Pharmacokinetics of tramadol and its major metabolite after intramuscular administration in piglets

Tramadol (T) is a centrally acting atypical opioid used for treatment of dogs. Piglets might experience pain following castration, tooth clipping and tail docking and experimental procedures. The aim of this study was to assess the pharmacokinetics of T and its active metabolite M1 in male piglets after a single intramuscular injection. Six healthy male piglets were administered T (5 mg/kg) intramuscularly. Blood was sampled at scheduled time intervals and drug plasma concentrations evaluated by a validated HPLC method. T plasma concentration was quantitatively detectable from 0.083 to 8 h. M1 was quantified over a shorter time period (0.083–6 h) with a Tmax at 0.821 h. The study demonstrated that piglets produce a larger amount of M1 compared with dogs, horses and goats. The human minimum effective concentration of M1 (40 ng/mL) was exceeded for over 3 h in piglets. If it is assumed to also apply to piglets, it could be speculated that the drug efficacy might exert its action over 3 h or longer. This assumption has to be confirmed by further specific pharmacokinetic/pharmacodynamic studies

Ex vivo evaluation of imatinib mesylate for induction of cell death on canine neoplastic mast cells with mutations in c-Kit exon 11 via apoptosis

Several studies of canine spontaneous mast cell tumours have described mutations in the c-kit proto-oncogene. These mutations produce a constitutively activated product and have been suggested to play a role in the malignant transformation of mast cells. We hypothesize that the selective tyrosine kinase inhibitor imatinib mesylate inhibits signal transduction and induces apoptosis when tested in cutaneous canine mast cell tumour samples positive for mutation in c-kit exon 11. Three-dimensional ex vivo cultures of canine grade II mast cell tumour treated with STI-571 at 48, 72, and 96 h and tested for signal transduction and apoptosis using appropriate assays were used. There was a progressive and significant increase in caspase-3 and TUNEL-positive mast cells compared to the untreated cultures. Additionally, a concurrent reduced expression of Ki67 and BCL-2 was observed. Furthermore, the treated cultures showed a marked reduction of Kit expression. Our results demonstrate that STI-571 induces Caspase-dependent apoptosis in a canine neoplastic mast cells possessing mutations in c-kit exon 11. © 2013 Springer Science+Business Media Dordrecht

Guaifenesin-Ketamine-Xylazine Infusion to Maintain Anesthesia in Mules Undergoing Field Castration

Many minor and surgical procedures can be performed in the field under sedation or general anaesthesia. Numerous drug combinations have been used for sedation, induction and maintenance. The purpose of this study was to determine if the combination of guaifenesin, ketamine and xylazine, commonly referred to as “triple drip”, produce safe and satisfactory total intravenous anaesthesia in mules undergoing field castration, premedicated with xylazine and induced with thiopental. Eight healthy adult intact male mules, aged 4 to 6 years and weighing 380 to 490, were anesthetized to performe field castration. Before anaesthesia a 14-gauge, 13–cm catheter was placed percutaneously in the external jugular vein. Mules were premedicated with 1.3 mg/kg xylazine IV and anaesthesia was then inducted with 6 mg/kg IV thiopental within 10 min after premedication, when the animals were at least moderately sedated. Additional xylazine was administered when the mules were inadequately sedated. Sedation was considered good when lowering of the head, drooping of the lower lip and drooping of the ears were present using a 4-point sedation score. Once the mules were recumbent, the infusion of guaifenesin (50 mg/ml) - ketamine (20 mg/ml) - xylazine (0.5 mg/ml) (GKX) was started to maintain general anaesthesia, approximately 1ml/kg/hr (based on monitoring eye signs, muscle relaxation of the neck, respiratory rate and pattern, and the responses to surgical stimulation. The spermatic cord of each testis was infiltrated with 5 ml of lidocaine to achieve local anaesthesia before the scrotum skin incision. The open technique of castration was applied to all mules for postoperative drainage. During anaesthesia heart rate (HR), respiratory rate RR), rectal temperature (RT) and hemoglobin saturation with oxygen (SpO2) were measured every 5 minutes. Times to sternal recumbency, lateral recumbency and standing were recorded. The data recorded were statistically analysed using simple one-way analysis of variance (ANOVA) and a pvalue>0.05 was considered significant. The qualities of anaesthesia were evaluated using induction, maintenance and recovery scores. The resultes suggest that the premedication using 1.3 mg/kg IV xylazine for mules undergoing thiopental anaesthesia was satisfactory and only one animal needed a supplemental dose of xylazine (0.3 mg/kg IV) to induce better sedation. The total IV amount of thiopental for induction was sufficient to achieve lateral recumbency in all animals. Furthermore, GKX provided adequate surgical plane of general anaesthesia to performe castration in all mules, without responses to the manuality or significant modification of HH, RR, RT, and SpO2 in comparison with the basal values and to maintain a satisfactory muscle relaxation. Recovery from anaesthesia was uneventful, smooth and clinically acceptable in all mules

