Tumour-cell-derived complement components C1r and C1s promote growth of cutaneous squamous cell carcinoma.

BACKGROUND: The incidence of epidermal keratinocyte-derived cutaneous squamous cell carcinoma (cSCC) is increasing worldwide. OBJECTIVES: To study the role of the complement classical pathway components C1q, C1r and C1s in the progression of cSCC. METHODS: The mRNA levels of C1Q subunits and C1R and C1S in cSCC cell lines, normal human epidermal keratinocytes, cSCC tumours in vivo and normal skin were analysed with quantitative real-time polymerase chain reaction. The production of C1r and C1s was determined with Western blotting. The expression of C1r and C1s in tissue samples in vivo was analysed with immunohistochemistry and further investigated in human cSCC xenografts by knocking down C1r and C1s. RESULTS: Significantly elevated C1R and C1S mRNA levels and production of C1r and C1s were detected in cSCC cells, compared with normal human epidermal keratinocytes. The mRNA levels of C1R and C1S were markedly elevated in cSCC tumours in vivo compared with normal skin. Abundant expression of C1r and C1s by tumour cells was detected in invasive sporadic cSCCs and recessive dystrophic epidermolysis bullosa-associated cSCCs, whereas the expression of C1r and C1s was lower in cSCC in situ, actinic keratosis and normal skin. Knockdown of C1r and C1s expression in cSCC cells inhibited activation of extracellular signal-related kinase 1/2 and Akt, promoted apoptosis of cSCC cells and significantly suppressed growth and vascularization of human cSCC xenograft tumours in vivo. CONCLUSIONS: These results provide evidence for the role of tumour-cell-derived C1r and C1s in the progression of cSCC and identify them as biomarkers and putative therapeutic targets in cSCC. What's already known about this topic? The incidences of actinic keratosis, cutaneous squamous cell carcinoma (cSCC) in situ and invasive cSCC are increasing globally. Few specific biomarkers for progression of cSCC have been identified, and no biological markers are in clinical use to predict the aggressiveness of actinic keratosis, cSCC in situ and invasive cSCC. What does this study add? Our results provide novel evidence for the role of complement classical pathway components C1r and C1s in the progression of cSCC. What is the translational message? Our results identify complement classical pathway components C1r and C1s as biomarkers and putative therapeutic targets in cSCC.

Matrix metalloproteinase-7 activates heparin-binding epidermal growth factor-like growth factor in cutaneous squamous cell carcinoma.

BACKGROUND: Tumour-specific expression of matrix metalloproteinase (MMP)-7 has been noted in cutaneous squamous cell carcinomas (SCCs) in patients with recessive dystrophic epidermolysis bullosa (RDEB). OBJECTIVES: To examine the potential role of MMP-7 in shedding of heparin-binding epidermal growth factor-like growth factor (HB-EGF) in RDEB-associated and sporadic SCCs. METHODS: Tissue microarrays of RDEB-associated SCC (n = 20), non-EB SCC (n = 60) and Bowen disease (n = 28) were immunostained for MMP-7, CD44 variant 3 (CD44v3) and HB-EGF. Shedding of HB-EGF was studied in vitro using two cutaneous SCC cell lines. RESULTS: Immunohistochemical analysis showed that HB-EGF was absent in tumour cells when MMP-7 and CD44v3 colocalized, and that the absence of HB-EGF was more pronounced in RDEB-associated SCCs than in non-EB SCCs. The loss of HB-EGF in MMP-7-CD44v3 double-positive areas was interpreted to indicate shedding and activation of HB-EGF; this was also detected in Bowen disease indicating its importance in the early phase of SCC development. Specific knockdown of MMP-7 expression in human cutaneous SCC cells by small interfering RNA inhibited shedding of HB-EGF and resulted in diminished activation of the EGF receptor (EGFR) and ERK1/2, and in reduced proliferation of SCC cells. CONCLUSIONS: These findings provide evidence for the role of MMP-7 in promoting the growth of cutaneous SCCs by shedding HB-EGF, and identify EGFR signalling as a potential therapeutic target in RDEB-associated SCC and unresectable sporadic cutaneous SCC.

Suppression of TGFß and Angiogenesis by Type VII Collagen in Cutaneous SCC.

