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S Craig

Publications and source records attributed to S Craig.

At least 55 records · Page 3Linked to original sources

Calculation of System International Sensitivity Index: how many calibrant plasmas are required?

AIMS: To determine the minimum number of calibrant plasmas required to assign an accurate System International Sensitivity Index (ISI). METHODS: Eighteen lyophilised adsorbed and eight frozen coumarin plasmas were despatched to 57 participating laboratories. Each centre determined the prothrombin time of all of the plasmas using their routine system of coagulometer and thromboplastin reagent and provided their system mean normal prothrombin time (MNPT). For each laboratory, the slope of the line relating local system results, including MNPT, to calibrated values obtained with BCT 441, RBT 1303 or RBT 90 was derived by orthogonal regression. The number of adsorbed calibrant plasmas included in the calculation was reduced, two at a time, from 18 to two. The validity of the System ISI was tested using the frozen coumarin plasmas. RESULTS: All systems tested gave higher International Normalised Ratios (INR) than those obtained manually with the reference thromboplastins. All systems therefore required correction. No changes were seen in the System ISI as the number of calibrant plasmas was reduced from 18 to 12. Below 12, inconsistent results were obtained with some systems and were dependent on plasma selection. CONCLUSIONS: Although System ISI could be assigned using as few as two calibrant plasmas, to avoid imprecise calibration, a minimum of 12 calibrant plasmas is recommended.

Calibration↗

Local INR correction: justification for a simplified approach.

AIMS: Errors in reporting International Normalised Ratios (INR) may be corrected by assignment of a System International Sensitivity Index (System ISI). This 57 centre study tests the validity of several procedures for INR correction. METHODS: Prothrombin times of eight lyophilised coumarin calibrants, a lyophilised normal pool calibrant, and eight frozen coumarin plasmas were determined at each centre. The calibrants were calibrated using international reference preparations. The eight frozen coumarin plasmas were calibrated in a four centre international exercise. The relations tested were: (a) the logarithm of local prothrombin time against the logarithm of reference prothrombin time; (b) reference INR against local prothrombin time; and (c) logarithm of reference INR against logarithm of local prothrombin time. These methods were analysed by both linear and orthogonal regression. RESULTS: All system groups required correction, the mean percentage deviation of the uncorrected data from the calibrated values was 19.0%. There was also considerable variation in INR, with the coefficient of variance (CV) ranging from 11.30 to 17.29%. Correction of INR was possible with all methods (CV reduced to < 7%). However, only when a plot of the logarithm of local prothrombin time against the logarithm of reference prothrombin time was fitted by orthogonal regression, or a plot of logarithm of reference INR against logarithm of local prothrombin time was fitted by either type of regression analysis, did the best fit line through the calibrant plasmas also pass close to the local mean normal prothrombin time. CONCLUSIONS: While INR correction may be achieved by all the above methods, that relating log reference INR to log local prothrombin time by linear regression analysis is the simplest to perform.

Anticoagulants↗

Implementation and process evaluation of a student "school-as-community" group. A component of a school-based drug abuse prevention program.

Little documentation exists regarding the functioning of formalized adolescent groups as drug abuse prevention agents. Two studies are described that were conducted at high schools whose students are at high risk for drug abuse. Twenty-one schools were randomly assigned to one of three conditions: (a) standard care, (b) classroom drug abuse education only, or (c) classroom plus school-as-community. Results of the first study indicated that the school-as-community component--which involved weekly meetings and periodic events at seven schools--was implemented as planned, drug abused focused, and perceived as productive in discouraging drug abuse. In the second study, staff in the classroom plus school-as-community condition self-reported involvement in the greatest number of community activities across the school year, compared with staff from the other two conditions. These two studies support the feasibility of formalized groups of high-risk youth to promote drug-free events.

Adolescent↗

A preterm infant with upper cervical spinal cord injury, following delivery with Kielland's forceps.

A report is presented of a male infant who was born at 34 weeks' gestation, following rotation and delivery using Kielland's forceps. The initial diagnosis of hypoxic ischaemic encephalopathy delayed recognition of an upper cervical spinal and cord injury, which was subsequently confirmed by magnetic resonance imaging. The infant never developed spontaneous respiration and died following withdrawal of intensive care at 21 days of age.

Adult↗

School-as-community activity selection at continuation high schools.

This paper describes two studies which together provide an example of an empirically based "school-as-community" development process used in a large-scale school-based drug prevention trial among youth at continuation high schools in California. First, we generated a list of schools' current community involvement from 96 school personnel and 144 students from 20 continuation high schools. Second, we reduced the number of activities indicated, with some discussion, to a list of 12 populars. Then, we administered this list to a total of 388 students from six continuation high schools who provided perceived quality ratings. We generated six categories of activities from the results of these two studies. These studies reveal a means to determine activities likely to be completed by this school system and a means to limit the variability of activities which could be manipulated in an experimental trial involving a school-as-community component.

