Pre-lysosomal divergence of alpha 2-macroglobulin and transferrin: a kinetic study using a monoclonal antibody against a lysosomal membrane glycoprotein (LAMP-1)

We have used electron microscopic immunocytochemistry to compare the distribution of LAMP-1, a marker for lysosomal membranes, with the intracellular localization of alpha 2-macroglobulin (alpha 2-M) and transferrin at various time points after their endocytosis into cultured NIH 3T3 cells. The purposes of this study were (a) to determine how soon endocytic ligands reach lysosomal organelles, (b) to examine whether the intermediate endocytic vesicles gained lysosomal markers gradually or in a precipitous, discrete event, and (c) to examine the relationship, if any, between the pathway of recycling ligands and lysosomes. At early time points (0-5 min) after initiation of endocytosis, most structures containing alpha 2-M labeled with colloidal gold (receptosomes) were not labeled by anti-LAMP-1 detected using ferritin bridge or peroxidase immunocytochemistry. At late time points (greater than or equal to 15 min), the structures containing alpha 2-M (lysosomes) were strongly labeled by anti-LAMP-1. In contrast, transferrin that was directly labeled with ferritin was mostly located in LAMP-1-negative structures at all time points studied. The proportion of alpha 2-M-gold containing vesicles strongly labeled for LAMP-1 roughly paralleled the proportion of alpha 2-M-gold-containing structures positive for cytochemically detectable acid phosphatase. Our data indicate that ligands such as transferrin that are internalized through coated pits and receptosomes, but not delivered to lysosomes, do not traverse a lysosomal organelle compartment as marked by LAMP-1 content. Ligands such as alpha 2-M that are destined for lysosomal delivery reach a LAMP-1-positive organelle compartment only after they traverse LAMP-1-negative, non-lysosomal vesicles previously described as receptosomes.     

The impact of diagnostic tests in evaluating patients with syncope

We reviewed the charts of 100 patients admitted to the hospital for evaluation of syncope. The charts were examined with special attention given to the causes of syncope, the frequency and benefit of diagnostic tests, and the relative cost of these tests. In 39 patients no etiology for syncope was found, and another 18 were felt to have had a vasovagal episode. Twelve patients had arrhythmias as the cause for syncope. Most of the patients underwent a variety of diagnostic tests including cardiac enzyme determinations, brain scans, electroencephalograms, head CAT scans, and Holter monitoring. In most instances, these tests added little useful information to the initial history and physical exam and were done at great expense to the patient. Our data suggest that extensive neurologic testing in patients with "routine" syncope is not warranted and that the focus of hospitalization should be to rule out potentially life-threatening arrhythmias.     

Internal proteins of bacteriophage T4D: Their characterization and relation to head structure and assembly

Three basic proteins of low molecular weight (about 8000, 10,000 and 18,000) were isolated from the T4D phage particle. Many molecules of each protein are located within the phage head, possibly in association with the DNA, and together with the proteins which form the head membrane comprise most of the head structural protein. The purified internal proteins were characterized by physicochemical and immunological techniques; a radio-immunoassay allowed measurement of their synthesis in phage infected bacteria. Each internal protein is synthesized at both early and late times after infection. Their structural genes are present in the phage genome, but do not appear to be among the known amber mutant-containing genes of T4D. No evidence was found to suggest that the internal proteins are formed from a common precursor molecule, nor are their origins related to those of the internal peptides; however, one of the internal proteins may be altered before its incorporation into the phage. Pulse-chase experiments with two of these proteins show that they are incorporated into certain defective T4D heads. Whether or not they are incorporated appears to depend on the degree of completion of these heads, perhaps with respect to DNA packaging.     

Effect of actinomycin D and guanidine on the formation of a ribonucleic acid polymerase induced by foot-and-mouth-disease virus and on the replication of virus and viral ribonucleic acid

The RNA-dependent RNA polymerase induced in baby-hamster kidney cells by infection with foot-and-mouth-disease virus can be detected as early as 60min. after infection, which is 60min. before viral RNA synthesis commences. The time at which the polymerase can first be detected coincides with the latest time at which actinomycin D (50mug./10(7) cells) or guanidine (1mg./10(7) cells) inhibits virus replication. However, by increasing the concentration of guanidine, viral replication can be inhibited later in the growth cycle, casting doubt on the validity of the hypothesis that guanidine acts specifically on the formation of the viral RNA polymerase.     

Growth rates in outbreak populations of the corallivorous gastropod Drupella cornus (Röding 1798) at Ningaloo Reef,Western Australia

Mark and recapture experiments were used to estimate the size-at-age relationship of the coral-eating gastropod Drupella cornus at Ningaloo Reef, Western Australia. Animals 15 mm long increased in length by 5.2 mm in six months at a site in an early stage of an outbreak, but by only 3.8 mm at a site within an established outbreak. Snails larger than 35 mm grew very little. Based on observed growth rates fitted to a Richards function growth equation, snails 28 mm long judged to be reproductively mature would be 2.5 to 3.5 years old, suggesting that local outbreaks represent no more than one or two generations of recruits. Indirect evidence indicates that growth is more rapid in the earlier than in later stages of outbreaking populations. The growth and timing of life-history transitions of D. cornus are not unusual among other muricid species.