Population based quantification of dendrites: evidence for the lack of microtubule-associate protein 2a,b in Purkinje cell spiny dendrites

The high molecular weight isoforms (a and b) of microtubule-associate protein 2 (MAP2a,b) are widely believed to be specific markers for neuronal somata and dendrites. We analyzed and quantified MAP2a,b stained dendrites of the cerebellar molecular layer using a novel approach that segmented and 3D reconstructed them, and the results have been compared with those obtained by other methods, including single-cell reconstruction and analysis of electron micrographs. Our results show that the molecular layer dendritic volume fraction is lower than in the neocortex (10% compared to neocortical 29%). The low total volume fraction of dendrites in the molecular layer is best explained by the majority of the afferents to the dendrites being from the very densely packed parallel fibers, which allows the dendritic fields of individual neurons to be smaller and more compact than in the cerebral cortex. However, the MAP2a,b dendritic volume fraction is even lower (5.2%) than the total volume fraction of dendrites in the molecular layer (10%). Analysis of the material shows that this difference between the two results is due to the unexpected finding that there were few MAP2a,b stained Purkinje cell spiny dendrites.     

Divergent Immune Responses to Mycobacterium avium subsp. paratuberculosis Infection Correlate with Kinome Responses at the Site of Intestinal Infection

Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne''s disease (JD) in cattle. M. avium subsp. paratuberculosis infects the gastrointestinal tract of calves, localizing and persisting primarily in the distal ileum. A high percentage of cattle exposed to M. avium subsp. paratuberculosis do not develop JD, but the mechanisms by which they resist infection are not understood. Here, we merge an established in vivo bovine intestinal segment model for M. avium subsp. paratuberculosis infection with bovine-specific peptide kinome arrays as a first step to understanding how infection influences host kinomic responses at the site of infection. Application of peptide arrays to in vivo tissue samples represents a critical and ambitious step in using this technology to understand host-pathogen interactions. Kinome analysis was performed on intestinal samples from 4 ileal segments subdivided into 10 separate compartments (6 M. avium subsp. paratuberculosis-infected compartments and 4 intra-animal controls) using bovine-specific peptide arrays. Kinome data sets clustered into two groups, suggesting unique binary responses to M. avium subsp. paratuberculosis. Similarly, two M. avium subsp. paratuberculosis-specific immune responses, characterized by different antibody, T cell proliferation, and gamma interferon (IFN-γ) responses, were also observed. Interestingly, the kinomic groupings segregated with the immune response groupings. Pathway and gene ontology analyses revealed that differences in innate immune and interleukin signaling and particular differences in the Wnt/β-catenin pathway distinguished the kinomic groupings. Collectively, kinome analysis of tissue samples offers insight into the complex cellular responses induced by M. avium subsp. paratuberculosis in the ileum and provides a novel method to understand mechanisms that alter the balance between cell-mediated and antibody responses to M. avium subsp. paratuberculosis infection.     

Kinetics of peripheral blood mononuclear cell mobilization with chemotherapy and/or granulocyte-colony-stimulating factor: implications for yield of hematopoietic progenitor cell collections

BACKGROUND: Peripheral blood progenitor cells (PBPCs) are commonly collected and used to reconstitute hematopoiesis after high-dose chemotherapy. However, strategies for optimal collection and assessment of leukapheresis components are not standardized. STUDY DESIGN and METHODS: Hematopoietic progenitor cell assays were performed on 369 leukapheresis components collected from 95 patients who had received doxorubicin-based chemotherapy and/or granulocyte-colony-stimulating factor (G-CSF). Precollection patient hematologic values, leukapheresis collection values, component hematopoietic progenitor cell assays, and patient outcome measures were summarized. The kinetics of mononuclear cell (MNC) and PBPC mobilization were assessed among four patient groups. RESULTS: Patient group was a significant predictor of the peripheral blood MNC count on the day of collection (p<0.0001), and that value was a significant predictor of granulocyte-macrophage– colony-forming unit (CFU-GM) yield (p<0.0001). This relationship between the peripheral blood MNC count on the day of collection and CFU- GM yield differed according to patient group (p<0.0001). CFU-GM made up a larger fraction of peripheral blood MNCs collected from patients who received chemotherapy plus G-CSF than collected from those who received G-CSF alone. Moreover, the peripheral blood MNC count and the corresponding CFU-GM yield increased significantly on consecutive days of collection in patient groups receiving chemotherapy and G-CSF but were unchanged or decreased in patients receiving G-CSF alone. CONCLUSION: The relationship between peripheral blood MNC count and leukapheresis component CFU-GM yield differed significantly between patients who received chemotherapy and G-CSF and those who received G- CSF alone for the mobilization of PBPCs. Patient peripheral blood MNC count and component CFU-GM yield are useful for both assessing and suggesting revisions to PBPC mobilization and collection strategies.     

