Methamphetamine intoxication in a dog: case report.

BACKGROUND: Methamphetamine abuse has undergone a dramatic worldwide increase, and represents a significant and global issue for public health. Incidents of methamphetamine intoxication and death in humans are relatively commonplace. Because of its increasing illicit availability, together with legitimate use in human medicine, accidental or intentional exposure to methamphetamine in dogs is becoming a more likely scenario. CASE PRESENTATION: A 3-year-old, 3.7 kg intact female Miniature Poodle who had been intentionally fed an unknown amount of a crystalline-like substance developed extreme agitation, seizures, tachycardia, hyperthermia, hypertension, disseminated intravascular coagulation (DIC), bloody diarrhea, and dilated pupils. Blood work revealed leukocytosis, erythropenia, lymphocytosis, thrombocytopenia, coagulation abnormalities, but all to a mild extent, together with mild elevation in both alanine aminotranferease (ALT) and alkaline phosphatase (ALKP), and a mild decreased in glucose. Radiologic diagnosis revealed generalized, severe distension of the stomach and small intestinal tract with air. Immunochromatographic screening tests and gas chromatography mass spectrometry analysis confirmed methamphetamine intoxication and revealed concentrations of methamphetamine in blood and urine of 0.32 µg/mL and 2.35 µg/mL respectively. The dog demonstrated progressive improvement after supportive care, with the high fever resolved over the initial 24 hours of hospitalization, and agitation was successfully controlled beyond 48 hours after initial hospitalization. Hemostatic abnormalities were progressive improved after heparin therapy and supportive care. By the sixth day of hospitalization the dog was clinically well, and all laboratory data had returned to normal with the exception of a mild elevateion of ALKP. CONCLUSION: To the authors' knowledge, this is the second case report of methamphetamine intoxication in dogs presented in veterinary practice in open literature so far. Although rare, methamphetamine intoxication should be considered as a differential diagnosis in dogs with a toxic substance ingestion history and with typical nervous and cardiovascular system symptoms. In such cases rapid diagnosis and aggressive intervention is important for prognosis. Blood methamphetamine concentration may be a helpful value for assessment of the severity of intoxication and prediction of clinical outcomes.

Methylseleninic acid restricts tumor growth in nude mice model of metastatic breast cancer probably via inhibiting angiopoietin-2.

BACKGROUND: Angiopoietin-2 (Ang-2) plays critical roles in vascular morphogenesis and its upregulation is frequently associated with various tumors. Previous studies showed that certain selenium compounds possess anti-tumor effects. However, the underlining mechanism has not been elucidated in detail. Plus, results of research on the anti-tumor effects of selenium compounds remain controversial. METHODS: We investigated levels of Ang-2 and vascular endothelial growth factor (VEGF) on the estrogen-independent bone metastatic mammary cancer (MDA-MB-231) cells in response to treatment by methylseleninic acid (MSeA), and further examined the effects of MSeA oral administration on xenograft mammary tumors of athymic nude mice by RT-PCR, Western, radioimmuno assay, and Immunohistochemistry. RESULTS: Treatment of MDA-MB-231 cells with MSeA caused significant reduction of Ang-2 mRNA transcripts and secretion of Ang-2 proteins by the cells. Level of VEGF protein was accordingly decreased following the treatment. Compared with the controls, oral administration of MSeA (3 mg/kg/day for 18 days) to the nude mice carrying MDA-MB-231 induced tumors resulted in significant reduction in xenograft tumor volume and weights, significant decrease in microvascular density, and promotion of vascular normalization by increasing pericytes coverage. As expected, level of VEGF was also decreased in MSeA treated tumors. CONCLUSIONS: Our results point out that MSeA exerts its anti-tumor effects, at least in part, by inhibiting the Ang-2/Tie2 pathway, probably via inhibiting VEGF.

Design and characterization of novel oxyntomodulin derivatives with potent dual GLP-1/glucagon receptor activation and prolonged antidiabetic effects.

