Ontario bulk milk somatic cell count reduction program: progress and outlook

The objective of this study was to describe the results of the ongoing somatic cell count (SCC) reduction program in Ontario, Canada and to provide an outlook regarding the possible scenarios beyond the final stages of the current reduction program. The data were from all of the approximately 9500 farms in the province of Ontario during the last 10 yr and included monthly data for kilograms of milk sold, mean milk component measurements, and milk quality measurements (bulk milk SCC, plate loop count, and inhibitor presence). Four of five steps in the control program had a significant impact on the mean monthly bulk milk SCC. The total decrease in mean SCC that was attributable to the program was approximately 80 x 10(3) cells/ml. The monthly occurrence of inhibitor violations increased significantly. Cross-classification of the annual mean bulk milk SCC and the incidence of inhibitor violations indicated that the incidence specifically increased for farms with a relatively high bulk milk SCC. In 1994, bulk milk SCC increased, primarily because of farms that had a mean bulk milk SCC between 150 x 10(3) and 450 x 10(3)/ml. The small relative contribution of farms with higher bulk milk SCC was due to their relatively low production and the small number of farms in those classes. The Ontario SCC reduction program was initially successful in the reduction of mean bulk milk SCC. Further reduction will require the active participation of farms with a relatively low bulk milk SCC. Not only is it necessary to penalize farms that exceed thresholds, but also an incentive to prevent farms with good udder health management from increasing bulk milk SCC is of great importance. The increase in inhibitor violations is of concern and requires further attention.     

The Homothorax homeoprotein activates the nuclear localization of another homeoprotein, extradenticle, and suppresses eye development in Drosophila

The Extradenticle (Exd) protein in Drosophila acts as a cofactor to homeotic proteins. Its nuclear localization is regulated. We report the cloning of the Drosophila homothorax (hth) gene, a homolog of the mouse Meis1 proto-oncogene that has a homeobox related to that of exd. Comparison with Meis1 finds two regions of high homology: a novel MH domain and the homeodomain. In imaginal discs, hth expression coincides with nuclear Exd. hth and exd also have virtually identical, mutant clonal phenotypes in adults. These results suggest that hth and exd function in the same pathway. We show that hth acts upstream of exd and is required and sufficient for Exd protein nuclear localization. We also show that hth and exd are both negative regulators of eye development; their mutant clones caused ectopic eye formation. Targeted expression of hth, but not of exd, in the eye disc abolished eye development completely. We suggest that hth acts with exd to delimit the eye field and prevent inappropriate eye development.     

Ex vivo generation of functional dendritic cells from mobilized CD34+ hematopoietic stem cells

Gene engineering to enhance tumour immunogenicity and elicit curative responses against established tumours and tumour recurrences has become an attractive prospect. Gene engineering enables new genes to be selectively inserted into the genome of a tumour cell, or the construction of new fusion plasmids coding tumour antigens and immunomodulatory molecules. The rationale behind current research is to enhance the immune recognition of tumour antigens through their association with the molecules on which immune recognition depends. The immunotherapy data obtained in many experimental tumour systems provide a realistic assessment of the potential and limits of this technological approach. Experimental vaccination of rodents has been shown to induce a significant immune memory, even against poorly immunogenic tumours, that can prevent tumour growth and cure initial metastases, but is poorly effective against established tumours. Its use in tumour prevention is a fresh dawning perspective.     

Immunoassayable beta-endorphin level in the plasma and CSF of heroin addicted and normal subjects before and after electroacupuncture

The present study was undertaken to evaluate if plasma or CSF beta-endorphin level can be induced to rise during the treatment of heroin addiction by electroacupuncture. Based on the examination of 30 addicts, we obtained no evidence indicating an increase of beta-endorphin level in either the plasma or the CSF after 30 min of acupuncture. In spite of this, the majority of the addicts experienced a reduction of withdrawal symptoms during treatment. Since electroacupuncture may only induce a highly localized secretion of beta-endorphin in the brain, our results cannot unequivocally exclude the possibility that this peptide is involved in mediating the action of acupuncture.     

Proteomic analysis of integral plasma membrane proteins

Efficient methods for profiling proteins integral to the plasma membrane are highly desirable for the identification of overexpressed proteins in disease cells. Such methods will aid in both understanding basic biological processes and discovering protein targets for the design of therapeutic monoclonal antibodies. Avoiding contamination by subcellular organelles and cytosolic proteins is crucial to the successful proteomic analysis of integral plasma membrane proteins. Here we report a biotin-directed affinity purification (BDAP) method for the preparation of integral plasma membrane proteins, which involves (1) biotinylation of cell surface membrane proteins in viable cells, (2) affinity enrichment using streptavidin beads, and (3) depletion of plasma membrane-associated cytosolic proteins by harsh washes with high-salt and high-pH buffers. The integral plasma membrane proteins are then extracted and subjected to SDS-PAGE separation and HPLC/MS/MS for protein identification. We used the BDAP method to prepare integral plasma membrane proteins from a human lung cancer cell line. Western blotting analysis showed that the preparation was almost completely devoid of actin, a major cytosolic protein. Nano-HPLC/MS/MS analysis of only 30 microg of protein extracted from the affinity-enriched integral plasma membrane preparation led to the identification of 898 unique proteins, of which 781 were annotated with regard to their plasma membrane localization. Among the annotated proteins, at least 526 (67.3%) were integral plasma membrane proteins. Notable among them were 62 prenylated proteins and 45 Ras family proteins. To our knowledge, this is the most comprehensive proteomic analysis of integral plasma membrane proteins in mammalian cells to date. Given the importance of integral membrane proteins for drug design, the described approach will expedite the characterization of plasma membrane subproteomes and the discovery of plasma membrane protein drug targets.     

Automatic heat-shrink sleeve applicator using the low-temperature catalytic combustor

Catalytic combustion for applying heat shrink sleeves (HSS) to a pipeline in KOGAS was investigated. We used a low temperature catalytic combustor in order to place HSS in the conduit. The surface temperature of the catalytic combustor maintained the conditions at which HSS can become well-adhesive at the conduit. Therefore, the automatic HSS construction provided the chance to establish gas pipeline safety by an automatic catalytic combustor device.