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C J Chapman

Publications and source records attributed to C J Chapman.

At least 19 recordsLinked to original sources
PubMed1992

Promoter-independent activation of heterologous virus gene expression by the herpes simplex virus immediate-early protein ICP27.

The herpes simplex virus (HSV) immediate-early protein ICP27 has been postulated to play an auxillary role in HSV. gene expression, augmenting or inhibiting the activation of different viral promoters by the other immediate-early proteins ICPO and ICP4. Here we show that ICP27 alone can up-regulate gene expression of a retroviral vector containing Moloney murine leukemia virus (MoMuLV) regulatory sequences. This is the first report of an effect of ICP27 on gene expression driven by heterologous virus regulatory sequences. The effect does not involve the region of ICP27 which inhibits gene activation of HSV early gene promoters by ICPO and ICP4, but rather is dependent on the same region of ICP27 as is required to augment the activation of HSV late gene promoters by ICPO and ICP4. This indicates that the two activation effects are likely to operate via the same mechanism. Activation by ICP27 is dependent on the 3' LTR and adjacent region of MoMuLV but is independent of the promoter in the 5' LTR which can be replaced by a heterologous promoter such as that of SV40 without affecting activation by ICP27. The significance of this effect for an understanding of the mechanism of action of ICP27 and its role in regulating the gene expression of HSV and potentially of other viruses is discussed.

3T3 Cells↗
PubMed1992

Fragile site Xq27.3 in a family without mental retardation.

Routine parental blood analysis for a couple undergoing prenatal diagnosis because of maternal age, revealed a 47,XXX karyotype in the mother and expression of the fragile site Xq27.3 in the father. Additional family studies show the fragile site in the father's sister and her two sons. There is no history of intellectual handicap in this family, nor of any physical manifestations of the Fra(X) mental retardation syndrome.

Adult↗
PubMed1992

Experience with direct molecular diagnosis of fragile X.

The utility of the pfxa3 probe for direct molecular diagnosis of the fragile X (FRAXA) has been established. This probe detects amplification of an unstable DNA element consisting of variable length CCG repeats. The size of the amplified fragment is correlated with phenotype and was determined using PstI digested DNA in family members. In 35 families with the fragile X, there was correspondence in 183 cases between the presence of an amplified unstable element and the presence of the fragile X chromosome independently determined by cytogenetics, position in the pedigree, or linked DNA markers flanking the fragile X. There was also correspondence in 124 cases between the presence of the normal 1.0 kb PstI fragment and absence of the fragile X chromosome independently determined by linked flanking markers. Six additional families considered to be isolated cases of 'fragile X' had been diagnosed before recognition of FRAXD. The pfxa3 probe confirmed the cytogenetic diagnosis in three families, the other three being rediagnosed as non-fragile X. A further two families had consistent expression of a different folate sensitive fragile site, FRAXE, close to FRAXA but not associated with fragile X syndrome and not detectable with the pfxa3 probe. Subsequent referrals were received from additional family members or from members of new families for whom carrier status had not been predetermined by linked markers. Direct pfxa3 diagnosis for the 135 females within these 222 additional cases was confirmed by dosage analysis with the control probe pS8.(ABSTRACT TRUNCATED AT 250 WORDS)

Artifacts↗
PubMed1992

T-cell tolerance induced in nude mice grafted with thymic epithelium.

