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Objective: To evaluate a urine dipstick test as a possible replacement for urine protein-to-creatinine (UPC) ratio for identifying proteinuria in dogs., Sample Population: 507 urine samples from adult dogs., Procedures: Urine dipstick, UPC ratio, specific gravity (USG), and sediment testing were performed on 507 samples. With UPC ratio as the reference criterion, diagnostic accuracy of the urine dipstick test was calculated for the entire data set and for urine samples grouped by USG (< or = 1.012 or > 1.012; < 1.030 or > or = 1.030). A UPC ratio < 0.2 was used to indicate absence of proteinuria., Results: The sensitivity of the urine dipstick test for detection of proteinuria was > 90% when 0 mg of protein/dL (a 0+ result) was used to indicate a negative test result, and the specificity ranged from 40% to 60%, depending on the USG. Sensitivity decreased to a range of 56% to 81% when 30 mg of protein/dL (a 1+ result) was used as the cutoff, depending on the USG, but the specificity increased to > 90%. The likelihood of correctly identifying nonproteinuric dogs was low when the USG was < or = 1.012, particularly when samples with a 1+ result were considered negative., Conclusions and Clinical Relevance: For dogs with a dipstick-test result of 1+ and USG < or = 1.012, proteinuria should be assessed by use of the UPC ratio; dogs with a USG value > 1.012 are likely nonproteinuric. When used together, the urine dipstick test and USG measurement were reliable as a rapid alternative to UPC ratio determination in dogs in this study.

Isolation of pancreatic islets is necessary to study the molecular mechanisms underlying beta-cell demise in diabetic cats. Six collagenase-based methods of isolation were compared in 10 cat pancreata, including single and double course of collagenase, followed or not by Ficoll centrifugation or accutase, and collagenase plus accutase. Morphometric analysis was performed to measure the relative area of islet and exocrine tissue. Islet specific mRNA transcripts were quantified in isolates by real-time PCR. The single and double course of collagenase digestion was successful in each cat and provided similar islet-to-exocrine tissue ratio. Quantities of insulin mRNA did not differ between the two methods. However, on histological examination either method yielded only approximately 2% of pure islets. The other methods provided disrupted islets or insufficient samples in 1-7 cats. Although pancreas digestion with single and double course of collagenase was superior, further studies are needed to improve islet isolation in cats.

Objective: To describe clinical characteristics, treatment, and outcome of dogs with inflammatory carcinoma (IC) and identify patient-, tumor-, and treatment-related factors associated with overall survival time., Design: Retrospective case series., Animals: 43 client-owned dogs., Procedures: Records of dogs with a clinical diagnosis of IC that had histologic evidence of dermal lymphatic invasion were reviewed. Data on clinical staging, treatment, toxicoses, response, and survival time were retrieved. Results-26 (60%) dogs had primary IC and 17 (40%) had secondary IC. Thirty-five (81%) dogs had distant metastases and 2 (5%) had local metastases at the time of initial examination. Six of 29 (21%) dogs had a coagulopathy. Sixteen (37%) dogs did not receive specific treatment for IC, 24 (56%) received medical treatment only, 2 (5%) underwent surgical excision and received medical treatment, and 1 (2%) underwent surgical excision only. Forty-one (95%) dogs had progressive disease, and 2 (5%) had stable disease. Mean survival time for all dogs was 60 days (range, 1 to 300 days). Dogs with a coagulopathy survived a significantly shorter time than did dogs without a coagulopathy (odds ratio, 0.28), and dogs that received medical treatment survived significantly longer than dogs that did not (odds ratio, 2.54)., Conclusions and Clinical Relevance: Results suggested that mammary IC is a biologically aggressive condition in dogs associated with a guarded prognosis. In addition, results suggested that medical treatment may improve outcome, thereby supporting its use in dogs with IC.

We report a 20-year-old male patient with an atypical Spitz tumor, located in the dorsal aspect of his left hand, and a positive sentinel axillary lymph node. After lymphadenectomy, 1 of 23 non-sentinel lymph nodes excised was found to contain small multiple deposits of large spindle atypical melanocytes. Reviewing the pertinent literature, 5 of 29 patients with atypical Spitz tumors and positive sentinel nodes who had undergone lymphadenectomy have shown non-sentinel node involvement (17.2%), a proportion similar to that reported in melanoma patients. The exact nature of atypical Spitz tumors and the interpretation of cell deposits detected in sentinel nodes are still debated; data regarding the non-sentinel lymph node involvement in patients with atypical Spitz tumors may contribute to better understand the real biological potential of such tumors.