Adolescent↗

Immediate impact of thirty-two drug use prevention activities among students at continuation high schools.

Those teenagers who are unable to remain in the regular school system for reasons including substance use are transferred to a continuation high school. Generic comprehensive social-influence-drug-use-prevention-activities are less likely to be effective for use with these at risk youth. Thus, both classroom and self-instruction (main mode of instruction at continuation high schools) versions of 16 activities derived from different theoretical sources were tested and ranked on immediate outcome variables. 388 students from six continuation high schools were provided with a pretest-activity-posttest "component study" protocol. The scores on perceived quality ratings were standardized and averaged to permit easy comparisons across lessons. While yielding similar knowledge changes, students who received the health educator led activity consistently reported higher scores on perceived quality. Social influence-oriented lessons, in general, were rated of relatively low perceived quality. The present approach assisted in selection of the lessons with the greatest overall immediate impact.

Adolescent↗

Accumulation of a sulphur-rich seed albumin from sunflower in the leaves of transgenic subterranean clover (Trifolium subterraneum L.).

A gene encoding a sulphur-rich, sunflower seed albumin (23% cysteine plus methionine) was modified to contain the promoter for the 35S RNA of cauliflower mosaic virus, in order to obtain leaf expression in transgenic plants. In addition, a sequence encoding an endoplasmic reticulum-retention signal was added to the 3' end of the coding region so as to stabilize the protein by diverting it away from the vacuole. The modified gene was introduced into subterranean clover (T. subterraneum L.) and its expression was detected by northern and western blots and by immunogold localization. The albumin was accumulated in the lumen of the endoplasmic reticulum, and, among six independent, transformed lines, it accumulated in the leaves of T0 transgenic plants at varying levels up to 0.3% of the total extractable protein. The level of accumulation of the sunflower albumin increased with increasing leaf age, and in the older leaves of the most highly expressing plants of the T1 generation it reached 1.3% of total extractable protein. Expression of the SSA gene was stable in the first and second generation progeny. These results indicate that there is potential for significantly improving the nutritional value of subterranean clover for ruminant animals such as sheep by expressing genes that code for sulphur-rich, rumen-stable proteins in leaves.

Albumins↗

Perinatal infection with Listeria monocytogenes.

Listeria monocytogenes has been increasingly recognized as a cause of intrauterine sepsis with associated perinatal wastage. The condition is mostly acquired through dietary intake and appropriate advice should be given to all pregnant women. The most common presentations in pregnancy include premature labour, an influenza-like illness and reduced fetal movements. In this report, we present a series of 24 cases of perinatal listeria infection presenting to either our obstetric or neonatal units and confirmed by the microbiology department of the hospital. In particular, we wish to highlight 3 cases in which antenatal diagnosis and aggressive therapy was associated with a successful outcome. Amongst the remaining 21 cases in which an antenatal diagnosis was not made, there were 5 perinatal deaths and 1 mid-trimester loss at 18 weeks. Clinicians must maintain a high index of suspicion for listeria, particularly in gravid patients who present with fever in the setting of a persistent 'flu-like' illness and premature labour. Once suspected, appropriate specimens for listeria culture should include blood, cervical swabs and midstream urine. Empirical antibiotic therapy with amoxicillin should be instituted while waiting for culture results in patients with possible Listeria monocytogenes sepsis.

Adult↗

The heterosexual human immunodeficiency virus type 1 epidemic in Thailand is caused by an intersubtype (A/E) recombinant of African origin.

