The methylation status of DNA derived from potato plants recovered from slow growth

The in vitro conservation of potato using tissue culture medium supplemented with the growth retardant mannitol causes morphological changes in the propagated material. These culture conditions seem to have an affect on the DNA extracted from the regenerated plants, when it is digested by the methylation sensitive restriction enzymes Hpa II/Msp I and Eco RII/Bst NI, compared to the control material. In most of these plants, there appears to be preferential methylation of nuclear domains that contain Eco RII/Bst NI recognition sites in contrast to those that contain Hpa II/Msp I sites. The refractory nature of the isolated DNA to these restriction enzymes was attributed to hypermethylation of genomic DNA and the ribosomal RNA genes. These findings indicate that methylation of DNA sequences may be an adaptive response to conditions of high osmotic stress. The importance of these results for the conservation of potato germplasm and international exchange is discussed.     

Inherited prion disease (PrP lysine 200) in Britain: two case reports.

OBJECTIVE--To identify cases of inherited prion diseases in Britain and to assess their phenotypic features. DESIGN--Screening study of patients suspected clinically to have Creutzfeldt-Jakob disease and other neurodegenerative diseases by prion protein gene analysis. SETTING--Biochemical research department. SUBJECTS--Patients suspected to have Creutzfeldt-Jakob disease and other neurodegenerative diseases. RESULTS--Two patients with symptoms characteristic of sporadic Creutzfeldt-Jakob disease were found to have inherited prion protein disease (PrP lysine 200), with a mutation at codon 200 of the prion protein gene. Both were homozygous at codon 129 of the gene. One patient was a man aged 58 of British descent while the other was of Libyan Jewish origin. CONCLUSION--Two foci of inherited prion disease are known, among Libyan Jews and in Slovakia. A separate British focus of the disease may also exist. Heterozygosity at codon 129 may lead to reduced penetrance of the mutation.     

THE TIME-COURSE OF PHOTOADAPTATION TO LOW-LIGHT IN PROROCENTRUM MARIAE-LEBOURIAE (DINOPHYCEAE)1

The estuarine dinoflagellate, Prorocentrum mariaelebouriae (Parke & Ballantine 1957) Faust 1974 undergoes increases in pigmentation and photosynthetic efficiency within several days of downward light shifts. These changes can be described by first-order kinetics, as has been reported previously for Chlorophyll (Chl) a in several phytoplankton species. The studies described in this paper were conducted with isolates of populations of Prorocentrum from the Chesapeake Bay. We determined rates of adaptation to low-light for cultures grown at a range of photon flux densities (I₀= 2.65–26.2 E.m^?2, d^?1, shifted to 6.3–7.0% I₀) at three temperatures (10°, 15°, and 20° C), bracketing the conditions this species experiences in situ. In this paper, I report the time-course of changes in α, P_max Chl a, peridinin, and I_k and first-order rate constants, K₁ for changes in α, Chl a and peridinin. cell^?1. K₁ for changes in α cell^?1 averaged 1.58 × 10^?2 h^?1 for conditions encompassing five light treatments and three temperatures; the corresponding mean for Chl a was 1.59 × 10^?2 h^?1. Increases in peridinin measured for five light treatments at 15° C showed a mean K₁ of 1.22 × 10^?2 h^?1, Average percent changes in per cell α, Chl a, and peridinin ranged from 0.4–4.0% h^?1 (10–90% d^?1) following exposure to low-light. Photoadaptive changes are important to Prorocentrum because in nature it occupies turbid waters (K_t≥ 0.5 m^?1) where the mixing depth often exceeds the depth of the photic layer. Cells are entrained beneath a seasonally-stable density discontinuity and are exposed to very low-light (< I E.m^?2.d^?1) for days to weeks during subpycnocline transport. The ability of this species to undergo changes in pigmentation and photosynthetic physiology confers increased efficiency of light harvesting and contributes to this species’survival in the estuary where it is an important component of the dinoflagellate flora.     

Rearrangement of lactose on sterilization

Summary Thermal sterilisation of a lactose-based medium resulted in rearrangement of substantial amounts of lactose to lactulose which was not fermented.     

The increasing complexities in the world of plastic surgery.

The increasing complexities in medicine, in medical education, and in plastic surgery seem to almost defy resolution. The best way to cope with these complexities, and to provide opportunity on an individual or a group basis, is by adherence to quality training program of the type which has served us well and in which the objective is constant. This implies the selection of high-quality candidates for training, and the control of resident flow, so that we may continue as a learned profession. History witnesses change--and time will bring new philsophies, new surgeons with changing values, and a public oriented to a different system of medical care. Shakespeare said, "when the day ends, the end will be known." I am confident that plastic surgery, with its traditional emphasis on quality and excellence, will long endure as a most learned profession.     

Mixture models and wavelet transforms reveal high confidence RNA-protein interaction sites in MOV10 PAR-CLIP data

The Photo-Activatable Ribonucleoside-enhanced CrossLinking and ImmunoPrecipitation (PAR-CLIP) method was recently developed for global identification of RNAs interacting with proteins. The strength of this versatile method results from induction of specific T to C transitions at sites of interaction. However, current analytical tools do not distinguish between non-experimentally and experimentally induced transitions. Furthermore, geometric properties at potential binding sites are not taken into account. To surmount these shortcomings, we developed a two-step algorithm consisting of a non-parametric two-component mixture model and a wavelet-based peak calling procedure. Our algorithm can reduce the number of false positives up to 24% thereby identifying high confidence interaction sites. We successfully employed this approach in conjunction with a modified PAR-CLIP protocol to study the functional role of nuclear Moloney leukemia virus 10, a putative RNA helicase interacting with Argonaute2 and Polycomb. Our method, available as the R package wavClusteR, is generally applicable to any substitution-based inference problem in genomics.     

Characterization of 12 microsatellite loci of the human MHC in a panel of reference cell lines

 The human genome contains a large number of interspersed microsatellite repeats which exhibit a high degree of polymorphism and are inherited in a Mendelian fashion, making them extremely useful genetic markers. Several microsatellites have been described in the HLA region, but allele nomenclature, a set of broadly distributed controls, and typing methods have not been standardized, which has resulted in discrepant microsatellite data between laboratories. In this report we present a detailed protocol for genotyping microsatellites using a semi-automated fluorescence-based method. Twelve microsatellites within or near the major histocompatibility complex (MHC) were typed in the 10th International Histocompatibility Workshop homozygous typing cell lines (HTCs) and alleles were designated based on size. All loci were sequenced in two HTCs providing some information on the level of complexity of the repeat sequence. A comparison of allele size obtained by genotyping versus that obtained by direct sequencing showed minor discrepancies in some cases, but these were not unexpected given the technical differences in the methodologies. Fluorescence-based typing of microsatellites in the MHC described herein is highly efficient, accurate, and reproducible, and will allow comparison of results between laboratories. Received: 10 May 1997 / Revised: 1 August 1997     

Synthesis,characterization, and cellular localization of a fluorescent probe of the antimalarial 8-aminoquinoline primaquine

Primaquine (PQ) is the only commercially available drug that clears dormant liver stages of malaria and blocks transmission to mosquito vectors. Although an old drug, much remains to be known about the mechanism(s) of action. Herein we develop a fluorescent tagged PQ to discover cellular localization in the human malaria parasite, Plasmodium falciparum. Successful synthesis and characterization of a primaquine-coumarin fluorescent probe (PQCP) demonstrated potency equivalent to the parent drug and the probe was not cytotoxic to HepG2 carcinoma cells. Cellular localization was found primarily in the cytosol of the asexual erythrocytic and gametocyte stages of parasite development.