Observations on the use of prostaglandin F(2alpha) as an abortifacient and effect of gonadotrophin-releasing hormone on ovarian activity after induced abortion during the breeding season in goats

Results of two experiments are described. In the first experiment, forty-one mixed-breed goats (does) with unknown gestation lengths were given 10 mg prostaglandin F(2alpha) (PGF(2alpha)) (2 doses of 5 mg each, 24 h apart) i.m. Blood samples were obtained before each treatment with PGF(2alpha) by jugular venipuncture, and plasma progesterone (P(4)) concentrations were determined by a nonextraction solid-phase radioimmunoassay. P(4) concentrations (ng/ml) were significantly decreased (15.47 vs 1.55, P<0.005) 24 h after the first injection of PGF(2alpha). A total of 63 fetuses was collected within 46.5 h following the first injection. Mean (+/- SE) crownrump lengths and body weights of 62 fetuses were 21.46 +/- 0.29 centimeters (cm) and 575.00 +/- 20.60 g, respectively. Based on these findings, the mean gestation length of these does was estimated to be 86.96 +/- 0.74 d. Thirty-one does retained their placenta for 12 to 72 h and were treated with a single injection of 5 mg PGF(2alpha) and 800 mg oxytetracycline i.m. Placental expulsion in all does occurred within 24 h posttreatment. The results of this study suggest that two doses of 5 mg PGF(2alpha) intramuscularly (i.m.) 24 h apart is an effective abortifacient at about 3 mo of pregnancy in does. In the second experiment, 38 does from the first experiment were divided in two groups of 19 each on Day 13 postabortion. Group A (treated) was given 50 ug GnRH i.m. while Group B (control) received 1 ml 0.9% saline i.m. Blood samples were obtained prior to treatment and on Day 23 postabortion and assayed for P(4) concentrations. There was no significant difference (P>0.10) in P(4) concentrations of samples obtained pre- and post-GnRH treatment. However, 14 of 19 and 12 of 19 in Groups A and B, respectively, exhibited estrus within 52 days following abortion. Twenty-six does were bred naturally and 17 became pregnant.     

Trypanosoma cruzi: mechanisms of intracellular calcium homeostasis.

Regulation of intracellular Ca2+ homeostasis was characterized in epimastigote forms of Trypanosoma cruzi using the fluorescence probe Fura-2. Despite an increase in extracellular Ca2+, [Ca2+]o, from 0 to 2 mM, cytosolic Ca2+, [Ca2+]i, increased only from 85 +/- 9 to 185 +/- 21 nM, indicating the presence of highly efficient mechanisms for maintaining [Ca2+]i. Exposure to monovalent Na+ (monensin)-, K+ (valinomycin, nigericin)-, and divalent Ca2+ (ionomycin)-specific ionophores, uncouplers of mitochondrial respiration (oligomycin), inhibitors of Na+/K(+)-ATPase (ouabain), and Ca(2+)-sensitive ATPase (orthovanadate) in 0 or 1 mM [Ca2+]o resulted in perturbations of [Ca2+]i, the patterns of which suggested both sequestration and extrusion mechanisms. Following equilibration in 1 mM [Ca2+]o, incubation with orthovanadate markedly increased [Ca2+]i, results which are compatible with an active uptake of [Ca2+]i by endoplasmic reticulum. In contrast, equilibration in 0 or 1 mM [Ca2+]o did not influence the relatively smaller increase in [Ca2+]i following incubation with oligomycin, suggesting a minor role for the mitochondrial compartment. In cells previously equilibrated in 1 mM [Ca2+]o, exposure to monensin or ouabain, conditions known to decrease the [Na+]o/[Na+]i gradient, upon which the Na+/Ca2+ exchange pathways are dependent, markedly increased [Ca2+]i. In a complementary manner, decreasing the extracellular Na+ gradient with Li+ increased [Ca2+]i in a dose-dependent manner. Finally, the calcium channel blockers verapamil and isradipine inhibited the uptake of Ca2+ by greater than 50%, whereas diltiazem, nifedipine, and nicardipine were ineffective. The results suggest that epimastigote forms of T. cruzi maintain [Ca2+]i by uptake, sequestration, and extrusion mechanisms, with properties common to eukaryotic organisms.     

Comparison of xylazine with tiletamine-zolazepam (Telazol) and xylazine-ketamine anesthesia in rabbits

Although widely used to provide short term anesthesia, ketamine-xylazine does not always produce satisfactory anesthesia. We compared the efficacy of ketamine-xylazine to tiletamine-zolazepam-xylazine for producing surgical anesthesia in rabbits. Four of six rabbits receiving ketamine-xylazine and all of the 12 animals given tiletamine-zolazepam-xylazine were anesthetized successfully. The mean surgical anesthesia time in the ketamine-xylazine group was 35 +/- 6 minutes as compared to the tiletamine-zolazepam-xylazine group, 72 +/- 8 minutes (p less than 0.05). There was no significant difference in the interval between the injection of the different anesthetic mixtures and the loss of either the righting reflex, the jaw reflex or the toe web pinch reflex. Respiratory rates and arterial oxygen partial pressure were higher in the ketamine-xylazine group (p less than 0.05). However, in both groups arterial blood pressure and arterial PO2 were lowered, while arterial PCO2 was elevated. No nephrotoxicity occurred. Tiletamine-zolazepam-xylazine provides effective surgical anesthesia in rabbits and in many cases may be preferable to conventional ketamine-xylazine regimen.     

