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J Campione-Piccardo

Publications and source records attributed to J Campione-Piccardo.

At least 19 recordsLinked to original sources
PubMed2003

Dengue viruses in the Caribbean. Twenty years of dengue virus isolates from the Caribbean Epidemiology Centre.

The first isolate of a dengue virus in the Americas was obtained in Trinidad and Tobago in 1953, and several dengue virus isolates were obtained in subsequent years. However, the systematic isolation and typing of dengue viruses in support of virus surveillance and outbreak investigations did not start until the creation of the Caribbean Epidemiology Centre (CAREC) in 1975. Since then, over two thousand viral isolates have been obtained and typed from many countries in the English and Dutch-speaking Caribbean. In this communication, virological data from 17 countries between 1977 and 1996 are presented and analyzed together for the first time with available epidemiological data. Types 1, 2 and 4 were isolated over the period, and geographic and temporal patterns in the distribution of the most prevalent strains are presented. The historical surveillance data is critically assessed. A temporal correlation with reported dengue incidence and rainfall data in Trinidad and Tobago is reported. Recent changes in epidemiological patterns are described, including reference to two large later outbreaks. Risk assessment of complicated forms of dengue virus infections in the Caribbean has been attempted, with some success. The importance of ongoing systematic surveillance is discussed.

Age Distribution↗
PubMed1999

Molecular diagnosis of dengue by reverse transcriptase-polymerase chain reaction.

The techniques of reverse transcriptase-polymerase chain reaction (RT-PCR) and subsequent PCR were employed in the analysis of serum samples from a range of patients from the Caribbean Epidemiology Centre (CAREC) member countries. Results were compared with those from viral isolation and immunofluorescence. In the second part of the study, ten serum samples were stored for one week under four sets of conditions: -20 degrees C, 4 degrees C, 25 degrees C, and thawed (20 degrees C) and frozen (-20 degrees C) daily. After one week of each treatment the samples were analysed by RT-PCR and PCR. 90.4% of results from PCR agreed with results from viral isolation (VI) and fluorescent antibody (FA) detection. All PCR positive samples originated from sera collected within five days of the date of onset of fever. Frozen, refrigerated and repeat freeze-thawed samples gave consistent positive results by RT-PCR. After storage at 25 degrees C, however, half the dengue-positive samples were negative by RT-PCR. The results indicate the sensitivity and reliability of this rapid technique and its applicability in the Caribbean. It provides a preliminary assessment of its advantages and limitations under certain conditions of serum collection and storage.

Dengue↗
PubMed1994

Measurement of nucleoside diphosphate kinase-Nm23 activity by anion-exchange high-performance liquid chromatography.

A first-order assay to detect the activity of nucleoside diphosphate kinase (NDP-kinase; EC 2.7.4.6) was developed. In this assay, the activity of NDP-kinase is measured using various deoxy- and ribonucleotide triphosphates as phosphate donors and dADP as phosphate acceptor. The enzyme activity is determined by quantifying, after anion-exchange HPLC, the amount of newly synthesized dATP. Contrary to the most common coupled enzymic assays or isotopic assays the use of different donor-acceptor pairs is not restricted. The resolution of the procedure described is limited only by the chromatographic separation of substrate and product pairs participating in the reaction.

Animals↗
PubMed1991

A highly conserved nucleotide string shared by all genomes of human papillomaviruses.

The nucleotide string TAAAACGAAAGT is the longest perfect homology shared by all sequenced human papillomavirus genomes. This nucleotide string, which was also found to be highly specific for human papillomavirus genomes, shares the same genomic position in all viral types (5' end of the E1 open reading frame) and putatively codes in every case for the same amino acids. One possible evolutionary model was used to estimate the probability of random occurrence of the nucleotide string in 10 human papillomavirus genomes. It assumed that the universal string had been subjected to the same mutation rate as the entire E1 open reading frame. The estimated probability was found to be very low, suggesting that the conservation of the string could not have resulted from random divergence and that its conservation among human papillomaviruses is likely to reflect the occurrence of biological constraints. It is speculated that this nucleotide string may be required to code for amino acids indispensable for the nuclear localization of E1-coded peptides or to bind cellular factors affecting viral replicative functions. Definitive evidence is expected to come from oligonucleotide-protein binding experiments and from site-directed mutagenesis of cloned HPV genomes. This motif, universal among human papillomaviruses, is being successfully used in the design of consensus primers from the early region.

Amino Acid Sequence↗
PubMed1989

Amplification of human papillomavirus DNA sequences by using conserved primers.

