Effect of tibial plateau angle < 5° on ground reaction forces in dogs treated with tibial plateau leveling osteotomy for cranial cruciate ligament rupture up to 6 months postoperatively.

Tibial plateau leveling osteotomy (TPLO) has been commonly performed in dogs with cranial cruciate ligament disease (CCLD) since the introduction by Slocum and Slocum (1993). To reduce cranial tibial thrust the TPLO technique aims for a postoperative tibial plateau angle (TPA) of 5-6.5°. In recent years studies have shown that a postoperative TPA below 5° could be beneficial regarding stifle stability or meniscal load. Dogs with CCLD that were treated with TPLO, were examined preoperatively, six weeks, three and six months postoperatively with gait analysis and grouped according to their postoperative TPA. The aims of study was (1) to evaluate if dogs with a postoperative TPA below 5° would have a faster limb function recovery up to six months postoperatively as measured objectively with ground reaction forces (GRFs) and (2) to determine whether the postoperative TPA correlates with the outcome measurements. Dogs with TPA <5° showed no faster limb function recovery postoperatively up to six months as measured with peak vertical force (PVF) or vertical impulse (VI) (p > 0.05). No correlation for the postoperative TPA <5° on GRFs was demonstrated. But the postoperative TPA showed a significant correlation with the symmetry indices of PVF (SIPVF) and VI (SIVI) for all dogs (>5° and <5° TPA together), indicating that with lower postoperative TPA dogs had a more symmetrical gait in hindlimbs SIPVF (r = 0.144, p < 0.05) and SIVI (r = 0.189, p < 0.01). The study indicates that a lower postoperative TPA could be beneficial regarding hindlimb symmetry indices of GRFs.

Effect of intra-articular platelet-rich plasma or hyaluronic acid on limb function recovery in dogs with TPLO for cranial cruciate ligament rupture: a randomised controlled trial.

OBJECTIVES: This study aimed to investigate the recovery of limb function following a single intra-articular injection of platelet-rich plasma or hyaluronic acid in dogs with cranial cruciate ligament rupture treated with tibial plateau levelling osteotomy compared to dogs receiving no injection intraoperatively. MATERIALS AND METHODS: Sixty-two dogs with cranial cruciate ligament rupture, body weights of 20 to 40 kg, and no other orthopaedic conditions were enrolled in this prospective, randomised, double-blind, controlled study at the small animal clinic at LMU Munich. All dogs underwent tibial plateau levelling osteotomy. Based on random allocation, they received either a single intra-articular injection of platelet-rich plasma, hyaluronic acid or no injection intraoperatively. Gait analysis, clinical examinations, radiography of the stifle joint for osteoarthritis progression and two validated owner questionnaires were compared among groups at three timepoints postoperatively (6 weeks, 3 and 6 months). Limb function was primarily assessed by measuring the ground reaction forces. RESULTS: At all times postoperatively, no differences were observed among groups regarding clinical examinations, osteoarthritis score values, ground reaction forces or owner questionnaires. All dogs showed significant improvement in limb function clinically, in all ground reaction forces and in the validated questionnaires. Osteoarthritis progressed minimally during rechecks in all dogs regardless of the additional injection or not. CLINICAL SIGNIFICANCE: All dogs treated with tibial plateau levelling osteotomy for cranial cruciate ligament rupture showed improvements in limb function. No additive effect on faster recovery was demonstrated with the additional intra-articular injection of platelet-rich plasma or hyaluronic acid. Addition of platelet-rich plasma/hyaluronic acid injections during tibial plateau levelling osteotomy is unnecessary considering the lack of benefit observed up to 6 months postoperatively.

Maternal undernutrition programs the apelinergic system of adipose tissue in adult male rat offspring.