Respiratory symptoms in preterm infants: burden of disease in the first year of life

Objective

While respiratory symptoms in the first year of life are relatively well described for term infants, data for preterm infants are scarce. We aimed to describe the burden of respiratory disease in a group of preterm infants with and without bronchopulmonary dysplasia (BPD) and to assess the association of respiratory symptoms with perinatal, genetic and environmental risk factors.

Methods

Single centre birth cohort study: prospective recording of perinatal risk factors and retrospective assessment of respiratory symptoms during the first year of life by standardised questionnaires. Main outcome measures: Cough and wheeze (common symptoms), re-hospitalisation and need for inhalation therapy (severe outcomes). Patients: 126 preterms (median gestational age 28.7 weeks; 78 with, 48 without BPD) hospitalised at the University Children''s Hospital of Bern, Switzerland 1999-2006.

Results

Cough occurred in 80%, wheeze in 44%, rehospitalisation in 25% and long term inhalation therapy in wheezers in 13% of the preterm infants. Using logistic regression, the main risk factor for common symptoms was frequent contact with other children. Severe outcomes were associated with maximal peak inspiratory pressure, arterial cord blood pH, APGAR and CRIB-Score.

Conclusions

Cough in preterm infants is as common as in term infants, whereas wheeze, inhalation therapy and re-hospitalisations occur more often. Severe outcomes are associated with perinatal risk factors. Preterm infants who did not qualify for BPD according to latest guidelines also showed a significant burden of respiratory disease in the first year of life.     

Feeding frequency for lactating cows: effects on digestion, milk production and energy utilization

The results are reported of four feeding experiments in which lactating cows were given fixed rations of hay and high-cereal concentrates at different meal frequencies. In Expt 1 the total ration was given in two and twenty-four meals daily and in Expts 2-4, the concentrates were given in two and five or six meals and the hay was given twice daily. The diets contained 600-900 g concentrate/kg. In all the experiments, more frequent feeding of these low-roughage diets reduced milk fat depression and increased milk fat yield. In each experiment the increase was greater with the diet containing the lower proportion of hay. There was no significant effect on milk yield, the protein or lactose contents of the milk or live-weight gain. Digestibility of dry matter, organic matter and energy was increased in one of the experiments but not in two others in which it was measured. More frequent feeding was calculated to increase the net energy secreted in milk and there was a tendency for it to increase the net energy in live-weight gain with diets containing 600 or 700 g concentrate/kg but to decrease it with diets containing 800 or 900 g concentrate/kg. These results are discussed in relation to theories of energy partition in lactating cows. It is concluded that at fixed feed intakes, the main response to increased meal frequency is likely to be a reduction in milk fat depression and that this will be confined to diets containing not more than about 200 g modified acid-detergent fibre/kg dry matter.     

Medullary carcinoma of thyroid gland in a girl aged 10 years

A healthy girl of 10 years presented with lumps on her tongue, shown on biopsy to be neuromas. She had had operations to her feet for pes cavus and was relatively long-limbed but had no other 'Marfanoid' features. She had a high plasma calcitonin level. At operation a normal sized thyroid gland was totally removed. It contained two discrete masses of malignant C-cells and diffuse foci elsewhere. The plasma calcitonin fell slowly to normal on follow-up but was noted to be rising 2 1/2 years later. We stress the importance of making this diagnosis as early as possible and mention briefly another child aged 15 months, similarly diagnosed and operated upon.