Cap-dependent and cap-independent translation: operational distinctions and mechanistic interpretations.
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Heat-Shock Proteins↗
Biomedical subjects
J E Reynolds
Publications and source records attributed to J E Reynolds.
Mcl-1, a member of the Bcl-2 family, delays apoptosis induced by c-Myc overexpression in Chinese hamster ovary cells.
Mcl-1, a protein increased early in the differentiation of human myeloblastic ML-1 cells, has sequence similarity to Bcl-2. In the present study, we determined whether Mcl-1 has functional similarity to Bcl-2 by testing its ability to inhibit apoptosis induced by c-Myc overexpression. This was carried out using Chinese hamster ovary 5AHSmyc cells which contain the human c-myc proto-oncogene under the control of a heat shock promoter. Heat treatment induces c-Myc overexpression and thus apoptosis as determined by internucleosomal DNA fragmentation. We transfected 5AHSmyc cells with mcl-1 and found that clones expressing the introduced Mcl-1 protein exhibited reduced DNA fragmentation. Mcl-1 was also capable of delaying the onset of cell death as judged by loss of membrane integrity, although it could not provide complete protection from c-Myc overexpression. Thus, Mcl-1 has functional homology to Bcl-2 in that Mcl-1 can enhance cell viability under conditions that otherwise cause apoptosis.
Animals↗
Dinucleotide repeat polymorphism at the CHRND locus.
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Base Sequence↗
Etoposide-induced apoptosis in human HL-60 cells is associated with intracellular acidification.
Apoptosis is a pathway of cell death characterized by internucleosomal digestion of genomic DNA. Such DNA digestion can be induced by both physiological stimuli and cytotoxic treatment with many anticancer agents. This digestion has generally been considered to be mediated by a Ca2+/Mg(2+)-dependent endonuclease that is activated by increases in intracellular Ca2+. However, we suggest that an alternate endonuclease, DNase II, may be a more likely candidate. In these studies, apoptosis was induced in human HL-60 cells by a 30-min incubation with the topoisomerase II inhibitor etoposide. DNA digestion characteristic of apoptosis began within 3 h of removal of etoposide. Morphological indication of apoptosis was observed concurrently. Only about 20% of the cells underwent apoptosis at this time; these appeared to be cells in S phase at the time of etoposide treatment. The remainder of the cells progressed to the G2 phase and arrested there for at least 48 h. Intracellular Ca2+ and pH were measured in individual cells by flow cytometry. No changes in intracellular Ca2+ were observed, but an acidification of up to 1 pH unit occurred in about 15% of the cells and correlated with the time course of appearance of DNA digestion. Cells were sorted on the basis of intracellular pH and only the acidic cells showed the morphology and DNA digestion characteristic of apoptosis. These results demonstrate the involvement of DNase II in apoptotic DNA digestion and suggest mechanisms of pH homeostasis as regulators of apoptosis.
Apoptosis↗
A method for recovery of high-molecular-weight DNA suitable for field-inversion gel electrophoresis from frozen tumor cells.
We describe a simple method for isolation of high-molecular-weight (high-mol-wt, > 800 kilobases) DNA from tumor cell lines frozen without dimethylsulfoxide (DMSO) or other protective agents. This method should be applicable to most frozen tissues and useful for molecular genetic studies such as field-inversion gel electrophoresis (FIGE), or cloning procedures that require access to intact, high-mol-wt DNA.
Cryopreservation↗
Molecular characterization of a 17q11.2 translocation in a malignant schwannoma cell line.
Malignant schwannomas are soft-tissue neoplasms that occur at increased frequency with germline alterations of the neurofibromatosis-1 (NF1) gene at 17q11.2. We report molecular and cytogenetic characterization of a malignant schwannoma cell line established from an individual affected with NF1. This cell line has a complex hyperdiploid karyotype with two cytogenetically identical der(13)t(13;17)(p11,q11.2) chromosomes. Using somatic cell hybrids, we mapped twelve chromosome-17 probes to either the der(13)t(13;17) chromosome or a small der(17) chromosome. Two chromosome-17p loci, including the p53 tumor suppressor gene, were present in the schwannoma cell line, but did not map to either of these chromosomes. Loss of heterozygosity studies indicated that the two der(13)t(13;17) chromosomes arose by duplication, presumably after the translocation event. The 17q11.2 translocation break-point maps distal to the NF1 gene, and may not disrupt its functioning. Although NF1 mRNA was detected in this cell line by polymerase chain reaction, Northern blot analysis revealed very little or none of the 13-kb mature NF1 transcript. This suggests that the single remaining allele of the NF1 gene contains a mutation that results in either greatly reduced transcription or message instability.
Adult↗
Astrometry of southern radio sources.
