Pneumococcal surface adhesin A antibody concentration in serum and nasopharyngeal carriage of Streptococcus pneumoniae in young African infants.
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Adhesins, Bacterial↗
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Evaluation of synthetic, M type-specific peptides as antigens in a multivalent group A streptococcal vaccine.
The recent development of emm gene sequence-based typing methodology has allowed group A streptococci (GAS) M serotype prevalence data to be determined. This information has been used to identify the components of a multivalent M protein peptide vaccine that could theoretically prevent most of the GAS-mediated diseases in the USA. In this study, we have evaluated in mice the immunogenicity and protective ability of multiple synthetic, M type-specific peptides, derived from the N-termini of three prevalent GAS serotypes (three peptides per serotype, total of nine peptides). At least one peptide, representing each of the three M types tested, was immunogenic. Five of the nine synthetic peptides tested, elicited an immune response in mice, and sera raised against four of the peptides, all possessed functional activity as demonstrated in a bactericidal assay. In vivo nasopharyngeal challenge experiments were carried out with peptides from the M1 (peptide M1-3) and M3 (peptide M3-2) proteins induced in vivo immune protection by reducing intranasal carriage. Reduction in colonization for M1-3 and M3-2 was 90% (P=0.02) and 66% (P<0.17), respectively. A reduction in colonization of 67% (P=0.03) was observed for M3-2 immunized mice when M43, a heterologous serotype, was used as the challenge strain. These results show the utility of synthetic, M type-specific peptides as antigens in a multivalent GAS vaccine.
Administration, Intranasal↗
Purification and characterization of Streptococcus pneumoniae palmitoylated pneumococcal surface adhesin A expressed in Escherichia coli.
All Streptococcus pneumoniae isolates tested to date express a species-common lipoprotein designated as pneumococcal surface adhesin A (PsaA). This protein is cell-associated, hydrophobic, immunogenic, and genetically conserved. It is currently under investigation as a potential component in third-generation pneumococcal vaccine formulations. To overcome the problem of low-level expression of native hydrophobic PsaA in S. pneumoniae, and also of the recombinant PsaA (rPsaA) in Escherichia coli, we generated a stable E. coli construct expressing functional palmitoylated rPsaA ( approximately 10 mg/l of fermentation culture) using Borrelia burgdorferi outer surface protein A (OspA, a hydrophobic lipoprotein) signal peptide. By Western blot analysis, the chimeric rPsaA ( approximately 34 kDa) was detected in the cell lysate using anti-PsaA antibodies. It was partially purified by extracting the cell pellet with PBS/Triton X(R)-114 buffers, followed by anion exchange filter chromatography. A trypsin digestion profile of rPsaA closely resembled that of the native protein, as revealed by SDS-PAGE/silver staining. Lipidation of rPsaA was confirmed by labeling recombinant E. coli cells with [(3)H] palmitic acid and analyzing the labeled E. coli cells by Western blotting coupled with autoradiography. Further, analysis of purified rPsaA by mass spectrometry (MALDI-TOF) revealed a heterogenous spectrum with a major peak (M+H)(+1) of mass 33,384 Da (theoretical mass of palmitoylated rPsaA=33,361 Da). Purified rPsaA was immunogenic in CBA/NCAHN-XID female mice following intranasal immunization with or without adjuvant, as determined by measurement of anti-PsaA serum IgG levels. These anti-PsaA antibodies reacted with both native and rPsaA polypeptides. Our data strongly suggest that E. coli-expressed rPsaA is palmitoylated and closely resembles the native protein in structure and immunogenicity. It was also observed to elicit measurable protection against nasopharyngeal carriage with S. pneumoniae.
Adhesins, Bacterial↗
Optimization of scanning transmission X-ray microscopy for the identification and quantitation of reinforcing particles in polyurethanes.
