Acute but transient neurological deterioration revealing adult polyglucosan body disease

Adult polyglucosan body disease (APBD) is a metabolic disorder usually caused by glycogen branching enzyme (GBE) deficiency. APBD associates progressive walking difficulties, bladder dysfunction and, in about 50% of the cases, cognitive decline. APBD is characterized by a recognizable leukodystrophy on brain MRI. We report here a novel presentation of this disease in a 35-year old woman who presented with an acute deterioration followed by an unexpected recovery. Enzymatic analysis displayed decreased GBE activity in leukocytes. Molecular analyses revealed that only one mutated allele was expressed, bearing a p.Arg515His mutation. This is the first observation reporting acute and reversible neurological symptoms in APBD. These findings emphasize the importance of searching GBE deficiency in patients presenting with a leukodystrophy and acute neurological symptoms mimicking a stroke, in the absence of cardiovascular risk factors.     

Immunodetection of Tau microtubule-associated protein in human sperm and testis

Dear Editor,
The cytosolic protein Tau is naturally present in human neurons, where it has a pivotal role in controlling microtubule stability. Hyperphosphorylation of Tau （observed during neurodegenerative diseases, such as Alzheimer＇s disease） impairs the protein＇s ability to bind microtubules. This results in microtubule disassembly and the formation of Tau aggregates, Tau protein is also widely expressed in peripheral tissues. In the male reproductive system, screening for Tau has focused solely on the rodent and bovine testis. In the present study, we used immunofluorescence and immunoenzymatic techniques （with a Tau-specific antibody） to investigate the presence of Tau protein in human ejaculated sperm and testicular tissue.     

Interleukin 6 in Intact and Injured Mouse Peripheral Nerves

The multifunctional cytokine interleukin 6 (IL-6) has direct growth, survival and differentiation effects on peripheral and central neurons. Furthermore, it can modulate the production by non-neuronal cells of other cytokines and growth factors, and thereby affect nerve cells indirectly. We have studied IL-6 expression and production in intact and injured peripheral nerves of C57/BL/6NHSD mice, which display the normal rapid progression of Wallerian degeneration. The IL-6 mRNA was detected in nerves degenerating in vitro or in vivo , but not in intact nerves. In vitro - and in vivo -degenerating nerve segments and neuroma nerve segments synthesized and secreted IL-6. The onset of IL-6 production was rapid and prolonged. It was detected as early as 2 h after injury and persisted for the entire period of 21 days tested after the injury. Of the non-neuronal cells that reside in intact and injured nerves, macrophages and fibroblasts were the major contributors to IL-6 production. We also studied IL-6 production in intact and injured nerves of mutant C57BL/6-WLD/OLA/NHSD mice, which display very slow progression of Wallerian degeneration. Injured nerves of C57BL/6-WLD/OLA/NHSD mice produced significantly lower amounts of IL-6 than did rapidly degenerating nerves of C57/BL/6NHSD mice.     

Toll-like receptor 9 signaling mediates the anti-inflammatory effects of probiotics in murine experimental colitis

BACKGROUND & AIMS: We tested whether the attenuation of experimental colitis by live probiotic bacteria is due to their immunostimulatory DNA, whether toll-like receptor (TLR) signaling is required, and whether nonviable probiotics are effective. METHODS: Methylated and unmethylated genomic DNA isolated from probiotics (VSL-3), DNAse-treated probiotics and Escherichia coli (DH5 alpha) genomic DNA were administered intragastrically (i.g.) or subcutaneously (s.c.) to mice prior to the induction of colitis. Viable or gamma-irradiated probiotics were administered i.g. to wild-type mice and mice deficient in different TLR or in the adaptor protein MyD88, 10 days prior to administration of dextran sodium sulfate (DSS) to their drinking water and for 7 days thereafter. RESULTS: Intragastric and s.c. administration of probiotic and E. coli DNA ameliorated the severity of DSS-induced colitis, whereas methylated probiotic DNA, calf thymus DNA, and DNase-treated probiotics had no effect. The colitis severity was attenuated to the same extent by i.g. delivery of nonviable gamma-irradiated or viable probiotics. Mice deficient in MyD88 did not respond to gamma-irradiated probiotics. The severity of DSS-induced colitis in TLR2 and TLR4 deficient mice was significantly decreased by i.g. administration of gamma-irradiated probiotics, whereas, in TLR9-deficient mice, gamma-irradiated probiotics had no effect. CONCLUSIONS: The protective effects of probiotics are mediated by their own DNA rather than by their metabolites or ability to colonize the colon. TLR9 signaling is essential in mediating the anti-inflammatory effect of probiotics, and live microorganisms are not required to attenuate experimental colitis because nonviable probiotics are equally effective.     

