Gender and age effects on risk factor-based prediction of coronary artery calcium in symptomatic patients: A Euro-CCAD study.

BACKGROUND AND AIMS: The influence of gender and age on risk factor prediction of coronary artery calcification (CAC) in symptomatic patients is unclear. METHODS: From the European Calcific Coronary Artery Disease (EURO-CCAD) cohort, we retrospectively investigated 6309 symptomatic patients, 62% male, from Denmark, France, Germany, Italy, Spain and USA. All of them underwent risk factor assessment and CT scanning for CAC scoring. RESULTS: The prevalence of CAC among females was lower than among males in all age groups. Using multivariate logistic regression, age, dyslipidaemia, hypertension, diabetes and smoking were independently predictive of CAC presence in both genders. In addition to a progressive increase in CAC with age, the most important predictors of CAC presence were dyslipidaemia and diabetes (ß = 0.64 and 0.63, respectively) in males and diabetes (ß = 1.08) followed by smoking (ß = 0.68) in females; these same risk factors were also important in predicting increasing CAC scores. There was no difference in the predictive ability of diabetes, hypertension and dyslipidaemia in either gender for CAC presence in patients aged <50 and 50-70 years. However, in patients aged >70, only dyslipidaemia predicted CAC presence in males and only smoking and diabetes were predictive in females. CONCLUSIONS: In symptomatic patients, there are significant differences in the ability of conventional risk factors to predict CAC presence between genders and between patients aged <70 and ≥70, indicating the important role of age in predicting CAC presence.

The coronary calcium score is a more accurate predictor of significant coronary stenosis than conventional risk factors in symptomatic patients: Euro-CCAD study.

AIMS: In this retrospective study we assessed the predictive value of the coronary calcium score for significant (>50%) stenosis relative to conventional risk factors. METHODS AND RESULTS: We investigated 5515 symptomatic patients from Denmark, France, Germany, Italy, Spain and the USA. All had risk factor assessment, computed tomographic coronary angiogram (CTCA) or conventional angiography and a CT scan for coronary artery calcium (CAC) scoring. 1539 (27.9%) patients had significant stenosis, 5.5% of whom had zero CAC. In 5074 patients, multiple binary regression showed the most important predictor of significant stenosis to be male gender (B=1.07) followed by diabetes mellitus (B=0.70) smoking, hypercholesterolaemia, hypertension, family history of CAD and age but not obesity. When the log transformed CAC score was included, it became the most powerful predictor (B=1.25), followed by male gender (B=0.48), diabetes, smoking, family history and age but hypercholesterolaemia and hypertension lost significance. The CAC score is a more accurate predictor of >50% stenosis than risk factors regardless of the means of assessment of stenosis. The sensitivity of risk factors, CAC score and the combination for prediction of >50% stenosis when measured by conventional angiogram was considerably higher than when assessed by CTCA but the specificity was considerably higher when assessed by CTCA. The accuracy of CTCA for predicting >50% stenosis using the CAC score alone was higher (AUC=0.85) than using a combination of the CAC score and risk factors with conventional angiography (AUC=0.81). CONCLUSION: In symptomatic patients, the CAC score is a more accurate predictor of significant coronary stenosis than conventional risk factors.

Coronary artery calcification correlates with the presence and severity of valve calcification.

AIM: To investigate the prevalence of coronary artery calcification (CAC) in symptomatic individuals with CT evidence for left heart valve calcification, aortic valve (AVC), mitral valve (MAC) or both. METHODS: This is a retrospective study of 282 consecutive patients with calcification in either the aortic valve or mitral annulus. Calcium scoring of the coronary artery, aortic and mitral valve was measured using the Agatston score. RESULTS: AVC was more prevalent than MAC (64% vs. 2.5%, p < 0.001), with 34% having both. Absence of CAC was noted in 12.7% of the study population. AVC + CAC were observed in 53.5%, MAC and CAC in 2.1%, and combined AVC, MAC and CAC in 31.6%. The median CAC score was higher in individuals with combined AVC+MAC, followed by those with AVC and lowest was in the MAC group. The majority (40%) of individuals with AVC had CAC score >400, and only in 16% had CAC = 0. The same pattern was more evident in individuals with AVC + MAC, where 70% had CAC score >400 and only 6% had CAC score of 0. These results were irrespective of gender. There was no correlation between AVC and MAC but there was modest correlation between CAC score and AVC score (r = 0.28, p = 0.0001), MAC (r = 0.36, p = 0.0001) and with combined AVC + MAC (r = 0.5, p = 0.0001). AVC score of 262 had a sensitivity of 78% and specificity of 92% for the prediction of presence of CAC. CONCLUSION: The presence and extent of calcification in the aortic valve or/and mitral valves are associated with severe coronary artery calcification.