This study investigated the ability of foetal thymic epithelium prepared by 24 degrees culture (24 degrees-TE) or treatment with deoxyguanosine (dGuo-TE), to induce tolerance in nude mice. Thymic chimeras were constructed in which the thymic epithelium differed from the host at both major histocompatibility complex (MHC) antigens and multiple minor histocompatibility antigens (mHa), or at mHa only. Peripheral CTL from nude mice receiving dGuo-TE disparate for mHa, or both MHC antigens and mHa, were uniformly tolerant of thymic mHa. CTL from nude mice grafted with 24 degrees-TE or dGuo-TE were tolerant of host MHC antigens, but the two treatments differed in the efficiency with which they induced tolerance to thymic MHC antigens. CTL responses specific for thymic MHC antigens could be generated in vitro from dGuo-TE grafted mice but not from those receiving 24 degrees-TE. The addition of concanavalin A (Con A) supernatant had no effect on the CTL tolerance observed in 24 degrees-TE grafted mice, suggesting that the lack of CTL responses was not due to tolerance in MHC class II restricted 'helper' cells. However, CTL responses against the thymic MHC antigens of dGuo-TE grafted mice displayed high sensitivity to blocking by anti-CD8 antibodies, indicating that these CTL were of low affinity. These results suggest that 24 degrees-TE induces tolerance in most thymic MHC-specific CTL precursors, whereas dGuo-TE induces tolerance only in CTL with high affinity for thymic MHC antigens. Therefore, 24 degrees-TE and dGuo-TE are both capable of inducing CTL tolerance, consistent with the previously reported acceptance of thymic donor-type skin grafts by nude recipients of dGuo-TE treatment. We conclude that MHC class I molecules on thymic epithelium play a role in negative selection of the developing T-cell repertoire.

Animals↗
PubMed1991

Effects of solute concentration on the entrapment of solutes in phospholipid vesicles prepared by freeze-thaw extrusion.

Phospholipid vesicles prepared by the freeze-thaw extrusion method contain internal solute concentrations which are much higher than the external values (entrapment ratios much greater than 1). This concentrating effect is a complex function of the total impermeant solute concentration in the medium used to prepare vesicles, the presence or absence of permeant solutes in the medium and the apparent competitive binding interactions between solutes and phospholipid. Increases in water phase solute concentration during freezing are thought to underlie the concentrating phenomenon, while osmotic pressure driven lysis of vesicles during thawing appears to limit its magnitude. By judicious selection of solute concentration and physical properties, further increases in the entrapment ratio should be obtainable, improving the usefulness of these vesicles as drug delivery vesicles and experimental systems.

Calcium↗
PubMed1991

Evidence of genetic recombination in Leishmania.

In the genus Leishmania there has been no convincing demonstration of genetic exchange, and it has been proposed that reproduction is clonal. However, preliminary characterization of two strains of Leishmania isolated from wild animals in a zoonotic focus of cutaneous leishmaniasis in the Eastern Province of Saudi Arabia, has suggested that they may represent hybrids of Leishmania major and Leishmania arabica. Evidence presented here strongly supports this hypothesis. Isoenzyme analysis and molecular karyotyping of cloned organisms indicated that the putative hybrids are distinct from other species of Leishmania, and possess characteristics of both L. major and L. arabica. Experiments using highly specific probes demonstrated that kinetoplast minicircle DNA from the putative hybrid contained L. major-specific, but not L. arabica-specific sequences. DNA fingerprinting data obtained using 6 genomic DNA probes were consistent in all cases with a L. major/L. arabica recombinant genotype, and implied both diploidy and allelic segregation. These observations suggest that sexual reproduction may generate genetic diversity within natural Leishmania populations.

Animals↗
PubMed1991

A recombinant HIV provirus is synergistically activated by the HIV Tat protein and the HSV IE1 protein but not by the HSV IE3 protein.

To study the effects of regulatory proteins encoded by herpes viruses on the HIV long terminal repeat (LTR) in the presence or absence of HIV-encoded regulatory products, we prepared a proviral construct containing 5' and 3' HIV LTR, but lacking the coding sequences of any HIV proteins. This construct allowed the effects of herpesvirus regulatory proteins on the HIV LTR to be assessed in a construct similar to the HIV provirus whilst also allowing their interactions with HIV-encoded regulatory proteins to be studied. In this system, the herpes simplex virus (HSV) protein IE1 (ICPO) but not the IE3 (ICP4) protein can activate the HIV LTR, whereas both proteins are active on a single plasmid-borne HIV LTR. Although the activation of the LTR by IE1 is strongly stimulated by the HIV Tat protein, it can also be observed in the absence of Tat, indicating that HSV infection via IE1 has the potential to activate an entirely silent, latent HIV provirus.