Characterization and Antibacterial Evaluation of Biodegradable Mannose‐Conjugated Fe‐MIL‐88NH2 Composites Containing Vancomycin against Methicillin‐Resistant Staphylococcus aureus Strains

The emergence of bacterial resistance has increased the economic burden of infectious diseases dramatically during the previous few decades. Multidrug resistance (MDR) is difficult to cure in both Gram‐negative and positive bacteria and is often incurable with traditional and broad-range antibiotics. Therefore, developing techniques to increase the antibacterial activity of therapeutic drugs is essential. Metal‐organic frameworks (MOFs) are extremely versatile hybrid materials made of metal ions coupled via organic bridging ligands. They have been widely used as an excellent vehicle for drug delivery due to their low toxicity, biodegradability, and structural stability upon loading and functionalization. The present study focused on the synthesis of mannose (MNS)‐coated MOFs with enhanced surface contact with S. aureus cells. The MNS coating on the surface of MOFs enhances their adherence to bacteria by binding to lectins present on the bacterial cell, resulting in improved VCM cellular penetration and activity against resistant bacteria. Various techniques, including atomic force microscopy, DLS, TGA, FT‐IR, and DSC, were employed to analyze MNS‐coated MOFs. They were also evaluated for their efficacy against resistant S. aureus. The results indicated that when VCM was loaded into MNS‐coated MOFs, their bactericidal activity rose dramatically, resulting in the greater suppression of resistant S. aureus. AFM investigation of S. aureus strains demonstrated total morphological distortion after treatment with MNS‐coated drug-loaded MOFs. The results of this work suggest that MNS‐coated MOFs may be effective in reversing bacterial resistance to VCM and open new pathways for improving antibiotic therapy for diseases associated with MDR

Effect of Thiamine Pyrophosphate (Bicarbossilasi®) Administration on the Exercising Horse Metabolism

Thiamine pyrophosphate (TPP) is the phosphorylated and active form of thiamine (Vitamin B1). We hypothesized that the administration of thiamine in its immediately available active form could provide the metabolic pathways for a supplement able to promote the metabolism of ketoacids and to reduce lactate accumulation in exercising horses. Ten horses were conditioned for 20 days to daily standardized exercise. All horses underwent a first “stress test” (ST) consisting in 1,200 meters at maximum speed on track, and were checked before and after for clinical and clinical pathology parameters. After the ST, the same animals were administered TPP (Bicarbossilasi®, ©Teknofarmas.p.a., Torino, Italy), 1 mg/Kg b.w., I.V., twice daily for seven days. At the end of treatment, a second stress test (STTPP) was performed. Post-exercise serum lactate concentration resulted in significantly lower levels (p< 0.05) after treatment with TPP (ST vs STTPP). These data suggest that supplementation with thiamine in its active form improves glucose metabolism and prevents lactate accumulation in muscle, enhancing aerobic capacity and metabolic pathways of glucose utilization during exercise