Ultrasonographic characteristics of soft tissue tumours in dogs.

OBJECTIVE: To identify and describe the ultrasonographic features of soft tissue tumours in dogs. PROCEDURE: Superficial soft tissue tumours of various histological types, including mast cell tumours (MCTs) and soft tissue sarcomas (STSs), were evaluated. Ultrasound was used to visualise internal characteristics of the tumour, including vascularity. Tumours were categorised according to size, shape, margin definition, tissue plane mobility, echogenicity, echotexture, acoustic shadowing or enhancement and vessel distribution. Objective measurements of intratumoural blood flow included velocities and maximal perfused cross-sectional area (fractional area). Logistic regression models incorporating a variety of data were used in an attempt to predict the histopathological type of tumours. RESULTS: The logistic regression model defined by the parameters echotexture, margin definition and presence of subcapsular vessels was highly predictive of MCTs (> 73%; P = 0.024). Several other trends, including a larger size for STSs and less vascularity for both MCTs and STSs, were observed, but did not reach statistical significance. CONCLUSION: This preliminary study has shown the potential diagnostic value of ultrasound in differentiating soft tissue tumours. However, at present, ultrasound cannot replace biopsy and histopathological evaluation for tumour diagnosis.

Spontaneous hepatic rupture in six cats with systemic amyloidosis.

Spontaneous hepatic rupture, secondary to the accumulation of hepatic amyloid, was diagnosed in six cats over a two-year period. Previous reports of feline hepatic amyloidosis have documented clusters of cases from breeding catteries. Most affected cats have been Siamese or a related breed and the disease is generally regarded as familial. In contrast, the cases presented here were sporadic, with relatives and other cats in the household not clinically affected. They included a Devon rex, a breed not previously reported with this condition, and a domestic shorthair. Clinical signs in three of these cases had, prior to referral, been misinterpreted as resulting from blunt trauma, immune-mediated haemolysis or a coagulopathy. Antemortem diagnostic features, including new data on the value of hepatic ultrasonography and fine-needle aspirate cytology, are reported. These cases illustrate how the course of this disease can vary between individuals and that, despite the dramatic underlying pathology, hepatic amyloidosis can present a diagnostic challenge and should be suspected in any young adult cat with consistent clinical signs, irrespective of breed or environment.

Metabotropic glutamate receptor activation causes a rapid redistribution of AMPA receptors.

Electrophysiology, immunostaining and time lapse imaging techniques were employed to study the mechanism of long-term depression (LTD) induced by DHPG, a specific group I metabotropic glutamate receptor (mGluR) agonist. Experiments were performed in primary hippocampal culture or in the CA1 area of acute rat hippocampal slices. In agreement with previous results by others, we show that DHPG (200 microM, 10 min) can induce LTD (DHPG-LTD) in acute slices, in the presence or absence of synaptic inhibition. In addition, in voltage clamp whole cell experiments we find that accompanying the reduction in the evoked excitatory postsynaptic current (EPSC), miniature EPSC amplitude and frequency are reduced. Similar results were obtained in cultured neurons. Immunostaining and time lapse imaging showed a long-lasting loss of AMPA receptors from the membrane surface of cultured neurons after DHPG treatment, which appears to occur in only a subset of the puncta. Further electrophysiological recordings on slices showed that blocking postsynaptic endocytosis by introducing a blocking peptide named D15 in recording pipettes abolished the DHPG-LTD. In conclusion, these data suggest that LTD induced by mGluR activation is due to a rapid removal of AMPA receptors from the postsynaptic membrane.

