Evaluation of reference genes for RT-qPCR gene expression analysis in Arabidopsis thaliana exposed to elevated temperatures.

Increases in environmental temperature are directly linked to the issue of climate change and are known to significantly disrupt plant growth and development. Studies of gene expression in plants commonly include RT-qPCR but the reliability of the method depends on the use of suitable reference genes for data normalization. Despite this, no reference genes have been validated specifically for experiments in Arabidopsis thaliana employing treatments with elevated temperature. Here, ten genes were selected for expression stability analysis based on the screening of available literature and microarray data from temperature-treated A. thaliana. Expression levels of candidate reference genes were measured in 12-day-old seedlings, rosette leaves and flower buds of 5-week-old A. thaliana plants exposed to five different temperatures (22°C, 27°C, 32°C, 37°C and 42°C) and their expression stabilities were assessed using four statistical algorithms (BestKeeper, geNorm, NormFinder and comparative ΔCq method). This study provides reliable reference genes for use in A. thaliana RT-qPCR expression analyses employing elevated temperature treatments, namely OGIO and PUX7 in seedlings, UBC21 and PUX7 in leaves, TIP41 and UBC21 in buds, and TIP41 and UBC21 in all three tissues combined. Orthologues of these genes can be of potential use in less studied plants, especially agricultural species heavily affected by climate change.

Scalable Production of Equine Platelet Lysate for Multipotent Mesenchymal Stromal Cell Culture.

Translation of multipotent mesenchymal stromal cell (MSC)-based therapies is advancing in human and veterinary medicine. One critical issue is the in vitro culture of MSC before clinical use. Using fetal bovine serum (FBS) as supplement to the basal medium is still the gold standard for cultivation of many cell types including equine MSC. Alternatives are being explored, with substantial success using platelet lysate-supplemented media for human MSC. However, progress lags behind in the veterinary field. The aim of this study was to establish a scalable protocol for equine platelet lysate (ePL) production and to test the ePL in equine MSC culture. Whole blood was harvested into blood collection bags from 20 healthy horses. After checking sample materials for pathogen contamination, samples from 19 animals were included. Platelet concentrates were prepared using a buffy coat method. Platelets, platelet-derived growth factor BB, and transforming growth factor ß1 concentrations were increased in the concentrates compared with whole blood or serum (p < 0.05), while white blood cells were reduced (p < 0.05). The concentrates were lysed using freeze/thaw cycles, which eliminated the cells while growth factor concentrations were maintained. Donor age negatively correlated with platelet and growth factor concentrations after processing (p < 0.05). Finally, all lysates were pooled and the ePL was evaluated as culture medium supplement in comparison with FBS, using adipose-derived MSC from four unrelated donor horses. MSC proliferated well in 10% FBS as well as in 10% ePL. However, using 5 or 2.5% ePL entailed highly inconsistent proliferation or loss of proliferation, with significant differences in generation times and confluencies (p < 0.05). MSC expressed the surface antigens CD90, CD44, and CD29, but CD73 and CD105 detection was low in all culture media. Adipogenic and osteogenic differentiation led to similar results in MSC from different culture media. The buffy coat method is useful to produce equine platelet concentrate with increased platelet and reduced white blood cell content in large scales. The ePL obtained supports MSC expansion similar as FBS when used at the same concentration (10%). Further investigations into equine MSC functionality in culture with ePL should follow.

Visual field deficits after epilepsy surgery: a new quantitative scoring method.

