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G Randall

Publications and source records attributed to G Randall.

At least 19 recordsLinked to original sources
PubMed2004

Transcription factor binding and induced transcription alter chromosomal c-myc replicator activity.

The observation that transcriptionally active genes generally replicate early in S phase and observations of the interaction between transcription factors and replication proteins support the thesis that promoter elements may have a role in DNA replication. To test the relationship between transcription and replication we constructed HeLa cell lines in which inducible green fluorescent protein (GFP)-encoding genes replaced the proximal approximately 820-bp promoter region of the c-myc gene. Without the presence of an inducer, basal expression occurred from the GFP gene in either orientation and origin activity was restored to the mutant c-myc replicator. In contrast, replication initiation was repressed upon induction of transcription. When basal or induced transcription complexes were slowed by the presence of alpha-amanitin, origin activity depended on the orientation of the transcription unit. To test mechanistically whether basal transcription or transcription factor binding was sufficient for replication rescue by the uninduced GFP genes, a GAL4p binding cassette was used to replace all regulatory sequences within approximately 1,400 bp 5' to the c-myc gene. In these cells, expression of a CREB-GAL4 fusion protein restored replication origin activity. These results suggest that transcription factor binding can enhance replication origin activity and that high levels of expression or the persistence of transcription complexes can repress it.

Amanitins↗
PubMed2001

A unique murine monoclonal antibody recognizing HLA-B53, B37, B51, B52, +/-B44.

Monoclonal antibodies have played an important role in studying the biochemistry of the HLA-Class I molecules. Some murine anti-HLA mAbs can identify configurations of HLA epitopes that have never been reported in human allosera. One of these configurations is identified by an IgM mAb designated as: BHA-1441. This antibody was produced using a lymphoblastoid cell line typed as: A*02, A*25; B*38, B*4402/4405; C*0501, C*07, BW4, as the immunogen. A lymphocytotoxicity test of this mAb over a panel of 109 frozen, 452 fresh and, later, 44 DNA typed T cells revealed its specificity as B53, 37, 51, 52, +/- 44. All of the antigens recognized by this mAb share the Bw4 motif at positions 81-83, except for the HLA-B37, which shares only 82L and 83R. Furthermore, while B37 and B44 cross-react due to the aspartic acid (D) substitution at position 156, the reactivity with B53, B5 (51,52), B37 and 60% of B44 cells, makes it unlikely that the target epitope could be due only to the primary amino-acid sequence. The antibody-binding site might involve changes in tertiary structure and peptides bound by the MHC. BHA-1441 is an interesting tool to study and type the HLA-B53 antigen and its cross-reactive epitopes.

Amino Acid Sequence↗
PubMed2001

Hepatitis C virus cell culture replication systems: their potential use for the development of antiviral therapies.

Hepatitis C virus is a significant public health problem. Current drug regimens have low efficacy against some hepatitis C virus genotypes, while no vaccine is available. The absence of an efficient cell culture system and an accessible small animal model to study hepatitis C virus replication and pathogenesis are major obstacles to the development of effective antiviral therapies. Studies of surrogate model systems, either related viruses or chimeric viruses containing part of the hepatitis C virus genome, have given insight into hepatitis C virus replication, in addition to being a powerful tool for drug discovery. The recent development of an efficient system for the initiation of replication in cell culture provides a viable screen for inhibitors of hepatitis C virus replication. It also brings us much closer to the ultimate goal of an infectious cell culture system for hepatitis C virus.

Antiviral Agents↗
PubMed2000

Clinical practice guidelines: the need for improved implementation strategies.

Although substantial resources have been spent developing clinical practice guidelines, little effort has been made to evaluate the effectiveness of their implementation. In the absence of effective implementation, CPGs will have minimal impact on changing behaviours. This article critically reviews the literature and identifies a number of reasons why these guidelines have not been implemented successfully. If CPGs are to have a positive impact upon clinical behaviour and practice, then more aggressive and positive implementation strategies are indicated. Five key ingredients for successful implementation of quality CPGs are identified and recommended.