Chromosomal mapping of brain-derived neurotrophic factor and neurotrophin-3 genes in man and mouse

Brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NTF3) are two new members of the nerve growth factor gene family, which play important roles in the development and maintenance of the vertebrate nervous system. Here we describe the assignments of the BDNF and NTF3 gene loci to human and mouse chromosomes and discuss the evolutionary relationship of human chromosomes 11 and 12. BDNF has been mapped to human chromosome 11p15.5-p11.2 and to mouse chromosome 2, and NTF3 to human chromosome 12p and mouse chromosome 6.     

Foam cell‐derived 4‐hydroxynonenal induces endothelial cell senescence in a TXNIP‐dependent manner

Vascular endothelial cell (VEC) senescence is considered an early event in the development of atherosclerotic lesions. Stressful stimuli, in particular oxidative stress, have been linked to premature senescence in the vasculature. Foam cells are a major source of reactive oxygen species and may play a role in the induction of VEC senescence; hence, we investigated their involvement in the induction of VEC senescence in a co‐culture transwell system. Primary bovine aortic endothelial cells, exposed to the secretome of THP‐1 monocyte‐derived foam cells, were analysed for the induction of senescence. Senescence associated β‐galactosidase activity and the expression of p16 and p21 were increased, whereas phosphorylated retinoblastoma protein was reduced. This senescent phenotype was mediated by 4‐hydroxnonenal (4‐HNE), a lipid peroxidation product secreted from foam cells; scavenging of 4‐HNE in the co‐culture medium blunted this effect. Furthermore, both foam cells and 4‐HNE increased the expression of the pro‐oxidant thioredoxin‐interacting protein (TXNIP). Molecular manipulation of TXNIP expression confirmed its involvement in foam cell‐induced senescence. Previous studies showed that peroxisome proliferator‐activated receptor (PPAR)δ was activated by 4‐hydroalkenals, such as 4‐HNE. Pharmacological interventions supported the involvement of the 4‐HNE‐PPARδ axis in the induction of TXNIP and VEC senescence. The association of TXNIP with VEC senescence was further supported by immunofluorescent staining of human carotid plaques in which the expression of both TXNIP and p21 was augmented in endothelial cells. Collectively, these findings suggest that foam cell‐released 4‐HNE activates PPARδ in VEC, leading to increased TXNIP expression and consequently to senescence.     

Regulation of Legionella phagosome maturation and infection through flagellin and host Ipaf

Legionella pneumophila is an intracellular bacterium that causes an acute form of pneumonia called Legionnaires' disease. After infection of human macrophages, the Legionella-containing phagosome (LCP) avoids fusion with the lysosome allowing intracellular replication of the bacterium. In macrophages derived from most mouse strains, the LCP is delivered to the lysosome resulting in Legionella degradation and restricted bacterial growth. Mouse macrophages lacking the NLR protein Ipaf or its downstream effector caspase-1 are permissive to intracellular Legionella replication. However, the mechanism by which Ipaf restricts Legionella replication is not well understood. Here we demonstrate that the presence of flagellin and a competent type IV secretion system are critical for Legionella to activate caspase-1 in macrophages. Activation of caspase-1 in response to Legionella infection also required host Ipaf, but not TLR5. In the absence of Ipaf or caspase-1 activation, the LCP acquired endoplasmic reticulum-derived vesicles, avoided fusion with the lysosome, and allowed Legionella replication. Accordingly a Legionella mutant lacking flagellin did not activate caspase-1, avoided degradation, and replicated in wild-type macrophages. The regulation of phagosome maturation by Ipaf occurred within 2 h after infection and was independent of macrophage cell death. In vivo studies confirmed that flagellin and Ipaf play an important role in the control of Legionella clearance. These results reveal that Ipaf restricts Legionella replication through the regulation of phagosome maturation, providing a novel function for NLR proteins in host defense against an intracellular bacterium.     

Antiviral and antimicrobial activities of three sesquiterpene lactones from Centaurea solstitialis L. ssp. solstitialis

Three sesquiterpene lactones (centaurepensin=chlorohyssopifolin A, chlorojanerin and 13-acetyl solstitialin A) isolated from the aerial parts of Centaurea solstitialis L. ssp. solstitialis (Asteraceae) were investigated for antimicrobial and antiviral activities. For the antimicrobial activity assessment, both standard and isolated strains of Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis, Staphylococcus aureus, Candida albicans and C. parapsilosis were employed by the microdilution method. Herpes simplex type-1, a DNA virus, and Parainfluenza, an RNA virus, were employed for the determination of the antiviral activity of these three sesquiterpene lactones using Vero cell lines. Ampicilline, ofloxocine, ketoconazole, fluconazole, acyclovir and oseltamivir were used as the reference drugs. 13-Acetyl solstitialin A displayed remarkable antibacterial activity against isolated strains of E. faecalis at 1 μg/ml concentration, which was close to the effective concentrations of ampicillin. The same compound also showed significant activity against the DNA virus, being as potent as the reference compound acyclovir at maximum and minimum concentrations of 16–<0.00006 μg/ml. This is the first report showing that 13-acetyl solstitialin A possesses significant antiviral activity.     

Functional characterisation of the regulation of CAAT enhancer binding protein alpha by GSK-3 phosphorylation of Threonines 222/226

Background  

Glycogen Synthase Kinase-3 (GSK3) activity is repressed following insulin treatment of cells. Pharmacological inhibition of GSK3 mimics the effect of insulin on Phosphoenolpyruvate Carboxykinase (PEPCK), Glucose-6 Phosphatase (G6Pase) and IGF binding protein-1 (IGFBP1) gene expression. CAAT/enhancer binding protein alpha (C/EBPα) regulates these gene promoters in liver and is phosphorylated on two residues (T222/T226) by GSK3, although the functional outcome of the phosphorylation has not been established. We aimed to establish whether CEBPα is a link between GSK3 and these gene promoters.