BACKGROUND: Anterior temporal lobectomy (ATL) as a treatment for drug-resistant temporal lobe epilepsy (TLE) frequently causes visual field deficits (VFDs). Reported VFD encompasses homonymous contralateral upper quadrantanopia. Its reported incidence ranges from 15 to 90%. To date, a quantitative method to evaluate postoperative VFD in static perimetry is not available. A method to quantify postoperative VFD, which allows for comparison between groups of patients, was developed. METHODS: Fifty-five patients with drug-resistant TLE, who underwent ATL with pre- and postoperative perimetry, were included. Temporal lobe resection length was measured on postoperative MRI. Percentage VFD was calculated for the total visual field, contralateral upper quadrant, or other three quadrants combined. RESULTS: Patients were divided into groups by resection size (< 45 and ≥ 45 mm) and side of surgery (right and left). We found significant higher VFD in the ≥ 45 vs. < 45 mm group (2.3 ± 4.4 vs. 0.7 ± 2.4%,p = 0.04) for right-sided ATL. Comparing VFD in both eyes, we found more VFD in the right vs. left eye following left-sided ATL (14.5 ± 9.8 vs. 12.9 ± 8.3%, p = 0.03). We also demonstrated significantly more VFD in the < 45 mm group for left- vs. right-sided surgery (6.7 ± 6.7 vs. 13.1 ± 7.0%, p = 0.016). A significant quantitative correlation between VFD and resection size for right-sided ATL was shown (r = 0.52, p < 0.01). CONCLUSIONS: We developed a new quantitative scoring method for the assessment of postoperative visual field deficits after temporal lobe epilepsy surgery and assessed its feasibility for clinical use. A significant correlation between VFD and resection size for right-sided ATL was confirmed.

Reticulocytosis in non-anaemic cats and dogs.

OBJECTIVE: To evaluate the proportion of blood samples diagnosed with reticulocytosis without anaemia in cats and dogs and report the aetiology and mortality rate of affected animals. MATERIALS AND METHODS: Retrospective multicentre study including haematological examination of 3956 cats and 11,087 dogs admitted to seven German veterinary clinics (2012 to 2014). The proportion of blood samples with reticulocytosis without anaemia was calculated, and after exclusion of multiple measurements of the same animal, clinical data were evaluated. Animals with reticulocytosis without anaemia were classified as healthy or diseased, and diseased patients were assigned to 12 disease groups. Pretreatment (i.e. non-steroidal anti-inflammatory drugs, glucocorticoids, dipyrone) was recorded. RESULTS: The proportion of blood samples with reticulocytosis without anaemia was 3·1% (124/3956) in cats and 4·4% (492/11,087) in dogs. Overall, 1·8% (2/111) of cats and 1·5% (7/458) of dogs with reticulocytosis without anaemia were healthy. Blood loss/anaemia, cardiac/respiratory disorders, gastrointestinal disorders and inflammatory disorders as well as cancer were the most frequent underlying diseases. Pretreatment was noted in 39·5% (43/111) of cats and 42·4% (194/458) of dogs. The mortality rate was 37·8% (42/111) in cats and 29·7% (136/458) in dogs with reticulocytosis without anaemia; the median survival time in non-survivors was 1 day (range: 0 to 376 days in cats, 0 to 444 days in dogs). CLINICAL SIGNIFICANCE: In both species, reticulocytosis without anaemia was observed in a low proportion of blood samples (dogs>cat). Though a bias towards sick animals is possible in our sample, reticulocytosis without anaemia was mainly seen in diseased animals and associated with a mortality rate of approximately one-third of patients.

Identification of Brucella spp. in feral swine (Sus scrofa) at abattoirs in Texas, USA.

Various tissues, nasal swabs, urine and blood samples were collected from 376 feral swine at two federally inspected abattoirs in Texas during six separate sampling periods in 2015. Samples were tested for Brucella spp. by culture and serology. Brucella spp. were cultured from 13.0% of feral swine, and antibodies were detected in 9.8%. Only 32.7% of culture-positive feral swine were also antibody positive, and 43.2% of antibody-positive feral swine were culture positive. Approximately, the same number of males (14.0%) and females (12.1%) were culture positive, and slightly more males (10.5%) than females (8.7%) were antibody positive. Our results indicate that serology likely underestimates the prevalence of feral swine infected, and that those who come in contact with feral swine should be aware of the symptoms of infection with Brucella spp. to ensure prompt treatment.

Continuous exposure of pancreatic cancer cells to dietary bioactive agents does not induce drug resistance unlike chemotherapy.