Guideline Adherence↗
PubMed2000

Herpes simplex virus 1 open reading frames O and P are not necessary for establishment of latent infection in mice.

Open reading frame (ORF) O and ORF P partially overlap and are located antisense to the gamma(1)34.5 gene within the domain transcribed during latency. In wild-type virus-infected cells, ORF O and ORF P are completely repressed during productive infection by ICP4, the major viral transcriptional activator/repressor. In cells infected with a mutant in which ORF P was derepressed there was a significant delay in the appearance of the viral alpha-regulatory proteins ICP0 and ICP22. The ORF O protein binds to and inhibits ICP4 binding to its cognate DNA site in vitro. These characteristics suggested a role for ORF O and ORF P in the establishment of latency. To test this hypothesis, two recombinant viruses were constructed. In the first, R7538(P-/O-), the ORF P initiator methionine codon, which also serves as the initiator methionine codon for ORF O, was replaced and a diagnostic restriction endonuclease was introduced upstream. In the second, R7543(P-/O-)R, the mutations were repaired to restore the wild-type virus sequences. We report the following. (i) The R7538(P-/O-) mutant failed to express ORF O and ORF P proteins but expressed a wild-type gamma(1)34.5 protein. (ii) R7538(P-/O-) yields were similar to that of the wild type following infection of cell culture or following infection of mice by intracerebral or ocular routes. (iii) R7538(P-/O-) virus reactivated from latency following explanation and cocultivation of murine trigeminal ganglia with Vero cells at a frequency similar to that of the wild type, herpes simplex virus 1(F). (iv) The amount of latent R7538(P-/O-) virus as assayed by quantitative PCR is eightfold less than that of the repair virus. The repaired virus could not be differentiated from the wild-type parent in any of the assays done in this study. We conclude that ORF O and ORF P are not essential for the establishment of latency in mice but may play a role in determining the quantity of latent virus maintained in sensory neurons.

Animals↗
PubMed1999

Region tracking on level-sets methods.

Since the work by Osher and Sethian on level-sets algorithms for numerical shape evolutions, this technique has been used for a large number of applications in numerous fields. In medical imaging, this numerical technique has been successfully used, for example, in segmentation and cortex unfolding algorithms. The migration from a Lagrangian implementation to a Eulerian one via implicit representations or level-sets brought some of the main advantages of the technique, i.e., topology independence and stability. This migration means also that the evolution is parametrization free. Therefore, we do not know exactly how each part of the shape is deforming and the point-wise correspondence is lost. In this note we present a technique to numerically track regions on surfaces that are being deformed using the level-sets method. The basic idea is to represent the region of interest as the intersection of two implicit surfaces and then track its deformation from the deformation of these surfaces. This technique then solves one of the main shortcomings of the very useful level-sets approach. Applications include lesion localization in medical images, region tracking in functional MRI (fMRI) visualization, and geometric surface mapping.

Algorithms↗
PubMed1997

The product of ORF O located within the domain of herpes simplex virus 1 genome transcribed during latent infection binds to and inhibits in vitro binding of infected cell protein 4 to its cognate DNA site.

The partially overlapping ORF P and ORF O are located within the domains of the herpes simplex virus 1 genome transcribed during latency. Earlier studies have shown that ORF P is repressed by infected cell protein 4 (ICP4), the major viral regulatory protein, binding to its cognate site at the transcription initiation site of ORF P. The ORF P protein binds to p32, a component of the ASF/SF2 alternate splicing factors; in cells infected with a recombinant virus in which ORF P was derepressed there was a significant decrease in the expression of products of key regulatory genes containing introns. We report that (i) the expression of ORF O is repressed during productive infection by the same mechanism as that determining the expression of ORF P; (ii) in cells infected at the nonpermissive temperature for ICP4, ORF O protein is made in significantly lower amounts than the ORF P protein; (iii) the results of insertion of a sequence encoding 20 amino acids between the putative initiator methionine codons of ORF O and ORF P suggest that ORF O initiates at the methionine codon of ORF P and that the synthesis of ORF O results from frameshift or editing of its RNA; and (iv) glutathione S-transferase-ORF O fusion protein bound specifically ICP4 and precluded its binding to its cognate site on DNA in vitro. These and earlier results indicate that ORF P and ORF O together have the capacity to reduce the synthesis or block the expression of regulatory proteins essential for viral replication in productive infection.