Impaired insulin sensitivity is increasingly recognised in cats, but sequences of genes involved in insulin-signalling are largely undetermined in this species. In this study, extended feline mRNA sequences were determined for the adiponectin, glucose transporter-1 (GLUT1), GLUT4, peroxisome proliferative activated receptor-gamma1 (PPARgamma1), PPARgamma2, plasminogen activator inhibitor-1 (PAI-1), monocyte chemoattractant protein-1 (MCP-1) and insulin receptor genes. Conserved dog-specific primers identified from human-dog mRNA alignments were used to amplify feline cDNA in the polymerase chain reaction (PCR). The feline sequences determined by this method were used to design feline-specific primers suitable for real-time PCR for quantification of gene expression in insulin sensitive tissues of healthy cats. Partial sequences of feline mRNAs had 86-95% identity with dog and human genes. Expression of adiponectin, GLUT1, GLUT4, PPARgamma1, PPARgamma2, PAI-1 and insulin receptor mRNA was detected and quantified in subcutaneous and visceral fat and skeletal muscle, whereas MCP-1 mRNA was detected in adipose tissue but not in skeletal muscle. Further characterisation of genes related to glucose metabolism in cats will provide additional insights into insulin-signalling mechanisms in this species.

Aims/hypothesis: In vitro studies point to a toxic effect of high glucose and non-esterified fatty acids on beta cells. Whether elevated levels of glucose and lipids induce beta cell loss in vivo is less clear. The domestic cat has recently been proposed as a valuable animal model for human type 2 diabetes because feline diabetes shows several similarities with diabetes in humans, including obesity-induced insulin resistance, impaired beta cell function, decreased number of beta cells and pancreatic amyloid deposition., Methods: We infused healthy cats with glucose or lipids for 10 days to clamp their blood concentrations at the approximate level found in untreated feline diabetes (glucose: 25-30 mmol/l; triacylglycerols: 3-7 mmol/l)., Results: Glucose and lipid levels were adequately targeted. Plasma non-esterified fatty acids were increased by lipid infusion 1.7-fold. A dramatic and progressive decline of plasma insulin levels was observed in glucose-infused cats beginning after 2 days of hyperglycaemic clamp. In contrast, plasma insulin concentration and glucose tolerance test were not affected by hyperlipidaemia. Compared with controls, glucose-infused cats had a 50% decrease in beta cells per pancreatic area. Apoptotic islet cells and cleaved caspase-3-positive beta cells were observed in glucose-infused cats only., Conclusions/interpretation: Sustained hyperglycaemia but not hyperlipidaemia induces early and severe beta cell dysfunction in cats, and excess glucose causes beta cell loss via apoptosis in vivo. Hyperglycaemic clamps in cats may provide a good model to study the pathogenesis of glucose toxicity in beta cells.

Many viruses have been identified in pericardial fluid and in tissue samples from humans with pericarditis by means of molecular diagnostics. In canine idiopathic pericardial effusion there is as yet no conclusive evidence to support the involvement of an infectious agent. This study was designed to investigate a possible relationship between idiopathic pericardial effusion in dogs and viruses most commonly encountered in humans affected with viral pericarditis. Coxsackievirus B3 RNA, influenza virus type A RNA, human adenovirus type 2 DNA, human cytomegalovirus DNA, and parvovirus B19 DNA were investigated using PCR on pericardial effusion samples and pericardial tissue specimens collected from 14 dogs with idiopathic pericardial effusion. PCR was also used to test for two bacteria, Borrelia burgdorferi and Chlamydia pneumoniae. The same microorganisms were also looked for in pericardial effusions or pericardial washes from 10 dogs with neoplastic pericardial effusion, and in samples collected from 10 dogs which died of a non-cardiac disease. One pericardial effusion sample from a dog with the idiopathic form of the disease tested positive for influenza virus type A and sequencing of the amplicon confirmed the PCR result. In another dog from the same group a cytomegalovirus was detected by PCR in the effusion, but sequencing showed this to be a false-positive result. The genomes of the microorganisms investigated were not detected in neoplastic effusions or pericardial washes. The results indicate that viral and bacterial DNA/RNA of relevance for human pericarditis is rare in pericardial samples from dogs with idiopathic pericardial effusion. The finding of influenza type A viral RNA in pericardial fluid from one dog with the idiopathic form of the disease warrants further investigation.