The morphology, size distributions, spatial distributions, and quantitative chemical compositions of co-polymer polyol-reinforcing particles in a polyurethane have been investigated with scanning transmission X-ray microscopy (STXM). A detailed discussion of microscope operating procedures is presented and ways to avoid potential artifacts are discussed. Images at selected photon energies in the C 1s, N 1s and O 1s regions allow unambiguous identification of styrene-acrylonitrile-based (SAN) copolymer and polyisocyanate polyaddition product-based (PIPA) reinforcing particles down to particle sizes at the limit of the spatial resolution (50 nm). Quantitative analysis of the chemical composition of individual reinforcing particles is achieved by fitting C 1s spectra to linear combinations of reference spectra. Regression analyses of sequences of images recorded through the chemically sensitive ranges of the C 1s, N 1s and O 1s spectra are used to generate quantitative compositional maps, which provide a fast and effective means of investigating compositional distributions over a large number of reinforcing particles. The size distribution of all particles determined by STXM is shown to be similar to that determined by TEM. The size distributions of each type of reinforcing particle, which differ considerably, were obtained by analysis of STXM images at chemically selective energies.
Microscopy, Electron, Scanning↗
An analysis of travel costs on transport of load and nest building in golden hamster.
We investigated the effects of travel costs on transporting nest material and nest-building activity in golden hamsters. Nest-deprived animals were submitted to run alleys 30, 90 and 180 cm long to access a source containing paper strips as nest material (Experiment 1) or were submitted to the same travel costs in 24-h experimental sessions (Experiment 2). We noted that increased travel costs were related to a decreased number of trips to the source, larger amounts (cm(2)) of nest material transported per trip (although total loads also decreased in longer alleys), longer intervals between trips, and increased time spent at the source and in nest building activity. Foraging efficiency (i.e. size of load divided by the time spent at the source) decreased as a function of travel costs, and animals transported their loads in two fundamental ways: in 30-cm alleys, they simply used their mouth to pull the paper strips, but in 90- or 180-cm alleys they transported the loads in their cheek pouches. The animals were faster when returning to the home-cage and their running speed (cm/s) increased as a function of the length of the alley, showing that animals are under different environmental pressures when searching for resources and subsequently running back with the load to the nest. Both male and female subjects were sensitive to travel costs, but males engaged in nest building activity more promptly and exhibited higher mean performances in most measures. We conclude that nest material is a good reinforcer, and our major results are in accordance with the predictions of microeconomic and optimal foraging theories.
Journal Article↗
Factors affecting age at first lambing in Yankasa ewes.
Data on 133 primiparous Yankasa ewes covering a period of 8 years (1983-1991) were analysed by least squares method for the effects of season, year, litter size, birth weight, weaning age and weaning weight on age at first lambing (AFL). Results showed the AFL (+/-SE) to be 533.1 +/- 10.3 days. Only the effect of season on AFL was significant (P < 0.01) while year of lambing, birth weight, litter size, weaning age and weaning weight did not affect AFL significantly (P > 0.01). AFL was 476.9 +/- 23.3, 523.9 +/- 19.2, 549.8 +/- 19.7 and 581.9 +/- 19.5 days for late wet, early dry, late dry and early wet season born ewes respectively. AFL was significantly (P < 0.01) lowest for ewes born in the late wet season and highest for ewes born in the early wet season. There was no significant (P > 0.01) difference in AFL between ewes born in the early dry season and ewes born in the late dry season, and between late dry and early wet season born ewes.
Animal Feed↗
Bilateral superior cervical sympathectomy and noise-induced, permanent threshold shift in guinea pigs.
The rich sympathetic innervation to the cochlea suggests its potential control of cochlear blood flow and activity during noise exposure, as part of the general and local stress sympathetic reaction evoked by noise. In a previous study, superior cervical sympathectomy prior to sound exposure in guinea pigs in an awake state, resulted in reduced temporary threshold shift. The present study was conducted to explore whether this potential protection would also be manifested in conditions producing permanent threshold shift (PTS). Thirty-six guinea pigs, divided into four groups of nine guinea pigs each, were sound exposed for 2 h in an awake state. Eighteen guinea pigs underwent superior cervical sympathectomy prior to sound exposure. Auditory brainstem thresholds were recorded prior to sound exposure, and then at 24 h, 1 and 6 weeks post-exposure. Results indicated a reduced PTS at 122 dB sound pressure level (SPL) exposure, suggesting a protective effect of the sympathectomy. However, at 125 dB SPL exposure, the protective effect was reduced.