The repeated treatment of cancer cells with chemo- or radiotherapy induces therapy resistance, but it was previously unknown whether the same effect occurs upon continuous exposure of cancer cells to diet-derived chemopreventive agents. We elucidated this interesting question in pancreatic ductal adenocarcinoma, which is a highly aggressive cancer entity with a marked resistance toward gemcitabine and other cytotoxic drugs. The isothiocyanate sulforaphane, present in cruciferous vegetables, and the polyphenol quercetin, present in many fruits and vegetables induced apoptosis and reduced viability in gemcitabine-sensitive BxPC-3 cells but not in non-malignant ductal pancreas cells and mesenchymal stromal cells. In turn, BxPC-3 cells were treated with increasing concentrations of gemcitabine, sulforaphane or quercetin for more than 1 year and the surviving subclones Bx-GEM, Bx-SF and Bx-Q were selected, respectively. While Bx-GEM cells acquired a total resistance, Bx-SF or Bx-Q cells largely kept their sensitivity as proved by MTT assay, annexin staining and FACS analysis. The evaluation of the self-renewal-, differentiation- and migration-potential by colony formation, differentiation or migration assays demonstrated that cancer stem cell features were enriched in gemcitabine-resistant cells, but decreased in sulforaphane- and quercetin-long time-treated cells. These results were confirmed by orthotopic xenotransplantation of cancer cells to the mouse pancreas, where Bx-GEM formed large, Bx-Q small and Bx-SF cells almost undetectable tumors. An mRNA expression profiling array and subsequent gene set enrichment analysis and qRT-PCR confirmed that tumor progression markers were enriched in Bx-GEM, but reduced in Bx-SF and Bx-Q cells. This study demonstrates that the continuous exposure of pancreatic cancer cells to sulforaphane or quercetin does not induce resistance in surviving cells but reduces tumorigenicity by inhibition of tumor progression markers. These results highlight that cancer cells may not adapt to the preventive and therapeutic effects of a regular fruit- and vegetable-based diet.

Hemostatic variables, plasma lactate concentration, and inflammatory biomarkers in dogs with gastric dilatation-volvulus.

OBJECTIVE: Prospective characterization of hemostastatic variables, plasma lactate concentration, and inflammatory biomarkers in dogs with gastric dilatation-volvulus (GDV). MATERIAL AND METHODS: Coagulation variables (platelets, prothrombin time [PT], activated partial thromboplastin time [aPTT], fibrinogen, antithrombin [AT], protein C [PC], protein S [PS], D-dimers), plasma lactate concentration and inflammatory biomarkers (C-reactive protein, white blood cell [WBC] count, lymphocyte and neutrophil numbers) were assessed in 20 dogs with GDV presented between 2011 and 2012. Blood was taken preoperatively and at days 1 and 3 postoperatively. The prognostic value of these variables before and after surgery was evaluated as well as the behavior of variables during the study. RESULTS: Overall, 7/20 (35%) dogs did not survive; two dogs (29%) were euthanized during surgery due to severe gastric necrosis and 5 (71%) dogs after surgery due to sepsis and disseminated intravascular coagulopathy. Prior to surgery, median plasma lactate concentration was significantly (p = 0.01) lower in survivors (6.2 mmol/l, range 1.9-9.7 mmol/l) when compared to non-survivors (11.8 mmol/l, range 7.5-16.2 mmol/l). In dogs dying after surgery, significantly higher plasma lactate concentration, coagulation times and D-dimer concentration were present as well as lower fibrinogen concentration and activity of PC and AT compared to survivors. At discharge, activity of AT, PC and PS were markedly below the reference interval in 6/13 (46%), 11/13 (85%), and 8/13 (62%) dogs, respectively. CLINICAL RELEVANCE: Only lactate plasma concentration was of preoperative prognostic value. After surgery, severe abnormalities of coagulation variables, especially the endogenous anticoagulants were present in most of the dogs. The severity of the abnormalities was associated with survival.

Computer Decision Support Changes Physician Practice But Not Knowledge Regarding Autism Spectrum Disorders.