The efficacy of the histological criteria currently used in the diagnosis of melanoma is still to be defined. We performed a quantitative analysis of 72 conventional (non-Spitzoid, nondesmoplastic) melanomas and 73 conventional melanocytic naevi, used as controls, for 13 histological diagnostic parameters (dimension >6 mm, asymmetry, poor circumscription, irregular and confluent nests, single melanocytes predominating, absence of maturation, suprabasal melanocytes, asymmetrical melanin, melanin in deep cells, cytological atypia, mitoses, necrosis and dermal lymphocytic infiltrate). Differences in the distribution of selected parameters between the two groups were investigated by Fisher's exact test; for each parameter sensitivity and specificity were calculated. Results showed that all parameters, except poor circumscription, seemed to be significantly associated with melanoma (P<0.05). Cytological atypia, dermal lymphocytic infiltrate, asymmetry, dimension >6 mm and absence of maturation showed a high sensitivity (>90%); absence of maturation, mitoses, necrosis, asymmetrical melanin, suprabasal melanocytes and melanin in deep cells showed a high specificity (>90%); irregular-confluent nests and single melanocytes predominating were poorly sensitive and poorly specific. In melanomas < or =2 mm, two additional parameters were sensitive (> or =90%): suprabasal melanocytes and single melanocytes predominating. We conclude that not all parameters showed to have the same diagnostic value. Absence of maturation and, limited to melanomas < or =2 mm, suprabasal melanocytes were the most discriminating (sensitive and specific) histological features. Cytological atypia, dimension >6 mm, suprabasal melanocytes and mitoses were additional reliable diagnostic features, showing a relatively high sensitivity and a relatively high specificity. Other useful features were dermal lymphocytic infiltrate and asymmetry (sensitive) and necrosis, asymmetrical melanin and melanin in deep cells (specific).

Purpose: To evaluate the tolerability and safety of a newly designed probe for trans-perineal microwave thermoablation (TPMT) of the prostate in patients with benign prostatic hyperplasia (BPH), and the in vivo microwave effects on prostatic tissue., Patients and Methods: Nine patients with obstructive BPH who were candidates for open prostatectomy were selected for this study. Under local anesthesia and transrectal ultrasound monitoring, all patients underwent a single standardized application of TPMT. The visual analog scale (VAS) and Short Form-36 health survey (SF-36) questionnaire were administered to each patient prior to, during, and 1 month after TPMT in order to evaluate pain and quality of life. Then the International Index of Erectile Function (IIEF-5) and International Prostate Symptom Score (IPSS) questionnaires were administered to each patient at baseline and 1 month after prostatectomy in order to evaluate sexual and urinary function, respectively. Then all patients were divided into three groups and underwent open prostatectomy 7, 15, and 30 days after TPMT, respectively. The prostatic adenomas were then evaluated by a pathologist., Results: No adverse events from TPMT treatment were noted. In particular, no patients reported local, pelvic, or abdominal pain during the procedure or subsequent alterations of defecation rhythm, ano-rectal/intestinal problems, or hematuria. No differences in quality of life or in sexual function were reported. The diameters of the lesions obtained with TPMT treatment ranged from 16 to 18.1 mm in all patients. Quasi-spheroid lesions with a well-defined area of complete coagulative necrosis were documented in all removed adenomas 7, 15, and 30 days after TMPT., Conclusions: The AMICA-PROBE is a safe, well-tolerated, and repeatable method to treat BPH with microwave thermotherapy. The spheroid lesions obtained demonstrated the maximal control over the radial and longitudinal coagulative effects of the therapy. Phase II studies are needed to further evaluate the efficacy of this new probe.