Animals↗
PubMed1990

General features in the stoichiometry and stability of ionophore A23187-cation complexes in homogeneous solution.

Existing literature describing the stoichiometry and stability of complexes between A23187 and divalent cations in solution has been extended to include additional transition series cations, the heavy-metal cations Cd2+ and Pb2+, plus seven lanthanide series trivalent cations. Stability constants of 1:1 complexes between the ionophore and the divalent cations vary by 6.2 orders of magnitude between Cu2+ and Ba2+ which are the strongest and weakest complexes, respectively. Considering alkaline-earth and first-series transition cations together, the pattern of stability constants obeys the extended Irving-Williams series as is seen with many nonionophorous liganding agents. Cd2+ and Pb2+ are bound with an affinity similar to those of Mn2+ and Zn2+, whereas the lanthanides are bound with little selectivity and slightly higher stability. Titration of the ionophore in the 10(-5) M concentration range with di- and trivalent cations gives rise first to complexes of stoichiometry MA2 and subsequently to MA as the metal concentration is increased. The second stepwise stability constants for formation of the MA2 species exceeds the first constant by approximately 10-fold. With lanthanides, heavy metals, and transition-metal cations, OH-, at near physiological concentrations, competes significantly with free ionophore for binding to the 1:1 complexes. This competition is not apparent when Ca2+ or Mg2+ are the central cations. Possible implications of the 1:1 complex selectivity pattern, the ionophore-hydroxide competitive binding equilibria, and potential ternary complexes involving 1:1 ionophore:cation complexes and other anions present in biological systems are discussed with respect to the ionophore's transport selectivity and biological actions.

Biological Transport↗
PubMed1990

A visual interface to computer programs for linkage analysis.

This paper describes a visual approach to the input of information about human families into computer data bases, making use of the GEM graphic interface on the Atari ST. Similar approaches could be used on the Apple Macintosh or on the IBM PC AT (to which it has been transferred). For occasional users of pedigree analysis programs, this approach has considerable advantages in ease of use and accessibility. An example of such use might be the analysis of risk in families with Huntington disease using linked RFLPs. However, graphic interfaces do make much greater demands on the programmers of these systems.

Computer Graphics↗
PubMed1990

Factors affecting solute entrapment in phospholipid vesicles prepared by the freeze-thaw extrusion method: a possible general method for improving the efficiency of entrapment.

It is often assumed that the internal solute concentrations of phospholipid vesicles are equal to those in the medium in which they were prepared, particularly when freeze-thaw cycles are employed during the procedure. Conditions are reported here which when used to prepare vesicles by the polycarbonate filter extrusion method, produce approximately 12- and approximately 7-fold higher internal concentrations of Ca2+ and sucrose, respectively, than exist in the external medium. Formation of these large gradients is dependent upon the use of freeze-thaw cycles during preparation, on the presence of tetraethylammonium perchlorate in the medium, and is independent of media pH across the region of pH 5-9. Gradient formation is antagonized by high concentrations of an impermeant solute (NaCl). It is proposed that gradients form because solutes are concentrated by exclusion from ice during freezing but that they are normally dissipated by osmotic lysis during thawing. The presence of a permeant solute such as tetraethylammonium perchlorate provides an alternative mechanism to balance osmotic pressure, thereby preserving the gradients of impermeable species.

Calcium↗
PubMed1990

A new species of Leishmania isolated from the great gerbil Rhombomys opimus.

