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J Furlong

Publications and source records attributed to J Furlong.

15 recordsLinked to original sources
PubMed1999

Sequence-tagged connectors: a sequence approach to mapping and scanning the human genome.

The sequence-tagged connector (STC) strategy proposes to generate sequence tags densely scattered (every 3.3 kilobases) across the human genome by arraying 450,000 bacterial artificial chromosomes (BACs) with randomly cleaved inserts, sequencing both ends of each, and preparing a restriction enzyme fingerprint of each. The STC resource, containing end sequences, fingerprints, and arrayed BACs, creates a map where the interrelationships of the individual BAC clones are resolved through their STCs as overlapping BAC clones are sequenced. Once a seed or initiation BAC clone is sequenced, the minimum overlapping 5' and 3' BAC clones can be identified computationally and sequenced. By reiterating this "sequence-then-map by computer analysis against the STC database" strategy, a minimum tiling path of clones can be sequenced at a rate that is primarily limited by the sequencing throughput of individual genome centers. As of February 1999, we had deposited, together with The Institute for Genomic Research (TIGR), into GenBank 314,000 STCs ( approximately 135 megabases), or 4.5% of human genomic DNA. This genome survey reveals numerous genes, genome-wide repeats, simple sequence repeats (potential genetic markers), and CpG islands (potential gene initiation sites). It also illustrates the power of the STC strategy for creating minimum tiling paths of BAC clones for large-scale genomic sequencing. Because the STC resource permits the easy integration of genetic, physical, gene, and sequence maps for chromosomes, it will be a powerful tool for the initial analysis of the human genome and other complex genomes.

Chromosome Mapping↗
PubMed1996

A survey of dairy cattle worm control practices in southeast Brazil.

A survey on the nematode control strategies utilized by dairy farmers in Southeastern Brazil, a region accounting for 46.4% of the national milk production, was conducted through interviews with farmers. To select the producers to be interviewed the region was split into 16 non-contiguous clusters, according to the level of milk production. A systematic sample was then selected in each of the clusters. The interview questionnaire consisted of one-way, multiple-choice and open-ended questions. Data collected were represented by numbers and digitized on a data base (Epi Info, version 5.01b) and analyzed. Out of the 89 farmers interviewed, 37.5% deworm their herd after clinical signs and 62.5% preventively. Generally, anthelmintics are applied from one to 12 times a year (average of 3.79 times a year) in all age categories of animals. Of the anthelmintics used in the last deworming, imidazothiazole was used exclusively by 17.1% of the farmers, benzimidazole by 9.8% and avermectin by 18.3%, while 55.8% used more than one anthelmintic class to deworm their animals. To choose the dosage, most farmers consult the product label (94.8%) and determine the volume to be applied, based on an estimate of the average body weight of each animal (62.9%). Improvements in the general appearance of the herd and weight gains of growing animals were observed by most farmers after deworming (87.3%). However, most of them (66.2%) recalled interrupting the use of some compound in the last few years, due to the detection of no improvement following treatment (32.7%), rising costs of the medication (28.6%), adverse reaction (8.2%), product not available at the time of purchase (4.1%) and decision to change the compound in use (10.2%). Most farmers (95.3%) intend to continue using the same control measures in the following year. Veterinarians play an important role in the farmer's choice to deworm their animals, as many seek advice from them. Therefore, programs aimed at technology transfer should include continuous updates on the subject, especially for veterinarians.

Animal Husbandry↗
PubMed1995

Flow cytometry in colon cancer: does flow cytometric cell cycle analysis help predict for short-term recurrence in patients with colorectal carcinoma?

The purpose of this prospective study was to determine whether tumor ploidy or S-phase fraction measurements were of prognostic value in predicting short-term recurrence for colorectal carcinoma. A total of 52 patients underwent curative resection of colorectal carcinoma. Fresh suspensions of tumor cells were used for flow cytometric analysis. Patients underwent follow-up for possible recurrence, which then was related to ploidy status, S-phase fraction measurement, adjuvant therapy received, and Dukes stage. Disease-free probability was determined using Kaplan-Meier actuarial curves for various subgroups of the study populations. Results revealed that elevated S-phase fraction (>16%) did predict for a higher probability of recurrence in those patients receiving no postoperative adjuvant treatment. Among patients receiving postoperative adjuvant therapy, however, a higher S-phase fraction predicted for a lower probability of recurrence. These data suggest that the S-phase fraction does predict for disease-free probability and also predict response to adjuvant therapy in patients with colorectal carcinoma.

Adenocarcinoma↗
PubMed1993

Herpes simplex virus type 1 ribonucleotide reductase large subunit: regions of the protein essential for subunit interaction and dimerization.

We have constructed a series of random N-terminal deletions of the large subunit (R1) of the herpes simplex virus type 1 ribonucleotide reductase. Deletions extended throughout the R1 gene open reading frame and, in total, 31 different truncated polypeptides were expressed in Escherichia coli using the T7 expression system. N-Terminal truncations were analyzed for their interaction with the small subunit (R2) of ribonucleotide reductase using a sensitive enzyme-linked immunosorbent assay (ELISA) method and for their ability to complement R2 in ribonucleotide reductase assays. Truncated proteins were also tested for homodimerization using gel-filtration chromatography. The results identified a region of R1 between amino acids 349 and 373 which was essential for subunit interaction. Proteins lacking up to 348 amino-terminal residues associated with R2 and complemented R2 in ribonucleotide reductase assays. Proteins commencing at amino acid 373 and beyond did not interact with R2 and were inactive in enzyme assays. Using a plasmid which expressed an N-terminal deleted protein commencing at amino acid 247, we constructed two defined C-terminal deletions to give proteins comprising amino acids 247-434 and 247-996 of R1. Neither of these truncated proteins bound R2 and we concluded that a second region between amino acids 996 and 1137 (the C-terminus) is required for interaction with R2. Gel-filtration studies indicated that deletion of the first 420 amino acids from R1 did not affect dimerization. However, deletions of 457 amino acids and larger gave proteins which existed as monomers.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1992

An autophosphorylating but not transphosphorylating activity is associated with the unique N terminus of the herpes simplex virus type 1 ribonucleotide reductase large subunit.