Animals↗
PubMed1997

Prospective randomized comparative study of bipolar electrocoagulation versus heater probe for treatment of chronically bleeding internal hemorrhoids.

BACKGROUND: Our purpose was to compare the efficacy, complications, failure rates, and crossovers of heater and bipolar probe treatments of chronically bleeding internal hemorrhoids. METHODS: Eighty-one patients (31 female, 50 male) with mean age of 53 years had large (grade 2 to 3) internal hemorrhoids with bleeding for a mean of 12 years, had failed medical management, and were randomized in a prospective study of anoscopic treatments to heater versus bipolar probes. Failure was defined as a major complication or failure to reduce the size of all internal hemorrhoids with three or more treatments. RESULTS: With similar background variables and no difference in treatment times, rectal bleeding and other symptoms were controlled in a shorter time with the heater probe than with the bipolar probe (77 versus 121 days). Five complications (fissures, bleeding, or rectal spasm) occurred with the bipolar probe, and two occurred with the heater probe. The heater probe caused more pain during treatments but had significantly fewer failures and crossovers. CONCLUSIONS: For patients who had failed medical management of chronically bleeding internal hemorrhoids, the techniques and complications of heater and bipolar probes were similar, but pain was more common, failures and crossovers were less frequent, and the time to symptom relief was shorter with the heater probe than with the bipolar probe.

Chronic Disease↗
PubMed1997

Identification of a 35-kilodalton serovar-cross-reactive flagellar protein, FlaB, from Leptospira interrogans by N-terminal sequencing, gene cloning, and sequence analysis.

During the screening of antibodies to pathogenic leptospires, a murine monoclonal antibody (designated M138) was found to react with various serovars. An antigen of approximately 35 kDa from Leptospira interrogans serovar pomona, which reacted strongly with M138, was characterized by N-terminal amino acid sequencing and identified as a flagellin, a class B polypeptide subunit (FlaB) of the periplasmic flagella. The gene encoding the FlaB protein, flaB, was amplified from the genomic DNA of several pathogenic serovars by PCR with a single pair of oligonucleotide primers, suggesting that FlaB is highly conserved among these serovars. Cloning and sequence analysis of flaB from serovar pomona revealed that it contains an 849-bp open reading frame with a G + C content of 46.88% which encodes a 283-amino-acid protein with a calculated molecular mass of 31.297 kDa and a predicted pI of 9.065. A sequence comparison of flagellin proteins revealed that the amino acid sequence is most variable in the central portion of the serovar pomona FlaB, which is believed to contain specific sequence information and which may thus be useful in the design of DNA or synthetic peptide probes suitable for the detection of infection with pathogenic leptospires.

Amino Acid Sequence↗
PubMed1997

Transcription of the derepressed open reading frame P of herpes simplex virus 1 precludes the expression of the antisense gamma(1)34.5 gene and may account for the attenuation of the mutant virus.

Open reading frame P (ORF P), located at the 3' terminus of the 8.5-kb DNA sequence transcribed during latency and almost completely antisense to the gamma(1)34.5 gene, is naturally repressed by infected cell protein 4 (ICP4), the major herpes simplex virus 1 regulatory protein. Earlier studies on cells infected with a mutant in which the expression of ORF P is derepressed have shown that (i) the accumulation of the alpha infected cell proteins 0 (ICP0) and 22 (ICP22), the products of spliced mRNAs, is reduced congruent with the binding of ORF P protein to p32, a component of the ASF/SF2 splicing factors, (ii) ORF P protein colocalizes with spliceosomes, (iii) both gamma(1)34.5 mRNA and protein are virtually undetectable, and (iv) the virus is attenuated on intracerebral inoculation in mice. We report the construction and characterization of two recombinant viruses: R7546, in which ORF P transcription was derepressed and the initiator methionine codon was replaced; and R7547, in which both mutations were repaired to the wild-type genotype. The mutations in R7546 do not alter the amino acid sequence of the gamma(1)34.5 gene. We report that (i) the reduction in the accumulation of gamma(1)34.5 mRNA and protein in cells infected with mutant viruses expressing derepressed ORF P genes reflects the effects of antisense transcription of ORF P rather than a function of ORF P protein, (ii) the attenuated phenotype of the viruses carrying derepressed ORF P genes is due largely to the absence of the gamma(1)34.5 protein, and (iii) the reduction in accumulation of ICP0 and ICP22 requires the expression of ORF P protein.