AIMS: To investigate the combination of dimerization and PEGylation to enhance the receptor activation and in vivo stability of Oxyntomodulin (OXM). MAIN METHODS: All LDM peptides were produced by using standard method of solid phase synthesis. The in vitro effects of LDM peptides were assessed by glucagon-like peptide-1 receptor (GLP-1R) and glucagon receptor (GcgR) binding test and Proteolytic stability test. Subsequently, saline, Liraglutide and three doses of LDM-3 treated groups were subjected to the evaluation of aute and long-term efficacy. KEY FINDINGS: Five long-acting OXM conjugates, termed LDM-1 to LDM-5, were designed using cysteine (Cys)-specific modification reaction including the activated PEG, bisMal-NH2, and OXM-Cys, and all prepared with high purity. LDM-3 exhibited greater GLP-1R and GcgR activation and ameliorative serum stability. In addition, LDM-3 was identified with enhanced insulinotropic and glycemic abilities in the gene knockout mice. The prolonged glucose-lowering effects of the LDM-3 were proved by hypoglycemic duration test and multiple oral glucose tolerance tests (OGTTs) in the diet-induced obesity (DIO) mice. Furthermore, the pharmacokinetic tests in Sprague Dawley (SD) rat and cynomolgus monkey exhibited the lifespans of LDM-3 at 90 nmol·kg-1 were 101.5 h and 119.4 h, respectively. Nevertheless, consecutive 8-week administration of LDM-3 improved the cumulative body weight gain, food intake, % HbA1c, glucose tolerance and the pancreatic of the obese mice. SIGNIFICANCE: LDM-3, as a dual GLP-1R and GcgR agonist, holds potential to be developed as a promising therapeutic candidate for both diabetes and obesity.

TLR4 signaling promotes the expression of VEGF and TGFbeta1 in human prostate epithelial PC3 cells induced by lipopolysaccharide.

Chronic inflammation promotes tumor development and progression, and Toll-like receptors (TLRs) may play an important role in this process. In this study, we found that human prostate epithelial PC3 cells constitutively express TLR4 in mRNA and protein level. lipopolysaccharide (LPS) promotes the expression and secretion of immunosuppressive cytokine TGFbeta(1) and proangiogenic factor VEGF in human prostate epithelial PC3 cells. We further elucidated that functionally activation of TLR4 is essential for the increased VEGF and TGFbeta(1) mRNA expression in the cells. In addition, after LPS stimulation, the increased expression of NF-(K)B p65 protein was also detected in human PC3 cells. Our results demonstrate that TLR4 expressed on human PC3 cells is functionally active, and may play important roles in promoting prostate cancer immune escape, survival, progression, and metastasis by inducing immunosuppressive and proangiogenic cytokines.

Phenotypic Characterization of Porcine IFNγ-Producing Lymphocytes in Porcine Reproductive and Respiratory Syndrome Virus Vaccinated and Challenged Pigs.

Porcine reproductive and respiratory syndrome (PRRS) continues to be one of the most important swine diseases worldwide. Interferon-γ (IFNγ)-mediated type I cell-mediated immune response plays an important role in protection from, and clearance of, PRRS virus (PRRSV). Several lymphocyte subsets including T-helper, CTLs, Th/memory cells, and γδ T lymphocytes were previously reported to produce IFNγ during PRRSV infection. However, the proportion and phenotypic characterization of these IFNγ-secreting lymphocytes have not been explored. In this study, IFNγ producted by different lymphocyte subsets was assessed by multi-color flow cytometry after vaccination with PRRSV modified live vaccine (PRRSV-MLV) and challenge with homogeneous or heterogeneous PRRSV. The results showed that T-helper cells were the major IFNγ-secreting cell population after PRRSV-MLV vaccination and PRRSV challenge. Additionally, the proportion of IFNγ producing Th/memory cells and γδ T cells increased after PRRSV challenge. This difference was accounted for an enhanced ability to produce IFNγ in Th/memory cells and an enlarged quantity of γδ T cells. The results presented here could contribute to our understanding of the roles of IFNγ in protective immunity against PRRSV infection and may be useful for assessment of cell-mediated immunity in vaccine tests.

Sevoflurane suppresses microglial M2 polarization.

Microglia can be polarized into the classical (M1) or alternative (M2) activation states in response to various stimuli. The M2 phenotype has its own set of receptor profiles, cytokine production, and chemokine secretion, of which arginase 1 (Arg1), chitinase 3-like 3 (Chi3l3, Ym1), and IL-10 have neuroprotective properties. Sevoflurane is one of the most commonly used volatile anesthetics in clinics. Previous studies have shown that sevoflurane promotes microglial M1 activation. However, it remains unclear whether sevoflurane regulates microglial M2 activation. In this study, we found that sevoflurane treatment abolished interleukin 4 (IL-4)-induced M2 microglial activation. Our results indicate that IL-4-mediated induction of the characteristic M2 marker genes and proteins Arg1, Ym1, and IL-10 was significantly attenuated by sevoflurane pretreatment in primary microglia. Microglial M2 polarization induced by incubation with culture supernatant from human umbilical cord mesenchymal stromal cells (HUC-MSCs) was abolished by treatment with 2% or 4% sevoflurane. Upregulation of SOCS1 and suppression of SOCS3 were shown to play a crucial role in the process of M2 microglial polarization. Our results also indicate that SOCS1 expression was induced by IL-4, but it was inhibited by pretreatment with sevoflurane. In contrast, IL-4 suppressed SOCS3 expression, which was restored by pretreatment with sevoflurane. Mechanistically, it was shown that sevoflurane suppresses STAT6 phosphorylation in primary microglia.