Fecal Protein Profile in Eight Dogs Suffering from Acute Uncomplicated Diarrhea before and after Treatment

Acute diarrhea is a very frequent condition affecting dogs; nevertheless, little is known about what happens in the GI tract during such conditions. Proteomics allows the study of proteins present in a specific biologic substrate, and fecal proteomic investigations have been recently implemented to study GI diseases in dogs. In the present study, the fecal protein profiles of eight dogs suffering from acute uncomplicated diarrhea at the time of inclusion was investigated for the first time, and then the same patients were followed, replicating two further evaluations at two subsequent time points (after 2 and 14 days from the first presentation), with the aim of gaining possible new insights regarding the pathologic changes in the gastrointestinal environment during such conditions. Two-dimensional gel electrophoresis (2-DE) was performed, followed by mass spectrometry. Nine spots, corresponding to four (groups of) proteins (i.e., albumin, alkaline phosphatase, chymotrypsin-C-like, and some immunoglobulins), showed significant differences at two or more of the three time points investigated, almost all behaving similarly and decreasing at T1 (2 days after the onset of the condition) and significantly increasing at T2 (14 days after the onset), mainly evidencing a reaction of the organism. Further studies including a greater number of patients and possibly different techniques are needed to confirm the present finding

Comparative study of 1H-NMR metabolomic profile of canine synovial fluid in patients affected by four progressive stages of spontaneous osteoarthritis

The study aimed to assess the metabolomic profile of the synovial fluid (SF) of dogs affected by spontaneous osteoarthritis (OA) and compare any differences based on disease progression. Sixty client-owned dogs affected by spontaneous OA underwent clinical, radiographic, and cytologic evaluations to confirm the diagnosis. The affected joints were divided into four study groups based on the Kallgreen-Lawrence classification: OA1 (mild), OA2 (moderate), OA3 (severe), and OA4 (extremely severe/deforming). The osteoarthritic joint's SF was subjected to cytologic examination and H-1-NMR analysis. The metabolomic profiles of the study groups' SF samples were statistically compared using one-way ANOVA. Sixty osteoarthritic joints (45 stifles, 10 shoulders and 5 elbows) were included in the study. Fourteen, 28, and 18 joints were included in the OA1, OA2, and OA3 groups, respectively (0 joints in the OA4 group). Metabolomic analysis identified 48 metabolites, five of which were significantly different between study groups: Mannose and betaine were elevated in the OA1 group compared with the OA2 group, and the 2-hydroxyisobutyrate concentration decreased with OA progression; in contrast, isoleucine was less concentrated in mild vs. moderate OA, and lactate increased in severe OA. This study identified different H-1-NMR metabolomic profiles of canine SF in patients with progressive degrees of spontaneous OA, suggesting H-1-NMR metabolomic analysis as a potential alternative method for monitoring OA progression. In addition, the results suggest the therapeutic potentials of the metabolomic pathways that involve mannose, betaine, 2-hydroxyisobutyrate, isoleucine, and lactate

Fecal proteome profile in dogs suffering from different hepatobiliary disorders and comparison with controls

In the present study, the fecal proteomes of clinically healthy dogs (HD = n. 10), of dogs showing clinical, ultrasonographic, and/or laboratory evidence of different hepatobiliary dysfunction (DHD = n. 10), and of dogs suffering from chronic hepatitis (CHD = n. 10) were investigated with an Ultimate 3000 nanoUPLC system, coupled to an Orbitrap Fusion Lumos Tribrid mass spectrometer. Fifty-two different proteins of canine origin were identified qualitatively in the three study groups, and quantitative differences were found in 55 proteins when comparing groups. Quantitatively, a total of 41 and 36 proteins were found differentially abundant in the DHD and CHD groups compared to the control HD, and 38 proteins resulted dysregulated in the CHD group as compared to the DHD group. Among the various proteins, differently abundant fecal fibronectin and haptoglobin were more present in the feces of healthy and DHD dogs than in chronic ones, leading us to hypothesize its possible diagnostic/monitoring role in canine chronic hepatitis. On the other hand, the trefoil factor 2 was increased in DHD dogs. Our results show that the analysis of the fecal proteome is a very promising field of study, and in the case of dogs suffering from different hepatobiliary disorders, it was able to highlight both qualitative and quantitative differences among the three groups included. Results need to be confirmed with western blotting and in further studies

