Urinary clusterin and cystatin B as biomarkers of tubular injury in dogs following envenomation by the European adder.

Diagnosing acute kidney injury remains a challenge since the established renal biomarkers, serum creatinine (sCr) and symmetric dimethylarginine (SDMA) reflect glomerular function and not tubular injury. Sensitive tubular markers such as urinary clusterin (uClust) and cystatin B (uCysB) have been proposed to detect AKI at an earlier stage. Since envenomation by the European adder (Vipera berus berus) could serve as a spontaneous disease model of AKI we investigated these new biomarkers in affected dogs. Concentrations of uClust and uCysB as well as sCr and SDMA were analyzed retrospectively in stored samples from 26 dogs with snake envenomation and 13 healthy controls. Higher concentrations of uClust (P < 0.012) and uCysB (P < 0.001) were observed in the snake-envenomed group. Normalization of uClust and uCysB to urinary creatinine did not alter the results. No differences were observed in sCr and SDMA between the snake-envenomed group and the healthy control group. Spearman rank correlation analysis revealed a strong association of uClust with uCysB in the snake-envenomed dogs (r = 0.75 P < 0.001) but not in the healthy controls. The high percentage of snake-envenomed dogs with increased uClust and uCysB concentrations in the absence of increased sCr and SDMA suggests renal tubular injury in the affected dogs. Larger prospective case-controlled studies are warranted to evaluate the clinical utility and prognostic value of these biomarkers.

Measurements of echocardiographic indices and biomarkers of kidney injury in dogs with chronic kidney disease.

Pathophysiological cardiac and renal interactions are termed cardiovascular-renal disorder (CvRD). Cardiovascular disease/dysfunction secondary to kidney disease (CvRDK), is a leading cause of death in human chronic kidney disease (CKD) patients. The presence and clinical impact of CvRDK in dogs with CKD is unknown. We hypothesized that echocardiographic measurements, and cardiac and renal biomarkers, will be altered in dogs with CKD and associated with survival. Eleven dogs with CKD (n = 6 IRIS stage 2, n = 5 IRIS stage 3) and without primary cardiac disease, plus 12 healthy age-matched control dogs, were recruited to this prospective observational study. Dogs underwent standard echocardiography, glomerular filtration rate (GFR) estimation by iohexol clearance, and measurement of plasma cardiac troponin I and N-terminal pro-B-type natriuretic peptide (NT-proBNP), plasma and urinary cystatin B, and urinary clusterin and neutrophil gelatinase-associated lipocalin (NGAL). Values were compared between groups, and their association with all-cause mortality explored. Dogs with CKD had significantly lower GFR and higher NT-proBNP, urinary cystatin B, clusterin, and NGAL, compared to controls (P < 0.05). Echocardiographic measurements were similar between dogs with CKD and controls. Median follow-up time was 666 days, during which six dogs with CKD died. Risk of death was associated with increasing age, serum total protein, and normalized left ventricular posterior wall thickness (LVPWDN) and decreasing bodyweight and packed cell volume. Although baseline differences in echocardiographic measurements were not evident between dogs with moderate CKD and controls, the presence of CvRDK was suggested by the association between LVPWDN and survival.

Reagentless biosensing using electrochemical impedance spectroscopy.

The use of electrochemical impedance spectroscopy (EIS) and the conducting polymer, poly (pyrrole), as an integrated recognition and transduction system for reagentless biosensor systems was demonstrated with two different systems. The first system being an immunoassay for detection of luteinising hormone (LH) with the antibody being entrapped with in the poly (pyrrole) matrix and the second, a construct for DNA hybridisation discrimination able to differentiate single- and double-stranded DNA based on the interaction of the DNA with poly (pyrrole).

Strategic issues in reliable sensing.

Amperometric enzyme biosensors must possess two important characteristics if they are to be successfully utilised as reliable monitoring devices. They must exhibit linearity over concentrations relevant to the target analyte, and they must avoid contamination, adverse reactions with the sample matrix response to interferents that react directly at the polarised working electrode surface. Covering polymeric membranes have provided a useful route to overcoming these problems. This report summarises successful modulation of membrane bulk as well as surface properties using surfactant-loaded diffusion limiting PVC and phenolic membranes, and the possible exploitation of direct response conducting poly (pyrrole) membrane loaded with affinity molecules through impedance spectroscopy.

Molecular patterning on carbon based surfaces through photobiotin activation.

We have demonstrated the site-specific adhesion of photobiotin as a method of producing protein micropatterns. These patterns were created by the selective UV irradiation of a thin film of deposited photobiotin. The UV activated areas of photobiotin were then developed using fluorescently labelled avidin. The size of pattern produced is an order of magnitude smaller than those previously reported by this method. The patterns were characterised, using atomic force microscopy (AFM) to determine their microstructure. It was found that the AFM could discriminate between the areas of protein immobilised to the surface through the activated photobiotin, and the bare substrate surface where the inactivated photobiotin had been removed during the washing process. The potential of these patterns as sensing surfaces is demonstrated through the creation of a spatially patterned immunosensing surface. In this case, a biotinylated antibody was bound to the surface and the pattern developed using a second antibody specific to the immobilised biotinylated antibody. This technique could thus provide a simple and efficient method of producing high density immunoassay systems.

[Pancreatic tumor: an unusual presentation of an occult breast carcinoma]. / Tumor de páncreas: presentación infrecuente de un carcinoma de mama oculto.