OBJECTIVE: To examine whether adding an autism module promoting adherence to clinical guidelines to an existing computer decision support system (CDSS) changed physician knowledge and self-reported clinical practice. METHODS: The CHICA (Child Health Improvement through Computer Automation) system, a CDSS, was enhanced with a module to improve management of autism in 2 of the 4 community pediatric clinics using the system. We examined the knowledge and beliefs of pediatric users using cross-sectional surveys administered at 3 time points (baseline, 12 months and 24 months post-implementation) between November 2010 and January 2013. Surveys measured knowledge, beliefs and self-reported practice patterns related to autism. RESULTS: A total of 45, 39, and 42 pediatricians responded at each time point, respectively, a 95-100% response rate. Respondents' knowledge of autism and perception of role for diagnosis did not vary between control and intervention groups either at baseline or any of the two post-intervention time points. At baseline, there was no difference between these groups in rates in the routine use of parent-rated screening instruments for autism. However, by 12 and 24 months post-implementation there was a significant difference between intervention and control clinics in terms of the intervention clinics consistently screening eligible patients with a validated autism tool. Physicians at all clinics reported ongoing challenges to community resources for further work-up and treatment related to autism. CONCLUSIONS: A CDSS module to improve primary care management of ASD in pediatric practice led to significant improvements in physician-reported use of validated screening tools to screen for ASDs. However it did not lead to corresponding changes in physician knowledge or attitudes.

[Extreme neutrophilia in cats - aetiology and prognosis]. / Extreme Neutrophilie bei der Katze - Ätiologie und Prognose.

OBJECTIVE: The aim of this retrospective study was to evaluate the aetiology and prognostic factors of extreme neutrophilia in cats. MATERIAL AND METHODS: Patient data over a 5-year period (January 2008  -  December 2013) were reviewed. Cats with a neutrophil count > 40 x 109/l were included. They were further assigned to four groups: "inflammation", "neoplasia", "immune-mediated diseases", "unknown aetiology". Clinical signs, rectal temperature, hospitalisation, duration of hospitalisation, survival, left-shift and toxicity of neutrophils were evaluated. RESULTS: In total, 28/5185 cats (0.5%) displayed extreme neutrophilia with a mean neutrophil count of 48.5 x 109/l (40.0-76.0 x 109/l). The most common aetiology was a severe inflammation, as seen in 16/28 cats (57%), whereby peritonitis (5/15 cats, 31%) predominated. In cats with neoplastic diseases (9/28 cats, 32%), intestinal neoplasia with subsequent peritonitis was the most common diagnosis (4/9 cats, 44%). Diseases of unknown aetiology (2/28 cats, 7%) and immune-mediated diseases (1/28, 3.6%) were rare. The most common clinical indications included lethargy, anorexia, fever, and gastrointestinal signs. Rectal temperature ranged between 33.9  °C and 40.2   °C, whereby in 2/24 cats (8%) hyperthermia (> 39.3°C) and in 5/24 cats (21%) hypothermia (< 38.0°C) was observed. Hospitalisation occurred in 21/28 cats (75%) with a median duration of 5.5 days (1-30 days). In 24/28 cats, a manual differential count was performed. A left-shift and toxicity of neutrophils were seen in 23/24 cats (96%) and 21/24 cats (88%), respectively. The overall median survival rate was 50%, whereby the survival rate was significantly lower in cats with neoplasia than in those with inflammatory diseases (22% vs. 56%, p < 0.0001). CLINICAL RELEVANCE: An extreme neutrophilia is rare. It is commonly caused by peritonitis due to foreign bodies or ruptured intestinal tumours (in particular, intestinal lymphomas) and is characterised by a high mortality.

Complexation of Cm(III) with the recombinant N-lobe of human serum transferrin studied by time-resolved laser fluorescence spectroscopy (TRLFS).