Based on the Developmental Origin of Health and Disease concept, maternal undernutrition has been shown to sensitize adult offspring to metabolic pathologies such as obesity. Using a model of maternal 70% food restriction in pregnant female rats throughout gestation (called FR30), we previously reported that obesity-prone adult male rat offspring displayed hyperleptinemia with modifications in leptin and leptin receptor messenger RNA (mRNA) levels in white adipose tissue (WAT). Apelin is a member of the adipokine family that regulates various aspects of energy metabolism and WAT functionality. We investigated whether apelin and its receptor APJ could be a target of maternal undernutrition. Adult male rat offspring from FR30 dams showed increased plasma apelin levels and apelin gene expression in WAT. Post-weaning high-fat diet led to marked increase in APJ mRNA and protein levels in offspring's WAT. We demonstrate that maternal undernutrition and post-weaning diet have long-term consequences on the apelinergic system of adult male rat offspring.

Perinatal nutrition programs the hypothalamic melanocortin system in offspring.

Epidemiological studies initially suggested that maternal undernutrition leading to low birth weight may predispose for long-lasting energy balance disorders. High birth weight due to maternal obesity or diabetes, inappropriate early postnatal nutrition, and rapid catch-up growth, may also sensitize to increased risk of obesity. As stated by the Developmental Origin of Health and Disease concept, the perinatal perturbation of fetus/neonate nutrient supply might be a crucial determinant of individual programming of body weight set-point. The hypothalamic melanocortin system composed of the melanocortin receptor 4, its agonist α-melanin-stimulating hormone (α-MSH), and its antagonist agouti-related protein (AgRP) is considered as the main central anorexigenic pathway controlling energy homeostasis. Studies in numerous animal models demonstrated that this system is a prime target of developmental programming by maternal nutritional manipulation. In rodents, the perinatal period of life corresponds largely to the period of brain maturation (i. e., melanocortin neuronal differentiation and development of their neural projections). In contrast, these phenomena essentially take place before birth in bigger mammals. Despite these different developmental time windows, altricial and precocial species share several common offspring programming mechanisms. Offspring from malnourished dams present a hypothalamic melanocortin system with a series of alterations: impaired neurogenesis and neuronal functionality, disorganization of feeding pathways, modified glucose sensing, and leptin/insulin resistance. Overall, these alterations may account for the long-lasting dysregulation of energy balance and obesity. Following maternal malnutrition, hormonal and epigenetic mechanisms might be responsible for melanocortin system programming in offspring.

Determination of and correlation between urine protein excretion and urine protein-to-creatinine ratio values during a 24-hour period in healthy horses and ponies.

OBJECTIVE-To determine whether urine protein-to-creatinine (UP:C) ratio assessment provides an estimate of urine protein excretion (UPE) over a 24-hour period in horses and ponies, establish a preliminary UP:C ratio reference range, and determine UP:C ratio variation over time in healthy equids. ANIMALS-11 female horses and 6 female ponies. PROCEDURES-Urine was collected from all equids at 4-hour intervals for 24 hours. Total 24-hour UPE (mg of protein/kg of body weight) and UP:C ratio were determined; these variables were also assessed in aliquots of urine collected at 4-hour intervals. On 2 additional days, urine samples were also obtained from 6 horses (1 sample/horse/d) to determine day-to-day variation in UP:C ratio. Correlation between 4-hour or 24-hour UPE and UP:C ratio values was assessed. Reference ranges for 24-hour UPE, 24-hour UP:C ratio, and 4-hour UP:C ratios were calculated as central 95th percentiles of observed values. RESULTS-Mean 24-hour UPE (4.28 +/- 2.99 mg/kg) and 24-hour UP:C ratio (0.0 to 0.37) had excellent correlation (R = 0.826; P < 0.001) in both horses and ponies; analysis of 4-hour data also revealed good correlation (R = 0.782; P < 0.001) with these variables. Calculated UPE and UP:C ratio reference ranges were similar to established ranges in other species. Day-to-day variability in UP:C ratio was minimal, and all results were within the reference range calculated by use of the 24-hour urine samples. CONCLUSIONS AND CLINICAL RELEVANCE-Assessment of the UP:C ratio appears to be a reliable method for estimating 24-hour UPE in horses and ponies.

