Restriction fragment length polymorphism at the HLA-C locus

We have used the HLA-C-specific DNA probe pC250 to investigate restriction fragment length polymorphism (RFLP) at the HLA-C locus. Genomic Southern blot hybridization included DNA prepared from a panel of homozygous typing cells representing serological specificities Cw1 to Cw8 and also from samples representing Cw blanks. Although many restriction nucleases failed to reveal any polymorphism, RFLPs were evident with Taq I, Pvu II, Bst XI, Nde 1, and Nci I in addition to the previously reported Eco RI. In the case of Bst XI, a unique RFLP defined a subset of serologically defined Cw blanks. Comparison of RFLP sizes with restriction fragment lengths obtained from the known HLA-Cw3 gene sequence permitted the localization of intragenic C locus RFLLs and the identification of a variable Taq I site in the second intron, a variable Nci I site near the end of the fourth exon, and a variable Pvu lI site in the fifth intron.     

A constitutive description of contracting papillary muscle and its implications to the dynamics of the intact heart

A simple mathematical model capable of simulating the major biomechanical attributes of contracting cardiac muscle is presented. This model is based on the phenomenological observations on heart muscle. The form of the equation can be readily extended to describe the pressure-volume-time-velocity of the intact heart as well, thus allowing a direct bridge between the dynamics of papillary muscle and the dynamics of intact heart. Parameters that are sensitive to inotropic state of the muscle can be obtained directly from the isometric tension-time record of the muscle or the isovolumic pressure-time record of the ventricle. These parameters have the potential to serve as quantitative measures of cardiac health.     

Long-wavelength fluorescence of tyrosine and tryptophan solutions

We report in this paper the presence of fluorescence bands of tryptophan and tyrosine solutions centered above 550 nm. This long-wavelength fluorescence is of much lower intensity, (0.4-2.7)%, than the UV fluorescence of these aromatic aminoacids. The basic characteristic of these fluorescence bands are: (a) tyrosine: lambda em = 600 nm with two excitation peaks centered at 453 nm and 550 nm (b) tryptophan: lambda em = 675 nm with two excitation peaks centered at 455 and 560 nm. It has been found that irradiation of tyrosine solutions with a potent UV lamp promotes an important increase of absorption at 310 nm and above 400 nm.     

Environmental factors influencing the occurrence of juvenile fish in the mangroves of Pagbilao,Philippines

128 fish species belonging to 54 families were collected from the mangroves of Pagbilao. Ambassis kopsi (Ambassidae) was the most abundant species.Fourteen environmental factors were correlated to the catch per unit time for some species. The occurrence of A. kopsi was positively correlated to nitrate in the water, carbon in the sediments and litter fall. Out of the 8 species investigated, 6 were positively correlated to litter fall. Three species showed positive correlation to phosphate in the water, two to organic carbon in the sediments, nitrate, silicate and pH, and one to salinity and carotenoids in water. The biomass of the total catch was correlated to carbon in the sediments and litter fall.     

U5 small nuclear ribonucleoprotein: RNA structure analysis and ATP-dependent interaction with U4/U6.

To understand how the U5 small nuclear ribonucleoprotein (snRNP) interacts with other spliceosome components, its structure and binding to the U4/U6 snRNP were analyzed. The interaction of the U5 snRNP with the U4/U6 snRNP was studied by separating the snRNPs in HeLa cell nuclear extracts on glycerol gradients. A complex running at 25S and containing U4, U5, and U6 but not U1 or U2 snRNAs was identified. In contrast to results with native gel electrophoresis to separate snRNPs, this U4/U5/U6 snRNP complex requires ATP to assemble from the individual snRNPs. The structure of the U5 RNA within the U5 snRNP and the U4/5/6 snRNP complexes was then compared. Oligonucleotide-targeted RNase H digestion identified one RNA sequence in the U5 snRNP capable of base pairing to other nucleic acid sequences. Chemical modification experiments identified this sequence as well as two other U5 RNA sequences as accessible to modification within the U5 RNP. One of these regions is a large loop in the U5 RNA secondary structure whose sequence is conserved from Saccharomyces cerevisiae to humans. Interestingly, no differences in modification of free U5 snRNP as compared to U5 in the U4/U5/U6 snRNP complex were observed, suggesting that recognition of specific RNA sequences in the U5 snRNP is not required for U4/U5/U6 snRNP assembly.     