The complexation of Cm(III) with the recombinant N-lobe of human serum transferrin (hTf/2N) is investigated in the pH range from 4.0 to 11.0 using TRLFS. At pH ≥ 7.4 a Cm(III) hTf/2N species is formed with Cm(III) bound at the Fe(III) binding site. The results are compared with Cm(III) transferrin interaction at the C-lobe and indicate the similarity of the coordination environment of the C- and N-terminal binding sites with four amino acid residues of the protein, two H2O molecules and three additional ligands (e.g. synergistic anions such as carbonate) in the first coordination sphere. Measurements at c(carbonate)tot = 0.23 mM (ambient carbonate concentration) and c(carbonate)tot = 25 mM (physiological carbonate concentration) show that an increase of the total carbonate concentration suppresses the formation of the Cm(III) hTf/2N species significantly. Additionally, the three Cm(III) carbonate species Cm(CO3)(+), Cm(CO3)2(-) and Cm(CO3)3(3-) are formed successively with increasing pH. In general, carbonate complexation is a competing reaction for both Cm(III) complexation with transferrin and hTf/2N but the effect is significantly higher for the half molecule. At c(carbonate)tot = 0.23 mM the complexation of Cm(III) with transferrin and hTf/2N starts at pH ≥ 7.4. At physiological carbonate concentration the Cm(III) transferrin species II forms at pH ≥ 7.0 whereas the Cm(III) hTf/2N species is not formed until pH > 10.0. Hence, our results reveal significant differences in the complexation behavior of the C-terminal site of transferrin and the recombinant N-lobe (hTf/2N) towards trivalent actinides.

A retrospective study of 1,098 blood samples with anemia from adult cats: frequency, classification, and association with serum creatinine concentration.

BACKGROUND: Frequency and classification of anemia in terms of regeneration status and erythrocyte indices are not well described in cats. OBJECTIVE: To determine frequency and regenerative status of anemia in samples from adult cats, to assess the sensitivity and specificity of macrocytosis and hypochromasia for detecting regenerative anemia (RA), and to evaluate the association of anemia with increased serum creatinine concentration (SC). STUDY POPULATION: Laboratory records from 30,503 blood samples from cats (2003-2011). METHODS: Clinicopathologic data reviewed retrospectively. Anemia defined as hematocrit (Ht) ≤27%, red blood cell count (RBC) ≤5.5 × 10(6)/µL and hemoglobin (Hb) ≤9.0 g/dL. RA defined by manual absolute reticulocyte count >50 × 10(3)/µL. Macrocytosis was defined as mean corpuscular volume (MCV) >55 fL and hypochromasia as mean corpuscular hemoglobin concentration (MCHC) <31 g/dL. Cutoff for increased serum creatinine concentration was 1.6 mg/dL. RESULTS: Overall, 1,098 of 30,503 blood samples (3.6%) from cats fulfilled criteria for anemia, 633 of 1,098 (57.7%) classified as nonregenerative (NRA) and 465 of 1,098 (42.3%) as regenerative. RBC, Ht, and Hb were significantly lower in the RA compared to NRA group (P < .05). Sensitivity and specificity of the combined high MCV and low MCHC to detect samples with RA were 19.5 and 90.7%. SC was increased in 572 of the 1,098 anemic samples (52.1%) and in 11,121 of 29,405 of nonanemic samples (37.8%). CONCLUSIONS AND CLINICAL IMPORTANCE: Majority of anemic samples were classified as NRA. Anemia was more severe in cats with RA. Erythrocyte indices were not sensitive indicators of RA.

Short-term glucocorticoid treatment causes spinal osteoporosis in ovariectomized rats.