The polymorphism Arg585Gln in the gene of the sterol regulatory element binding protein-1 (SREBP-1) is not a determinant of ketosis prone type 2 diabetes (KPD) in Africans.

AIM: Ketosis prone type 2 diabetes (KPD) is an atypical form of diabetes described mainly in people of sub-Saharan African origin. Its pathogenesis is unknown, although we have previously described a high prevalence of glucose-6-phosphate-dehydrogenase (G6PD) deficiency in patients with KPD. However, 50% of these deficient patients lacked the G6PD gene mutation. The isoforms of the transcription factor sterol regulatory element binding protein 1 (SREBP-1) are known to stimulate G6PD gene expression, and some polymorphisms in the SREBP-1 gene (SREBF-1) have been described only in Africans. We investigated one of these, the Arg585Gln polymorphism, in a candidate gene approach for KPD. METHODS: We examined the presence of the Arg585Gln polymorphism in SREBF-1 in 217 consecutive unrelated Africans [73 patients with KPD, 80 with classical type 2 diabetes (T2D) and 64 nondiabetic subjects]. Patients underwent clinical and biochemical evaluations, and were assessed for G6PD activity and insulin secretion (glucagon test). RESULTS: There were no differences in frequency of the Arg585Gln polymorphism and the 585Gln allele among the three groups (allele frequency: KPD: 0.089, T2D: 0.031, nondiabetic group: 0.070; P=0.1). When the 585Gln allele frequency was compared separately between patients with KPD and those with T2D, it was significantly higher in the former (P=0.032). There was no difference between carriers and noncarriers of the 585Gln allele regarding G6PD activity and insulin secretion. CONCLUSION: The results of this exploratory study show that the polymorphism Arg585Gln in SREBF-1 is not associated with the KPD phenotype. Further studies in larger populations are needed to confirm our findings.

Single nucleotide polymorphisms of protein tyrosine phosphatase 1B gene are associated with obesity in morbidly obese French subjects.

AIMS/HYPOTHESIS: The development of insulin resistance may contribute to the occurrence and progression of the metabolic syndrome associated with obesity. Components contributing to the insulin pathway and its regulation are good candidates for the molecular study of metabolic syndrome pathogenesis. Protein tyrosine phosphatase 1B (PTP 1B) is an important negative regulator of insulin. We investigated whether PTP 1B SNPs are associated with obesity and obesity-related traits as well as global metabolic syndrome in morbidly obese subjects. METHODS: Untranslated and coding regions of the PTP 1B gene were screened in groups of non-diabetic and diabetic obese subjects and in non-obese subjects. Unrelated morbidly obese ( n=711) and non-obese ( n=427) French Caucasian subjects were genotyped for a case-control study. RESULTS: Six SNPs were identified: two rare variants were located in 5'UTR (-109 C>T and -69 C>T), two in the intronic regions (IVS3+38 G>T and IVS5+3666delT) and two have been described previously (P303P in exon 8 and P387L in exon 9). A case-control study showed an association between the frequent IVS5+3666delT SNP and obesity ( p=0.02). In the obese group, associations between PTP 1B SNPs and features of dyslipidaemia were found. P303P was associated with lower apolipoprotein A1 levels ( p=0.05) whereas P387L was associated with higher triglyceride ( p=0.0003), apolipoprotein B ( p=0.09) and lipoprotein a concentrations ( p=0.006). CONCLUSIONS/INTERPRETATION: Our results support the hypothesis that the PTP 1B gene contributes to the polygenic basis of obesity. PTP 1B SNPs may interact with environmental factors to induce more severe phenotypes, e.g. atherogenic dyslipidaemia, in morbidly obese subjects.

Profile identification of disease-associated humoral antigens using AMIDA, a novel proteomics-based technology.