Propofol attenuates LPS-induced tumor necrosis factor-α, interleukin-6 and nitric oxide expression in canine peripheral blood mononuclear cells possibly through down-regulation of nuclear factor (NF)-κB activation.

Sepsis is a major cause of mortality in intensive care medicine. Propofol, an intravenous general anesthetic, has been suggested to have anti-inflammatory properties and able to prevent sepsis induced by Gram-positive and Gram-negative bacteria by down-regulating the gene expression of pro-inflammatory cytokines. However, propofol's anti-inflammatory effects upon canine peripheral blood mononuclear cells (PBMCs) have not yet been clarified. Here, we isolate canine PBMCs and investigate the effects of propofol on the gene expressions of both lipopolysaccharide (LPS)-induced interleukin-6 (IL-6) and tumor necrosis factor (TNF)-α and upon the production of nitric oxide (NO). Through real-time quantitative PCR and the Griess reagent system, we found that non-cytotoxic levels of propofol significantly inhibited the release of NO and IL-6 and TNF-α gene expression in LPS-induced canine PBMCs. Western blotting revealed that LPS does significantly increase the expression of inducible NO synthase (iNOS) protein in canine PBMCs, while pretreatment with propofol significantly decreases the LPS-induced iNOS protein expression. Propofol, at concentration of 25 µM and 50 µM, also significantly inhibited the LPS-induced nuclear translocation of nuclear factor (NF)-κB p65 protein in canine PBMCs. This diminished TNF-α, IL-6 and iNOS expression, and NO production was in parallel to the respective decreased NF-κB p65 protein nuclear translocation in the LPS-activated canine PBMCs pretreated with 25 µM and 50 µM propofol. This suggests that non-cytotoxic levels of propofol pretreatment can down-regulate LPS-induced inflammatory responses in canine PBMCs, possibly by inhibiting the nuclear translocation of the NF-κB p65 protein.

Ginsenoside Re as an adjuvant to enhance the immune response to the inactivated rabies virus vaccine in mice.

The inactivated rabies virus vaccine (RV) is a relatively expensive vaccine, prone to failure in some cases. Ginsenoside Re (Re) is a saponin isolated from Panax ginseng, and has an adjuvant property. Here the adjuvant effect of Re to improve the immune response to the RV is evaluated in mice. ICR mice were immunized with saline, 2.50mg/kg Re, 20µl RV, 100µl RV, or 20µl of RV adjuvanted with Re (1.25, 2.50 or 5.00mg/kg). Different time points after boosting, we measured serum antibodies in blood samples and separated splenocytes to detect lymphocyte proliferation and the production of IL-4, IL-10, IL-12, and IFN-γ in vitro. We also compared immunizations containing 20µl RV and 20µl RV adjuvanted with Re (5.00mg/kg) for the expression of CD4(+) and CD8(+) T-cell subsets at different time points. Results indicated that co-administration of Re significantly enhanced serum antibody titers, increased the CD4(+):CD8(+) ratio, and enhanced both proliferation responses and IL-4, IL-10, IL-12 and IFN-γ secretions. Both Th1 and Th2 immune responses were activated. The supplementation of the Re (5.00mg/kg) to 20µl of RV significantly amplified serum antibody responses and Th1/Th2 responses inducing similar protection as did 100µl of RV. This suggests that Re could be used to reduce the dose, and therefore the cost, of the RV to achieve the same effective protection. Re merits further studies for use with vaccines of mixed Th1/Th2 immune responses.

Sodium selenite inhibits the expression of VEGF, TGFbeta(1) and IL-6 induced by LPS in human PC3 cells via TLR4-NF-(K)B signaling blockage.

Lipopolysaccharide (LPS)-induced TLR4-NF-(K)B signaling plays an important role in the development of prostatic tumors from chronic bacterial prostatic infection. Although many studies support the role of selenium in protecting against the development of prostate cancer secondary to chronic prostatitis, the mechanism of action remains unclear. The aim of our study was to investigate whether selenium inhibits the LPS-induced TLR4 signaling pathway in human prostate cancer PC3 cells. Using real-time quantitative PCR and ELISA analysis, we found that pretreatment with selenium (0.5-5uM) inhibited the LPS-induced expression of TGFbeta(1) and VEGF and production of these cytokines and IL-6 by PC3 cells, but did not alter the expression of TLR4 mRNA. Further experiments using Western blot showed that selenium at 3 and 5uM significantly inhibited the translocation of the NF-(K)B p65 subunit to the nucleus in LPS-stimulated PC3 cells. Our results suggest that low doses of selenium may protect the prostate from prostatitis-induced cancer by inhibiting nuclear translocation of the NF-(K)B and the subsequent production of the immunosuppressive cytokine TGFbeta(1), proangiogenic factor VEGF and pro-inflammatory factor IL-6.