SUB-CHROMATID REARRANGEMENTS IN TRILLIUM ERECTUM. I. ORIGIN AND NATURE OF CONFIGURATIONS INDUCED BY IONIZING RADIATION

Wilson , G. B. (Michigan State U., East Lansing.), A. H. Sparrow , and Virginia Pond . Subchromatid rearrangements in Trillium erectum. I. Origin and nature of configurations induced by ionizing radiation. Amer. Jour. Bot. 46(4): 309–316. Illus. 1959.—Microsporocytes of Trillium erectum were x-irradiated with 25 r at various stages of meiotic prophase and at first metaphase. Analysis of these cells at the following first and second anaphase revealed that post-pachytene irradiation produces 2-side-arm bridges which are indicative of half-chromatid exchanges. The occurrence of these bridges and knowledge of the structure and spatial relationship of chromatid strands in T. erectum have led to certain conclusions regarding the target and the number of strands broken by a single event: (1) the most likely target for primary effects is the 4 associated half-chromatids of a half-bivalent. The results of irradiation experiments suggest that the half-bivalent is effectively as well as structurally quadripartite at stages following pachytene. (2) Consideration of the configurations which would result from breakage and rejoining of 2, 3 or all 4 strands of the half-bivalent indicates that only 2 of the 4 half-chromatids are broken by a single event. Exchanges between 2 half-chromatids of sister chromatids will produce two recognizable types of 2-side-arm bridges: one with a true dicentric half-chromatid and one in which the bridge results merely from an interlocking of coils. Whether a 2-side-arm bridge appears at first or second meiotic anaphase is determined by the position and number of chiasmata between the point of breakage and the kinetochore. No 2-side-arm bridges have been detected at microspore anaphase following meiotic prophase irradiation. The types of configurations which might be expected at microspore metaphase as a result of broken 2-side-arm bridges are noted.     

A scalable, fully automated process for construction of sequence-ready barcoded libraries for 454

We present an automated, high throughput library construction process for 454 technology. Sample handling errors and cross-contamination are minimized via end-to-end barcoding of plasticware, along with molecular DNA barcoding of constructs. Automation-friendly magnetic bead-based size selection and cleanup steps have been devised, eliminating major bottlenecks and significant sources of error. Using this methodology, one technician can create 96 sequence-ready 454 libraries in 2 days, a dramatic improvement over the standard method.     

The Nova Scotia (type D) form of Niemann-Pick disease is caused by a G3097-->T transversion in NPC1.

Niemann-Pick type D (NPD) disease is a progressive neurodegenerative disorder characterized by the accumulation of tissue cholesterol and sphingomyelin. This disorder is relatively common in southwestern Nova Scotia, because of a founder effect. Our previous studies, using classic linkage analysis of this large extended kindred, defined the critical gene region to a 13-cM chromosome segment between D18S40 and D18S66. A recently isolated gene from this region, NPC1, is mutated in the majority of patients with Niemann-Pick type C disease. We have identified a point mutation within this gene (G3097-->T; Gly992-->Trp) that shows complete linkage disequilibrium with NPD, confirming that NPD is an allelic variant of NPC1.     

Xenopus laevis transgenesis by sperm nuclear injection

The stable integration of transgenes into embryos of the frog Xenopus laevis is achieved using the procedure described here. Linear DNA containing the transgene is incorporated randomly into sperm nuclei that have had their membranes disrupted with detergent treatment. Microinjection of these nuclei into unfertilized eggs produces viable embryos that can be screened for activity of the transgene. The proportion of embryos that harbor the transgene varies from 10 to 40% of the total number of surviving embryos. Multiple copies of the transgene can integrate as a concatemer into the sperm genome, and more than one site of DNA integration might occur within resulting animals. Germ cell transmission of the transgene is routine and the procedure is well suited to the production of transgenic reporter frog lines. One day should be allocated for the preparation of the sperm nuclei, which are stored as aliquots for future use. The transgenesis reaction and egg injection take one morning.     

Lipolysis of ApoC-II deficient very low density lipoproteins: enhancement of lipoprotein lipase action by synthetic fragments of apoC-II.

Enzymic hydrolysis of triacylglycerol has been studied with very low density lipoproteins from an individual with a genetically determined absence of apoC-II, the activator apoprotein for lipoprotein lipase. Normal rates of ester cleavage by purified bovine milk lipoprotein lipase can be achieved $\text{in}\text{vitro}$ with native apoC-II and by three shorter synthetic peptides, apoC-II(55–78), apoC-II(50–78) and apoC-II(43–78), which contain part of the carboxyl terminal third of the native apoprotein. At 0.5 μM concentration, all peptides produced a 7-fold activation. ApoC-II(43–78), but not apoC-II(50–78) or apoC-II(55–78), could bind VLDL as shown by separation of unbound ¹²⁵I peptides and the lipoproteins. Thus, residues 43–50 of apoC-II are part of a lipid binding region. High affinity binding of apoC-II peptides to the lipoprotein substrate is not obligatory for activation of lipoprotein lipase.     

OBSERVATIONS ON CHYTRIDIACEOUS PARASITES OF PHANEROGAMS. XXVII. A FURTHER STUDY OF PHYSODERMA HYDROCOTYLIDIS VIÉGAS AND TEIXEIRA

A further study of Physoderma hydrocotylidis from California on its host Hydrocotyle ranunculoides gives further details on its structure and development. EM micrographs reveal that the notably thin wall of the resting spore consists of an outer, an intermediate, and an inner layer. The complete wall thickness from measurements of these EM preparations is only 0.5 μm, the thinnest of any known Physoderma resting spore. Such resting spores germinate readily in 2–5 h by the dehiscence of a broad cap and formation of a protruding endosporganium. Zoospores from the latter infect epidermal host cells and produce either the endobiotic, polycentric rhizoidal system with tenuous filaments, turbinate cells and eventually resting spores, or a hitherto unknown epibiotic, monocentric, rhizidiaceous sporangial stage. Zoospores from the latter may all bear colorless lipid globules or all faintly orange-colored ones.     

Fluorescent pigments of the retinal pigment epithelium and age-related macular degeneration.

The major hydrophobic fluorophore of the retinal pigment epithelium (RPE) is A2E, a pyridinium bis-retinoid derived from all-trans-retinal and phosphatidyl-ethanolamine. The accumulation of fluorophores such as A2E is implicated in the pathogenesis of age-related macular degeneration (AMD), a disease associated with the deterioration of central vision and a leading cause of blindness in the elderly. Recent chemical and biological studies have provided insight into the synthesis and biosynthesis of A2E, the spectroscopic properties of this pigment, and the role of A2E and RPE cell death.     

Successful immunotherapy with matrix metalloproteinase-derived peptides in adjuvant arthritis depends on the timing of peptide administration

We have recently found that matrix metalloproteinases (MMPs) are targets for T-cell and B-cell reactivity in experimental arthritis. In the present article, we investigate whether modulation of MMP-specific T-cell responses could influence the course of adjuvant arthritis (AA). Lewis rats were treated nasally with MMP peptides prior to or after AA induction. Administration of the MMP-10 or the MMP-16 peptide prior to AA induction reduced the arthritic symptoms. In contrast, administration of the MMP-10 peptide after AA induction aggravated the arthritic symptoms. The present study shows the possible usefulness of MMP peptides for immunotherapy. However, a clear understanding of proper timing of peptide administration is crucial for the development of such therapies.