Fractional excretion of electrolytes and neutrophil gelatinase-associated lipocalin as early biomarkers of acute kidney injury in dogs with acute pancreatitis.

OBJECTIVES: To determine if fractional excretion of urinary electrolytes and neutrophil gelatinase-associated lipocalin could detect different degrees of kidney injury in dogs with naturally occurring acute pancreatitis. MATERIALS AND METHODS: We included dogs with acute pancreatitis. Dogs with a history of kidney disease, urinary tract infection, dogs which received potentially nephrotoxic drugs and dogs managed with haemodialysis were excluded. Acute kidney injury was diagnosed if there was an acute onset of clinical signs, haemato-chemical results compatible with acute kidney injury. Students or staff-owned dogs were selected to build the healthy group. RESULTS: The study population was composed of 53 dogs: acute pancreatitis with AKI (n=15), acute pancreatitis alone (n=23), and healthy dogs (n=15). In dogs with acute pancreatitis and AKI, all the FEs of urine electrolytes were significantly higher than dogs with acute pancreatitis alone and healthy ones. Dogs with acute pancreatitis alone had higher uNGAL/uCr than healthy dogs (median 54 ng/mg vs. 0.1 ng/mg) and lower compared to AP-AKI patients (54 ng/mg vs 209 ng/mg). CLINICAL SIGNIFICANCE: Some fractional excretion of electrolytes are increased acute kidney injury dogs, however their role in the early detection of renal injury in acute pancreatitis dogs remains doubtful. On the contrary, urinary neutrophil gelatinase-associated lipocalin had higher concentrations in dogs with acute pancreatitis with or without acute kidney injury compared to healthy controls, suggesting that it may be used as an early marker of renal tubular damage in acute pancreatitis dogs.

Comparison of clinicopathological patterns of renal tubular damage in dogs with acute kidney injury caused by leptospirosis and other aetiologies.

In humans, leptospiral acute kidney injury (AKI) is characterised by tubulointerstitial involvement and renal electrolyte losses, impacting clinical presentation and case management. The aim of this study was to evaluate urine chemistry findings in dogs with leptospirosis in order to identify characteristic patterns of tubular damage associated with this disease. Dogs with intrinsic AKI caused by leptospirosis and by other aetiologies were prospectively enrolled. Clinical and clinicopathological variables, including serum and urine chemistry, fractional excretion (FE%) of electrolytes, and urinary neutrophil gelatinase-associated lipocalin (NGAL), were evaluated in both groups and compared statistically. Dogs with leptospirosis (n = 38) had significantly higher serum creatinine concentration than dogs with AKI caused by other aetiologies (n = 37). Serum potassium and glucose concentrations were comparable between groups. Dogs with leptospiral AKI had significantly higher FE of potassium (median 100%, range 20-480 vs. median 68%, range 5-300; P = 0.048), as well as higher magnitude of glucosuria (urine glucose to creatinine ratio, median 0.64, range 0-26 vs. median 0.22, range 0-13; P = 0.023) and frequency of positive glucose dipstick reaction (59% vs. 18%; P = 0.002), than dogs with AKI of other aetiologies. Additional markers of tubular damage considered in this study, including FE of other electrolytes and urinary NGAL, did not differ between groups. In conclusion, when compared to other aetiologies of intrinsic AKI, canine leptospirosis was characterised by increased glucosuria and kaliuresis.

The effect of chronic kidney disease on the urine proteome in the domestic cat (Felis catus).