A case of cholestasis due to a synchronous pancreatic head metastasis from an occult lobular breast carcinoma is presented. The patient had a clinical and radiological picture compatible with a pancreatic head primary tumor with cholestasis and ascites. Ultrasonographically guided fine needle aspiration cytology demonstrated a metastatic breast lobular cancer (positive for cytokeratin AE1 and AE3, cytokeratin 7 and epithelial membrane antigen and negative for cytokeratin 20, CA 19.9, CA 125, CEA and estrogenic receptors). The same cytologic findings were observed in skin and subcutaneous armpit nodules. Clinical and radiological breast examination was unable to demonstrate any tumor in the breast. Pancreatic metastases are rare events and the majority of them are secondary to renal and lung cancer and rarely to breast cancer. In these latter cases, metastases are usually disclosed after a disease-free interval of months or years between primary tumor resection and recognition of the pancreatic tumor. Synchronous presentation is extremely rare. Metastases of epithelial origin are uncommon in pancreas and generally are first misdiagnosed as primary pancreatic cancer. Fine needle aspiration is a useful tool for the differential diagnosis in patients with widespread disease.

Tumor de páncreas: presentación infrecuente de un carcinoma de mama oculto / Pancreatic tumor: an unusual presentation of an occult breast carcinoma

Se presenta un caso de tumor de cabeza de páncreas con colestasis debido a una metástasis de un carcinoma lobular oculto de mama. El cuadro es interpretado inicialmente como un carcinoma de páncreas diseminado, con ascitis, colestasis e implantes peritoneales. La biopsia de lesiones de piel y celular subcutánea que resultan en metástasis de carcinoma lobular de mama, confirmado con inmunomarcación (positiva para citoqueratinas clon AE1 y AE3, citoqueratina 7 y antígeno epitelial de membrana y negativa para citoqueratina 20, CA 19.9, CA 125, CEA y receptores estrogénicos) hacen a la presunción de metástasis pancreática. Ésta presunción se confirma mediante PAAF ecoguiada. Las metástasis en páncreas de tumores primitivos de origen epitelial son infrecuentes y en general se presentan como imágenes nodulares indistinguibles de los tumores primitivos. Se observan habitualmente en casos de diseminación masiva por carcinoma de riñon o de pulmón siendo raras las metástasis de carcinoma de mama, las cuales se presentan habitualmente con el antecedente de cáncer de la misma. El diagnóstico del tumor pancreático debe ser efectuado por anatomía patológica. Este es un caso atípico en su forma de presentación, dada la aparición de colestasis debida a un tumor metastático a nivel de cabeza de páncreas sincrónico con un carcinoma lobular de mama oculto.

Tumor de páncreas: presentación infrecuente de un carcinoma de mama oculto / Pancreatic tumor: an unusual presentation of an occult breast carcinoma

Se presenta un caso de tumor de cabeza de páncreas con colestasis debido a una metástasis de un carcinoma lobular oculto de mama. El cuadro es interpretado inicialmente como un carcinoma de páncreas diseminado, con ascitis, colestasis e implantes peritoneales. La biopsia de lesiones de piel y celular subcutánea que resultan en metástasis de carcinoma lobular de mama, confirmado con inmunomarcación (positiva para citoqueratinas clon AE1 y AE3, citoqueratina 7 y antígeno epitelial de membrana y negativa para citoqueratina 20, CA 19.9, CA 125, CEA y receptores estrogénicos) hacen a la presunción de metástasis pancreática. Esta presunción se confirma mediante PAAF ecoguiada. Las metástasis en páncreas de tumores primitivos de origen epitelial son infrecuentes y en general se presentan como imágenes nodulares indistinguibles de los tumores primitivos. Se observan habitualmente en casos de diseminación masiva por carcinoma de riñon o de pulmón siendo raras las metástasis de carcinoma de mama, las cuales se presentan habitualmente con el antecedente de cáncer de la misma. El diagnóstico del tumor pancreático debe ser efectuado por anatomía patológica. Este es un caso atípico en su forma de presentación, dada la aparición de colestasis debida a un tumor metastático a nivel de cabeza de páncreas sincrónico con un carcinoma lobular de mama oculto. (Au)

Tumor de páncreas: presentación infrecuente de un carcinoma de mama oculto. / [Pancreatic tumor: an unusual presentation of an occult breast carcinoma]

A case of cholestasis due to a synchronous pancreatic head metastasis from an occult lobular breast carcinoma is presented. The patient had a clinical and radiological picture compatible with a pancreatic head primary tumor with cholestasis and ascites. Ultrasonographically guided fine needle aspiration cytology demonstrated a metastatic breast lobular cancer (positive for cytokeratin AE1 and AE3, cytokeratin 7 and epithelial membrane antigen and negative for cytokeratin 20, CA 19.9, CA 125, CEA and estrogenic receptors). The same cytologic findings were observed in skin and subcutaneous armpit nodules. Clinical and radiological breast examination was unable to demonstrate any tumor in the breast. Pancreatic metastases are rare events and the majority of them are secondary to renal and lung cancer and rarely to breast cancer. In these latter cases, metastases are usually disclosed after a disease-free interval of months or years between primary tumor resection and recognition of the pancreatic tumor. Synchronous presentation is extremely rare. Metastases of epithelial origin are uncommon in pancreas and generally are first misdiagnosed as primary pancreatic cancer. Fine needle aspiration is a useful tool for the differential diagnosis in patients with widespread disease.