Purpose: We evaluated the impact of epirubicin perioperative instillation in improving subsequent bacillus Calmette-Guerin instillation efficacy in high risk patients with nonmuscle invasive bladder cancer., Materials and Methods: Between January 2005 and June 2007, 161 patients affected by high risk nonmuscle invasive bladder cancer were enrolled in this prospective, randomized, controlled, double-blind study. A total of 80 patients were assigned to group A (perioperative epirubicin 80 mg/50 ml normal saline) plus delayed bacillus Calmette-Guerin instillations (5 x 108 colony-forming units in 50 ml saline) and 81 to group B (delayed bacillus Calmette-Guerin alone). The main outcome measures were time to first recurrence and recurrence rate. All data obtained from a median followup of 15.3 months in group A and 14.8 months in group B, were analyzed., Results: At the end of followup 46 of 80 patients in group A (57.5%) had no evidence of disease, just like 41 of 81 in group B (50.6%). No statistical difference was observed between the 2 groups in terms of recurrence rate (p = 0.82) or time to first recurrence (p = 0.095). Kaplan-Meier analysis of recurrence showed no significant differences between group A and group B (p = 0.0952). On multivariate analysis the early single dose instillation of epirubicin was not indicated as an independent prognostic factor (HR 0.50, 95% CI 0.32-1.18)., Conclusions: The present study showed no statistically significant differences in terms of disease-free time and recurrence rate between high risk patients with nonmuscle invasive bladder cancer who had undergone perioperative epirubicin instillation plus delayed bacillus Calmette-Guerin and those who had undergone delayed bacillus Calmette-Guerin alone.

Objective: To identify prognostic factors in cats with injection-site sarcomas (ISSs)., Design: Retrospective case series., Animals: 57 cats with ISSs., Procedures: Medical records of cats were reviewed with regard to sex, age, anatomic site of tumor, tumor size, histologic grade, excision of a primary tumor versus excision of a recurrent ISS, use of excision alone versus excision plus adjuvant therapy, local tumor recurrence, and development of distant metastasis to predict overall survival time (ie, time from tumor excision to death)., Results: In univariate analyses, local recurrence and development of distant metastasis were significantly associated with survival time in cats. On multivariate analysis, development of distant metastasis remained a significant prognostic factor. Histologic grade was associated with distant metastasis, with cats having grade 3 tumors being significantly more likely to develop metastasis than cats with grade 1 and 2 tumors. Factors associated with local recurrence of ISSs were not identified., Conclusions and Clinical Relevance: The development of distant metastasis, which may occur later during the course of the disease, was identified as a prognostic factor for overall survival time in cats with ISSs. In addition, cats with histologic grade 3 ISSs should be considered for further interventional studies with chemotherapy to prevent the high rate of distant metastasis.

Purpose: Several potential markers have been investigated to improve the noninvasive diagnosis of recurrent superficial bladder carcinoma. We evaluated the role of the interleukin-6/10 ratio as a prognostic marker of recurrence in patients with intermediate risk superficial bladder carcinoma., Materials and Methods: A total of 65 consecutive urological patients seen in the office, including 41 with intermediate risk superficial bladder carcinoma and 24 controls, were selected for this prospective study. Five urine samples for urinary cytology and interleukin analyses were collected from each subject at baseline, and 3, 6, 9 and 12 months after surgery, respectively. Interleukin-6 and 10 were determined in urine by the Quantikine solid phase interleukin-6 and 10 enzyme-linked immunosorbent assay, respectively. Sensitivity, specificity, and positive and negative predictive values of the method were calculated., Results: At baseline sample collection the interleukin-6/10 ratio was not statistically different between patients and controls (p = 0.58). Interleukin-6/10 was statistically different between patients with vs without recurrence 3 (0.009 vs 0.408), 6 (0.011 vs 0.268), 9 (0.012 vs 0.288) and 12 months (0.009 vs 0.302) after pre-transurethral bladder tumor resection (each p <0.001). Multivariate analysis indicated that interleukin-6/10 was an independent prognostic factor of recurrence (HR 3.62, 95% CI 2.80-4.92, p <0.001). Test sensitivity and specificity were 0.83% (95% CI 0.57-0.95) and 0.76% (95% CI 0.45-0.93), respectively., Conclusions: The current study highlights the feasible role of the interleukin-6/10 ratio for predicting intermediate risk superficial bladder carcinoma recurrence. However, clinical trials with a greater number of patients are needed to consider its use in clinical urological practice.