Hippocampal synaptic transmission and plasticity are preserved in myosin Va mutant mice.

Recent studies have identified myosin Va as an organelle motor that may have important functions in neurons. Abundantly expressed at the hippocampal postsynaptic density, it interacts with protein complexes involved in synaptic plasticity. It is also located in presynaptic terminals and may function to recruit vesicles in the reserve pool to the active zone. Dilute-lethal mice are spontaneous myosin Va mutants and have severe neurological symptoms. We studied hippocampal physiology at CA3-CA1 excitatory synapses in dilute-lethal mutant mice to test the hypothesis that myosin Va plays a role in pre- or postsynaptic elements of synaptic transmission. In all assays performed, the mutant synapses appeared to be functioning normally, both pre- and postsynaptically. These data suggest that myosin Va is not essential for the synaptic release machinery, postsynaptic receptor composition, or plasticity at this synapse, but does not exclude significant roles for myosin Va in other cell types nor potential compensation by other myosin V isoforms.

The role of brain-derived neurotrophic factor receptors in the mature hippocampus: modulation of long-term potentiation through a presynaptic mechanism involving TrkB.

The neurotrophin BDNF has been shown to modulate long-term potentiation (LTP) at Schaffer collateral-CA1 hippocampal synapses. Mutants in the BDNF receptor gene trkB and antibodies to its second receptor p75NTR have been used to determine the receptors and cells involved in this response. Inhibition of p75NTR does not detectably reduce LTP or affect presynaptic function, but analyses of newly generated trkB mutants implicate TrkB. One mutant has reduced expression in a normal pattern of TrkB throughout the brain. The second mutant was created by cre-loxP-mediated removal of TrkB in CA1 pyramidal neurons of this mouse. Neither mutant detectably impacts survival or morphology of hippocampal neurons. TrkB reduction, however, affects presynaptic function and reduces the ability of tetanic stimulation to induce LTP. Postsynaptic glutamate receptors are not affected by TrkB reduction, indicating that BDNF does not modulate plasticity through postsynaptic TrkB. Consistent with this, elimination of TrkB in postsynaptic neurons does not affect LTP. Moreover, normal LTP is generated in the mutant with reduced TrkB by a depolarization-low-frequency stimulation pairing protocol that puts minimal demands on presynaptic terminal function. Thus, BDNF appears to act through TrkB presynaptically, but not postsynaptically, to modulate LTP.

Perirenal pseudocysts in 26 cats.

OBJECTIVE: To evaluate clinical features, anatomical location, nature of pseudocyst fluid, results of surgical treatment and links with underlying renal disease in cats with perirenal pseudocysts. DESIGN: A retrospective study of 26 affected cats, including 8 treated surgically. RESULTS: Nineteen (73%) affected cats were male. The median age was 11 years. Most presented for abdominal enlargement and had varying degrees of renal dysfunction on presentation. Thirteen cats (50%) had bilateral pseudocysts. The pseudocyst fluid was a transudate or modified transudate in all cases. All surgically treated cats had subcapsular perirenal pseudocysts. Associated renal lesions were identified in all cats that had renal biopsies or detailed ultrasonographic examinations. Surgery relieved clinical signs but did not stop progression of renal disease. Cats survived a median of 9 months after surgery and survival was correlated statistically to degree of azotaemia at presentation. Percutaneous drainage of pseudocysts was ineffective in controlling long-term fluid accumulation. CONCLUSIONS: Subcapsular perirenal pseudocysts are formed in cats by accumulation of transudate between the capsule and parenchyma of the kidney as a result of underlying parenchymal disease. Pseudocyst formation can occur at variable stages of renal dysfunction. Resection of the pseudocyst wall is usually effective in eliminating signs but does not stop progression of renal disease. The prognosis for cats with pseudocyst formation is related to the degree of renal dysfunction at time of diagnosis.

Effects of reduced vesicular filling on synaptic transmission in rat hippocampal neurones.