Alternative Splicing↗
PubMed1996

Phenotypic properties of herpes simplex virus 1 containing a derepressed open reading frame P gene.

Open reading frame P (ORF P) maps in the viral DNA sequences transcribed during latency and is located antisense to the gamma 1 34.5 gene. Earlier studies have shown that the expression of ORF P is repressed by an infected cell protein no. 4 binding site straddling the transcription initiation site. We have made monospecific polyclonal antibodies to the protein and constructed a virus, designated ORF P++, in which the infected cell protein no. 4 binding site has been mutagenized, thereby allowing full expression of an unmodified ORF P gene from its natural promoter. We report the following findings. (i) The native protein forms multiple bands on denaturing polyacrylamide gels suggestive of extensive processing and aggregation of the protein; (ii) the protein accumulates in the nucleus in rod-shaped structures perpendicular to the axis of attachment of the infected cell to the solid matrix; (iii) the virus was highly attenuated on inoculation into mice by the intracerebral or ocular route, and virus was not recovered upon explantation of trigeminal ganglia; (iv) although protein synthesis was not prematurely shut off in the human neuroblastoma cell line SK-N-SH, gamma 1 34.5 protein was not detected in immunoblasts. Analyses of electrophoretically separated denatured RNAs indicated that in cells infected with the ORF P++ virus, there was a large increase in the amount of ORF P RNA and a corresponding decrease in the amount of gamma 1 34.5 RNA. We conclude that either the overproduction of ORF P protein blocks the expression of some herpes simplex virus 1 genes or derepression of the transcription of ORF P has a negative effect on the transcription of the antisense gamma 1 34.5 RNA.

Animals↗
PubMed1994

A controlled study of ranitidine for the prevention of recurrent hemorrhage from duodenal ulcer.

BACKGROUND: Hemorrhage is the most common complication of duodenal ulcer disease, but there is little information about the effectiveness and safety of long-term maintenance therapy with histamine H2-receptor blockers. METHODS: We conducted a double-blind study in patients with endoscopically documented hemorrhage from duodenal ulcers. Patients were randomly assigned to maintenance therapy with ranitidine (150 mg at night) or placebo and were followed for up to three years. Endoscopy was performed at base line (to document that the ulcers had healed), at exit from the study, and when a patient had persistent ulcer symptoms unrelieved by antacids or had gastrointestinal bleeding. Symptomatic relapses without bleeding were treated with ranitidine; if the ulcer healed within eight weeks, the patient resumed taking the assigned study medication. RESULTS: The two groups were similar at entry, which usually occurred about three months after the index hemorrhage. After a mean follow-up of 61 weeks, 3 of the 32 patients treated with ranitidine had recurrent hemorrhage, as compared with 12 of the 33 given placebo (P < 0.05). Half the episodes of recurrent bleeding were asymptomatic. One patient in the ranitidine group withdrew from the study because of asymptomatic thrombocytopenia during the first month. CONCLUSIONS: For patients whose duodenal ulcers heal after severe hemorrhage, long-term maintenance therapy with ranitidine is safe and reduces the risk of recurrent bleeding.

Double-Blind Method↗
PubMed1993

Burst-promoting activity in the serum of patients with chronic renal failure undergoing long-term hemodialysis.