We report on a protein kinase function encoded by the unique N terminus of the herpes simplex virus type 1 (HSV-1) ribonucleotide reductase large subunit (R1). R1 expressed in Escherichia coli exhibited autophosphorylation activity in a reaction which depended on the presence of the unique N terminus. When the N terminus was separately expressed in E. coli and partially purified, a similar autophosphorylation reaction was observed. Importantly, transphosphorylation of histones and of proteins in HSV-1-infected cell extracts was also observed with purified R1 and with truncated R1 mutants in which most of the N terminus was deleted. Ion-exchange chromatography was used to separate the autophosphorylating activity of the N terminus from the transphosphorylating activity of an E. coli contaminant protein kinase. We propose a putative function for this activity of the HSV-1 R1 N terminus during the immediate-early phase of virus replication.

Adenosine Triphosphate↗
PubMed1991

The large subunit of herpes simplex virus type 1 ribonucleotide reductase: expression in Escherichia coli and purification.

The open reading frame of the large subunit (R1) of herpes simplex virus type 1 (HSV-1) ribonucleotide reductase has been positioned downstream of the phage T7 gene 10 promoter in the expression vector, pET. Transformation of this recombinant plasmid into Escherichia coli BL21 DE3 cells containing the T7 RNA polymerase, under the control of the lac UV5 promoter, allows expression of the subunit on induction of the T7 RNA polymerase by isopropyl thiodigalactoside. The expressed protein is soluble and can be purified with yields up to 0.5 mg of R1 per litre of bacterial culture. The subunit can complement R2 produced in BHK cells or E. coli to give specific activities comparable to that produced in BHK cells infected with HSV-1. Enzyme activity reconstituted from E. coli-expressed R1 and R2 is inhibited by the nonapeptide YAGAVVNDL with an IC50 comparable to that obtained with enzyme extracted from BHK cells infected with HSV-1. Results suggest that the E. coli produced enzyme is a good source of protein for further structural and functional studies.

Animals↗
PubMed1991

Purification and characterization of the herpes simplex virus type 1 ribonucleotide reductase small subunit following expression in Escherichia coli.

The herpes simplex virus type 1 (HSV-1) gene encoding the ribonucleotide reductase (RR) small subunit (R2) was cloned as an unfused and intact open reading frame into a T7 RNA polymerase expression system in Escherichia coli. The expressed product was recovered from bacteria in soluble form and constituted 7% of the soluble protein. Protein purification yielded 3.5 mg of 95% pure R2 per litre of bacterial culture. The correct composition of the purified protein was verified by amino acid analysis and N-terminal sequencing. The isoelectric point of the protein was 5.3. Atomic emission spectroscopy indicated that the iron content of the E. coli-expressed R2 was 0.2 to 0.5 atoms of iron per R2 protomer as compared with a theoretical maximum value of 2. The E. coli-expressed HSV-1 R2 existed as a combination of a stable dimer and monomer. Combination of the E. coli-expressed R2 with the E. coli-expressed large subunit (R1) gave an active holoenzyme. Thus, the T7 expression system provides a rich source of enzymically active HSV-1 RR.

Amino Acids↗
PubMed1987

New Marfanoid syndrome with craniosynostosis.

We report on a patient with various connective tissue abnormalities suggesting a distinctive disorder combining some features of the Marfan syndrome with craniosynostosis and other anomalies. A comparison is made with the Marfan syndrome and other phenotypically similar conditions.

Adolescent↗
PubMed1981

Detailed anatomy of the extensor mechanism at the proximal aspect of the finger.

To explain the unique action of the lateral bands during flexion and extension of the finger, a study was undertaken to examine the extensor mechanism at the level of the proximal interphalangeal joint. The study was carried out in three stages-histologic, anatomic, and dynamic. The histologic sections showed no elastic tissue present to account for the expansile and contractile movement of the lateral bands. The anatomic and dynamic studies showed that the extensor mechanism at the level of the proximal aspect of the finger was composed of an intricate, layered, crisscross fiber pattern which altered its geometric arrangement as the finger was flexed and extended. It is our opinion that the action of the crisscross fibers may govern both the action and degree of displacement of the lateral bands during flexion and extension of the finger.

Cadaver↗
PubMedDate not supplied

Prenatal diagnosis of trisomy 21 and X/XX sex chromosome mosaicism.

Double aneuploidy involving Down syndrome and Turner syndrome is a rare chromosomal abnormality presumed to occur with a frequency of about 1 in 2 million births. Twenty-one cases of this combined anomaly have been reported and two infants were born with this anomaly after a mistake in prenatal diagnosis. We report the first prenatal diagnosis of Down syndrome combined with Turner mosaicism and suggest that this polysyndrome may be more common than previously estimated. We, therefore, wish to alert cytogenetic laboratories performing prenatal diagnoses of the potential risks of misdiagnosis of this polysyndrome if banding is not performed and if a sufficient number of mitotic cells are not analysed.

Adult↗