Since 1989, human immunodeficiency virus type 1 (HIV-1) has spread explosively through the heterosexual population in Thailand. This epidemic is caused primarily by viruses classified as "subtype E", which, on the basis of limited sequence comparisons, appear to represent hybrids of subtypes A (gag) and E (env). However, the true evolutionary origins of "subtype E" viruses are still obscure since no complete genomes have been analyzed, and only one full-length subtype A sequence has been available for phylogenetic comparison. In this study, we determined full-length proviral sequences for "subtype E" viruses from Thailand (93TH253) and the Central African Republic (90CR402) and for a subtype A virus from Uganda (92UG037). We also sequenced the long terminal repeat (LTR) regions from 16 virus strains representing clades A, C, E, F, and G. Detailed phylogenetic analyses of these sequences indicated that "subtype E" viruses do indeed represent A/E recombinants with multiple points of crossover along their genomes. The extracellular portion of env, parts of vif and vpr, as well as most of the LTR are of subtype E origin, whereas the remainder of the genome is of subtype A origin. The possibility that the discordant phylogenetic positions of "subtype E" viruses in gag- and env-derived trees are the result of unusual rates or patterns of evolution was also considered but was ruled out on the basis of two lines of evidence: (i) phylogenetic trees constructed for synonymous and nonsynonymous substitutions yielded the same discordant branching orders for "subtype E" gag and env gene sequences, thus excluding selection-driven evolution, and (ii) multiple crossovers in the viral genome are most consistent with the copy choice model of recombination and have been observed in other documented examples of HIV-1 intersubtype recombination. Thai and CAR "subtype E" viruses exhibited the same pattern of A/E mosaicism, indicating that the recombination event occurred in Africa prior to the spread of virus to Asia. Finally, all "subtype E" viruses were found to contain a distinctive two-nucleotide bulge in their transactivation response (TAR) elements. This feature was present only in viruses which also contained a subtype A 5' pol region (i.e., subtype A viruses or A/D and A/E recombinants), raising the possibility of a functional linkage between the TAR region and the polymerase. The implications of epidemic spread of a recombinant HIV-1 strain to viral natural history and vaccine development are discussed.

Africa↗

Molecular cloning and analysis of functional envelope genes from human immunodeficiency virus type 1 sequence subtypes A through G. The WHO and NIAID Networks for HIV Isolation and Characterization.

Present knowledge of human immunodeficiency virus type 1 (HIV-1) envelope immunobiology has been derived almost exclusively from analyses of subtype B viruses, yet such viruses represent only a minority of strains currently spreading worldwide. To generate a more representative panel of genetically diverse envelope genes, we PCR amplified, cloned, and sequenced complete gp160 coding regions of 35 primary (peripheral blood mononuclear cell-propagated) HIV-1 isolates collected at major epicenters of the current AIDS pandemic. Analysis of their deduced amino acid sequences revealed several important differences from prototypic subtype B strains, including changes in the number and distribution of cysteine residues, substantial length differences in hypervariable regions, and premature truncations in the gp41 domain. Moreover, transiently expressed glycoprotein precursor molecules varied considerably in both size and carbohydrate content. Phylogenetic analyses of full-length env sequences indicated that the panel included members of all major sequence subtypes of HIV-1 group M (clades A to G), as well as an intersubtype recombinant (F/B) from an infected individual in Brazil. In addition, all subtype E and three subtype G viruses initially classified on the basis of partial env sequences were found to cluster in subtype A in the 3' half of their gp41 coding region, suggesting that they are also recombinant. The biological activity of PCR-derived env genes was examined in a single-round virus infectivity assay. This analysis identified 20 clones, including 1 from each subtype (or recombinant), which expressed fully functional envelope glycoproteins. One of these, derived from a patient with rapid CD4 cell decline, contained an amino acid substitution in a highly conserved endocytosis signal (Y721C), as mediated virus entry with very poor efficiency, although they did not contain sequence changes predicted to alter protein function. These results indicate that the env genes of primary HIV-1 isolates collected worldwide can vary considerably in their genetic, phylogenetic, and biological properties. The panel of env constructs described here should prove valuable for future structure-function studies of naturally occurring envelope glycoproteins as well as AIDS vaccine development efforts targeted against a broader spectrum of viruses.

Amino Acid Sequence↗

BB-10010: an active variant of human macrophage inflammatory protein-1 alpha with improved pharmaceutical properties.

The stem cell inhibitor, macrophage inflammatory protein-1 alpha (MIP-1 alpha) or LD78, protects multipotent hematopoietic progenitors in murine models from the cytotoxic effects of chemotherapy. Clinical use of human MIP-1 alpha during chemotherapy could therefore lead to faster hematologic recovery and may allow dose intensification. We have also shown that human MIP-1 alpha causes the rapid mobilization of hematopoietic cells, suggesting an additional clinical use in peripheral blood stem cell transplantation. However, the clinical evaluation of human MIP-1 alpha is complicated by its tendency to associate and form high molecular weight polymers. We have produced a variant of rhMIP-1 alpha, BB-10010, carrying a single amino acid substitution of Asp26 > Ala, with a reduced tendency to form large polymers at physiologic pH and ionic strength. This greatly increases its solubility, facilitating its production and clinical formulation. We confirmed the potency of BB-10010 as a human MIP-1 alpha-like agonist in receptor binding, calcium mobilization, inhibition of colony formation, and thymidine suicide assays. The myeloprotective activity of BB-10010 was shown in a murine model of repeated chemotherapy using hydroxyurea. BB-10010 is therefore an ideal variant with which to evaluate the therapeutic potential of recombinant human MIP-1 alpha.

