Synthesis and characterisation of chitosan-graft-poly(OEGMA) copolymers prepared by ATRP

Two synthetic routes, “grafting-from” and “grafting-to” the chitosan backbone, were investigated to prepare chitosan-graft-poly(OEGMA) copolymers by ATRP. The copolymers were characterised by ¹H NMR and FT-IR spectroscopy, elemental analyses and GPC. Estimates of the degree of grafting were quantitatively found from integration of the ¹H NMR spectra. The GPC data indicated that the hydrodynamic volumes of the polymers were significantly altered by the incorporation of poly(OEGMA). A comparison of the two synthetic methods showed differences in the ratio of synthetic and natural polymers present in the resulting combs. The “grafting-to” synthetic route was preferable as the polymers were of higher purity and more defined.     

Characterization of a human orphon 28 S ribosomal DNA

We have isolated clones in which two regions of the human genome are represented, each containing an orphon: a dispersed copy of 28S rDNA. Nucleotide (nt) sequence analysis established that one of these, H28S-O1, corresponds to nt 3627-4105 of human 28S rDNA, but in a mutated form. The orphon was flanked on one side by a portion of the L1Hs long interspersed repeat family of the human genome. Although H25S-O1 is not flanked by the terminal direct repeats characteristic of transposed DNA, it is possible that it is a processed pseudogene.     

Changes in the state of subunit association of lactate dehydrogenase from Bacillus stearothermophilus

Time-resolved measurements of the fluorescence anisotropy of an extrinsic dye-group attached to lactate dehydrogenase from B. stearothermophilus revealed that the rotational correlation time of the enzyme at low concentrations is 55 ns, while at high enzyme concentrations or in the presence of fructose 1,6-bisphosphate (Fru-1,6-P2) the correlation time increases to 95 ns. These correlation times are consistent with a change in Mr from 85 000 +/- 12 000 (dimer) to 150 000 +/- 22 000 (tetramer) and show that the tetrameric state can be induced either by raising the protein concentration or by the addition of the ligand. We have confirmed this change in molecular weight by gel-filtration experiments. In the ligand-induced tetramer, two Fru-1,6-P2 molecules are bound.     

An analysis of 12 years of craniomaxillofacial surgery in Toronto

Over 12 years, 1092 patients underwent 2019 craniomaxillofacial procedures in Toronto. Overall mortality was 0.64 percent, and that directly related to surgery was 0.18 percent. Major complications developed in 14.3 percent of patients, but not all had permanent sequelae. Infection was the greatest problem, occurring in 5.3 percent of the major operations. The annual incidence of complications dropped to 5.4 percent as our experience increased. Certain types of deformity, particularly those in patients with orbital hypertelorism, Treacher-Collins syndrome, or Crouzon's or Apert's syndrome, require more than one operation to achieve maximum correction. Our procedures have undergone many changes, the main one being a reduced operating time.     

Mapping of 12 translocation breakpoints in the Xp21 region with respect to the locus for Duchenne muscular dystrophy

Over 20 females have been reported to carry reciprocal X; autosome translocations with breakpoints in Xp21 and to suffer from Duchenne muscular dystrophy (DMD). We have positioned nine of these breakpoints with respect to the Duchenne gene by mapping probes from the DMD region against a panel of somatic cell hybrids, each containing one of the translocation chromosomes from a different female patient; further information has also been obtained by in situ hybridization, including the breakpoint location in a tenth DMD patient. We have also characterized two translocation breakpoints that lie in the same chromosomal region but which are not associated with the expression of DMD. All the DMD-associated translocation breakpoints examined lie at several sites within the DMD locus and between the two non-DMD breakpoints.     

A C-terminal signal prevents secretion of luminal ER proteins

Proteins that permanently reside in the lumen of the endoplasmic reticulum (ER) must somehow be distinguished from newly synthesized secretory proteins, which pass through this compartment on their way out of the cell. Three luminal ER proteins whose sequence is known, grp78 ("BiP"), grp94, and protein disulphide isomerase, share the carboxy-terminal sequence Lys-Asp-Glu-Leu (KDEL). We show that deletion (or extension) of the carboxyl terminus of grp78 results in secretion of this protein when it is expressed in COS cells. Conversely, a derivative of chicken lysozyme containing the last six amino acids of grp78 fails to be secreted and instead accumulates in the ER. We propose that the KDEL sequence marks proteins that are to be retained in the ER and discuss possible retention mechanisms.     

Changes in Escherichia coli cells starved in seawater or grown in seawater-wastewater mixtures

Some metabolic modifications of Escherichia coli cells during starvation in seawater were studied in laboratory microcosms. The apparent die-off of this bacterium under such conditions, as observed by comparing the enumeration of CFU in conventional freshwater media and direct epifluorescence counts, was partially prevented when cells were previously grown in salted organic medium or on seawater-wastewater agar. beta-Galactosidase activity of starved cells disappeared gradually with time, even though some other enzymatic activities, such as that of alkaline phosphatase, increased. Moreover, some modifications of sensitivity to antibiotics, heavy metals, and bacteriophages in seawater- and wastewater-grown cells suggested that the cells undergo structural changes under natural marine conditions. These results provide additional experimental data indicating the possible active adaptation of E. coli cells to seawater.