Bioresorbable new terpolymers of L-lactide, glycolide, and trimethylene carbonate with different compositions were synthesized via ring-opening polymerization reaction of the cyclic monomers using low-toxicity zirconium(IV) acetylacetonate as initiator. The thermal and mechanical properties were investigated by means of thermogravimetry, differential scanning calorimetry, stress-strain measurements, and dynamical mechanical analysis. The glass transition temperature of the terpolymers changes with composition from 12 to 42 degrees C in a predictable manner. All terpolymers display shape memory properties and, after undergoing 100% deformation, they recover the permanent shape in a time frame of seconds. Terpolymers with high L-lactide content show a glass transition in the range of 38-42 degrees C, recovery temperature close to body temperature, and good recovery ratio (>0.89). Low-toxicity bioresorbable terpolymers with shape memory properties are promising new materials for biomedical applications.

A 13-year-old Siamese cat was presented for investigation of lethargy and progressive abdominal enlargement. Serum chemistry revealed severe reduction of total and ionised serum calcium. The omentum appeared hyperechoic with scattered hypoechoic foci on abdominal ultrasound examination. Elevated serum parathormone and low fractional excretion of calcium excluded a parathyroid disorder and renal loss of the electrolyte. During laparotomy the omentum appeared opaque, white and firm. Post-mortem examination revealed that the thoracic and subcutaneous fat was also affected. Histopathology confirmed the diagnosis of pansteatitis with diffuse calcium soaps formation. While, severe hypocalcaemia is occasionally seen in cats, the association with pansteatitis has not been reported previously. In man, a cause-and-effect relationship between calcium soaps and hypocalcaemia is recognised, though the association is rare.

Case Description: A 6-month-old domestic shorthair cat was evaluated because of acute lethargy., Clinical Findings: Severe nonregenerative anemia and thrombocytopenia were identified. Cytologic examination of a bone marrow aspirate revealed selective erythroid and mega-karyocytic aplasia and a high number of apparently normal small lymphocytes. Infectious agents implicated in feline hematologic disorders were excluded on the basis of serologic tests or PCR amplification, including FeLV, Ehrlichia canis, Anaplasma phagocytophilum, Mycoplasma haemofelis, Candidatus Mycoplasma haemominutum, and Candidatus Myco-plasma turicensis., Treatment and Outcome: A 10-day course of prednisolone administration did not improve the hematologic disorder. Administration of human polyclonal immunoglobulins preceded increased reticulocyte count by 3 days. A second bone marrow examination confirmed restoration of erythroblasts and megakaryocytes. After 1 relapse, the disease was successfully controlled with prednisolone for > 3 years., Clinical Relevance: Immune-mediated bone marrow aplasia is rare in cats and usually affects only erythrocyte progenitors. Concomitant involvement of erythroid and megakaryocytic cell lines can be successfully treated via immunosuppressive therapy. Human immunoglobulins seem to be well tolerated in cats; however, proof of a beneficial effect requires further study.

Twenty-two dogs and cats with symptomatic renal or hepatic cysts that had undergone ultrasound-assisted drainage and alcoholization were retrospectively evaluated. Common presenting complaints were anorexia, reluctance to move, and vomiting. Abdominal pain was observed in all cases. Systemic hypertension was identified in four dogs and four cats with renal cysts. Cyst drainage and alcoholization were achieved without complications in 19 animals, and all clinical signs resolved after the procedure. In three cases, transient bleeding was observed during alcoholization, and the procedure was interrupted. Blood pressure normalized in the four dogs with renal cysts, but it remained elevated in the four cats.

Objective: To determine results of cytologic examination of fine-needle aspirates and impression smears of gastrointestinal tract tumors in dogs and cats., Design: Retrospective case series., Animals: 38 dogs and 44 cats with histologically confirmed gastrointestinal tract tumors., Procedures: Results of cytologic examination of fine-needle aspirates (n = 67) or impression smears (31) were compared with the histologic diagnosis, and extent of agreement was classified as complete, partial, none, or undetermined., Results: For 48 of the 67 (72%) fine-needle aspirates, there was complete or partial agreement between the cytologic and histologic diagnoses. For 12 (18%) aspirates, the extent of agreement could not be determined because the cytologic specimen was considered unsatisfactory. For 29 of the 31 (94%) impression smears, there was complete agreement between the cytologic and histologic diagnoses, and for 2 (6%), there was partial agreement. None of the impression smears were considered unsatisfactory. Proportion of samples with complete agreement and proportion of samples with complete or partial agreement were significantly higher for impression smears than for fine-needle aspirates., Conclusions and Clinical Relevance: Results suggest that there was moderate agreement between results of cytologic examination of fine-needle aspirates from dogs and cats with gastrointestinal tract neoplasia and the definitive histologic diagnosis. The agreement between results of cytologic examination of impression smears and the histologic diagnosis appeared to be higher.