Amino Acid Sequence↗

Two-year behavior outcomes of project towards no tobacco use.

The article presents 2-year follow-up data from a school-based tobacco use prevention project designed to test the effectiveness of 3 primary components in social influence programs. The components either teach refusal skills, awareness of social value misperceptions, or physical consequences. Curricula were tested with a randomized experiment involving 48 junior high schools. These data suggested that (a) a physical-consequences curriculum is successful at attenuating increases in adolescent smokeless tobacco use, (b) cigarette experimentation may be attenuated by various approaches, and (c) a comprehensive program with all 3 components was necessary to attenuate increases in weekly use of both forms of tobacco. These results also indicate that school-based tobacco use interventions can be effective at least 2 years postprogram, after students make their transition to high school.

Adolescent↗

Differences between specificities of IgE and IgG4 antibodies: studies using recombinant chain 1 and chain 2 of the major cat allergen Felis domesticus (Fel d) I.

IgE- and IgG4 antibodies were compared for reactivity with recombinant chain 1 and chain 2 of the cat allergen Felis domesticus (Fel d) I. Recombinant chain 1 and chain 2 were coupled to sepharose and tested in IgE- and IgG4 radioallergosorbent test (RAST) experiments. Substantial IgE- and IgG4 binding was found. The fraction of Fel d I-specific antibody that bound to the recombinant chains was calculated. For chain 1, the mean value of this fraction was 0.30 for IgE and 0.23 for IgG4 (P = 0.05). For chain 2, the mean value of this fraction was 0.19 for IgE and 0.13 for IgG4 (P = 0.02). These results indicate that differences in fine specificity exist between IgE and IgG4 antibodies. Moreover, these findings support our results with chemically prepared peptides derived from these two chains and suggest that the B cells producing IgE antibodies are more likely to recognize a less 'native' form of Fel d I, compared with IgG4.

Adolescent↗

Continuation high schools: youth at risk for drug abuse.

Students at alternative high schools may be at substantial risk for drug abuse. The present article provides a general overview of the drug use-related context of continuation high schools in southern California. A total of 144 students and ninety-six staff were interviewed from twenty continuation high schools. The interview data revealed that continuation school students show high levels of substance use. However, only 20 percent of the students report that they received any drug abuse prevention programming. Also, students at continuation high schools aspired to a productive life after high school including continued education. Thus, these youth may still be amenable to preventive educational interventions which deter them from drug use and help them to fulfill their future goals.

Adolescent↗

Purification of human basophils by density and size alone.

Basophils typically account for approximately 1% of the white cells in peripheral blood. We have developed a unique method for purifying basophils from whole blood of normal subjects to at least 95% purity. Basophils are separated from other cell types by density-dependent sedimentation in Percoll and cell sorting, based solely on their size and granularity. The mean overall yield ranged from 5% to 28%. The procedure is typically completed within 4 h. The highly purified basophils obtained are functionally competent and morphologically intact. They release histamine in response to Fc epsilon RI-mediated stimulation, express Fc epsilon RI and BSP-1 ligand as analyzed by flow cytometry, and exhibit the known characteristic ultrastructural features of basophils by electron microscopy. This procedure avoids positive-selection antibodies that might perturb receptors on basophils or negative-selection antibodies that might activate other cell types, and can be used to obtain basophils for studies in vitro.

Basophils↗

Genetic variation of HIV type 1 in four World Health Organization-sponsored vaccine evaluation sites: generation of functional envelope (glycoprotein 160) clones representative of sequence subtypes A, B, C, and E. WHO Network for HIV Isolation and Characterization.

As part of the WHO Network for HIV Isolation and Characterization, we PCR amplified, cloned, and sequenced gp120 and gp160 genes from 12 HIV-1 isolates collected in four WHO-sponsored vaccine evaluation sites (Brazil, Rwanda, Thailand, Uganda). Envelope clones were derived from PBMC-grown isolates obtained from asymptomatic individuals within 2 years of seroconversion. Analysis of their deduced amino acid sequences identified all but one to contain an uninterrupted open reading frame. Transient expression and biological characterization of selected gp160 constructs identified six clones to encode full length and functional envelope glycoproteins. Phylogenetic analysis of their nucleotide sequences revealed that they represent HIV-1 subtypes A, B, C, and E. Since current knowledge of HIV-1 envelope immunobiology is almost exclusively derived from subtype B viruses, these reagents should facilitate future envelope structure, function and antigenicity studies on a broader spectrum of viruses. This should assist in the design and evaluation of effective vaccines against HIV-1.

AIDS Vaccines↗