Acyl sulfonamide anti-proliferatives. Part 2: activity of heterocyclic sulfonamide derivatives

The anti-proliferative activity of acylated heterocyclic sulfonamides is described in Vascular Endothelial Growth Factor-dependent Human Umbilical Vascular Endothelial Cells (VEGF-HUVEC) and in HCT116 tumor cells in a soft agar diffusion assay.     

Identification and characterization of the immunodominant rat HER-2/neu MHC class I epitope presented by spontaneous mammary tumors from HER-2/neu-transgenic mice

The HER-2/neu (neu-N)-transgenic mice are a clinically relevant model of breast cancer. They are derived from the parental FVB/N mouse strain and are transgenic for the rat form of the proto-oncogene HER-2/neu (neu). In this study, we report the identification of a MHC class I peptide in the neu protein that is recognized by CD8(+) T cells derived from vaccinated FVB/N mice. This 10-mer was recognized by all tumor-specific FVB/N T cells generated regardless of the TCR Vbeta region expressed by the T cell or the method of vaccination used, establishing it as the immunodominant MHC class I epitope in neu. T cells specific for this epitope were able to cure FVB/N mice of transplanted neu-expressing tumor cells, demonstrating that this is a naturally processed peptide. Altered peptide analogs of the epitope were analyzed for immunogenicity. Vaccination with dendritic cells pulsed with a heteroclitic peptide provided FVB/N and neu-N mice with increased protection against tumor challenge as compared with mice immunized with dendritic cells loaded with either wild-type or irrelevant peptide. Discovery of this epitope allows for better characterization of the CD8(+) T cell responses in the neu-N mouse model in which neu-specific tolerance must be overcome to produce effective antitumor immunity.     

Crystal structure and evolution of a prokaryotic glucoamylase

The first crystal structures of a two-domain, prokaryotic glucoamylase were determined to high resolution from the clostridial species Thermoanaerobacterium thermosaccharolyticum with and without acarbose. The N-terminal domain has 18 antiparallel strands arranged in beta-sheets of a super-beta-sandwich. The C-terminal domain is an (alpha/alpha)(6) barrel, lacking the peripheral subdomain of eukaryotic glucoamylases. Interdomain contacts are common to all prokaryotic Family GH15 proteins. Domains similar to those of prokaryotic glucoamylases in maltose phosphorylases (Family GH65) and glycoaminoglycan lyases (Family PL8) suggest evolution from a common ancestor. Eukaryotic glucoamylases may have evolved from prokaryotic glucoamylases by the substitution of the N-terminal domain with the peripheral subdomain and by the addition of a starch-binding domain.     

A comparison of the suitabilities of rectal,gut, and insulated axilla temperatures for measurement of the circadian rhythm of core temperature in field studies

Eight healthy males were studied for a total of 13 subject-days to assess if gut (from an ingested pill) and axilla (from a thermally insulated skin probe) temperatures would act as a substitute for rectal temperature in field studies of the circadian rhythm of core temperature. Subjects slept and went about their activities, indoors and outdoors, normally. Regular recordings (at 6min intervals) were made of temperatures from the three sites. In addition, activity was measured (by a sensor on the nondominant wrist) so that the raw temperature data could be “purified,” that is, corrected for the direct effects of sleep and activity. Inspection of the raw data indicated that there was a close parallelism between rectal and gut temperatures, but that the parallelism between rectal and insulated axilla temperatures was less reliable. This parallelism was supported by initial calculations of the correlations between rectal and gut temperatures (high and positive) and between rectal and insulated axilla (lower, though still positive) temperatures. Calculation of the limits of agreement between the parameters of the cosine curves fitted to the raw data confirmed that the rectal and gut temperatures were far closer with regard to acrophase and amplitude than were rectal and insulated axilla temperatures (-0.31±0.89 vs. +0.75±6.03 h and +0.002±0.116 vs. +0.083±0.625°C, respectively). After purification of the temperature data, the limits of agreement for the cosine parameters acrophase and amplitude still indicated that there was a closer agreement between rectal and gut temperatures than between rectal and insulated axilla temperatures (-0.30±1.12 vs. +0.58±6.69 h, and +0.007±0.116 vs. +0.104±0.620°C, respectively). Part of the explanation of this difference was the unreliable relationships between temperature changes in insulated axilla temperature and bursts of activity and going to bed. It is concluded that, whereas gut temperature is a viable alternative to rectal temperature (from the viewpoints of both user acceptability and the reliability of data obtained), insulated axilla temperature, though acceptable to subjects, is unreliable from an experimental viewpoint.     