Animals↗
Central markers of body fat distribution are important predictors of plasma lipids in elderly men and women.
We determined the contribution of body fat distribution, peak VO2, fat mass, and dietary intake to variation in plasma lipids in elderly individuals. Volunteers were a healthy cohort of older Caucasian women (n = 75, mean age +/- SD, 72 +/- 5 years) and older men (n = 101, 72 +/- 5 years). We determined fat mass from underwater weighing, fat patterning from waist circumference, as well as peripheral and truncal skinfolds, exercise capacity from peak VO2, and dietary intake from three-day food diaries. Plasma lipid levels were measured in the fasting state and included total cholesterol, high density lipoprotein (HDL-C), low density lipoprotein (LDL-C), and fasting triglycerides. Older women weighted less than older men, but had higher fat mass, truncal, and peripheral skinfolds. Waist circumference and peak VO2 were lower in older women than older men. Older women had higher total cholesterol (217 +/- 31 vs. 197 +/- 30; p < 0.01), HDL-C (54 +/- 12 vs. 49 +/- 14; p < 0.05), and LDL-C (133 +/- 26 vs. 121 +/- 27; p < 0.01) when compared with older men. No gender differences were noted in fasting triglycerides. Truncal skinfolds were the best predictor of plasma lipids in older men, accounting for between 9% and 30% (r2) of the variation in plasma lipids. Similarly, in older women, central markers of fatness (i.e., waist circumference and truncal skinfolds) were the best predictors of plasma lipids (r2 = 3% to 24%). Total fat mass, peak VO2 and dietary intake were not independent predictors of plasma lipids in older men and women. Indices of central body fatness, rather than total fat mass, peak VO2 or dietary intake are stronger predictors of plasma lipids in healthy older men and women.
Adipose Tissue↗
Effects of exercise and cardiac rehabilitation on cardiovascular outcomes.
Cardiac rehabilitation was originally conceived to counteract the deconditioning and comorbidities associated with prolonged bed rest after a myocardial infarction. Contemporary cardiac rehabilitation has taken a more comprehensive approach, with a broader range of participating patients. Relevant cardiovascular outcomes of cardiac rehabilitation can be classified as primary clinical outcomes, intermediate clinical outcomes, and quality-of-life outcomes. In this article, the effects of exercise training alone and, more importantly, the value of comprehensive cardiac rehabilitation are reviewed from the point of view of individual cardiovascular outcomes.
Coronary Disease↗
Exercise rehabilitation of older patients with cardiovascular disease.
As the population of elderly patients with cardiovascular disease continues to increase, much research needs to be done with the goal of maintaining physical functioning and personal independence in this population. It is of particular importance to determine whether training programs can improve physical functioning in the most severely disabled older coronary patients. Effects of cardiac rehabilitation programs on other outcome measures, including psychosocial outcomes, lipid levels, insulin levels, and body composition require better study. Finally, the economic benefits of cardiac rehabilitation in the older coronary patients has received little attention, although early reports are promising. In summary, the older population with coronary disease is characterized by high rates of disability. Exercise training has been demonstrated to be safe and to improve strength, aerobic fitness capacity, endurance and physical function. It remains to be seen whether exercise training can reverse or prevent disability in a broad population of older patients with cardiovascular disease. If successful, cardiac rehabilitation programs will pay great medical, social, and economic dividends in this population.
Aged↗
Gender differences in cardiac rehabilitation.
It is now widely recognized that comprehensive cardiac rehabilitation has substantial benefits in patients with cardiovascular disease. Despite the fact that cardiovascular disease is the leading cause of death for women in the United States with more than 250,000 deaths each year, most studies of cardiac rehabilitation have not included a substantial number of women. By making cardiac rehabilitation programs more responsive to the needs and goals of female coronary patients, participation rates and compliance rates should increase with a favorable impact on morbidity, mortality, and quality of life for women with cardiovascular diseases.