Anemia in patients with chronic renal failure (CRF) is multifactorial, and while the majority will respond to the paternal administration of recombinant human erythropoietin (r-HuEPO), a role for coexistent plasma inhibitors and stimulators, such as burst-promoting activity (BPA), remains controversial. To evaluate the latter possibility, eight individuals with CRF on long-term hemodialysis, were studied before (mean hemoglobin 58.4 +/- 8.0 g/l and after 12 weeks of r-HuEPO therapy (mean hemoglobin 100.4 +/- 18.3 g/l). In vitro erythroid cultures using erythroid burst forming unit (BFU-E) and erythroid colony forming unit (CFU-E) assays were performed, plating 5 x 10(4) bone marrow mononuclear cells and comparing growth in heat-inactivated autologous serum with AB serum. Using Step III sheep erythropoietin (Connaught Laboratories, Willowdale, Ontario, Canada) (n = 4), mean BFU-E pre-therapy were 89.7 +/- 75.1, CFU-E were 418.5 +/- 150.6, whereas the corresponding figures in AB serum were 2.5 +/- 2.9 and 197 +/- 94.19, p = 0.1, p = 0.01, respectively. Similarly, with r-HuEPO (EPOCONN: Connaught Laboratories, Willowdale, Ontario, Canada) (n = 4), mean BFU-E were 145.25 +/- 103.3 in autologous serum and 31.0 +/- 56.75 in AB serum (p = 0.04). As controls, erythroid progenitors from two normal donors yielded 69 and 61 BFU-E colonies in autologous serum and 103 and 42 in AB serum; the corresponding CFU-E were 52 and 235 versus 136 and 137.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1992

Distribution of dysplasias and early invasive carcinoma in Barrett's esophagus.

To assess the anatomic relationships between areas of dysplasia and "early" carcinoma, we evaluated histologically the entire mucosal surfaces of seven esophagectomy specimens resected for high-grade dysplasia or early invasive (intramucosal and submucosal) carcinoma. We developed surface area maps and assessed the various degrees of dysplasia or carcinoma at 10 equidistant points. Our analysis shows an equal likelihood of high-grade dysplasia and/or early invasive carcinoma occurring throughout the length of Barrett's epithelium. Foci of carcinoma appear within fields of Barrett's epithelium and adjacent to areas of dysplasia, supporting a dysplasia-carcinoma sequence. The amount of dysplastic epithelium appears related to the surface area of Barrett's epithelium present. However, we found no association between the extent of dysplasia and the likelihood of finding carcinoma. This study supports the current standard of practice for clinical surveillance of patients with Barrett's esophagus by uniformly distributed endoscopic biopsy of the complete length. In addition, the presence of any degree of dysplasia may be an indication for close clinical follow-up.

Aged↗
PubMed1992

In vivo growth hormone (GH) response to human GH-releasing factor (GRF) or somatostatin (SRIF) in foetal pigs.

The short-term effect of hypothalamic GRF and SRIF on the pituitary release of GH at different stages of gestation has been studied. In the present experiment eighteen gilts were used, six at each of 66, 88 and 110 days of gestation. Ventral laparotomy was performed under general anaesthesia and a section of uterus was exteriorized. Blood samples were obtained from the umbilical vein of three foetuses per gilt just prior to the injection of each foetus with either saline, 5 micrograms/kg of hGRF (1-44)NH2 or 50 micrograms/kg of SRIF into the umbilical vein. Additional blood samples were obtained 15, 30, 45 and 60 min post-injection. Serum samples were radioimmuno-assayed for GH (porcine). There was a treatment by gestational age interaction (P less than 0.01) on mean GH concentrations, area under the GH curve and GH peaks. While treatments had no effect (P greater than 0.1) on GH variables at 66 days of gestation, the area under the GH curve was slightly increased by GRF (P = 0.14) at day 88 and all GH variables were significantly increased (P less than 0.01)) by GRF at 110 days of gestation. There was a quadratic effect of time post-injection on GH concentrations at 88 (P less than 0.05) and 110 (P less than 0.001) days of gestation. There was no effect of SRIF injection (P greater than 0.1) on GH concentrations at any gestational age. In conclusion, the foetal pituitary responsiveness to GRF develops with foetal age and is maximal at the end of gestation, whereas there is no short-term response to a bolus of SRIF at any stage of gestation.

Animals↗
PubMed1989

The effects of the anti-tumor agent mezerein on the cytotoxic capacity and oxidative metabolism of human blood cells.