The consequence of reduced uptake of neurotransmitters into synaptic vesicles on synaptic transmission was examined in rat hippocampal slices and culture using bafilomycin A1 (Baf), a potent and specific blocker of the vacuolar-type (V-type) ATPase, which eliminates the driving force for the uptake of both glutamate and GABA into synaptic vesicles. After incubation with Baf, both the amplitude and frequency of GABAergic miniature inhibitory postsynaptic currents (mIPSCs) were reduced in the slice preparation. Similar effects were seen with glutamatergic miniature excitatory postsynaptic currents (mEPSCs) and GABAergic mIPSCs from cultured neurons. This result indicates that vesicular content is reduced by Baf. The dramatic reduction in the frequency of mPSCs could result either from the exocytosis of empty vesicles or from a mechanism which prevents the exocytosis of depleted vesicles. Vesicle cycling was directly examined using confocal imaging with FM 1-43. In the presence of Baf, vesicles could still be endocytosed and they were released at the same probability as from control untreated synapses. Prolonged high-frequency electrical stimulation of synapses in culture failed to alter the amplitude of mEPSCs, suggesting that the filling of vesicles is rapid compared to the rate of vesicle recycling during repetitive synaptic stimulation. Profound release of glutamate with alpha-latrotoxin did cause a small, but reproducible, reduction in quantal size. These results indicate that decreasing the amount of glutamate and GABA in synaptic vesicles reduces quantal size. Furthermore, the probability of vesicle exocytosis appears to be entirely independent of the state of filling of the vesicle. However, even during high-frequency action potential-evoked release of glutamate, quantal size remained unchanged.

Concurrent Aelurostrongylus abstrusus infection and salmonellosis in a kitten.

A 14-week-old kitten had a history of vomiting, diarrhoea and pyrexia, all of which resolved without treatment. Three weeks later the kitten developed a violent non-productive dry cough. Thoracic radiographs revealed pneumothorax and nodular alveolar disease. Aelurostrongylus abstrusus larvae and intracellular Gram-negative bacilli were seen in bronchial wash fluid and pleural exudate, and Salmonella Typhimurium was cultured from both fluids but not from faeces. Therapy included unilateral closed-tube thoracostomy, enrofloxacin and fenbendazole. Historical signs were compatible with gastrointestinal salmonellosis and secondary broncho-pneumonia. Seeding of the lungs with salmonellae may have occurred as a result of migration of A abstrusus from a gastro-intestinal tract residually infected or colonised by S Typhimurium. Alternatively, the development of lungworm infection in the cat may have activated quiescent S Typhimurium pulmonary granulomata from bacteraemia secondary to gastro-intestinal salmonellosis. Two years after diagnosis the cat was reportedly in good health.

Intestinal obstruction by trichobezoars in five cats.

Between 1997 and 1999, five domestic crossbred cats (four long haired, one short haired) presented with a palpable abdominal mass and were shown to have small intestinal trichobezoars at laparotomy or necropsy. Hair balls were associated with partial or complete intestinal obstruction and were situated in the proximal jejunum to distal ileum. In four cats obstructions were simple, while the remaining cat had a strangulating obstruction. Three of the cats were 10 years or older, and two were less than 4 years. In the three older cats abdominal neoplasia was suspected and investigations were delayed or declined in two of these cats because of a perceived poor prognosis. Predisposing factors identified in this series of cats included a long-hair coat, flea allergy dermatitis, inflammatory bowel disease and ingestion of non-digestible plant material. This report shows that the ingestion of hair is not always innocuous and that intestinal trichobezoars should be considered in the differential diagnoses of intestinal obstruction and intra-abdominal mass lesions, particularly in long-haired cats.

A role for cAMP in long-term depression at hippocampal mossy fiber synapses.

Mossy fiber synapses on hippocampal CA3 pyramidal cells, in addition to expressing an NMDA receptor-independent form of long-term potentiation (LTP), have recently been shown to express a novel presynaptic form of long-term depression (LTD). We have studied the mechanisms underlying mossy fiber LTD and present evidence that it is triggered, at least in part, by a metabotropic glutamate receptor-mediated decrease in adenylyl cyclase activity, which leads to a decrease in the activity of the cAMP-dependent protein kinase (PKA) and a reversal of the presynaptic processes responsible for mossy fiber LTP. The bidirectional control of synaptic strength at mossy fiber synapses by activity therefore appears to be due to modulation of the cAMP-PKA signaling pathway in mossy fiber boutons.