We describe AMIDA (autoantibody-mediated identification of antigens), a novel target identification technology based on the immunoprecipitation of disease-specific antigens by autologous serum antibodies followed by two-dimensional electrophoretic separation, and their identification via mass spectrometry. Twenty-seven potential carcinoma antigens were identified including proteins of hitherto unknown function. Validation of one of the identified antigens, cytokeratin 8, revealed its de novo expression in hyperplastic tissue, gradual overexpression with increasing malignancy, and ectopic localization on the cell surface. Furthermore, a strong prevalence of CK8-specific antibodies occurred in the serum of cancer patients already at early disease stages. In situ hybridization for one marker of unknown function, KIAA1273/TOB3, demonstrated its strong overexpression in head and neck carcinomas, thus making it a likely tumor antigen candidate. Eventually, AMIDA could foster significant improvements for the diagnosis and therapy of human diseases eliciting a humoral immune response, and allows for the rapid identification of new target molecules.

Chiral resolution of pantoprazole sodium and related sulfoxides by complex formation with bovine serum albumin in capillary electrophoresis.

The separation of enantiomers of pantoprazole sodium, omeprazole and lansoprazole by capillary zone electrophoresis using bovine serum albumin (BSA) as the chiral selector is described. Baseline separation of the three structurally related drugs was obtained after optimization of the most important experimental parameters. For this purpose, influences such as BSA concentration, pH and concentration of 1-propanol as organic modifier on the separation were investigated. Increasing concentrations of BSA improved the chiral resolution but lowered the sensitivity of the detection system. Discrimination of the enantiomers was observed only in a narrow pH range of 7-8. An optimum of pH 7.4 was a good compromise in terms of enantio-resolution and peak shape. 1-Propanol when added to the buffer system, improved the peak shape of the analytes and the resolution. The optimized method has been validated for pantoprazole sodium and is useful for routine analysis.

Recognition of depression.

Depression in the terminally ill has never been examined systematically. Frequently depression has been perceived as an inevitable part of illness. The purpose of the present study was to develop an instrument (the Mood Evaluation Questionnaire) to measure depression among terminally ill patients. The MEQ and the Geriatric Depression Scale were completed by 27 hospice patients. A Modified Karnofsky score and index of somatic complaints were obtained. There was no correlation among the Modified Karnofsky, the number of somatic complaints, and the level of depression. However the MEQ and GDS were highly correlated (p < .01). For several reasons, the MEQ appears to be an effective instrument to explore the incidence of depression in the terminally ill.

Demand and continuous flow intermittent mandatory ventilation systems.

A mechanical lung was used to evaluate the pressure and flow characteristics of four demand and two continuous flow intermittent mandatory ventilation (IMV) systems. The amount of negative pressure required to initiate inspiratory flow and peak expiratory resistance were measured. The inspiratory pressure required to initiate flow in the demand mode was also compared to pressures generated in the assist mode. In addition, the peak expiratory resistance was measured with four commercially available exhalation valves. Results showed that the ventilator manometer measuring internal machine pressures significantly underestimated the amount of negative pressure required to open the demand valve (p less than 0.01). There are major differences in the flow and pressure characteristics among demand and continuous flow IMV systems. Systems that impose high inspiratory elastic threshold loads and expiratory flow resistive loads may have a deleterious effect on the mechanics of breathing, and thereby limit weaning success and eventually impair the recovery of certain patients in respiratory failure. The basic methodology, especially the simple technique of inserting an aneroid manometer in line next to a patient's ET tube, for measuring proximal negative inspiratory force (NIF test) can be easily applied to any and all ventilators at any practitioner's individual institution.

Role of glutathione in gastric mucosal cytoprotection.

Exogenous thiol compounds have been reported to protect the stomach from ethanol-induced necrotic lesions. The gastric mucosa contains high levels of an endogenous thiol, glutathion (GSH). Because of the known role of glutathione in protecting against hepatic injury, its role in gastric mucosal cytoprotection was of interest. By use of an animal model for acute gastric injury from ethanol, a close parallel relation between depletion of endogenous mucosal GSH and induction of mucosal protection was demonstrated. Surprisingly, mucosal protection varied inversely with the level of mucosal GSH obtained after treatment with specific GSH-depleting agents (diethyl maleate and cyclohexene-1-one). Depletion of gastric mucosal GSH was associated with an increase in the mucosal content of prostaglandins 6-keto F1 alpha and F2 alpha but not E2. The protective effect induced by GSH-depleting agents was partially reversed by indomethacin in some but not all studies. Although GSH depletors increased gastric juice volume, protection with these agents persisted after the volume and mucosal GSH had returned to control levels and also was not reversed by increasing the dose of ethanol threefold to overcome a possible dilutional effect. We conclude that, contrary to apparent predictions, depletion of endogenous gastric GSH protects the stomach from acute ethanol-induced injury. Although the mechanism of this protection is unknown, a mediation by endogenous release of prostaglandins seems to play a minor role since diethyl maleate was protective even in indomethacin-treated animals.