Chronic kidney disease (CKD) is a major cause of mortality in cats, but sensitive and specific biomarkers for early prediction and monitoring of CKD are currently lacking. The present study aimed to apply proteomic techniques to map the urine proteome of the healthy cat and compare it with the proteome of cats with CKD. Urine samples were collected by cystocentesis from 23 healthy young cats and 17 cats with CKD. One-dimensional sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (1D-SDS-PAGE) was conducted on 4-12% gels. Two-dimensional electrophoresis (2DE) was applied to pooled urine samples from healthy cats (n = 4) and cats with CKD (n = 4), respectively. Sixteen protein bands and 36 spots were cut, trypsin-digested and identified by mass spectrometry. 1D-SDS-PAGE yielded an overall view of the protein profile and the separation of 32 ± 6 protein bands in the urine of healthy cats, while CKD cats showed significantly fewer bands (P < 0.01). 2-DE was essential in fractionation of the complex urine proteome, producing a reference map that included 20 proteins. Cauxin was the most abundant protein in urine of healthy cats. Several protease inhibitors and transport proteins that derive from plasma were also identified, including alpha-2-macroglobulin, albumin, transferrin, haemopexin and haptoglobin. There was differential expression of 27 spots between healthy and CKD samples (P < 0.05) and 13 proteins were unambiguously identified. In particular, increased expression of retinol-binding protein, cystatin M and apolipoprotein-H associated with decreased expression of uromodulin and cauxin confirmed tubular damage in CKD cats suggesting that these proteins are candidate biomarkers.

New horizon in dialysis depuration: Characterization of a polysulfone membrane able to break the 'albumin wall'.

The uremic syndrome is attributed to the progressive retention of a large number of toxins, which under normal conditions are excreted by the healthy kidneys. Standard dialytic membranes do not purify middle-high molecular weight toxins. Haemodiafiltration with endogenous reinfusion coupled with a highly permeable membrane could break the limit of the 'albumin wall' improving the dialytic depuration without loss of important nutrients. The aim of this study was to evaluate the performance of a new polysulfone membrane, Synclear 0.2, to remove uremic molecules. Surface Enhanced Laser Desorption Ionization-Time of Flight was employed to evaluate the proteomic profile of ultrafiltrate and Electrospray Ionization-Quadruple-ToF coupled with on-chip elution was used for proteins identification. A high and specific permeability for middle-high molecular weight molecules was revealed by mass spectrometry for the investigated membrane. The identified proteins are mostly uremic toxins: their relative abundance, estimated in the ultrafiltrate by exponentially modified protein abundance index, showed a high purification efficiency of the new membrane when compared with conventional ones. In conclusion, Synclear 0.2, used as convective membrane in hemodiafiltration with endogenous reinfusion treatment, permits to break the 'albumin wall', clearing middle-high molecular weight uremic toxins, improving the dialytic treatment purification efficiency.

Nitinol Carbofilm coated stents for peripheral applications: study in the porcine model.

PURPOSE: Testing the safety and foreign body reaction (FBR) of a nitinol self-expandable carbon-coated stent system in the porcine animal model at different follow-up (FU) periods. METHODS: Fifteen minipigs received 30 carbon-coated self-expandable nitinol stents in iliac arteries. Explants were carried out at 7 (3 animals), 30 (4 animals), 90 (4 animals) and 180 (4 animals) day FU, for evaluation of acute, sub-acute and chronic biological response to the implanted devices. Histological, immunohistochemical, histomorphometric and scanning electron microscopy (SEM) analyses were performed to assess inflammatory reaction, endothelialization process, neointimal growth and cellular composition. RESULTS: Thirty stents were successfully implanted. No mural thrombi were observed at gross examination or by angiography. Histologically no significant inflammatory reaction was detected: the stents appeared covered by a thin monolayer of endothelial cells even at 7 day FU. The neointima presented homogeneous growth and moderate thickness after 30, 90 and 180 days explants (0.38+/- 0.36 mm, 0.33+/- 0.30 mm, 0.27+/- 0.25 mm respectively). Internal and external elastic laminae were intact in 95% of stented arteries. Histological data validations of vessel endothelialization was obtained with SEM for the seven day follow-up group. CONCLUSIONS: This study showed good remarkable technical performances, minimal FBR and biocompatibility comparable with other available pre-clinical experimentations.

Detection of micro- and nano-sized biocompatible particles in the blood.

The research deals with new scanning electron microscopic evaluations of the interface between blood and explanted temporary vena cava filters from patients affected by blood disorders. The biological tissues adherent to the filter and the small thrombi formed in vivo were detached from the metallic structure of the device, fixed, dehydrated and prepared for the histological and the electron microscopy. The analyses showed that both samples (thrombus and newly formed tissue) contained foreign, in some cases nano-sized, bodies. The chemistry of these particles was different and varied, and unusual compounds containing non-biocompatible elements like bismuth, lead, wolfram, tungsten were also detected. The interaction between these debris travelling in the blood stream and the blood itself leads to suspect that the formation of the thrombus can originate from these inorganic and inert foreign bodies that act as triggering agent of the blood coagulation.