Effects of PKA and PKC on miniature excitatory postsynaptic currents in CA1 pyramidal cells.

Protein kinases play an important role in controlling synaptic strength at excitatory synapses on CA1 pyramidal cells. We examined the effects of activating cAMP-dependent protein kinase or protein kinase C (PKC) on the frequency and amplitude of miniature excitatory postsynaptic currents (mEPSCs) with perforated patch recording techniques. Both forskolin and phorbol-12,13-dibutryate (PDBu) caused a large increase in mEPSC frequency, but only PDBu increased mEPSC amplitude, an effect that was not observed when standard whole cell recording was performed. These results support biochemical observations indicating that PKC, similar to calcium/calmodulin-dependent protein kinase II, has an important role in controlling synaptic strength via modulation of AMPA receptor function, potentially through the direct phosphorylation of the GluR1 subunit.

Activity differentially regulates the surface expression of synaptic AMPA and NMDA glutamate receptors.

Distinct subtypes of glutamate receptors often are colocalized at individual excitatory synapses in the mammalian brain yet appear to subserve distinct functions. To address whether neuronal activity may differentially regulate the surface expression at synapses of two specific subtypes of ionotropic glutamate receptors we epitope-tagged an AMPA (alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid) receptor subunit (GluR1) and an NMDA (N-methyl-D-aspartate) receptor subunit (NR1) on their extracellular termini and expressed these proteins in cultured hippocampal neurons using recombinant adenoviruses. Both receptor subtypes were appropriately targeted to the synaptic plasma membrane as defined by colocalization with the synaptic vesicle protein synaptophysin. Increasing activity in the network of cultured cells by prolonged blockade of inhibitory synapses with the gamma-aminobutyric acid type A receptor antagonist picrotoxin caused an activity-dependent and NMDA receptor-dependent decrease in surface expression of GluR1, but not NR1, at synapses. Consistent with this observation identical treatment of noninfected cultures decreased the contribution of endogenous AMPA receptors to synaptic currents relative to endogenous NMDA receptors. These results indicate that neuronal activity can differentially regulate the surface expression of AMPA and NMDA receptors at individual synapses.

Structural and functional characterisation of human sulfotransferases.

The human aryl sulfotransferases HAST4 and HAST4v vary by only two amino acids but exhibit markedly different affinity towards the sulfonate acceptor p-nitrophenol and the sulfonate donor 3'-phosphoadenosine-5'-phosphosulfate (PAPS). To determine the importance of each of these amino acid differences, chimeric constructs were made of HAST4 and HAST4v. By attaching the last 120 amino acids of HAST4v to HAST4 (changing Thr235 to Asn235) we have been able to produce a protein that has a Km for PAPS similar to HAST4v. The reverse construct, HAST4v/4 produces a protein with a Km for PAPS similar to HAST4. These data suggests that the COOH-terminal of sulfotransferases is involved in co-factor binding.

Quantitative ultrasonography of the liver in cats during obesity induction and dietary restriction.

A study was designed to evaluate quantitative ultrasonographic characteristics of a diffuse parenchymal disease in an experimental model of subclinical fatty infiltration of the liver using diet-induced obesity and dietary restriction in the cat. Ultrasound images of livers were quantitatively analysed by a video signal analysis technique before, during and at the end of obesity induction, and then during and following dietary restriction. Attenuation and brightness (backscatter coefficient) were correlated with hepatic lipid content obtained from the livers by surgical and ultrasound-guided percutaneous biopsy. Attenuation and backscatter increased as hepatic lipid content increased. Both attenuation and backscatter significantly correlated with the hepatic lipid content (P=0.002 and P=0.02 respectively). This model of subclinical fatty infiltration of the liver in the cat demonstrates that hepatic lipid content increases as a consequence of obesity and of severe dietary restriction. Infiltration of the liver with fat may be evaluated non-invasively using quantitative ultrasonography by the video signal analysis technique. This method of image analysis may prove useful for the evaluation of diffuse parenchymal organ disease.