Objectives: Host genetic factors, including the IL1 gene cluster, play a key role in determining the long-term outcome of Helicobacter pylori infection. The aim of the study was to investigate the relationship between selected IL1 loci polymorphisms and gastric cancer risk in an Italian population., Methods: In a case-control study we compared the IL1B-31 and IL1B+3954 biallelic and IL1RN pentaallelic variable number of tandem repeats (VNTR) polymorphisms in 185 gastric cancer patients and 546 controls randomly sampled from the general population of an area at high gastric cancer risk (Tuscany, Central Italy)., Results: Genotype frequencies of the IL1B-31 T/C, IL1B+3954 C/T, and IL1RN polymorphisms among our population controls were in Hardy-Weinberg equilibrium. In multivariate analyses, no increase in gastric cancer risk was observed for the IL1B-31*C- and IL1B+3954*T- carriers; a significant 50% increase emerged for IL1RN*2 allele carriers (OR = 1.49; 95% CI: 1.01-2.21). Analyses based on combined genotypes showed also that the association with IL1RN*2 allele was limited to two-variant allele carriers who were also homozygous for the IL1B-31*T allele (OR = 2.23; 95% CI: 1.18-4.23) with a statistically significant interaction between these two genotypes (p= 0.043). Haplotype analysis showed an increased risk for the haplotype IL1RN*2/IL1B-31*T., Conclusions: Our results suggest that host genetic factors (such as the IL1RN and the IL1B-31 polymorphisms) interact in the complex process of gastric carcinogenesis in this high-risk Italian population. Overall, this effect appears more modest than previously reported in other populations, supporting the hypothesis that other still-to-be-defined factors are important in gastric carcinogenesis. These findings might be due to a haplotype effect.

Our objective was to evaluate possible differences in the number of glomeruli and length of renal biopsies collected in canine subjects by two different types of biopsy needles: a semiautomatic 18-gauge Trucut and an automated 18-gauge Jamshidi modified (Biopince). One hundred biopsy samples obtained from dogs of different ages and gender affected by different nephropathies were evaluated retrospectively. All animals were biopsied using one of the two different needles. Biopsies were performed under ultrasound guidance and evaluated by a single pathologist. Statistical analysis was performed to evaluate possible differences in the number of glomeruli and length of renal biopsies collected comparisons were determined between subgroups of dogs with or without the identification of renal interstitial infiltrates and/or fibrosis. Neither the mean difference of the number of glomeruli nor the length of tissue sample collected with the different needles was significantly different. Likewise, the average biopsy length did not differ in dogs with or without renal interstitial infiltrate in animals biopsied with either biopsy needle. Both the Biopince and the Trucut devices provide diagnostically adequate biopsy renal specimens using ultrasound-guidance.

Hepatic abscesses are rare and difficult to diagnose in dogs and cats. Ultrasonographic examination is essential to thoroughly examine hepatic abnormalities, and it may also help in the localization of hepatic abscesses. In this retrospective study, five dogs and one cat with focal hepatic lesions compatible with a hepatic abscess were treated with percutaneous ultrasound-assisted drainage and alcoholization using 95% ethanol. The procedure was performed rapidly under injectable anesthesia and provided excellent results in all animals. No complications or relapses were noted for 120 days following the procedure. The technique of percutaneous ultrasound-assisted drainage and alcoholization of hepatic abscesses in the dog and cat is also described.