PURPOSE: In humans, glucocorticoid-induced osteoporosis is the most common cause of medication-induced osteoporosis. Recent clinical data suggest that glucocorticoid therapy increases the risk of vertebral fractures within a short treatment period. Therefore, this study aimed at investigating vertebral bone in a rat model of glucocorticoid-induced postmenopausal osteoporosis. METHODS: Fifty Sprague-Dawley rats were randomly assigned into three groups: 1) untreated controls, 2) Sham-operated group, and 3) ovariectomized rats treated with glucocorticoid (dexamethasone) for 3 months (3M) after recovery from bilateral ovariectomy. Osteoporotic bone status was determined by means of the gold standard dual energy X-ray absorptiometry (DEXA) scan. Vertebral bodies were examined using µCT, histological analysis, mRNA expression analysis, and biomechanical compression testing. Further systemic effects were studied biochemically using serum marker analysis. RESULTS: Dexamethasone treatment showed at 3M a significantly lower bone mineral density in ovariectomized rats compared to Sham-operated control (p < 0.0001) as analyzed in vivo by DEXA. Furthermore, Z scores reached levels of -5.7 in the spine indicating sever osteoporotic bone status. Biomechanical testing of compression stability indicated a lower functional competence (p < 0.0001) in the spine of treated rats. µCT analysis showed significant reduction of bone volume density (BV/TV%; p < 0.0001), significantly enhanced trabecular spacing (Tb.Sp; p < 0.0001) with less trabecular number (Tb.N; p < 0.001) and complete loss of trabecular structures in glucocorticoid-treated ovariectomized rats. Histological analysis by osteoblast and osteoclast activities reflected a higher bone catabolism reflected by osteoclast counts by TRAP (p < 0.019) and lower bone catabolism indicated by ALP-stained area (p < 0.035).Serum analysis showed a significant increase in osteocalcin (p < 0.0001), osteopontin (p < 0.01) and insulin (p < 0.001) at 3M. Expression analysis of molecular markers in the vertebral body revealed lower expression in tenascin C in the OVX-steroid animals at 3M. CONCLUSIONS: Short-term glucocorticoid treatment of ovariectomized rats indicates according to DEXA standards a severe osteoporotic bone status in vertebral bone. Nonetheless, dysfunctional bone anabolism and enhanced bone catabolism are observed. Alterations of bone extracellular matrix proteins that correlate to inferior mechanical stability and affected microstructure were noticed and suggest further investigation. Treatment with dexamethasone was also seen to affect insulin and osteopontin levels and thus osteoblast function and maturation. This described animal model presents a recapitulation of clinically obtained data from early phase glucocorticoid-induced osteoporosis observed in patients.

[Radio-induced breast morphoea: an unrecognized complication]. / Morphée du sein radio-induite : une complication méconnue.

Morphoea is a localized scleroderma. Since the 1990s, it is described as a complication after irradiation for breast cancer. This complication is unrecognized and underdiagnosed. Irradiation seems to be a factor inducing an autoimmune process. Clinicians should be aware in case of an erythema in a treated area, appearing on average one year after the end of the irradiation. Histology alone can prove the diagnosis and rule out differential diagnoses: principally mastitis (carcinomatous, infectious), chronic radiodermatitis or radiation recall. Treatment of this rare complication is not consensual; it is most often topical steroids. The evolution is marked by a gradual decrease from the initial inflammation. Sclerosis persists, more or less substantially.

Thermal inactivation of a single strain each of serotype O26:H11, O45:H2, O103:H2, O104:H4, O111:H⁻, O121:H19, O145:NM, and O157:H7 cells of Shiga toxin-producing Escherichia coli in wafers of ground beef.

For each of two trials, freshly ground beef of variable fat content (higher: 70:30 %lean:%fat; lower: 93:7 %lean:%fat) was separately inoculated with ca. 7.0 log CFU/g of a single strain of Escherichia coli serotypes O26:H11, O45:H2, O103:H2, O104:H4, O111:H⁻, O121:H19, O145:NM, and O157:H7. Next, ca. 3-g samples of inoculated beef were transferred into sterile filter bags and then flattened (ca. 1.0 mm thick) and vacuum sealed. For each temperature and sampling time, three bags of the inoculated wafers of beef were submerged in a thermostatically controlled water bath and heated to an internal temperature of 54.4°C (130°F) for up to 90 min, to 60°C (140°F) for up to 4 min, or to 65.6°C (150°F) for up to 0.26 min. In lower fat wafers, D-values ranged from 13.5 to 23.6 min, 0.6 to 1.2 min, and 0.05 to 0.08 min at 54.4, 60.0, and 65.6°C, respectively. Heating higher fat wafers to 54.4, 60.0, and 65.6°C generated D-values of 18.7 to 32.6, 0.7 to 1.1, and 0.05 to 0.2 min, respectively. In addition, we observed reductions of ca. 0.7 to 6.7 log CFU/g at 54.4°C after 90 min, ca. 1.1 to 6.1 log CFU/g at 60.0°C after 4 min, and 0.8 to 5.8 log CFU/g at 65.6°C after 0.26 min. Thus, cooking times and temperatures effective for inactivating a serotype O157:H7 strain of E. coli in ground beef were equally effective against the seven non-O157:H7 Shiga toxin-producing strains investigated herein.

Characterisation of changes in the haemostasis system in dogs with thrombosis.