The polymerase chain reaction has potential for use in the detection of small amounts of human papillomavirus (HPV) viral nucleic acids present in clinical specimens. However, new HPV types for which no probes exist would remain undetected by using type-specific primers for the polymerase chain reaction before hybridization. Primers corresponding to highly conserved HPV sequences may be useful for detecting low amounts of known HPV DNA as well as new HPV types. Here we analyze a pair of primers derived from conserved sequences within the E1 open reading frame for HPV sequence amplification by using the polymerase chain reaction. The longest perfect homology among HPV sequences is a 12-mer within the first exon of E1M. A region of conserved amino acids coded by the E1 open reading frame allowed the detection of another highly conserved region about 850 base pairs downstream. Two 21-mers derived from these conserved regions were used to amplify sequences from all HPV DNAs used as templates. The amplified DNA was shown to be specific for HPV sequences within the E1 open reading frame. DNA from HPVs whose sequences were not available were amplified by using these two primers. HPV DNA sequences in clinical specimens could also be amplified with the primers.

Animals↗
PubMed1989

Plasmid library for the transcription of RNA probes complementary to the entire genome of the human immunodeficiency virus type 1 (HIV-1).

A plasmid (pBH10R3) containing a 9-kb Sst I fragment of HIV-1 (clone BH-10) inserted in pSP64, an in vitro expression vector, has been used for the transcription of anti-sense HIV-1 RNA. With this system, the transcripts obtained in vitro were not usually full length (1 to 2 kb long) and they predominantly span the 3' end ORF and ENV regions of the viral genome. We have rearranged the HIV-1 genomic sequences with respect to the SP6 promoter in the pSP64 vector and have obtained a series of new constructs allowing the expression in vitro of RNA transcripts complementary to other regions in the HIV-1 genome, including the 5' end of the ENV region as well as the TAT, POL, and GAG regions. In fact, the combined use of these constructs as templates for in vitro transcription allows the production of RNA probes spanning the entire viral genome. Compared with the 1- to 2-kb probes mentioned above, the combined use of such probes results in a several-fold increase in the sensitivity of molecular hybridization for the detection of HIV-1 nucleic acid sequences. Also, these constructs enable the preparation of RNA probes that have the potential to detect restriction polymorphisms throughout the HIV-1 genome.

Gene Library↗
PubMed1988

Modification of DNA ends and detection of restriction enzyme recognition sequences in their ligated junctions.

A program has been developed for the modelling of modifications in DNA ends, for the construction of ligated junctions, and for the analysis in these junctions of new restriction enzyme recognition sequences. This program allows the analysis of restriction enzyme specificities in ligated junctions of cohesive or blunt DNA ends. Cohesive ends are considered in their natural configuration or after modification by possible blunt-ending procedures. The program also allows the modelling of partial filling-in for 5'-single-stranded ends. This program has proven useful for the design of sequences with new restriction sites or to predict or confirm the sequence of junctions created by the ligation of modified ends.

Base Sequence↗
PubMed1988

Algorithms for determining the fate of sites and domain boundaries in computer simulations of recombinant DNA procedures.

Structural and functional features within genomic sequences are best described by their position within the genomic structure. Cleavage sites can be conveniently described by single positions but genomic domains require the position of their two boundaries. The handling of these positions simultaneously to sequence manipulations in computer simulations of recombinant DNA procedures greatly improves the understanding of the resulting recombinant constructs. In addition, the algorithms describing the fate of domain boundaries can be used for the handling of nucleotide sequences in dynamic database environments handled by languages like Prolog which are particularly suitable for artificial intelligence implementations. This communication describes a set of algorithms for the automatic updating of single sites and double domain boundaries in linear and circular models for computer simulation of recombinant DNA procedures.

Algorithms↗
PubMed1987

Restricted replication of herpes simplex virus type 1 in murine embryonal carcinoma cells.

Herpes simplex virus type 1 (HSV-1) has a broad host range but the KOS strain of HSV-1 did not replicate efficiently in murine embryonal carcinoma (EC) cells. The yield of infectious HSV-1 from EC cells was 100- to 1000-fold lower than that from fibroblast cell lines of mouse, monkey or human origin. The thymidine kinase (TK) gene of HSV-1 is expressed early during the infectious cycle. The levels of TK mRNA and of TK activity in infected EC cells were only two- to threefold lower than levels from infected fibroblast cells. Infected EC cells supported replication of about half as much HSV-1 DNA as did fibroblast cells. The reduced yield of infectious virus was consistent with a paucity of virions in infected EC cells examined by electron microscopy, suggesting a major block late during the HSV-1 infectious cycle. We isolated a variant strain of HSV-1, called KOSEC, which replicated as efficiently in EC cells as in mouse fibroblasts. KOSEC infected EC and fibroblast cells, synthesized more TK mRNA, more TK enzyme, and more HSV-1 DNA than did the same cells infected with the KOS stain. Both HSV-1 strains induced similar levels of synthesis of gD, an early viral glycoprotein. By co-infection of EC cells with the KOS and KOSEC virus, both the elevated virus yield and the elevated TK synthesis seen in KOSEC-infected cells appeared to be recessive. Apparently a viral mutation that affects expression of some early viral functions can also overcome the EC cell restriction to HSV-1 replication.

Animals↗
PubMed1987

[Influence of cell-cell interaction upon differentiation of murine embryonal carcinoma cells].