An overview is presented of a number of astrometry and astrophysics programs based on radio sources from the Parkes 2.7 GHz catalogues. The programs cover the optical identification and spectroscopy of flat-spectrum Parkes sources and the determination of their milliarc-second radio structures and positions. Work is also in progress to tie together the radio and Hipparcos positional reference frames. A parallel program of radio and optical astrometry of southern radio stars is also under way.
Astronomical Phenomena↗
Gross and microscopic anatomy of the kidney of the West Indian manatee, Trichechus manatus (Mammalia: Sirenia).
Examination of the gross and microscopic anatomy of the kidney of the West Indian manatee, Trichechus manatus, revealed that: (1) the medulla is about 6 times thicker than the cortex; (2) juxtamedullary glomeruli have a mean diameter 1.3 times greater than that of cortical glomeruli; (3) juxtamedullary glomeruli have 1.7 times as much volume as cortical glomeruli; (4) there are about twice as many cortical glomeruli as juxtamedullary glomeruli per square millimeter of cortical tissue, and (5) the vasa recta are closely juxtaposed to the thin loops of Henle in the outer medulla. Many of these results suggest an enhanced urine-concentrating ability in this species.
Animals↗
Erythropoiesis and granulopoiesis in the West Indian manatee, Trichechus manatus (Mammalia: Sirenia).
The West Indian manatee, Trichechus manatus, has amedullary, pachyostotic long bones. Since marrow cavities of the long bones typically are the locations of hemopoiesis in adult mammals, the manatee has evolved an alternative primary site of hemopoiesis. Histological examinations of spleens, livers, kidneys, and vertebral bodies indicate that the last-named location is the main site of erythropoiesis and granulopoiesis in T. manatus. This conclusion is based on the presence in the vertebrae of bone-marrow-containing diagnostic cell types including erythroblasts, megakaryocytes, and myelocytes, which are the precursor cells of erythrocytes, platelets, and granular leukocytes, respectively. No developing lymphocytes were found, so that the location of lymphopoiesis remains unknown.
Animals↗
Fetal hydrocephalus: a case of ventricular-amniotic shunting.
The case we have reported describes the management of fetal hydrocephalus using an intrauterine ventricular-amniotic shunt, which unfortunately was not recovered after delivery. To prevent this potential complication, we suggest such shunts be made radiopaque. The encouraging results in this case are attributed to the efforts of an interdisciplinary team and suggest that vascular accidents may well prove to be amenable to treatment by this new procedure.
Adult↗
Direct fetal therapy: the multidisciplinary team concept.
A multidisciplinary medical and surgical team is essential to direct fetal therapy. This group of skilled specialists is usually available only in university hospitals. Through the use of recently developed technology and a high degree of skill, the team composed of nonphysician and physician members can detect, evaluate, and occasionally treat affected patients. Through research, education,aand communication, the team endeavors to meet the need for direct fetal therapy in life-threatening malformations.
Congenital Abnormalities↗
A note on the duodenum of the West Indian manatee (Trichechus manatus), with emphasis on the duodenal glands.
The anatomy of the duodenum of the West Indian manatee has both macroscopic and microscopic features that are unusual when compared to other mammalian forms. Macroscopically, the voluminous duodenal ampulla and the paired duodenal diverticula are distinctive. The general microscopic structure of the manatee duodenum is not unusual, but the duodenal glands secrete an acid mucin (sialomucin). The cells of these glands appear to be intermediate between classical serous and mucous cell types. These cells also contain granules with regions of high peripheral electron density. The overall structure of the manatee duodenum most strongly resembles that of the dugong, another member of the order Sirenia.
Animals↗
Analysis of locus heterogeneity in Waardenburg syndrome types 1 and 2 using highly informative microsatellite markers.
We performed linkage and locus heterogeneity analyses of Waardenburg syndrome (WS) types 1 and 2 using 9 DNA markers from 2q35-q37, including two highly polymorphic microsatellites very closely linked to the PAX3 candidate gene. None of 5 WS type 2 (WS2) families showed linkage to the PAX3 candidate region. We localized the marker D2S102 to less than 1 cM from PAX3 (lod = 33.7, theta = 0), but a complete absence of crossovers prevented determining whether it maps distal or proximal to PAX3. Study of 14 WS type 1 (WS1) families yielded a maximum lod score of 27.81 at PAX3, theta f = 0.010, theta = 0.007 assuming homogeneity. However, we found significant evidence of locus heterogeneity, with one family initially classified as WS1 unlinked to the PAX3 region. Reevaluation of the clinical features of this family revealed atypical morphology of inner canthi. This produced the appearance of dystopia canthorum and high W-index scores. While our one unlinked WS1 family exhibits atypical canthal morphology, our type 1 families with classic dystopia appear to be homogeneously linked to PAX3. These and other findings identify precautions that need to be addressed before using PAX3-linked markers for diagnostic purposes.
Chromosome Mapping↗