Absorption of histamine from the gastrointestinal tract of dogs in vivo

1. In anaesthetized dogs, various amounts of [(14)C]histamine were introduced into the lumen of a ligated intestinal loop or of the ligated stomach and the absorption of this histamine was studied by determining the radioactive histamine in the venous blood coming from the ligated part.2. After the introduction of 5-5000 mug [(14)C]histamine, into a loop of jejunum, or of 50 mug into a loop of duodenum, ileum or colon, radioactive histamine was detected in all eight successive 15 min samples of venous blood collected during 2 hr. The percentage recovery of the [(14)C]histamine in the blood during this period varied between 0.04 and 3.7.3. After the introduction of 10 mg [(14)C]histamine into the stomach, radioactive histamine was detected in all samples of gastric venous blood collected at various times during the following 4 hr.4. After the introduction of 50-5000 mug [(14)C]histamine into a loop of jejunum, radioactive histamine was also detected in the general arterial blood.5. When a jejunal loop was perfused through its artery with a dextransaline solution, the absorption of [(14)C]histamine from the lumen into the venous effluent was much greater than when the blood supply was kept intact.6. A large part of the [(14)C]histamine introduced into an intestinal loop was inactivated or destroyed either in the lumen or the wall since only a part was recovered in the venous blood, contents and wall of the loop at the end of 2 hr. When different amounts of [(14)C]histamine were introduced into a jejunal loop the recovery was shown to be dependent on the dose. With 5 mug it amounted to about 1% whereas with 5000 mug to 29-42%. The recovery of the [(14)C]histamine introduced into a perfused jejunal loop was greater.7. In dogs and cats large amounts of free histamine were found in the contents of the stomach 2 hr after a meat meal, and much smaller amounts in the contents of loops from the small intestine. The amounts found in the contents of loops from the colon varied greatly.8. In eighteen commercial dog and cat foods the free histamine contents were found to vary over fiftyfold, from 0.06 to 3.5 mg/100 g.9. The significance of the results is discussed in relation to the role of histamine as a humoral agent in the ;gastric' and ;intestinal phases' of gastric secretion.     

Interaction of pre-programmed control and natural stretch reflexes in human landing movements

Pre-programmed mechanisms of motor control are known to influence the gain of artificially evoked stretch reflexes. However, their interaction with stretch reflexes evoked in the context of unimpeded natural movement is not understood. We used a landing movement, for which a stretch reflex is an integral part of the natural action, to test the hypothesis that unpredicted motor events increase stretch reflex gain. The unpredicted event occurred when a false floor, perceived to be solid, collapsed easily on impact, allowing the subjects to descend for a further 85 ms to a solid floor below. Spinal stretch reflexes were measured following solid floor contact. When subjects passed through the false floor en route to the solid floor, the amplitude of the EMG reflex activity was double that found in direct falls. This was not due to differences in joint rotations between these conditions. Descending pathways can modify H- and stretch-reflex gain in man. We therefore manipulated the time between the false and real floor contacts and hence the time available for transmission along these pathways. With 30 ms between floors, the enhancement of the reflex was extinguished, whereas with 50 ms between floors it reappeared. This excluded several mechanisms from being responsible for the doubling of the reflex EMG amplitude. It is argued that the enhanced response is due to the modulation of reflex gain at the spinal level by signals in descending pathways triggered by the false platform. The results suggest the future hypothesis that this trigger could be the absence of afferent signals expected at the time of false floor impact and that salient error signals produced from a comparison of expected and actual sensory events may be used to reset reflex gains.     

Chromosome content and ultrastructure of radiation-induced micronuclei

Unrepaired or misrepaired radiation damage in mammalian chromosomescan result in micronucleus formation at the first cell division.This represents loss of genomic information which may causecell death. To improve our understanding of the mechanism ofradiation-induced micronucleus formation, we characterized micronucleusultrastructure and identified the origin of micronucleus DNA.Immunofluorescence microscopy showed that micronuclei were structurallysimilar to main nuclei since they contained nuclear lamins Aand C and were encapsulated by a network of vimentin intermediatefilaments. The contents of radiation-induced micronuclei werecharacterized using fluorescence in situ hybridization to probefor DNA originating from chromosomes 2, 7, 11 and 16. We postulatedthat if incorporation of DNA into micronuclei were random, thenthe probability of chromosomal DNA in micronuclei would be relatedto the target, i.e. chromosome size. Our results demonstratedthat incorporation of DNA from smaller chromosomes (11 and 16)was not different from expected values but incorporation ofDNA from the larger chromosomes (2 and 7) was significantlygreater than expected. Not all chromosomes in the human genome,therefore, were equally susceptible to genomic loss by micronucleusencapsulation. In conclusion, radiationinduced micronuclei havesimilar structural characteristics to main nuclei, chromosomedamage and/or repair after ionizing radiation may be non-random,and micronucleus formation may reflect this variability. ³To whom correspondence should be addressed     

Properties of a Hemolysin Produced by Group B Streptococci

A hemolysin that appears to be responsible for the zones of beta-hemolysis surrounding colonies of group B streptococci (Streptococcus agalactiae) on blood agar plates has been isolated and partially purified. No soluble hemolysin was detectable in the supernatants of streptococcal cultures grown in several types of media. However, hemolytic activity was detected when streptococci were incubated with erythrocytes, and soluble hemolysin was isolated when bacterial suspensions were incubated in the presence of a variety of agents, including calf serum, albumin, Tween 80, and starch. Glucose and other fermentable carbohydrates stimulated hemolysin production, and metabolic inhibitors greatly reduced the titer of hemolysin that could be recovered, suggesting that cellular metabolism is necessary for hemolysin production or release. The soluble hemolysin was concentrated by ammonium sulfate precipitation and partially purified by gel filtration. Agents known to inhibit other streptococcal hemolysins, including phospholipids, trypan blue, proteases, and cholesterol, were tested for their effect on the group B hemolysin. Only the phospholipids inhibited hemolysin activity. The group B streptococcal hemolysin appears to be similar to, but distinct from, streptolysin S produced by Streptococcus pyogenes.     