The case records of 58 German Shepherds (GS group) affected by mitral valve prolapse (MVP) and/or mitral valve regurgitation (MR), and 49 dogs weighing < 15 kg (D group), affected by chronic valvular disease (CVD) were reviewed. The dogs of the GS group were presented more often without a detectable heart murmur (p < 0.01), and less frequently with a high intensity heart murmur (p < 0.01). Atrial fibrillation (AF) was more common in the GS group (p < 0.001). MVP associated with mitral valve thickening was more common in the D group (p < 0.001). Fractional shortening (FS) was lower (p < 0.0001) and end-systolic volume index (ESV-I) was increased (p < 0.0001) in the GS group, whereas end-diastolic volume index (EDV-I) did not differ between the 2 groups. Prevalence and severity of pulmonary hypertension were similar in the 2 groups. Dogs with mitral valve disease weighing more than 20 kg had a 5.8 higher chance of developing decreased FS, increased ESV-I, AF and ventricular arrhythmias. In the GS group, the decreased FS and increased ESV-I were not associated with the presence of AF or ventricular arrhythmias (p > 0.05). It appears that GS may be affected both by mitral valve prolapse and mitral insufficiency. It also appears that a comparatively large proportion of GS shows no major mitral valve thickening or MVP, but still presents with significant mitral regurgitation, possibly suggesting a different cause for the important incompetence observed in most cases.

The purpose of this retrospective study was to investigate the relationship among proteinuria consisting of immunoglobulin free light chains (FLCs), renal histopathologic findings, and routine markers of renal function in 11 dogs exposed to Leishmania infantum (n = 8), Ehrlichia canis (n = 2), and Babesia canis (n = 1). FLC proteinuria was suspected based on identification of a 22- to 27-kDa band by sodium dodecyl sulfate-agarose gel electrophoresis (SDS-AGE) and later confirmed by immunofixation electrophoresis. SDS-AGE identified an isolated band of 22-27 kDa in 8 dogs, whereas the remaining 3 had a 22- to 27-kDa band and an additional band of 67-72 kDa. The median urine protein-to-urine creatinine ratio was 0.37 (range, 0.11-2.24) and increased ratios were found in 6 dogs (54.5%) (reference value, <0.7). All dogs underwent histologic examination of renal percutaneous biopsy specimens and determination of serum creatinine and urea concentrations. Tissue samples for light microscopy were stained with hematoxylin-eosin, periodic acid-Schiff, Goldners trichrome, and methenamine silver. In the study group, the glomerular tufts, mesangium, tubulointerstitium, and vessels appeared unaffected. The median serum creatinine concentration in these 11 dogs was 1.3 mg/dL (range, 0.8-1.5 mg/dL; reference range, 0.6-1.5 mg/dL), whereas the concentration for urea was 28 mg/dL (range, 22-52 mg/dL; reference range, 20-50 mg/dL). All dogs had normal renal morphology and had normal serum creatinine and urea concentrations, suggesting that immunoglobulin FLC may be detected in the urine of dogs exposed to L. infantum, E. canis, and B. canis without any apparent structural or functional renal derangement.

Objective: To evaluate results of SDS-agarose gel electrophoresis (AGE) of urinary proteins for use in defining glomerular and tubulointerstitial derangements, investigate patterns of high-molecular-weight (HMW) proteins for differentiating among glomerular disorders, and assess low-molecular-weight (LMW) proteins as markers of severity of tubulointerstitial disease in dogs., Animals: 49 dogs with increased serum creatinine concentrations or abnormal renal protein loss., Procedure: Urinary proteins were examined by use of SDS-AGE and differentiated on the basis of molecular weight. The HMW proteins (> or = 69 kd) were considered indicative of glomerular origin, whereas LMW proteins (< 69 kd) were of tubular origin. Renal specimens were examined by use of light microscopy. Glomerular and tubulointerstitial lesions were differentiated by use of the classification for the World Health Organization and semiquantitative grading, respectively., Results: Sensitivity of SDS-AGE was 100% for detection of glomerular lesions and 92.6% for tubulointerstitial lesions; specificity was 40% and 62.5%, respectively. Although HMW urinary proteins were not significantly associated with the type of glomerular lesion, LMW urinary proteins were significantly associated with the grade of tubulointerstitial damage. Detection of 12- or 15-kd proteins or both was highly indicative of a severe tubulointerstitial lesion., Conclusions and Clinical Relevance: SDS-AGE of urinary proteins in dogs represents a noninvasive test with high sensitivity for identifying glomerular and tubulointerstitial damage, but low specificity limits its validity as a stand-alone test to differentiate between glomerular and tubulointerstitial lesions. The test is particularly useful for identifying dogs with advanced tubulointerstitial disease but cannot be used to characterize glomerular disorders.