OBJECTIVE: To prospectively assess (2008 to 2009) all phases of coagulation including thromboelastography in dogs with thrombosis to determine if thrombosis is associated with hypercoagulable states. METHODS: Coagulation reaction in dogs with thrombosis (n = 7) diagnosed by diagnostic imaging or histopathology was compared with 56 control dogs. Dogs pretreated with antiplatelet and anticoagulation drugs were excluded. Thromboelastographic G-values >10 · 3 Kdyn/cm(2) were used to define a hypercoagulable state. RESULTS: Compared with the controls, there was a significantly higher mean platelet component indicating lower platelet activation status (17 · 99 ±1 · 36 versus 20 · 48 ±2 · 04 g/dL, P = 0 · 0004), increased thromboelastographic G-value (6 · 4 ±1 · 6 versus 13 · 1 ±6 · 2 Kdyn/cm(2) , P = 0 · 0029), activated partial thromboplastin time, fibrinogen, D-dimers (all: P < 0 · 0001) as well as decreased antithrombin (P = 0 · 0049), factor VIII (P < 0 · 0001), protein C (P < 0 · 0001), protein S (P = 0 · 0373) and activated protein C-ratio (P = 0 · 0013). On the basis of thromboelastographic G-value, three of six thromboelastographic tracings were classified as normocoagulable and three as hypercoagulable. CLINICAL RELEVANCE: In dogs with thrombosis, both normo- and hypercoagulable states are present and activated protein C resistance is common.

Automated and visual analysis of commercial urinary dipsticks in dogs, cats and cattle.

Two dipsticks developed for human use were evaluated for routine urinalysis and for detection of proteinuria in dogs (n=101), cats (n=50) and cattle (n=100). The aims were to determine their diagnostic usefulness in dogs, cats and cattle and to compare automated versus visual methods of reading. Results obtained with automated reading correlated better with reference methods than visual reading. Correlation with the reference methods was good to excellent for automated estimation of creatinine (dog: r(s)=0.86, cat: r(s)=0.83, cattle: r(s)=0.87) and pH (dog: r(s)=0.96, cat: r(s)=0.91, cattle: rs=0.94). The correlation was good for protein (dog: r(s)=0.88, cat: r(s)=0.91), glucose (cat: r(s)=0.83) and urine protein:creatinine (UPC) ratio (dog: r(s)=0.75, cat: r(s)=0.89). Estimation of proteinuria in cattle and pyuria in cats lacked specificity and detection of isosthenuria lacked sensitivity in all species. Semiquantitative estimation of UPC ratio was specific (100% and 91.2% at a cut-off of 0.2 in cats and 0.4 in dogs, respectively).

[Bacterial susceptibility testings of the lower airways of diseased dogs]. / Bakterienisolate aus dem unteren Respirationstrakt von erkrankten Hunden und deren aktuelle Resistenzsituation.

OBJECTIVE: In this retrospective study over 5 years the bacterial flora of the deep airways of diseased dogs was sampled using bronchioalveolar lavage and its in-vitro susceptibility to numerous antimicrobial agents was analysed. MATERIAL AND METHODS: Susceptibility testing of bacterial isolates from 84 dogs performed from 2004 to 2009 were evaluated and compared to data collected in 1999/2000. RESULTS: The 99 bacterial isolates comprised Pasteurella spp. (27.3%), Bordetella bronchiseptica (20.2%), Staphylococcus spp. (18.2%), Escherichia coli (15.2%), Klebsiella spp. (8.1%), Pseudomonas spp. (7.0%), and Streptococcus spp. (4.0%). Lavage samples of eight dogs revealed simultaneous occurrence of more than one species. The majority of the tested Bordetella bronchiseptica-isolates were susceptible to fluoroquinolones as well as tetracycline, doxycycline and polymyxin B. The number of amoxicillin/clavulanic acid- and chloramphenicol-resistant isolates decreased compared to 1999/2000. The increase of chloramphenicol-susceptible isolates was statistically significant. The tested Staphylococcus-isolates revealed a susceptibility to fluoroquinolones, amoxicillin, cephalexin, doxycycline, and polymyxin B. Compared to the earlier study, a major portion exhibited susceptibility to chloramphenicol and tetracycline. None of the Klebsiella spp. tested in 1999/2000 was resistant to enrofloxacin, whereas only 62.5% of the isolates examined during 2004-2009 were susceptible. A susceptibility of all tested Klebsiella -isolates was detectable with respect to polymyxin B. Regarding E.coli , none of the tested antibiotics revealed an in-vitro activity against all of the tested isolates. The increase of amoxicillin/clavulanic acid-resistant isolates was statistically significant. CONCLUSION AND CLINICAL RELEVANCE: The study demonstrates the benefits of bronchioalveolar lavage as a diagnostic tool to ensure a responsible utilisation of antibiotics. Most of the tested bacterial isolates were susceptible to fluoroquinolones.