Leishmania turanica n.sp., found infecting the desert rodent Rhombomys opimus in the southern territories of the USSR and the Mongolian People's Republic, is described. This parasite exists sympatrically with L. major and L. gerbilli in R. opimus and is the predominant species. A total of 284 isolates of L. turanica from R. opimus, 3 from naturally infected Phlebotomus andrejevi and 1 from P. papatasi were characterized and found to be clearly distinguishable on isoenzyme and nuclear DNA characteristics from all other Old World taxa of Leishmania.

Animals↗
PubMed1990

Three cases of partial trisomy 7q owing to rare structural rearrangements of chromosome 7.

Three cases of partial trisomy 7q are described. One case had duplication of region 7q22.1----q31.2 owing to a de novo direct intra-arm intrachromosomal duplication. The other two cases, first cousins, were trisomic for 7q34----qter, resulting from recombination within the inserted segment of a dir ins(7;17)(q34;q23.1q25.3)mat. All three cases had a number of the already recorded manifestations of partial trisomy 7q, namely strabismus, low set ears, depressed nasal bridge, small nose, hypotonia, and mental retardation.

Adult↗
PubMed1990

IgG antibody and delayed-type hypersensitivity responses in nude mice grafted with thymic epithelium.

This study compared the ability of foetal thymic epithelium depleted of lymphocytes and dendritic cells, by low temperature or deoxyguanosine (dGuo) treatment in organ culture, to reconstitute T-cell function in nude mice. It is shown that renal capsule grafts of either type could promote the development of functional T lymphocytes in the periphery, as judged by in vivo assays. Both syngeneic and allogeneic thymic epithelium endowed nude mice with the capacity to mount IgG antibody and delayed-type hypersensitivity (DTH) responses to the T-dependent antigen ovalbumin (OVA). Functional reconstitution was accompanied by the appearance of Thy-1-bearing cells in the spleens of thymic grafted nude mice. The results from allogeneically grafted recipients show that a substantial population of peripheral T cells was present that collaborated with B cells and other antigen-presenting cells (APC) which do not express major histocompatibility complex (MHC) molecules of the thymus donor haplotype.

Animals↗
PubMed1989

Familial distal trisomy 8(q24.13----qter).

Trisomy for the distal part of the long arm of chromosome 8(q24.13----qter) is described in three sibs. The anomaly arose as an adjacent 1 meiotic segregation from a balanced reciprocal translocation t(1;8)(q44; q24.13)mat.

Adult↗
PubMed1987

Partial monosomy 12p13.1----13.3.

We describe a 27 month old female child with partial monosomy for the short arm of chromosome 12: 46,XX,del(12)(p13.1----p13.3). She differs from the eight cases described by others, in that she is less severely affected. Her main clinical features are developmental delay, protruding tongue, strabismus, slightly unusual facies, slight micrognathia, and speech delay.

Child, Preschool↗
PubMed1986

Studies on three rare fragile sites. 2q13, 12q13, and 17p12 segregating in one family.

Three fragile sites 2q13, 12q13, and 17p12 were found in one family. In the index case, who was first investigated in 1969 for low birth weight and bilateral inguinal hernia, three tissues were examined, blood, marrow, and skin. Three of the family have been reinvestigated after 17 years. Cultures for sister chromatid exchange (SCE) and the effects of aphidicolin, fluorodeoxyuridine (FUdR), bromodeoxyuridine (BrdU), and methotrexate on the frequency of the fragilities were studied. The mother of the index case who is an obligate carrier for the fragile 2q13 does not express it in folate/thymidine deficient medium. Further studies on her using a lymphoblastoid cell line, showed that there was a reduced level of fragility of 12q13 and 17p12 in B-lymphocytes compared to T-lymphocytes. Excess thymidine and FUdR when added to the lymphoblastoid cell line did not induce the 2q13. These studies also confirm the induction of a range of common fragile sites by treatment with aphidicolin, showing in addition homozygosity for at least 3p14, 6q26, 16q23, and Xp22. There were no detectable increases in the SCE rate between individuals with fragile sites and the five controls tested. There was no history of cancer or phenotypic abnormalities in the family.

Adolescent↗