Apple ( Malus x domestica Borkh.) sequences sharing homology with known resistance genes were cloned using a PCR-based approach with degenerate oligonucleotide primers designed on conserved regions of the nucleotide-binding site (NBS). Sequence analysis of the amplified fragments indicated the presence of at least 27 families of NBS-containing genes in apple, each composed of several very similar or nearly identical sequences. The NBS-leucine-rich repeat homologues appeared to include members of the two major groups that have been described in dicot plants: one possessing a toll-interleukin receptor element and one lacking such a domain. Genetic mapping of the cloned sequences was achieved through the development of CAPS and SSCP markers using a segregating population of a cross between the two apple cultivars Fiesta and Discovery. Several of the apple resistance gene homologues mapped in the vicinity, or at least on the same linkage group, of known loci controlling resistance to various pathogens. The utility of resistance gene-homologue sequences as molecular markers for breeding purposes and for gene cloning is discussed.

The immunophenotype of peripheral blood lymphocytes was investigated in 23 dogs diagnosed with idiopathic pericardial effusion in order to provide information about a possible role of the immune system in this pathology. Flow cytometric analysis showed a significant reduction in nearly all lymphocyte subsets examined and a strong, significant (P < 0.001) reduction of the CD4 subset, which gave rise to a significantly lower CD4/CD8 ratio. Our data suggest that an imbalance in the immune system is present during the course of the disease, preferentially affecting the T helper cell response.

Flax fibers are investigated as reinforcing agents for biodegradable polyesters (Bionolle and poly(lactic acid) plasticized with 15 wt.-% of acetyltributyl citrate, p-PLLA). The composites are obtained either by high temperature compression molding fiber mats sandwiched between polymer films, or by batch mixing fibers with the molten polymer. Fibers in composites obtained by the latter method are much shorter (140-200 microm) than those of the mats (5,000 microm). Flax fibers are found to reinforce both p-PLLA and Bionolle (i.e. tensile modulus and strength increase) when composites based on fiber mats are investigated. Conversely, analogous composites obtained by batch mixing show poor mechanical properties. The observed behavior is attributed to the combined effect of fiber length and fiber-matrix adhesion. If flax fibers with a modified surface chemistry are used, the strength of short fiber composites is seen to improve significantly because the interface strengthens and load is more efficiently transferred. Appropriate surface modifications are performed by heterogeneous acylation reactions or by grafting poly(ethylene glycol) chains (PEG, molecular weight 350 and 750). The highest tensile strength of p-PLLA composites is reached when PEG-grafted flax fibers are used, whereas in the case of Bionolle the best performance is observed with acylated fibers.

In the period 1997-2001, 466 sentinel lymph nodes from 342 lymphatic basins in 322 melanoma patients were examined at the Health Unit of Florence. The lymphatic mapping was performed through pre-operative lymphoscintigraphy using technetium-labelled nano-colloid, intradermal injections of vital blue dye and intra-operative gamma-probe. The examined patients were 182 females and 140 males. Sentinel lymph node was one in 65.2% of cases; two sentinel lymph nodes were detected in 27% of cases and more than 2 sentinel nodes were detected in 7.8% of cases. Melanoma metastases in one or more sentinel lymph nodes were found in 61/322 patients (18.9%). Lymphatic basins resulted to be involved by melanoma metastases were 64/342 (18.7%); sentinel lymph nodes containing metastatic melanoma deposits were 73/466 (15.6%). No metastasis was found in patients with melanoma thickness < or = 1 mm. One or more positive sentinel lymph nodes were found in 7.5% of patients with melanoma thickness > 1.00 and < or = 1.50 mm, in 27.7% of patients with melanoma > 1.50 and < or = 3.00 mm, in 38.2% of patients with melanoma > 3.00 and < or = 4.00, and in 60.7% of patients with melanoma > 4.00 mm. Frozen section analysis of sentinel lymph nodes, performed in 59/61 patients with nodal metastases, detected nodal involvement in 21 patients (35.6%). Metastases were identified by routine hematoxylin-eosin staining in 57/64 positive lymphatic basins; in 7 cases (11%) metastases were detected by immunohistochemical stainings (S100 and HMB-45). A nodal nevus was found in 3/466 sentinel lymph nodes (0.6%). Our data are analyzed and compared to previously data of the literature. The value of frozen section analysis and the major problems in the diagnosis of melanoma micrometastases in sentinel lymph nodes are discussed. The importance of the sentinel node biopsy for the detection of occult metastases and for the correct staging of melanoma patients are stressed, according to the new TNM melanoma classification.