Mezerein, the most active antitumor compound isolated from the daphne species of plants, has a structural similarity to phorbol myristate acetate (PMA), the major active compound isolated from croton oil. PMA is known to have tumor promoting activity and is a potent inflammatory agent. Mezerein has similarly been reported to have potent inflammatory properties but appears to be a weaker tumor promoter than PMA. While the effect of PMA on the function and metabolism of human blood cells has been extensively studied, there is little similar information concerning mezerein. Therefore, in these studies, we have compared the capacities of mezerein and PMA to activate the cytotoxic capacity and oxidative metabolism of human granulocyte (PMNs), monocyte, lymphocyte, and mononuclear cell (lymphocytes and monocytes) cultures in vitro. Mezerein stimulated the oxidative metabolism of PMNs in an identical manner to PMA as indicated by a burst in the activity of the HMPS pathway, the production of H2O2, hydroxyl radical and stable oxidants. Mezerein also stimulated the release of thromboxane B2 from PMNs. Both compounds activated the oxidative metabolism of monocytes but not the oxidative metabolism of lymphocytes. The enhanced oxidative metabolism of the phagocytic cells was associated with an increased cytotoxicity against human red cells which are sensitive to oxidant damage but not against the NK resistant Raji lymphoblast cell line or the SW1116 colon tumor cell line. Of interest is that mezerein did not augment significantly the minimal cytotoxic capacity (NK activity) of mononuclear cells, monocytes or freshly isolated lymphocyte cultures against the tumor cell targets used in our experiments. However, lymphocyte cultures preincubated for 15 hours with mezerein had a marked enhancement of cytotoxicity against the tumor targets. This activation was not observed in similarly treated mononuclear cell cultures suggesting a suppressor activity of the monocytes. Our data suggest that the potent inflammatory activity of mezerein similar to PMA, may be related to its capacity to activate the oxidative and arachidonic metabolism of phagocytic cells. In addition, the capacity of mezerein to activate the cytotoxic capacity of lymphocytes may relate to its reported in vivo antitumor activity.

Antineoplastic Agents, Phytogenic↗
PubMed1988

Comparison of low-power YAG laser and BICAP tumor probe for palliation of esophageal cancer strictures.

The purposes of this study were (a) to determine the applicability of endoscopic palliation for patients with esophagogastric cancer strictures in a referral center, and (b) to compare the efficacy and safety of the BICAP tumor probe with the neodymiumyttrium-aluminum-garnet (YAG) laser for such palliation. Forty-two consecutive patients with weight loss and obstructive symptoms from an unresectable, malignant esophageal stricture were referred for endoscopic palliation. Fourteen patients did not meet the criteria for YAG laser or BICAP tumor probe treatment and other therapies were recommended. Twenty-eight patients were treated, the first 14 with low-power YAG laser and the last 14 with BICAP tumor probe. All patients had coagulation of malignant strictures in one session. Treated patients were similar in background variables and stricture lengths but twice as much thermal energy was needed for the YAG laser as the BICAP tumor probe treatment. Treatment results were not statistically different during the median follow-up and survival of 16 wk. As minor complications, either pain or edema requiring dilatation was more common in the YAG laser-treated group than the BICAP tumor probe group. Treatment-related esophageal strictures developed in 21% of patients treated with YAG laser. A fistula developed in 1 patient with noncircumferential cancer in the BICAP tumor probe group. Compared with only the intake of liquids before treatment, 86% of patients could eat a soft or solid diet after initial treatment with BICAP tumor probe or YAG laser. Our conclusions were that for BICAP tumor probe and YAG laser, endoscopic palliation efficacy and safety for circumferential esophageal cancer strictures were similar. The advantages of using the BICAP tumor probe were portability, lower equipment costs, and the ability to treat submucosal, long, or high esophageal cancer strictures in one session. Treatment with YAG laser was safer than BICAP tumor probe for exophytic, noncircumferential cancers because the laser could be directed endoscopically. Use of the BICAP tumor probe is not recommended for noncircumferential esophagogastric cancer strictures.

Adenocarcinoma↗