Prevalence of Swimming Puppy Syndrome in 2,443 Puppies during the Years 2006–2012 in Thailand

The purpose of this study was to report on the prevalence of swimming puppy syndrome (SPS) and investigate predisposing factors. Data were recorded from 2,443 puppies (1,183 males and 1,260 females) in Thailand, October 2006–September 2012, including breed, sex, number of puppies per litter, type of nest floor, number of affected limbs, and occurrence of pectus excavatum. Fifty-two puppies (2.13%) were diagnosed with SPS. The breed most frequently affected was English Bulldog (8.33%). There was no significant difference (P>0.05) between presence and absence of disease based on sex, breed, and nest floor type. The number of puppies per litter was associated with SPS; puppies from smaller litters (1.92 ± 1.12) had a higher prevalence of the disease (P<0.01) than puppies from larger litters (3.64 ± 2.24). Moreover, 15.38% of puppies with affected limbs showed signs of pectus excavatum (8/52); this clinical sign was more prevalent (P<0.01) in puppies with all four limbs affected with SPS

Functional analysis of genes during bovine preimplantation embryo development

This study, the RNA interference (RNAi) approach was applied to suppress the expression of the maternal (C-mos) and embryonic (Oct-4) transcripts in bovine oocytes and embryos, respectively using microinjection of sequence specific double-stranded RNA (dsRNA). For this 435 bp C-mos and 341 bp Oct-4 dsRNA were synthesised and microinjected into the cytoplasm of immature oocytes and zygotes, respectively. In experiment 1, immature oocytes were categorized into three groups: those injected with C-mos dsRNA, water (RNase-free), and uninjected controls. In experiment 2, in vitro produced zygotes were categorized into three groups: those injected with Oct-4 dsRNA, water (RNase-free) and uninjected controls. The developmental phenotypes, the level of mRNA and protein expression were investigated after treatment in both experiments. Microinjection of C-mos dsRNA has resulted in reduction of C-mos transcript (70%) and protein after maturation compared to the water injected and uninjected controls (P Funktionelle Analyse von Genen in boviner preimplantativer Entwicklung In dieser Arbeit wurde der RNA interference (RNAi) Ansatz angewendet, um die Expression maternaler C-mos und embryonaler Oct-4 Transkripte in bovinen Oozyten bzw. Embryos mittels Mikroinjektion sequenzspezifischer doppelsträngiger RNA (dsRNA) zu unterdrücken. Hierzu wurden 435 bp C-mos und 341 bp Oct-4 dsRNA synthetisiert und in das Zytoplasma immaturer Oozyten bzw. Zygoten mikroinjiziert. In Experiment 1 wurden die immaturen Oozyten in drei Gruppen eingeteilt: mit C-mos dsRNA injizierte, mit Wasser (RNase-freiem) injizierte und uninjizierte Kontrollen. In Experiment 2 wurden die in vitro produzierten Zygoten in drei Gruppen eingeteilt: mit Oct-4 injizierte, mit Wasser (RNase-freiem) injizierte und uninjizierte Kontrollen. Entwicklungsphänotypen, mRNA Level und Protein Expression wurden nach der Behandlung in beiden Experimenten untersucht. Die Mikroinjektion von C-mos dsRNA resultierte in einer Reduktion von C-mos Transkript (70%) und Protein nach Maturation verglichen mit mit Wasser injizierten und uninjizierte Kontrollen (P < 0.01). 60% der mit C-mos dsRNA injizierten Oozyten zeigten Extrusion des ersten Polrkörpers verglichen mit 50% der mit Wasser injizierten und 44% der uninjizierten Kontrollen. Überdies zeigten einzig mit C-mos dsRNA injizierte Oozyten spontane Aktivierung. Die Mikroinjektion von Zygoten mit Oct-4 dsRNA resultierte ebenfalls in einer Reduktion von Oct-4 Transkriptabundanz (72%) und Protein im Blastozystenstadium verglichen mit den uninjizierten Kontrollzygoten (P < 0.01). Teilungs-, Morula- und Blastozystenrate waren nicht signifikant unterschiedlich zwischen den 3 Behandlungsgruppen. Ferner wurde eine signifikante Reduktion der Anzahl der Zellen der inneren Zellmasse in Oct-4 dsRNA injizierten Embryos beobachtet im Vergleich zu den anderen Gruppen. Diese Resultate demonstrieren, dass sequenzspezifische dsRNA zum Knockdown maternaler oder embryonalen Transkripte in der bovinen Embryogenese genutzt werden um die Funktion von Genen zu untersuchen

Articular Cartilage Gene Expression after Coxofemoral Joint Luxation in the Dog

This study examined the relationship between days of hip luxation and the expression of various mRNA. Twenty-six articular cartilages were used in the experiment: 3 samples were from normal dogs and 23 samples were collected from the femoral heads of hips that had been luxated for different lengths of time. Ten mRNA, including nonapoptotic genes (AGG, COL2A1, MMP-3, HAS-1, HAS-2, and TIMP-1) and apoptotic genes (BAX, BCL-2, CAS-3, and CAS-9), were studied for their expression using real-time PCR. We found very high correlation between expression level and luxation days (r2>0.9) in COL2A1, MMP-3, HAS-1, HAS-2, TIMP-1, BAX, and CAS-9, while the others (AGG, BCL-2, and CAS-3) also showed high correlation (r2=7–9). And we found a significant difference (P<0.05) in the expression of transcripts depending on the number of luxation days. In conclusion, a delay in joint reduction may increase the chances of development of osteoarthritis

