Single-stranded phage DNAs containing thymine glycols were prepared by oxidation with osmium tetroxide (OsO4) and were used as templates for DNA synthesis by E. coli DNA polymerase I. The induction of thymine glycol lesions in DNA, as measured by immunoassay, quantitatively accounted for an inhibition of in vitro DNA synthesis on modified templates. Analysis of termination sites for synthesis by DNA polymerase I (Klenow fragment) showed that DNA synthesis terminated at most template thymine sites in OsO4-treated DNA, indicating that incorporation occurred opposite putative thymine glycols in DNA. Nucleotides 5' and 3' to putative thymine glycol sites affect the reaction, however, since termination was not observed at thymines in the sequence 5'-CTPur-3'. Conversion of thymine glycols to urea residues in DNA by alkali treatment caused termination of DNA synthesis one nucleotide 3' to template thymine sites, including thymines in the 5'-CTPur-3' sequence, showing that the effect of surrounding sequence is on the elongation reaction by DNA polymerase rather than differential damage induction by OsO4.

In the present study we employed light microscopic immunocytochemical techniques in order to investigate the temporal response of choline acetyltransferase (ChAT) and nerve growth factor receptor (NGFr) within hypoglossal motoneurons following unilateral transection or crushing of the XII nerve or after intraneural injections of ricin into the nerve. In control rats (i.e., sham operated) virtually all the motoneurons of the XII nucleus displayed intense immunolabeling for ChAT and were devoid of NGFr immunoreactivity. As early as 3 days post-operative the intensity and the number of ChAT-labeled neurons were reduced on the axotomized side compared to the non-lesioned side. This decrease was maximal approximately two weeks post-operative when virtually no ChAT-labeled cells were present on the lesioned side. In contrast, no loss of hypoglossal neurons was found using Nissl stains. This absence of ChAT immunolabeling persisted for several days, yet by 30 days many of the motoneurons had begun to re-express the enzyme. In contrast to the decrease in ChAT immunoreactivity, transection of the XII nerve also resulted in the expression of NGFr immunoreactivity within the lesioned motoneurons. This response was detected as early as one day post-operatively and continued throughout all time points thus far examined including times after many of the motoneurons had begun to re-express ChAT. Crushing of the XII nerve effected the expression of ChAT and NGFr in a manner comparable to, yet less intense than, that observed following transection. Ricin injected directly into the XII nerve resulted in the loss of hypoglossal motoneurons as demonstrated both in immunohistochemical and Nissl-stained tissue preparations. The cell loss was readily apparent 3 days post-operatively, and ChAT immunoreactivity permanently disappeared. NGFr immunolabeling was seen only in scattered surviving neurons but not in ricin poisoned cells. The possible mechanisms underlying the differential expression of ChAT and NGFr are discussed.

The incidence of NMSC in the province of New Brunswick is similar to that reported from 1973 through 1987 in the province of British Columbia, higher than those reported in most parts of Europe, and lower than all published rates in the United States and Australia. Owing to the inability of the registry to account for tumor multiplicity, the actual annual number of all NMSC lesions in this population is likely much higher.

No abstract

A gene in chromosome region 13q14 has been identified as the human retinoblastoma susceptibility (RB) gene on the basis of altered gene expression found in virtually all retinoblastomas. In order to further characterize the RB gene and its structural alterations, we examined genomic clones of the RB gene isolated from both a normal human genomic library and a library made from DNA of the retinoblastoma cell line Y79. First, a retiction and exon map of the RB gene was constructed by al overlapping genomic clones, yielding three contiguous regions ("contig') of 150 kilobases total length separated by two gaps. At least 20 exons were identified ip genomic clones, and these were provisionally numbered. Second, two overlapping genomic clones that demonstrated a DNA deletion of exons 2 through 6 from one RB allele were isolated from the Y79 library. To confirm and extend this result, a unique sequence probe from intron 1 was used to detect similar and possibly identical heterozygous deletions in genomic DNA from three retinoblastoma cell lines, thereby explaining the origs of their shortened RB mRNA htansripts. The same probe detecte genomic rearrangements in fibroblasts from two hereditary retinoblastoma patients, indicating that intron 1 includes a frequent site for mutations conferring predisposition to retinoblastona. Third, this probe also detete a polymorphic site for BamEil with allele frequencies near 0.5/0.5.Identification of commonly mutated regions will contribute significantly to genetic diagnosis in retinoblastoma patients and families.Retinoblastoma is an intraocular cancer of early childhood that arises from the developing retina. In a substantial proportion of cases, susceptibility to retinoblastoma can be inherited from a parent who was previously cured of the tumor (1,2 Recently, a gene in band 13q14 encoding an mRNA of 4.7 kilobases (kb) was identified as the RB gene based on tumor-specific alterations in expression and its apparent recessive nature at the cellular level (10). cDNA segments representing the RB gene transcript have been cloned (10-12) and sequenced (10). All hereditary and nonhereditary retinoblastomas examined to date have demonstrated altered RB gene expression: RB mRNA transcripts are either markedly reduced in quantity or are abnormal in length (10-12). About 40% of retinoblastomas show DNA deletions detectable by RB cDNA probes. Antibodies generated against the RB gene product, ppllORn, show complete absence of this nucleophosphoprotein in five out of five retinoblastomas, whereas it was easily detected in all normal or neoplastic cells containing a normal RB mRNA transcript (13). These data further strengthen the hypothesis that the absence of functional RB protein is potentially oncogenic.Given the different patterns of genetic alteration observed in retinoblastomas, it is evident that the RB gene is subject to a variety of mutational mechanisms, the details of which are as yet unknown. In order to further characterize the RB gene and some of its mutations, we construc...

Development of best practice treatment guidelines for NMSC in New Brunswick would improve future health care efficiencies, and standard protocols for registering new cases of NMSC in Canada would strengthen surveillance and reporting capacity.

A vascular thrombotic lesion localized to the rat sensorimotor cortex was produced following intravenous injection of the photosensitive dye rose bengal, and its activation with a small beam of high-intensity white light focused to the skull overlaying the sensorimotor cortex. In the sensorimotor cortex at various times after the triggering event, two contiguous brain regions with different degree(s) of neuronal damage can be distinguished: (1) a primary thrombotic ischemic core where the majority of cells are dead and (2) a penumbra region surrounding the core lesion in which a slower progressive neuronal degeneration is occurring. Importantly, in both brain regions the neuronal degeneration is associated with the activation and persistent translocation of protein kinase C (PKC) as indicated by an increase in 4-beta-3H-phorbol-12,13-dibutyrate (3H-PDBu) binding. Moreover, the demonstration that in the area penumbra the neuronal degeneration and the persistent translocation of PKC can be inhibited by a pretreatment with dizocilpine (i.e., MK-801) indicates that the dynamics of the progression of the neuronal degeneration are maintained by glutamate accumulating in the extraneuronal fluids. MK-801 additionally prevents the transcriptional activation of several immediate-early genes (IEGs) (e.g., c-fos) and their cognate third nuclear messenger (i.e., c-Fos) expression present in the hemisphere ipsilateral to the lesion. On the other hand, LIGA4 and LIGA20 derivatives of GM1 lysoganglioside reduce the membrane translocation of PKC and the neuronal damage in the penumbra area, but fail to change the increase of IEG expression in the cortex ipsilateral to the lesion.

This case supports the safety and efficacy of topical tacrolimus in patients with steroid-refractory OLP associated with chronic hepatitis C.