The diurnal rhythm in isometric muscular performance differs with eumenorrheic menstrual cycle phase

The aim of this study was to examine the effect of the interaction of circamensal and diurnal rhythms in temperature upon the production of maximal voluntary muscle force. Ten eumenorrheic females (mean age: 24 +/- 3 yr mean body mass: 58.4 +/- 6.9 kg) participated in the experiment at both 06:00 and 18:00h at the mid-point of both the follicular and luteal phases of the menstrual cycle. Subjects performed tasks of maximal isometric lifting strength (MILS) at knee height, and endurance time (t) for lifting 45% of MILS, upon an isometric lift dynamometer. Body temperature was elevated at 18:00h and in the luteal phase by 0.52 +/- 0.4 and 0.26 +/- 0.35 degrees C, respectively. The amplitude of the diurnal variation in temperature was blunted by 0.3 degrees C within the luteal phase. Maximal isometric performance was elevated by 8% at 18:00h in the luteal phase of the cycle (p < 0.05 interaction for MILS) but unaffected by time of day in the follicular phase. Endurance time was unaffected by time or phase (p > 0.05). It should be noted that the classic diurnal rhythm in maximal voluntary isometric muscle force may not be evident in all phases of the female menstrual cycle.     

Genomewide linkage analysis of quantitative spirometric phenotypes in severe early-onset chronic obstructive pulmonary disease

Chronic obstructive pulmonary disease (COPD) is a common, complex disease associated with substantial morbidity and mortality. COPD is defined by irreversible airflow obstruction; airflow obstruction is typically determined by reductions in quantitative spirometric indices, including forced expiratory volume at 1 s (FEV(1)) and the ratio of FEV(1) to forced vital capacity (FVC). To identify genetic determinants of quantitative spirometric phenotypes, an autosomal 10-cM genomewide scan of short tandem repeat (STR) polymorphic markers was performed in 72 pedigrees (585 individuals) ascertained through probands with severe early-onset COPD. Multipoint variance-component linkage analysis (using SOLAR) was performed for quantitative phenotypes, including FEV(1), FVC, and FEV(1)/FVC. In the initial genomewide scan, significant evidence for linkage to FEV(1)/FVC was demonstrated on chromosome 2q (LOD score 4.12 at 222 cM). Suggestive evidence was found for linkage to FEV(1)/FVC on chromosomes 1 (LOD score 1.92 at 120 cM) and 17 (LOD score 2.03 at 67 cM) and to FVC on chromosome 1 (LOD score 2.05 at 13 cM). The highest LOD score for FEV(1) in the initial genomewide scan was 1.53, on chromosome 12, at 36 cM. After inclusion of 12 additional STR markers on chromosome 12p, which had been previously genotyped in this population, suggestive evidence for linkage of FEV(1) (LOD score 2.43 at 37 cM) to this region was demonstrated. These observations provide both significant evidence for an early-onset COPD-susceptibility locus on chromosome 2 and suggestive evidence for linkage of spirometry-related phenotypes to several other genomic regions. The significant linkage of FEV(1)/FVC to chromosome 2q could reflect one or more genes influencing the development of airflow obstruction or dysanapsis.     

Quality control of CD4+ T-lymphocyte enumeration: results from the last 9 years of the United Kingdom National External Quality Assessment Scheme for Immune Monitoring (1993-2001)

The human immunodeficiency virus (HIV) global epidemic has necessitated the routine enumeration of T-lymphocyte subsets, which has created a need for external quality assurance (EQA). The United Kingdom National External Quality Assessment Scheme (UK NEQAS) for Immune Monitoring provides EQA for 296 laboratories in 40 countries. In 1993, UK NEQAS developed and incorporated into its program stabilized whole blood that enables the accurate monitoring of laboratory performance. Overall, the mean interlaboratory coefficient of variation (CV) for percentage CD4(+) T-lymphocyte subset enumeration has fallen from 15% to less than 5%, as a direct result of the increased use of CD45/ side scatter (SSC) gating. Laboratories using alternative gating strategies (i.e., CD45/CD14 or forward scatter [FSC]/SSC) were about 7.4 times more likely to fail an EQA exercise. Furthermore, the adoption of single-platform technology resulted in a reduction of the overall mean interlaboratory CV for absolute CD4(+) T lymphocytes from 56% (prior to the widespread use of single-platform technology) to 9.7%. Individual laboratory deficiencies were also identified using a performance monitoring system and, through re-education by collaboration with the coordinating center, satisfactorily resolved. In conclusion, during the last 9 years, the UK NEQAS for Immune Monitoring program has highlighted the significant technological advances made by laboratories worldwide that undertake lymphocyte subset enumeration.