BACKGROUND: Individuals with severe generalized recessive dystrophic epidermolysis bullosa (RDEB), an inherited blistering disorder caused by mutations in the COL7A1 gene, develop unexplained aggressive squamous cell carcinomas (SCC). Here we report that loss of type VII collagen (Col7) in SCC results in increased TGFß signaling and angiogenesis in vitro and in vivo. METHODS: Stable knockdown (KD) of Col7 was established using shRNA, and cells were used in a mouse xenograft model. Angiogenesis was assessed by immunohistochemistry, endothelial tube-forming assays, and proteome arrays. Mouse and zebrafish models were used to examine the effect of recombinant Col7 on angiogenesis. Findings were confirmed in anonymized, archival human tissue: RDEB SCC tumors, non-EB SCC tumors, RDEB skin, normal skin; and two human RDEB SCC cell lines. The TGFß pathway was examined using immunoblotting, immunohistochemistry, biochemical inhibition, and siRNA. All statistical tests were two-sided. RESULTS: Increased numbers of cross-cut blood vessels were observed in Col7 KD compared with control xenografts (n = 4 to 7 per group) and in RDEB tumors (n = 21) compared with sporadic SCC (n = 24, P < .001). Recombinant human Col7 reversed the increased SCC angiogenesis in Col7 KD xenografts in vivo (n = 7 per group, P = .04). Blocking the interaction between α2ß1 integrin and Col7 increased TGFB1 mRNA expression 1.8-fold and p-Smad2 levels two-fold. Increased TGFß signaling and VEGF expression were observed in Col7 KD xenografts (n = 4) compared with control (n = 4) and RDEB tumors (TGFß markers, n = 6; VEGF, n = 17) compared with sporadic SCC (TGFß markers, n = 6; VEGF, n = 21). Inhibition of TGFß receptor signaling using siRNA resulted in decreased endothelial cell tube formation (n = 9 per group, mean tubes per well siC = 63.6, SD = 17.1; mean tubes per well siTßRII = 29.7, SD = 6.1, P = .02). CONCLUSIONS: Type VII collagen suppresses TGFß signaling and angiogenesis in cutaneous SCC. Patients with RDEB SCC may benefit from anti-angiogenic therapy.

Krypton-81m in the physiologic assessment of coronary arterial stenosis in man.

Fifteen patients with frequent anginal chest pain underwent diagnostic cardiac catheterization. After coronary arteriography a specially designed cardiac catheter was seated in the aortic root, permitting the continuous infusion of krypton-81m into the right and left aortic sinuses. A gamma camera, areas of interest and a visual display unit were used to record images and the regional myocardial equilibrium of activity before, during and after a standarized atrial pacing test. The unique physical properties of krypton-81m allowed the continuous imaging and recording of moment to moment changes in regional myocardial perfusion. This investigation revealed that when the coronary arteriogram was normal or revealed lumonal stenosis of less than 50 percent, regional myocardial perfusion was uniform at rest and during stress. Two patients with a previous history of myocardial infarction had defects of regional perfusion at rest and during stress. Krypton scintigraphy demonstrated reversible regional defects in myocardial perfusion during stress in seven patients with greater than 70 percent stenosis of one or more coronary arteries. Alterations in regional myocardial perfusion occurred within 30 seconds of the start of atrial pacing in all the patients and preceded the onset of electrocardiographic signs of ischemia or chest pain.

The clinical importance of magnetic resonance imaging versus computed tomography in malignant pleural mesothelioma.

There is no standard therapy for malignant pleural mesothelioma (MPM), but recent reports have shown that extensive surgery combined with chemo- and radiotherapy prolongs the survival of selected patients with early stage disease. This emphasises the need for accurate staging procedures at diagnosis and reliable imaging methods to assess response to treatment. Computed tomography (CT) of the chest has been the standard imaging method for these purposes for the last decade, but it is limited in its ability to demonstrate accurately the platelike growth pattern of MPM within the thorax due to the partial volume effect on curved surfaces. In order to define the value of magnetic resonance imaging (MRI) in the imaging of MPM, we have compared the findings from 26 parallel paired CT and MRI scans of mesothelioma patients at various stages of the disease. MRI showed tumour spread into the interlobar fissures, tumour invasion of the diaphragm and through the diaphragm, and invasion of bony structures better than CT. Invasion of the chest wall and mediastinal soft tissue and tumour growth into the lung parenchyma were equally well seen on both imaging methods. CT was better for detecting the inactive pleural calcifications. MRI is a sensitive detector of the characteristic growth pattern and extension of MPM and we recommend its use more widely for the clinical management of MPM especially when evaluating tumour resectability and in research protocols when an accurate evaluation of disease extent is essential.