Fecal Proteome Profile in Dogs Suffering from Different Hepatobiliary Disorders and Comparison with Controls

In the present study, the fecal proteomes of clinically healthy dogs (HD = n. 10), of dogs showing clinical, ultrasonographic, and/or laboratory evidence of different hepatobiliary dysfunction (DHD = n. 10), and of dogs suffering from chronic hepatitis (CHD = n. 10) were investigated with an Ultimate 3000 nanoUPLC system, coupled to an Orbitrap Fusion Lumos Tribrid mass spectrometer. Fifty-two different proteins of canine origin were identified qualitatively in the three study groups, and quantitative differences were found in 55 proteins when comparing groups. Quantitatively, a total of 41 and 36 proteins were found differentially abundant in the DHD and CHD groups compared to the control HD, and 38 proteins resulted dysregulated in the CHD group as compared to the DHD group. Among the various proteins, differently abundant fecal fibronectin and haptoglobin were more present in the feces of healthy and DHD dogs than in chronic ones, leading us to hypothesize its possible diagnostic/monitoring role in canine chronic hepatitis. On the other hand, the trefoil factor 2 was increased in DHD dogs. Our results show that the analysis of the fecal proteome is a very promising field of study, and in the case of dogs suffering from different hepatobiliary disorders, it was able to highlight both qualitative and quantitative differences among the three groups included

Immunosuppressive Effects of Thallium Toxicity in Nile Tilapia Fingerlings: Elucidating the Rescue Role of Astragalus membranaceus Polysaccharides

This study evaluated the immunotoxic effects of thallium (Tl) in Nile tilapia fingerlings and the recovery role of dietary Astragalus membranaceus polysaccharides (ASs). An 8-week experiment was designed where 180 fishes were randomly and equally assigned in triplicates into the six groups: the control group (CNT) was reared in unpolluted water and fed a commercial diet, two groups were fed a well-balanced commercial diet plus 1.5 and 3.0 g AS/kg diet (AS0.15 and AS0.30), respectively, the fourth group was exposed to a sublethal dose of Tl (41.9 μg l−1) [equal to 1/10 of 96-h lethal concentration 50 (LC50)], and the last two groups were fed 0.15 and 0.3% AS, respectively, and concurrently exposed to Tl (41.9 μg l−1) (AS0.15+Tl and AS0.30+Tl). Fish hematobiochemical parameters, serum immunity [nitric oxide, total immunoglobulin M (IgM) levels, and lysozyme activity], transcription of hepatic interferon-g (IFN-g), interleukin-1b (IL-1b), and tumor necrosis factor-a (TNF-a), and resistance to Aeromonas hydrophila (A. hydrophila) were assessed. Hematobiochemical parameters and serum immune indices were significantly decreased in the fish group exposed to sublethal Tl concentration compared to the CNT group. Furthermore, Tl exposure significantly induced overexpression of IL-1b, TNF-a, and IFN-g genes (4.22-, 5.45-, and 4.57-fold higher, respectively) compared to CNT values. Tl exposure also increased the cumulative mortality (%) in Nile tilapia challenged with A. hydrophila. Remarkably, the groups fed AS0.15+Tl and AS0.30+Tl significantly ameliorated all the aforementioned parameters, but did not reach CNT values. Our findings suggest the possible immunomodulating roles of dietary AS in recovering the immunotoxic effects of Tl in Nile tilapia. We can conclude that dietary AS would be useful for maintaining the immunity of Nile tilapia fingerlings