Cardiac Rehabilitation↗
Task force #3--getting results: who, where, and how? 33rd Bethesda Conference.
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Cardiovascular Diseases↗
Prevalence, predisposing factors, and prognosis of clinically unrecognized myocardial infarction in the elderly.
OBJECTIVES: This study was designed to determine the prevalence of unrecognized myocardial infarction (UMI), as well as risk factors, and to compare prognosis after detection of previously UMI to that after recognized myocardial infarction (RMI). BACKGROUND: Past studies revealed that a significant proportion of MIs escape recognition, and that prognosis after such events is poor, but the epidemiology of UMI has not been reassessed in the contemporary era. METHODS: The Cardiovascular Health Study (CHS) database, composed of individuals > or =65, was queried for participants who, at entry, demonstrated electrocardiographic evidence of a prior Q-wave MI, but who lacked a history of this diagnosis. The features and outcomes of this group were compared to those of individuals with prevalent RMI. RESULTS: Of 5,888 participants, 901 evidenced a past MI, and 201 (22.3%) were previously unrecognized. The independent predictors of UMI were the absence of angina and the absence of congestive heart failure (CHF). Six-year mortality did not significantly differ between the two groups. CONCLUSIONS: 1) In the elderly, UMI continues to represent a significant proportion of all MIs; 2) associations with angina and CHF may reflect complex neurological issues, but they also may represent diagnosis bias; 3) these individuals can otherwise not be distinguished from those with recognized infarctions; and 4) mortality rates after UMI and RMI are similar. Future studies should address screening for UMI, risk stratification after detection of previously UMI, and the role of standard post-MI therapies.
Aged↗
American Cancer Society urban demonstration projects: models for successful intervention.
In 1989, the American Cancer Society funded three Community Cancer Detection, Education and Prevention Demonstrations Projects for socioeconomically disadvantaged populations in Miami, Harlem, and West Oakland. Each project has demonstrated that by using a multifunction, multisite cancer control model with an action theory that integrates individual and structural influences, many barriers to screening and follow-up can be minimized.
Aftercare↗
The impact of oestradiol on ages at onset in schizophrenia reevaluated through Klinefelter syndrome.
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Adult↗
Predicting the HER2 status of breast cancer from basic histopathology data: an analysis of 1500 breast cancers as part of the HER2000 International Study.
The tests that are currently available for the measurement of overexpression of the human epidermal growth factor-2 (HER2) in breast cancer have shown considerable problems in accuracy and interlaboratory reproducibility. Although these problems are partly alleviated by the use of validated, standardised 'kits', there may be considerable cost involved in their use. Prior to testing it may therefore be an advantage to be able to predict from basic pathology data whether a cancer is likely to overexpress HER2. In this study, we have correlated pathology features of cancers with the frequency of HER2 overexpression assessed by immunohistochemistry (IHC) using HercepTest (Dako). In addition, fluorescence in situ hybridisation (FISH) has been used to re-test the equivocal cancers and interobserver variation in assessing HER2 overexpression has been examined by a slide circulation scheme. Of the 1536 cancers, 1144 (74.5%) did not overexpress HER2. Unequivocal overexpression (3+ by IHC) was seen in 186 cancers (12%) and an equivocal result (2+ by IHC ) was seen in 206 cancers (13%). Of the 156 IHC 3+ cancers for which complete data was available, 149 (95.5%) were ductal NST and 152 (97%) were histological grade 2 or 3. Only 1 of 124 infiltrating lobular carcinomas (0.8%) showed HER2 overexpression. None of the 49 'special types' of carcinoma showed HER2 overexpression. Re-testing by FISH of a proportion of the IHC 2+ cancers showed that only 25 (23%) of those assessable exhibited HER2 gene amplification, but 46 of the 47 IHC 3+ cancers (98%) were confirmed as showing gene amplification. Circulating slides for the assessment of HER2 score showed a moderate level of agreement between pathologists (kappa 0.4). As a result of this study we would advocate consideration of a triage approach to HER2 testing. Infiltrating lobular and special types of carcinoma may not need to be routinely tested at presentation nor may grade 1 NST carcinomas in which only 1.4% have been shown to overexpress HER2. Testing of these carcinomas may be performed when HER2 status is required to assist in therapeutic or other clinical/prognostic decision-making. The highest yield of HER2 overexpressing carcinomas is seen in the grade 3 NST subgroup in which 24% are positive by IHC.