[Evaluation of the biocompatibility of grafts for bone defects]. / Valutazione della biocompatibilità del fisiograft per difetti ossei.

BACKGROUND: This paper examines a new material composed of polylactic and polyglycolic acids mixed with destrane easy to handle in bone defect reconstruction in periodontology and implantology: its name is fisiograft. METHODS: According to the European directives, fisiograft was checked for some biological tests selected for its specific use. In particular, tests of cytotoxicity, genotoxicity, sensitization and implantation in rabbit's femurs were carried out. RESULTS: The results show that the composite material is not cytotoxic when it is in direct and indirect contact with murine fibroblasts. It is not genotoxic and not allergenic. The implantation in rabbit, already after 30 days, shows that around the material there is a rapid bone growth and the material is reabsorbing. The degradation is suitable for its clinical use. CONCLUSIONS: Fisiograft showed to be biocompatible, easy to handle and its degradation kinetics in bone is appropriate.

In vivo evaluation of an experimental dental HA-Ti implant.

A new design of Titanium dental implant, coated with Titanium oxide and with a Hydroxyapatite apex, was experimented in a Beagle dog's jaw for one year to evaluate the biocompatibility of the materials used and the biomechanical efficiency of the design. The explanted prosthesis and the surrounding bone were embedded in methylmethacrylate resin and the sections observed under Scanning Electron Microscopy (SEM), with an X-ray microprobe to check the interface with bone and the occurrence of ionic release. The results showed that the pin was well accepted by the bone. New bone grew on the metal surface developing a mechanical bond with the prosthesis. No ionic release of the metal was seen.

Calcification of chemically treated bovine pericardium.

One of the most important problems arising in cardiac bioprostheses made with bovine pericardium and, more generally, with biologically-derived tissues is tissue calcification. The present study assessed four chemical treatments on patches of bovine pericardium, intended to avoid or minimize calcification. Pericardium specimens were treated with: A) 0.5% glutaraldehyde; B) 0.5% glutaraldehyde + 4% formaldehyde; C) same as A, but with a further neutralization treatment; D) acylation of fresh bovine pericardium. Circular samples of 1 cm diameter were subcutaneously implanted in the abdominal region of three groups of six rats. The explants were retrieved after 2, 4 and 8 weeks. The calcium content and the histological results showed better behaviour for C and D samples than with the commonly used fixation methods (A and B). The lowest calcification was observed with treatment D, even though its morphological structures were somewhat modified with homogenation of collagen bundles. Among the glutaraldehyde-based treatments, treatment C appears to be the most promising because the pericardium shows slower calcium accumulation with a diffusive pattern.

Physical characterization of acrylic bone cement cured with new accelerator systems.

In the attempt to substitute dimethyl-p-toluidine (DMPT), a toxic tertiary aryl-amine accelerator, into the formulation of acrylic cements, less toxic accelerator systems are developed. These systems consist of benzoyl peroxide (BPO) and unsaturated tertiary-aryl-amines, such as acryloyl- (ANP) and methacryloyl-(MNP) N-phenylpiperazine, which can be chemically incorporated in the polymerizing resin or, at least, result in less leaching from cured materials. In this work compressive mechanical properties and ageing tests for colour stability of acrylic cement cured with BPO and ANP or MNP have been considered. For compressive tests, cylindrical specimens were cured with BPO and equivalent molecular amounts of DMPT, ANP and MNP. Compressive yield stress (sigma y), strain at yield (epsilon y, %) and elastic modulus (E) gave very similar results for samples cured with DMPT and ANP, and slightly lower results for samples cured with MNP. In colour stability tests, the samples (disks of 1.5 cm diameter) were exposed to UV light at different irradiation times (up to 42 h). The evaluation of the colour change was performed with a digital analyser for images, and observed under scanning electron microscopy. From the obtained results, ANP appeared to be the best candidate as accelerator in the preparation of biomedical acrylic resins and composites.