Improving the Quality of EDTA-treated Blood Specimens from Mice

Nonterminal blood sampling in laboratory mice is a very common procedure. With the goal of improving animal welfare, different sampling sites and methods have been compared but have not achieved a consensus. Moreover, most of these studies overlooked the quality of blood specimens collected. The main preanalytical concern with EDTA-treated blood specimens for hematology analyses is platelet aggregation, which is known to cause analytical errors. Our objective was to find a nonterminal blood sampling method with minimal adverse effects on mice and few or no platelet aggregates. We tested and compared 2 collection sites, 4 sampling methods, and 3 antithrombotic drugs in 80 C57BL6/j male and female mice by evaluating platelet aggregates on blood smears and platelet, WBC, and RBC counts. In addition, the blood collection process was carefully evaluated, and adverse effects were recorded. Platelet aggregation was lower in specimens collected from the jugular vein than from the facial vein, with no effect of the sampling device or the presence of an antithrombotic additive. Highly aggregated specimens were significantly associated with lower platelet counts, whereas aggregation had no effect on WBC or RBC counts. Adverse events during sampling were significantly associated with more numerous platelet aggregates. The jugular vein is thus a satisfactory sampling site in mice in terms of both animal welfare and low platelet aggregation. Using antithrombotic agents appears to be unnecessary, whereas improving sampling conditions remains a key requirement to ensure the quality of EDTA-treated blood specimens from mice.

Industrial and academic research often require hematology analyses of mouse blood. Consequently, many terminal and nonterminal techniques have become available for blood sampling in mice.^{12,21,27,40,42,53} Preanalytical variation in clinical pathology is known to be a major issue.^5,45,49 Although the effects of the blood sampling method on animal welfare have been the subject of many preanalytical hematology and biochemical analyses,^{1,6,8,9,15,16,18,24-26,36,47,50-52,54} no agreement has been reached regarding the optimal method for nonterminal blood collection in mice and, to our knowledge, only a few investigations^1,8,15,16,18 have addressed the quality of the resulting blood specimens.Our own experience of hematology measurements from nonterminal mouse EDTA-blood specimens is that some specimens show both visible clots and platelet aggregation, the latter being detected only from microscopic examination of blood smears.³³ Whereas specimens with visible clots can be eliminated, microscopic platelet aggregates can also interfere with hematology analyses or cause analytical errors, as has been reported in other species including cats.^13,22,31,39 These abnormalities require repeat sampling when possible; otherwise, the number of validated results is decreased. EDTA-treated mouse blood is especially prone to platelet aggregation and clotting.^14,28,43 This characteristic leads to errors in platelet counts (pseudothrombocytopenia) and possible misidentification of platelet aggregates as eosinophils, resulting in false leukocytosis and eosinophilia.¹⁴ In vitro platelet aggregation in mice is due to high platelet counts^34,43 and is influenced by numerous preanalytical factors including the sampling method, collection site, specimen processing, anticoagulant used, the blood:anticoagulant ratio, the mouse strain and genetic alterations.^19,28,30,43 The literature on the influence of preanalytical factors on the quality of CBC analyses in mice is scant,⁴³ and no agreement has yet been reached regarding the optimal method for nonterminal blood collection in mice. In humans and various animal species, platelet aggregation can be reduced by adding platelet aggregation inhibitors that act at different steps of aggregation. To our knowledge, the addition of such inhibitors to mouse whole blood has not been tested as a means to improve the quality of mice EDTA-treated blood specimens.The aim of this study was therefore to identify the best preanalytical conditions for nonterminal blood collection in mice, based on animal welfare, scores of platelet aggregation, and platelet, RBC, and WBC counts. The hypotheses we tested were that 1) adding an antithrombotic drug (or multiple such drugs) to the EDTA-treated blood specimen would prevent or at least significantly lower platelet aggregation, 2) the site and the method of collection influence in vitro platelet aggregation, and 3) high-quality blood sampling is a key to reducing platelet aggregation in blood specimens.     

Effects of propranolol or paracetamol on lidocaine concentrations in serum and tissues.

PURPOSE: The goal of this study was to determine the interaction of lidocaine after paracetamol or propranolol administration. MATERIALS AND METHODS: Group A rats received a mixture of cold lidocaine and (14)C lidocaine into the masseter muscle. Group B rats received lidocaine and paracetamol 7.5 mg/kg orally, whereas group C received lidocaine intramuscularly and propranolol 1.5 mg/kg orally. Five consecutive doses were administered. The levels of these drugs were estimated in serum and the total concentration and the percent protein binding of lidocaine in tissues (liver and mandible) were determined. The rats were killed 2 hours after the last dose. RESULTS: The results suggest that lidocaine concentrations in serum were significantly increased after either paracetamol or propranolol administration. Additionally, the combined therapy of propranolol and lidocaine resulted in a significant decrease in the total concentration and the percent protein binding of lidocaine in tissues. CONCLUSIONS: The coadministration of lidocaine with paracetamol or propranolol interferes with the metabolic profile, resulting in pharmacokinetic interactions that may be significant for the determination of the correct dose of lidocaine in clinical applications.