Adult↗
Engrailed (Gln50-->Lys) homeodomain-DNA complex at 1.9 A resolution: structural basis for enhanced affinity and altered specificity.
BACKGROUND: The homeodomain is one of the key DNA-binding motifs used in eukaryotic gene regulation, and homeodomain proteins play critical roles in development. The residue at position 50 of many homeodomains appears to determine the differential DNA-binding specificity, helping to distinguish among binding sites of the form TAATNN. However, the precise role(s) of residue 50 in the differential recognition of alternative sites has not been clear. None of the previously determined structures of homeodomain-DNA complexes has shown evidence for a stable hydrogen bond between residue 50 and a base, and there has been much discussion, based in part on NMR studies, about the potential importance of water-mediated contacts. This study was initiated to help clarify some of these issues. RESULTS: The crystal structure of a complex containing the engrailed Gln50-->Lys variant (QK50) with its optimal binding site TAATCC (versus TAATTA for the wild-type protein) has been determined at 1.9 A resolution. The overall structure of the QK50 variant is very similar to that of the wild-type complex, but the sidechain of Lys50 projects directly into the major groove and makes several hydrogen bonds to the O6 and N7 atoms of the guanines at base pairs 5 and 6. Lys50 also makes an additional water-mediated contact with the guanine at base pair 5 and has an alternative conformation that allows a hydrogen bond with the O4 of the thymine at base pair 4. CONCLUSIONS: The structural context provided by the folding and docking of the engrailed homeodomain allows Lys50 to make remarkably favorable contacts with the guanines at base pairs 5 and 6 of the binding site. Although many different residues occur at position 50 in different homeodomains, and although numerous position 50 variants have been constructed, the most striking examples of altered specificity usually involve introducing or removing a lysine sidechain from position 50. This high-resolution structure also confirms the critical role of Asn51 in homeodomain-DNA recognition and further clarifies the roles of water molecules near residues 50 and 51.
Animals↗
Mechanisms of hepatic very low-density lipoprotein overproduction in insulin resistance.
An important complication of insulin-resistant states, such as obesity and type 2 diabetes, is an atherogenic dyslipidemia profile characterized by hypertriglyceridemia, low plasma high-density lipoproteins (HDL) cholesterol and a small, dense low-density lipoprotein (LDL) particle profile. The physiological basis of this metabolic dyslipidemia appears to be hepatic overproduction of apoB-containing very low-density lipoprotein (VLDL) particles. This has focused attention on the mechanisms that regulate VLDL secretion in insulin-resistant states. Recent studies in animal models of insulin resistance, particularly the fructose-fed hamster, have enhanced our understanding of these mechanisms, and certain key factors have recently been identified that play important roles in hepatic insulin resistance and dysregulation of the VLDL secretory process. This review focuses on these recent developments as well as on the hypothesis that an interaction between enhanced flux of free fatty acids from peripheral tissues to liver, chronic up-regulation of de novo lipogenesis by hyperinsulinemia and attenuated insulin signaling in the liver may be critical to the VLDL overproduction state observed in insulin resistance. It should be noted that the focus of this review is on molecular mechanisms of the hypertriglyceridemic state associated with insulin resistance and not that observed in association with insulin deficiency (e.g., in streptozotocin-treated animals), which appears to have a different etiology and is related to a catabolic defect rather than secretory overproduction of triglyceride-rich lipoproteins.
Animals↗