Beta 1 and beta 2 adrenergic agonists in exocrine pancreatic secretion in the rabbit

The effect of Dobutamine (a beta 1-adrenergic agonist) and Terbutaline (a beta 2-adrenergic agonist) on exocrine pancreatic secretion was studied in anaesthetized rabbits, simultaneously controlling pancreatic blood flow and blood pressure. The secretion of fluid and ions (bicarbonate, sodium and potassium) was unaffected by the infusion of Dobutamine (8 micrograms.kg-1.min-1) or Terbutaline (10 micrograms.kg-1.min-1). Neither were pancreatic blood flow or mean blood pressure altered. Dobutamine or Terbutaline depress the function of the acinar cells, amylase secretion being more affected by the action of Terbutaline. The results show that beta 1 and beta 2-adrenergic stimulation has no effect on the ductular cells but does decrease the secretion by the acinar cells.     

The endogenous lectins of the chick blastoderm are present in association with an apolipoprotein in distinct organelles and in the extracellular matrix

Summary Affinity purified preparations of the galactose-binding lectin from gastrulating chick blastoderms consist of three main polypeptides. Two of these have been identified as the 14 kD and 16 kD galactose-binding lectins. A third one migrates in SDS-PAGE gels with a relative molecular weight of 6,500±500 and has been identified as an apolipoprotein (Apo) of plasma very low density lipoproteins, Apo-VLDL-II. We have studied the localization of these polypeptides using immunofluorescence and ultrastructural immunocytochemistry with peroxidase and protein-A gold. The 14 kD lectin occurs in the intracellular yolk where it is mainly present within the electron lucent component. The 16 kD is also present in the intracellular yolk platelets, but tends to predominate in the electron-dense component. In addition, the 16 kD lectin is also present in pleiomorphic yolk-associated organelles and in the extracellular matrix. Apo-VLDL-II is also localized in the electron-lucent component of the yolk platelet and in the extracellular matrix. Our results suggest that the lectin(s) are associated with Apo-VLDL-II in the yolk platelet, and may subsequently become externalized.     

Temporal relationship between nerve-stump-length-dependent changes in the autophosphorylation of a cyclic AMP-dependent protein kinase and the acetylcholine receptor content in skeletal muscle

The acetylcholine receptor (AChR) content and the autorphosphorylation of the regulatory subunit of cyclic AMP-dependent protein kinase type II (R-II) were evaluated in rat soleus muscles at 24, 30 and 66 hr after surgical denervation by cutting the nerve at a short distance (short-nerve-stump) and at a long distance (long-nerve-stump) from the muscle. AChR content was based on the specific binding of [¹²⁵I]alpha-bungarotoxin (BUTX); changes in the autophosphorylation of R-II were based upon the predominant in vitro³²P-phosphorylation of a 56-Kd soluble protein in cytosolic fractions of solei. The AChR content and the³²P-autophosphorylation of R-II were increased in samples from short-nerve-stump solei, but not from long-nerve-stump solei, after a denervation-time of 30 hr. This nerve-stump-length dependency indicates that the two denervation effects are not related to the immediate halt of impulse-evoked muscle contractility. Furthermore, the results show that alterations in the³²P-autophosphorylation of R-II occurred before, as well as whenever, increases in the AChR content were found. Speculatively, this temporal relationship may be significant with respect to the potential role of R-II in gene expression.Abbreviations ACh acetylcholine - AChR acetylcholine receptor(s) - BUTX alpha-bungarotoxin - Kd kilodalton - PAGE polyacrylamide gel electrophoresis - R-II regulatory subunit of cyclic AMP-dependent protein kinase type II - SDS sodium dodecyl sulfate     

Label-fracture of cell surfaces by replica staining

We introduce replica-staining label-fracture, a method for the cytochemical mapping of membrane surfaces. This method is a corollary of the rationale of label-fracture (Pinto da Silva and Kan, 1984: J Cell Biol 99:1156). After freeze-fracture the exoplasmic halves of the membrane remain attached to the replica. We show that cytochemical labeling of cell surfaces can be performed by direct post-fracture staining of freeze-fracture replicas. This new variant of label-fracture leads to miniaturization of labeling procedures and allows standardization of labeling conditions and simultaneous processing of different specimens.