A computer-interfaced photometer and systematic spacing of duplicates to control within-plate enzyme-immunoassay variation

A Multiskan photometer for reading microtiter plate enzyme immunoassays was linked with a time sharing computer to facilitate control of assay variation and analysis of results. The interface that converted photometer output to RS-232-C format required changes to divide the output into segments short enough for input to the computer. To measure within-plate variation and investigate how the method of allocating sample duplicates to plate wells may affect the estimation of sample variance, uniformity tests were conducted with 47 plates. Coefficients of variation (CV) among wells within-plates ranged from 4.6 to 20.7% and in two-thirds of the plates exceeded 10%. Duplicates allocated to adjacent wells (method 1) gave consistently higher CV for sample means than duplicates allocated to opposite plate quadrants (method 2). In general, the CV by method 2 was about 30% smaller than that by method 1. Analysis of variance confirmed the effectiveness of the quadrant pattern of duplicate allocation as a method of controlling variation that arises from well position effects.     

Cloning, expression, and mapping of the Staphylococcus aureus alpha-hemolysin determinant in Escherichia coli K-12.

A fragment of Staphylococcus aureus DNA encoding the alpha-hemolysin determinant was cloned from strain Wood 46 by inserting Sau3A-generated genomic DNA fragments between the BamHI sites of the lambda replacement vector L47.1. Phages expressing alpha-hemolysin were detected by overlaying plaques formed from several thousand independent recombinant phage with erythrocytes and looking for zones of hemolysis. One phage expressing alpha-hemolysin was purified and named lambda w alpha 3. This was subsequently shown to contain a 10.2-kilobase pair insert of S. aureus DNA. A 7.6-kilobase pair HindIII fragment encoding the alpha-hemolysin was subcloned from lambda w alpha 3 into the plasmid vector pACYC184 to form the hybrid plasmid pDU1148. Escherichia coli K-12 cells harboring pDU1148 synthesized a low level of alpha-hemolysin which remained associated with the cells and was not secreted into culture supernatants. When the same strain was stabbed onto blood agar plates, no zones of hemolysis were detected after overnight growth at 37 degrees C but hemolysis developed if the plates were left at room temperature for 48 h. By introducing specific deletions or Tn5 insertions into plasmid pDU1148, the alpha-hemolysin gene was mapped to a region within a 3.3-kilobase pair EcoRI-HindIII fragment which was subcloned onto the vector plasmid pBR322. A specific enzyme-linked immunosorbent assay with peroxidase-labeled rabbit anti-alpha-hemolysin antibodies was used to measure the levels of alpha-hemolysin antigen expressed in E. coli K-12 cells harboring pDU1148 or a variety of pDU1148::Tn5 and pDU1148 deletion mutants.     

RhoC is dispensable for embryogenesis and tumor initiation but essential for metastasis

The Rho proteins are Ras-related guanosine triphosphatases (GTPases) that function in cytoskeletal reorganization, cell migration, and stress fiber and focal adhesion formation. Overexpression of RhoC enhances the ability of melanoma cells to exit the blood and colonize the lungs. However, in vivo confirmation of RhoC's role in metastasis has awaited a RhoC-deficient mouse model. Here we report the generation of RhoC-deficient mice and show that RhoC is dispensable for embryonic and post-natal development. We demonstrate that loss of RhoC does not affect tumor development but decreases tumor cell motility and metastatic cell survival leading to a drastic inhibition of metastasis.     

Gingipain adhesin domains mediate Porphyromonas gingivalis adherence to epithelial cells

Porphyromonas gingivalis, a Gram-negative oral anaerobe, interacts with epithelium lining the gingival sulcus. Continuing our studies on the role of gingipain cysteine proteinases in P. gingivalis adherence to epithelial cells, we showed that antibody raised to the recombinant adhesin domain of arg-gingipain A blocked bacterial attachment, providing new additional evidence that P. gingivalis adherence to epithelial cells is mediated by gingipain adhesin peptides.     

IgA, IgM and IgG anti-M. leprae antibodies in babies of leprosy mothers during the first 2 years of life.

IgA, IgM and IgG anti-M. leprae antibody activity was estimated by solid phase radioimmunoassay in repeated serum samples from cord sera to sera taken 2 years after birth from 29 babies of mothers with lepromatous leprosy (Group 1) and 16 babies of mothers with tuberculoid leprosy and non-leprosy control mothers (Group 2). IgA anti-M. leprae antibody activity could be detected in 30% and IgM anti-M. leprae antibody activity in 50% of cord sera from Group 1, but not in any of the cord sera from Group 2. After birth, there was a significantly higher increase of IgA and IgM anti-M. leprae antibody activity in sera taken 3-6 months after birth from babies of Group 1 compared to Group 2, but the IgA and IgM activity in sera taken after 6 months of age showed the same increase in the two groups. IgG anti-M. leprae antibody activity showed a marked decrease in sera from both Groups 1 and 2 taken 3-6 and 6-9 months after birth compared to the activity in the cord sera. No increase of the IgG activity could be demonstrated even in sera taken 15-24 months after birth in any of the two groups. These findings are discussed in relation to possible transfer of M. leprae bacilli across the placenta, the influence of M. leprae and other mycobacteria exposure on the antibody activity, the poor IgG anti-M. leprae antibody response and subclinical leprosy infection in babies exposed to leprosy below 2 years of age.