[Stereotactic radiotherapy of intracranial benign tumors]. / Radiothérapie en conditions stéréotaxiques des tumeurs bénignes intracrâniennes.

Stereotactic radiotherapy can be delivered in one fraction or in multiple fractions schedule. It is used in benign tumours such as meningiomas, mainly localized in the base of the skull, for acoustic schwannoma and pituitary tumours. Whatever the tumour, results with the Gamma Knife(®) are the most numerous, but those obtained by linear accelerators, adapted or dedicated, are comparable. The peripheral dose is preferred to the dose delivered to the isocentre. One fraction stereotactic irradiation should be proposed in small lesions and fractionated treatment for tumours larger. Whatever the tumour, the results are satisfactory with a control rate of 90%. However, this value reflects a disparity assessment, radiological stability for meningiomas, radiological stability and preservation of useful hearing in schwannoma and radiological stability and a decrease in hormonal secretions for pituitary adenomas. Overall complication rates are low. In total, the treatment of benign lesions with stereotactic irradiation gives satisfactory results with few complications.

Effect of experimental endotoxemia on thrombelastography parameters, secondary and tertiary hemostasis in dogs.

BACKGROUND: Thrombelastography (TEG) and indicators of secondary and tertiary hemostasis might be altered in dogs with endotoxemia. HYPOTHESIS: Endotoxemia influences measures of coagulation in dogs. ANIMALS: Ten healthy cross-bred dogs. MATERIAL AND METHODS: Prospective laboratory study between controls (n = 5) receiving 0.9% saline IV and the study group (n = 5) treated with low-dose lipopolysaccharide (0.02 mg/kg IV). Physical examination and sampling for measurement of leukocytes, platelets, and coagulation variables were performed at time points 0, 1, 4, and 24 hours. Coagulation variables included kaolin-activated TEG, 1-stage prothrombin time (OSPT), activated partial thromboplastin time (aPTT), fibrinogen, factor VIII, antithrombin, protein C, protein S, activated protein C (APC)-ratio calculated from aPTT with and without presence of APC), and D-Dimers. RESULTS: Endotoxemia-induced clinical signs included lethargy (n = 5/5), diarrhea (n = 4/5), emesis (n = 4/5), and abdominal pain (2/5). After 1 hour there was severe leukopenia (2.5 ± 0.7 × 10(9)/L; mean ± SD, P < .0001) and a 2.2-fold increase in D-Dimers (0.81 ± 0.64 mg/L, P < .0001). After 4 hours there was hyperthermia (40.3 ± 0.4°C, P < .0001) and increases in OSPT (10.5 ± 2.7 seconds, P < .0001), aPTT (16.7 ± 5.2 seconds, P = 0.002). A significant decrease in fibrinogen (1.5 ± 1.0 g/L, P = 0.001), protein C (31 ± 33%, P < .0001), protein S (63 ± 47%, P < .0001), TEG α (58 ± 19, P = .007), and TEG maximal amplitude (50 ± 19 mm, P = .003) was seen compared with the controls. APC-ratio rose significantly (2.5 ± 0.2, P < .0001) without exceeding the reference interval (n = 4/5). CONCLUSION AND CLINICAL IMPORTANCE: D-Dimers are the earliest indicator for endotoxemia-associated coagulation abnormalities followed by decreased protein C concentration. APC-ratio and TEG were not good screening variables.