical skills. If the site of biopsy is carefully chosen, this simple technique provides tissue specimens of adequate size and quality for accurate histology and is a most useful adjunct in the management of periocular tumours.