The use of amphipathic maleimides to study membrane-associated proteins

A series of amphiphilic polymethylenecarboxymaleimides has been synthesized for use as sulfhydryl reagents applicable to membrane proteins. Physical properties of the compounds which are relevant to their proposed mode of action have been determined. By comparing rates of reaction in aqueous and aprotic solvents, the compounds have been shown to react exclusively with the thiolate ion. The effects of the reagents on three membrane-associated proteins are reported, and in two cases a comparative study has been made of the effects on the proteins in the absence of membranes. A mechanism is proposed whereby the reagents are anchored at the lipid/water interface by the negatively charged carboxyl group, thus siting the reactive maleimide in a plane whose depth is defined by the length of the reagent. Supporting evidence for this model is provided by the inability of the reagents to traverse membranes, and variation of their inhibitory potency with chain length when the proteins are embedded in the membrane, but not when extracted into solution. As examples of general use of the reagents to probe sulfhydryl groups in membrane proteins, the reagents have been used to (a) determine the depths in the membrane at which two populations of sulfhydryl groups occur in the mitochondrial phosphate transporter; (b) locate a single sulfhydryl associated with the active site ofD--hydroxybutyrate dehydrogenase in the inner mitochondrial membrane; (c) examine sulfhydryl groups in theD-3-glyceraldehyde phosphate dehydrogenase associated with the human red blood cell membrane.     

Acquired resistance in dogs to repeated infestation with Ixodes scapularis (Acari: Ixodidae) reduces tick viability and reproductive success

The purpose of this study was to determine whether dogs develop acquired resistance to adult Ixodes scapularis infestation in an experimental model. Five dogs were each infested with ten mating pairs of ticks every week for 7 consecutive weeks, another five dogs were each infested with ten mating pairs once every 2 weeks for 10 weeks and four dogs served as controls not exposed to ticks. All ticks were allowed to feed to repletion and were collected only after dropping from the host. Several variables were measured to determine the extent of blood feeding success. Regression analysis indicated that the engorgement success, survival and mean tick engorgement weight declined with repeated infestation in both groups of dogs (p<0.05). Tick oviposition as well as the F1 viability declined with each successive infestation in both groups. These results suggest that repeated infestation with I. scapularis elicits a protective immune response against tick feeding and could serve as a limiting factor in the spread and transmission of Borrelia burgdorferi.     

Observations on the morphology of submaxillary gland cells isolated in a density gradient,and the demonstration of interacinar bridges

Summary Methods are presented for the recovery of separate fractions of rabbit submaxillary acinar and striated duct cells from acini and duct segments, by means of density gradient centrifugation in glycerol-sucrose mixtures.Incomplete acinar fragmentation discloses the presence of acellular, nonstaining interacinar structures, referred to as interacinar bridges. The effectiveness of trypsin in liberating the acinar cells is ascribed to the disruption of intercellular secretory capillaries and interacinar bridges.The basal processes of the isolated striated duct cells correspond with both the striations observed with optical microscopy and the infolded cytoplasmic compartments revealed with electron microscopy. The presence of clearly discernible spaces between the basal processes is compatible with the concept of intermembrane dilations.     

Biosynthesis of calystegines: 15N NMR and kinetics of formation in root cultures of Calystegia sepium

Calystegines are nortropane alkaloids bearing between three and five hydroxyl groups in various positions. [15N]Tropinone was administered to root cultures of Calystegia sepium and the incorporation into calystegines was followed. Increase of label in calystegines was measured by one-dimensional 15N NMR and inverse-detected 2D NMR techniques. The results show that tropinone and pseudotropine are metabolites in the biosynthetic pathway of calystegines. The velocity of calystegine accumulation was followed kinetically by transfer of root cultures from 15N-enriched medium to 14N-medium and analysis by GC-MS. A constant calystegine formation with no interference by excretion or degradation was observed. A biosynthetic rate for individual calystegines at each time point was calculated, the maximum was 0.4 mg/day/g of biomass. This allowed the velocity of individual biosynthetic steps to be estimated.     

Mold colonization during use of preservative-treated and untreated air filters, including HEPA filters from hospitals and commercial locations over an 8-year period (1996–2003)

High efficiency particulate arrestance (HEPA; 99.97% efficient at 0.3 m) filters, filters with ASHRAE particulate arrestance rating of 90–95% at 1 m (90–95% filters), and lower efficiency cellulosic-polyester filters from air conditioning systems in hospitals and commercial buildings were removed from the systems and examined microscopically for mold colonization. Cellulosic-type filters from systems with water entrainment problems typically were colonized, or became colonized upon incubation in moisture chambers. Species of Acremonium, Aspergillus, and Cladosporium were most common. With air filters of all types, treatment of filter media with an antimicrobial preservative tended to reduce or delay colonization. Mold colonization of HEPA and 90–95% filters was observed most often on the load surfaces, but two untreated HEPA filters were permeated with fungi, one with Aspergillus flavus, the other with Cladosporium sp. Air filters in heating, ventilating, and air conditioning (HVAC) systems, particularly those with chronic or periodic exposure to moisture, may serve as point sources for indoor molds.     

La fertilité chez le blessé médullaire. Effet du niveau de la lésion médullaire

The majority of spinal cord injured males cannot procreate naturally due to anejaculation as well as abnormal sperm characteristics. Treatment of this impaired fertility must be associated with treatment of neurogenic urinary tract disorders, bowel dysfunction and spasticity. The level of the spinal lesion and the spinal cord injury syndrome influence the possibility of inducing reflex ejaculation by penile vibratory stimulation and sperm quality.