Embryonal carcinoma (EC) cells are the pluripotent stem cells of teratocarcinoma. The aggregates of P19, a mouse EC cell line, undergo differentiation and rapidly lose its colony-forming ability in culture with retinoic acid (RA) or DMSO. Loss of plating efficiency is used to assess the rate of drug-induced differentiation. When exposed respectively to DMSO or RA, the isolated EC cell mutants (D3 and RAC65) did not differentiate but the gap junctions could be formed. In RA-treated co-aggregates of RAC65 and O1A1, each cell line gave responses independent of the presence of the other cell line. However, in DMSO-treated co-aggregates of P19 and D3, D3 cells remained undifferentiated and P19 cell differentiation was much poorer than that in the absence of D3 cells. The results suggest that cell-cell interaction be present in the early stage of DMSO-induced differentiation and absent in the early stage of RA-induced differentiation.

Animals↗
PubMed1986

An integrated software system for microcomputer management of recombinant DNA data.

A database-oriented system (pCP123) is described for the manipulation of recombinant DNA data. This system was developed within the context of an integrated software package with spreadsheet, database, graphing and programming capabilities. The system includes two databases, one of sites and another of regions, coordinately handled by a series of macro-programs operated from four user-define menus. A distinctive feature of the system is the possibility of handling both ends of defined functional or structural regions in situations of simulated deletions or insertions.

Computers↗
PubMed1986

Software library with applications in virology and molecular biology.

A menu-driven interactive package of computer programs has been developed to help in calculations commonly required in laboratories using virological and molecular biological techniques. Two of the programs use two of the most accurate methods available for determining viral titers, several programs were designed to optimize experimental parameters and others help in the analysis of recombinant DNA data. All of the programs use specially developed original algorithms which in some cases are based also on equations originally derived. The programs were written in Microsoft BASIC using mostly hardware independent commands and functions and should run without major modifications in most microcomputers with BASIC interpreters or compilers. The programs are menu-driven and fully interactive.

Computers↗
PubMed1985

Cell-cell interaction can influence drug-induced differentiation of murine embryonal carcinoma cells.

When cultured in the presence of either retinoic acid (RA) or dimethyl sulfoxide (DMSO), aggregates of the P19 line of mouse embryonal carcinoma (EC) cells differentiate and the spectrum of cell types formed depends on the drug dose. It is shown here the EC cells rapidly lose their colony-forming ability when cultured as aggregates in the presence of DMSO. This loss of plating efficiency (PE) also occurs rapidly following RA treatment. Loss of PE has been used as a quantitative procedure for assessing the rate of drug-induced differentiation. The relationship between drug dose and loss of PE is much steeper for DMSO than for RA, suggesting that these two drugs affect different stages of the differentiation decision-making apparatus. Mutant EC cell lines (D3 and RAC65) do not differentiate in the presence of drug-inducers (DMSO and RA, respectively). Neither differentiation-deficient mutant has an altered ability to form gap junctions. When D3 and P19 cells were mixed within the same DMSO-treated aggregates, the D3 cells remained undifferentiated and the P19 cells differentiated much less efficiently than if they were cultured in the absence of the D3 cells. When RAC65 and P19 cells were mixed in RA-treated aggregates, each cell responded to the drug as though the other were absent. Thus RA behaves as a cell-autonomous inducer of differentiation, whereas DMSO-induced differentiation seems to be mediated by interactions between neighboring cells.

Animals↗
PubMed1985

Commitment in a murine embryonal carcinoma cell line during differentiation induced by retinoic acid.

Murine embryonal carcinoma (EC) cells are induced to differentiate when cultured in the presence of retinoic acid (RA). Whereas the EC cells have a high plating efficiency, the differentiated cells have little or no colony-forming ability under the same conditions. We have assumed that the loss of colony-forming ability following exposure of EC cells to RA corresponds to the irreversible commitment of EC cells to differentiate. We found that uncommitted EC cells persist in RA-treated aggregates of EC cells and that the proportion of EC cells stabilizes at a level inversely related to the RA concentration. Both experimental evidence and mathematical modelling results are consistent with the interpretation that there is a dynamic equilibrium achieved by a balance between the processes of EC cell proliferation and differentiation. Since different cell types are induced by different RA concentrations, our results suggest that the commitment to differentiate is not related in any simple way to the developmental program which ensues.

Animals↗
PubMed1985

Analysis of world-wide age-specific data for the association of cervical cancer with infections by herpes simplex virus type 2.

World-wide incidence data obtained from the scientific literature are used to expand to age-specific incidence rates the observation that the incidence of invasive cervical cancer in many human populations is related to the incidence of infections with herpes simplex virus type 2 (HSV-2) in the same community. The data for the incidence of the tumor were obtained from international registries. Data for prevalence of HSV-2 antibodies were used to evaluate the incidence of past primary infections with the virus and a model was used to generate the required age-specific data. Given the nature of the data used and the assumptions implied by the analysis, the conclusions remain speculative. They nevertheless constitute the best available assessment of the age distributions of cancer cases related and unrelated to the virus, and of the time elapsing from the initial virus infection to the development of an invasive tumor.

Adolescent↗