[Studies on the method of nucleic acid precipitation with metallic ions].
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Biomedical subjects
Publications and source records attributed to L Liu.
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We have studied the development of thyroid hormone dependence for growth and differentiation of rat skeletal tissues. The radius, tail tip or xiphoid process from 1-day-old rats was transplanted in its entirety under the kidney capsule of euthyroid or hypothyroid (TX) adult syngeneic virgin rats and grown there for 3, 6, 9, 12 or 15 days. Pieces of the sternum or calvarium were treated similarly. Growth of transplanted radius and sternum in TX hosts showed significant retardation by day 3, and that of transplanted calvarium, tail tip and xiphoid showed significant inhibition by day 6 post transplantation. Tissue differentiation of all of the transplants was inhibited in TX hosts and the appearance of epiphyseal ossification centers was significantly delayed. Injection of Propylthiouracil (PTU) in solvent into the neonatal rats from day 1 to day 15 post partum inhibited tail length increase by day 7, and body weight gain by day 9, as compared to the growth of solvent injected littermate pups. Our results indicate that the growth and differentiation of endochondral bones are more dependent on TH levels than are those of membranous bones and cartilage. Endochondral bones develop TH dependence for growth and differentiation before day 4 post partum, while the membranous bones and cartilage develop this dependence at the end of the first week.
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We studied the biological activity of a recently characterized 31-amino acid inhibin-like peptide (ILP) using a synthetic preparation. While the material yielded a single component on chromatography, amino acid sequence analysis suggested that only 30% of the molecules possess the complete structure. Bioactivity was tested in vitro using whole pituitaries from 25 day-old male Sprague-Dawley rats. Pituitaries were incubated with 500 ng/ml of the ILP preparation or vehicle alone for 60 min, followed by a 3 h exposure to 2 ng/ml of luteinizing hormone releasing hormone (LHRH) or its diluent. In the ILP-incubated pituitary media, no significant suppression of basal FSH and LH release or stimulated FSH release was observed, while a significant increase in stimulated LH release was seen (p less than 0.05). An in vivo bioassay study was performed on 38 day-old male Sprague-Dawley rats. Test animals were injected with 0.1, 1, 10 or 20 mcg of ILP immediately after castration, and 10 and 24 h later. Control animals received bovine serum albumin or vehicle alone. There were no statistically significant differences in serum LH or FSH concentrations taken at 30 h after castration between the ILP-treated rats and controls. Possible reasons for our inability to demonstrate inhibin bioactivity (selective FSH suppression) with this ILP preparation include: Only about 30% of the peptides had the complete amino acid sequence. Other peptides with deletions might have acted as antagonists, thus obscuring the inhibin bioactivity. The 31-amino acid ILP may not be the authentic inhibin molecule. We suggest that further studies are needed to determine the true identity of inhibin.
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A novel H-2 gene in which the first external (N) domain of the H-2Ld antigen was replaced with that of the H-2Dd antigen was constructed and introduced into L cells. A transformant expressing the products of the hybrid gene was studied for binding to monoclonal antibodies specific for H-2Ld and H-2Dd antigens. It was found that serological determinants are distributed both in the N (Dd) and Cl (Ld) domains. Determinants recognized by allospecific cytotoxic T lymphocytes (CTLs) and virus-specific CTLs also mapped to the N and Cl domains. Determinants recognized by vesicular stomatitis virus (VSV)-specific effect cells, however, were not present on the recombinant molecule. These results show that a recombinant gene of two H-2 antigens in which the first external domain has been reshuffled can express a functional H-2 antigen that can then be used to map serological and CTL determinants to specific domains.
The spatio-temporal expression of specific binding sites for FITC-labeled peanut agglutinin (Arachis hypogaea) in frozen sections of whole embryonic heads of the chick between embryonic day 2 (E2) and embryonic day 8 (E8) is described. PNA-binding sites are present already at E2 on the external limiting membrane and the ventricular boundary of the neural wall. With progressing differentiation fluorescent bands are formed from the outside of the neural wall radiating inside, forming second layers inside the CNS at later stages. The visual system (retina and tectum) shows remarkably little PNA-binding activity until about E7/8, a time when binding sites in the other brain areas are already most abundant. In the telencephalon a fuzzy distribution of fluorescence is found. A possible significance of this finding for fiber guidance from the retina to the tectum is briefly discussed. Local high activities can be observed in different parts of the brain. Intense bands of PNA-fluorescence on parts of the epidermis, along the eye cups, around the fore- and hindbrain and in the medial part of the mesenchyme delineate early biochemical compartments foreshadowing the succeeding ossification.
The first clinically and biochemically documented case of recurrent thyrotoxicosis after I-131 induced hypothyroidism in a patient with Graves' disease is reported. Two months after the administration of 9.2 mCi of I-131, the subject developed hypothyroidism. One month later, the patient became euthyroid. Then, nine months following ablation, the patient again developed thyrotoxicosis. A second dose of I-131 of 12.5 mCi was required to finally produce permanent hypothyroidism. This case illustrates the recurrence of hypothyroidism after what had seemed to have been adequate I-131 radiation.
Epipodophyllotoxins are an important new class of anticancer agents which include the compounds VM-26 (teniposide) and VP-16 (etoposide). The mechanism of action of these drugs appears to involve production of DNA single- and double-strand breaks by virtue of a temperature-sensitive interaction between drug and a heat-labile intranuclear component. We now report evidence indicating that type II topoisomerase is the likely intracellular target for the DNA strand-breaking effects of the epipodophyllotoxins. Both VM-26 and VP-16 stimulate site-specific DNA cleavage by a highly purified calf thymus type II topoisomerase. VM-26 is 5- to 10-fold more potent than VP-16 in this assay, a difference that is also seen when DNA strand breaks are assayed in isolated nuclei of mouse leukemia cells following drug exposure. Furthermore, a similar potency difference exists with respect to cytotoxicity. Equilibrium dialysis experiments using [3H]VP-16 indicate that the drug does not bind to DNA. Thus, we suggest that the epipodophyllotoxins exert their anti-cancer effects by "poisoning" type II topoisomerase without binding to DNA. In this regard, their actions may be analogous to those of nalidixic acid in bacteria.
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FITC-labeled peanut agglutinin (PNA), a lectin from peanut (Arachis hypogaea) which binds specifically to D-galactose, was used as a probe to trace the distribution of PNA-lectin receptors during early development of chick retina. We found that specific fluorescence within the inner plexiform layer (IPL) and the outer plexiform layer (OPL) appears at different developmental stages. The specific fluorescence of the IPL first appears near the central area of the ventro-posterior quadrant of the 7-day retina, then extends progressively towards the periphery, reaching the ora serrata earlier on the posterior than on the anterior side. The development of the specific fluorescence of the OPL follows the same scheme, but it appears later (starting at day 9) than that of the IPL and it is less pronounced. The spatio-temporal patterns of the expression of the lectin receptors show relationships to those of withdrawal of proliferating cells from mitosis, and of other events related to differentiation.
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