Effect of digital edge-enhancement on the visibility of normal parenchymal lung markings.

Digital 'conventional-like' (C-L) and edge-enhanced (E-E) posteroanterior chest roentgenograms of 42 healthy individuals were ranked twice (interval of at least 5 days) in the order of increasing lung parenchymal markings (a total of four rankings). This was done by three radiologists, two residents, one medical student and one radiographer. There was a good general consistency of rankings for both the C-L and E-E images. The correlation coefficients were best (median 0.600) (P < 0.05) between the consequent rankings of C-L images compared to the subsequent rankings of E-E images (median 0.440) and to the various combinations between the rankings of both kinds of images. Subtle differences in normal lung parenchyma could, therefore, generally (five of the seven observers) best be demonstrated in C-L images, but two observers managed best with the E-E images.

Observed lung markings in normal chest roentgenograms.

The digital chest posterior-anterior roentgenograms of 42 healthy individuals were ranked twice (interval of at least 5 days) in the order of increasing lung parenchymal markings. The evaluations were made by three radiologists, two residents, a medical student and a radiographer. All observers regardless of their radiological experience showed good intraobserver correlations between their two subsequent rankings (p < 0.05-0.001). The interobserver agreement on rankings was generally poor, even if the radiologists were considered. Radiologic training seemed to eliminate the influence of false leading factors (the object's respiration and body constitution). "The golden standard of evaluation" (= the added-up rankings by the radiologists) did not correlate either with the patient's ages (19-54 years) or smoking habits (0-45 pack-years, mean 4.2).

Computed radiography and image workstation from the radiologist's point of view.

A computed radiography system (Digiscan, Siemens) connected to an image workstation (Siemens) has been used for 1 1/2 years in our department. The image quality is good and it has been possible to reduce radiation dose by about 30% without any appreciable loss of image quality. The image workstation has been used in cases where image postprocessing is considered to be useful. For routine reporting of X-ray images the workstation is too slow although the image quality is comparable to that of the computed radiography film.

Transformation-specific matrix metalloproteinases (MMP)-7 and MMP-13 are expressed by tumour cells in epidermolysis bullosa-associated squamous cell carcinomas.

BACKGROUND: Patients with recessive dystrophic epidermolysis bullosa (RDEB) have an increased risk of developing rapidly progressive and metastatic cutaneous squamous cell carcinomas (SCC). It is unclear why these SCC behave more aggressively than sporadic SCC. Matrix metalloproteinases (MMP) are a family of endopeptidases that contribute to growth, invasion and metastasis of SCC. The role of MMP in RDEB-associated SCC is not known. OBJECTIVES: To investigate the expression of MMP-7, MMP-13 and MMP-9 in RDEB-associated SCC in comparison with sporadic SCC and Bowen's disease. METHODS: Immunohistochemical analysis of 25 RDEB-associated SCC, 61 sporadic SCC and 28 sporadic lesions of Bowen's disease was carried out using monoclonal antibodies for MMP-7, MMP-9, MMP-13 and E-cadherin and syndecan-1. RESULTS: MMP-7 was detected in all RDEB-associated SCC, in tumour cells within the invasive edge, where E-cadherin and syndecan-1 were markedly diminished or absent. MMP-7 expression was also observed in 98% of sporadic SCC and in 68% of Bowen's diseases. MMP-7 staining was significantly stronger in RDEB-associated SCC than in sporadic SCC, and was most abundant in poorly differentiated tumours. MMP-13 was detected in tumour cells in 96% of RDEB-associated SCC and in all sporadic cutaneous SCC. MMP-9 was detected in the inflammatory cells in all SCC examined. CONCLUSIONS: These results identify MMP-7 and MMP-13 as tumour cell-specific markers for SCC progression and as potential therapeutic targets in RDEB-associated SCC. The pattern of immunolabelling suggests that MMP-7 may shed E-cadherin and syndecan-1 from the SCC cell surface.

Intra-abdominal abscess in a patient with tumour necrosis factor receptor-associated periodic syndrome.

Tumour necrosis factor (TNF) receptor-associated periodic syndrome (TRAPS) is an autoinflammatory disorder characterized by periodic attacks of fever and inflammation, due to mutations in the gene coding for the TNF type I receptor (TNFRSF1A). A 16-year-old patient with the diagnosis of TRAPS was admitted to hospital because of fever and abdominal pain. Initially, the symptoms were interpreted as manifestations of another TRAPS attack, but the patient's condition worsened, despite treatment with corticosteroids and antibiotics. A repeated computer tomography revealed an intra-abdominal abscess, which necessitated urgent surgical intervention. This case stresses the importance of differential diagnostic vigilance when dealing with patients with rare genetic diseases.