The effect of doxycycline on canine hip osteoarthritis: design of a 6-months clinical trial

Twenty-five dogs were included in a randomized, double-blind trial to assess the efficacy of doxycycline (DOX) orally administered twice a day at 4 mg/kg/day (n = 12) for the treatment of osteoarthritis of the hip. Chondroitin sulfate (CS; 525 mg/day) was used as a positive control (n = 13). Dogs were re-examined monthly for 6 months after initiation of treatment. The assessment protocol included clinical score, radiographic findings and serum osteoarthritis biomarkers. Dogs treated with DOX showed statistically significant improvements (p < 0.05) in lameness, joint mobility, pain on palpation, weight-bearing and overall score at 2, 6, 4, 4 and 4 months, respectively, after treatment. Biomarker levels of CS-WF6 epitope and hyaluronan were significantly increased and decreased (p < 0.05) at 2 and 3 months after treatment compared to pretreatment. These results showed that DOX had a positive therapeutic effect in dogs with osteoarthritis

Heart rate change during aquatic exercise in small, medium and large healthy dogs

Abstract This study was divided into two experiments. For the first experiment, heart rates during exercise of 21 healthy male dogs were recorded. The animals were brought to swim every 2 days, 8 times in all, for 10 minutes. Heart rates were measured every minute using a pulse watch. For the second experiment, 134 healthy adult dogs were categorized into three groups: small (41), medium (51) and large breed (42). Their heart rates were measured every minute for 34 minutes after the 5 th swimming. In the first experiment, the heart rates during the 1 st -4 th swimming were significantly higher (p&lt;0.05) than during the 5 th 8 th swimming. In the second experiment, the heart rates were significantly different (p&lt;0.05) between small, medium and large dogs. No correlations were found between weight, age, and heart rate. From the results of the second experiment, we were able to formulate an equation for predictable heart rate of each group of dogs (small, medium and large dogs). From the results, we recommend that the limits on the length of time spent for aquatic exercise should be 15-30 minutes, depending on the breed (size) of dog. Moreover, there should be trainers observing each dog to prevent over-exercise. Keywords: dog, heart rate, swimmin

Determination of two fluoroquinolones and their combinations with hyaluronan effect in in vitro canine cartilage explants

Background Previous studies reported the effect of enrofloxacin (Enro) and marbofloxacin (Mar) on cell death and alteration of the key genes involved in catabolic and anabolic processes and demonstrated the beneficial effects of hyaluronan (HA) combined with fluoroquinolones (FQs) on primary canine chondrocytes. This study further determines the effects of these treatments on canine cartilage explants in both normal and interleukin-1 beta (IL-1β)-stimulated conditions. Methods We examined sulfate glycosaminoglycan (s-GAG) release, uronic acid (UA) content, and safranin-O staining, as well as the expression patterns of inflammatory, extracellular matrix (ECM) component and enzymes. Results Enro treatment alone effectively stimulated proteoglycan anabolism by increasing UA content and glycosaminoglycans (GAGs) in normal and pre-IL-1β-stimulated explant, whereas Mar showed opposite results. The combination of HA and FQs increased s-GAG release and UA content in normal explants in addition to effective down-regulated expression of MMP3. HA reduced the adverse effects of Mar by enhancing UA and GAG contents in both normal and pre-IL-1β-explants. Moreover, HA effectively induced HAS1and ACANup-regulation and reduced MMP9, TNF, PTGS2,and NFKB1 expression for a long term. Discussion Our results suggest the direct effects of Enro and Mar may selectively stimulate the conditioned explants to express MMP-codinggenes and promote gene expression involved in matrix production, pro-inflammatory cytokines, and cell degradation in different directions. HA successfully reduced the adverse effects of FQs by enhancing s-GAG and UA contents and down-regulated expression of MMPs

In vitro suppression of the MMP-3 gene in normal and cytokine-treated human chondrosarcoma using small interfering RNA

<p>Abstract</p> <p>Background</p> <p>Matrix metalloproteinase (MMPs) synthesized and secreted from connective tissue cells have been thought to participate in degradation of the extracellular matrix. Increased MMPs activities that degrade proteoglycans have been measured in osteoarthritis cartilage. This study aims to suppress the expression of the <it>MMP-3 </it>gene in <it>in vitro </it>human chondrosarcoma using siRNA.</p> <p>Methods</p> <p>Cells were categorized into four groups: control (G.1); transfection solution treated (G.2); negative control siRNA treated (G.3); and <it>MMP-3 </it>siRNA treated (G.4). All four groups were further subdivided into two groups - treated and non-treated with IL-1β- following culture for 48 and 72 h. We observed the effects of gene suppression according to cell morphology, glycosaminoglycan (GAG) and hyaluronan (HA) production, and gene expression by using real-time polymerase chain reaction (PCR).</p> <p>Results</p> <p>In IL-1β treated cells the apoptosis rate in G.4 was found to be lower than in all other groups, while viability and mitotic rate were higher than in all other groups (<it>p </it>< 0.05). The production of GAG and HA in G.4 was significantly higher than the control group (<it>p </it>< 0.05). <it>MMP-3 </it>gene expression was downregulated significantly (<it>p </it>< 0.05).</p> <p>Conclusion</p> <p><it>MMP-3 </it>specific siRNA can inhibit the expression of <it>MMP-3 </it>in chondrosarcoma. This suggests that <it>MMP-3 </it>siRNA has the potential to be a useful preventive and therapeutic agent for osteoarthritis.</p