Adult respiratory distress syndrome: risk with common predispositions.

A 1-year survey of patients in three hospitals identified 936 patients who had one predisposition and 57 who had several predispositions to the adult respiratory distress syndrome. From the total predisposed population of 993 patients, 68 subsequently developed the syndrome. An additional 20 patients developed the syndrome from causes other than eight identified predispositions, to bring the total of patients studied to 88. A highly significant difference (p less than 0.0001) was found in the incidence rates of the syndrome between patients with one and several predispositions (5.8 versus 24.6 per 100 patients). Within 72 hours of onset of predisposition, 89.5% of patients who developed the syndrome had been intubated and placed on mechanical ventilation. Fifty-seven of the 88 patients (64.8%) with the syndrome died. By the 14th day 90% of deaths had occurred. There were no age- or sex-specific differences in either incidence or mortality rates. Case fatality rates of the syndrome were high in all predisposed groups.

Rapid oxidation in vitro of endogenous and exogenous glutathione in bile of rats.

Biliary excretion of glutathione has recently been described but poorly characterized. Controversy has existed concerning the relative contribution of oxidized and reduced glutathione to total glutathione efflux from the liver into bile. We found that bile, unlike liver cytosol or buffer, had the unique ability to oxidize GSH rapidly (t 1/2 = 5 min) to the disulfide form by a nonenzymatic, O2, and pH-dependent chemical reaction inhibited by only certain chelating agents. Significant oxidation of GSH occurred during the collection of bile samples resulting in significant time-dependent alterations in the ratio of biliary GSH to GSSG. Thus the preponderance of GSSG in bile in the normal state reported by others represents a postexcretory in vitro artifact. Inhibition of oxidation by acidification of bile during its collection established the true contribution of GSH and GSSG to total biliary efflux.

Disseminated North American blastomycosis. Occurrence with clinical manifestations of adrenal insufficiency.

Adrenal involvement in cases of disseminated North American blastomycosis occurs frequently, but clinically substantive adrenal insufficiency is rare. An unusual case of disseminated blastomycosis with clinical manifestations of adrenal insufficiency was found. One should be alert to the symptoms of adrenal insufficiency that may develop in patients with disseminated mycoses.

Comparative studies of instrumental and bioassay methods for the analysis of herbicide residues.

A study of the quantitative analysis of herbicide residues by both chemical and bioassay methods in soils is presented. Field and laboratory residue trials were carried out with a representative member of the following groups of herbicides: ureas, triazines, diphenylethers, phenoxyacetic acids, and dithiophosphates. Representative samples were taken at different time intervals, and degradation curves were established both by chemical methods and by two types of bioassay. Chemical analysis either separated active ingredient and metabolites by chromatographic techniques or compresied total residues. Bioassays were performed using either monocotyledons and dicotyledons or algae. The results obtained by chemical and bioassay analysis for the degradation rates of chlorotoluron, ametryn, 2,4-D and C 19490 showed a correlation coefficient of 0.914, indicating that the two methods gave almost identical results. Especially with the highly adsorbed urea and triazine herbicides, the uptake of biologically active material by test plants was slightly less than the solvent-extractable parent compound plus its metabolities, and so the absolute level of residues obtained by bioassay was lower. In the case of fluorodifen, the correlation between the methods was not established. The bioassay showed higher residues and slower degradation than chemical analysis. Various factors which could explain this anomalous result are discussed.