Late haemodynamic response to metrizamide and ioxaglate in canine renal angiography. Preliminary report.

The renal blood flow as examined in six dogs for 2 h after contrast agent injection into the renal artery using the dye dilution method. After injection of an ionic contrast agent (iodamide) there was an initial vasodilatation and later constriction, but the blood flow was back to normal after 10-15 min. Ioxaglate caused a similar initial response, but after a relatively large dose the blood flow remained below normal for 2 h. Metrizamide caused an immediate reduction in the blood flow, which remained constantly reduced for 2 h. A total cessation of the renal blood flow was seen in two cases.

99mTc-DTPA and 99mTc-DMSA renal gamma imaging in the surveillance of patients with conduit urinary diversion.

We studied renal anatomy and function using 99mTc-2-3 dimercaptosuccinic acid (DMSA) and 99mTc-diethylenetriaminepentaacetic acid (DTPA) in 27 patients with conduit urinary diversion. In this condition, free ureteral reflux is often associated with bacteriuria, and these factors are thought to precipitate progressive renal deterioration. Gamma-camera images provided valuable information concerning the structure of the renal parenchyma, the function of individual kidneys and possible ureteral obstruction, thus helping us to decide whether or not to instigate further treatment. The information gained using renal gamma imaging with 99mTc-DTPA and 99mTc-DMSA was complementary and partly overlapping. We preferred the use of 99mTc-DTPA because of its ability to visualise the ureters and the region of ureteroconduit anastomosis. Using diuretic medication, we were able to differentiate true ureteral obstruction from atony in 9 patients using 99mTc-DTPA.

Fractures on chest radiographs in detection of alcoholism.

The prevalence of fractures seen on chest radiographs among alcoholics varies in various series from 36.0% to 8.7% while that for controls remains below 5%. The aim of our study was to find out whether the selection of the alcoholic subjects could account for this four-fold variation. We therefore chose two groups of alcoholics, group 1 consisting of 76 mainly unmarried or divorced lower class alcoholics who had attended the emergency department while intoxicated, and group 2 consisting of 108 mainly married middle class alcoholics with alcoholic liver cirrhosis. The control group consisted of 98 randomly chosen hospital patients. There was no significant difference between group 1 and group 2 in the prevalence of fractures detectable with chest radiography (25% and 21.3% respectively). 8.2% of the controls also had fractures. The fracture prevalence did not correlate with socioeconomic group. Married alcoholics had significantly less thoracic fractures than unmarried, divorced or widowed alcoholics as a group. The sensitivity and specificity of the fractures visible on chest radiograph as a marker for alcoholism is not sufficient to warrant its use in clinical or research work.

The value of urethrocystography in the investigation of patients with spinal cord injury.

In a review of 127 urethrocystograms carried out on 92 males with spinal cord injury the following conclusions were drawn. (1) In the presence of vesico-ureteric reflux or urethral reflux into the male adnexa, bladder outlet obstruction should be suspected. (2) Lesions of the anterior urethra develop easily as the result of indwelling catheterisation in patients lacking sensation. A retrograde urethrogram is necessary to demonstrate these lesions. (3) Severe trabeculation of the bladder wall can be a sign of high-pressure outlet obstruction. (4) Dilatation of the bladder neck in patients with detrusor sphincter dyssynergia may lead to total urinary incontinence after sphincterotomy. (5) Urethrocystography is not a valid investigation to detect bladder stones.

Upper urinary tract changes in patients with spinal cord injury.

Two hundred and six intravenous urograms on 119 patients with spinal cord injury were reviewed and the findings correlated with the clinical data. Fifty (42%) of 119 patients had pathological changes in their upper urinary tracts. The most common feature was impaired renal emptying. Patients with normal and pathological upper tracts had similar findings according to the number of positive urine cultures during the first post-injury year, but in the follow-up those with pathological urograms showed bacteriuria significantly more often. Febrile urinary tract infections at least once a year were encountered in the follow-up of 40% of the patients with pathological urograms, as compared with 8% with normal urograms. All patients with severe renal changes had impaired emptying from the kidneys. This supports the view that the basic patho-physiological mechanism leading to upper tract deterioration in patients with spinal cord injury is a functional or mechanical obstruction of the lower urinary tract. This should be treated actively before irreversible renal changes develop.