Essential genes from Arctic bacteria used to construct stable,temperature-sensitive bacterial vaccines

All bacteria share a set of evolutionarily conserved essential genes that encode products that are required for viability. The great diversity of environments that bacteria inhabit, including environments at extreme temperatures, place adaptive pressure on essential genes. We sought to use this evolutionary diversity of essential genes to engineer bacterial pathogens to be stably temperature-sensitive, and thus useful as live vaccines. We isolated essential genes from bacteria found in the Arctic and substituted them for their counterparts into pathogens of mammals. We found that substitution of nine different essential genes from psychrophilic (cold-loving) bacteria into mammalian pathogenic bacteria resulted in strains that died below their normal-temperature growth limits. Substitution of three different psychrophilic gene orthologs of ligA, which encode NAD-dependent DNA ligase, resulted in bacterial strains that died at 33, 35, and 37 °C. One ligA gene was shown to render Francisella tularensis, Salmonella enterica, and Mycobacterium smegmatis temperature-sensitive, demonstrating that this gene functions in both Gram-negative and Gram-positive lineage bacteria. Three temperature-sensitive F. tularensis strains were shown to induce protective immunity after vaccination at a cool body site. About half of the genes that could be tested were unable to mutate to temperature-resistant forms at detectable levels. These results show that psychrophilic essential genes can be used to create a unique class of bacterial temperature-sensitive vaccines for important human pathogens, such as S. enterica and Mycobacterium tuberculosis.     

The effect of glucosamine and/or chondroitin sulfate on the progression of knee osteoarthritis: a report from the glucosamine/chondroitin arthritis intervention trial

OBJECTIVE: Osteoarthritis (OA) of the knee causes significant morbidity and current medical treatment is limited to symptom relief, while therapies able to slow structural damage remain elusive. This study was undertaken to evaluate the effect of glucosamine and chondroitin sulfate (CS), alone or in combination, as well as celecoxib and placebo on progressive loss of joint space width (JSW) in patients with knee OA. METHODS: A 24-month, double-blind, placebo-controlled study, conducted at 9 sites in the United States as part of the Glucosamine/Chondroitin Arthritis Intervention Trial (GAIT), enrolled 572 patients with knee OA who satisfied radiographic criteria (Kellgren/Lawrence [K/L] grade 2 or grade 3 changes and JSW of at least 2 mm at baseline). Patients with primarily lateral compartment narrowing at any time point were excluded. Patients who had been randomized to 1 of the 5 groups in the GAIT continued to receive glucosamine 500 mg 3 times daily, CS 400 mg 3 times daily, the combination of glucosamine and CS, celecoxib 200 mg daily, or placebo over 24 months. The minimum medial tibiofemoral JSW was measured at baseline, 12 months, and 24 months. The primary outcome measure was the mean change in JSW from baseline. RESULTS: The mean JSW loss at 2 years in knees with OA in the placebo group, adjusted for design and clinical factors, was 0.166 mm. No statistically significant difference in mean JSW loss was observed in any treatment group compared with the placebo group. Treatment effects on K/L grade 2 knees, but not on K/L grade 3 knees, showed a trend toward improvement relative to the placebo group. The power of the study was diminished by the limited sample size, variance of JSW measurement, and a smaller than expected loss in JSW. CONCLUSION: At 2 years, no treatment achieved a predefined threshold of clinically important difference in JSW loss as compared with placebo. However, knees with K/L grade 2 radiographic OA appeared to have the greatest potential for modification by these treatments.     

Evaluation of the S-type of alpha-1-antitrypsin as an in vivo and in vitro inhibitor of neutrophil elastase

S-type alpha 1-antitrypsin (alpha 1AT) is a deficiency haplotype that differs from the common normal M1 (val213) alpha 1AT haplotype by a single amino acid (glu264 to val264). To evaluate the adequacy of the antineutrophil elastase protection associated with the S homozygous state, alpha 1AT plasma and lung epithelial lining fluid (ELF) levels and antineutrophil elastase function were analyzed in 9 PISS subjects. The plasma alpha 1AT levels of SS subjects were intermediate between that of M1M1 and ZZ subjects (p less than 0.001, all comparisons) and the plasma neutrophil elastase inhibitory capacity paralleled the differences in alpha 1AT concentration (p less than 0.001, all comparisons). The association rate constant for neutrophil elastase of the purified S protein was less than that of the normal molecule (S-type, 7.1 +/- 0.1 X 10(6) M-1 s-1; M1-type, 9.6 +/- 0.2 X 10(6) M-1 s-1; p less than 0.001), but much greater than that for the Z molecule (p less than 0.001). Exposure of the purified S protein to increasing oxidant burdens resulted in a dose-dependent reduction in the ability of the molecule to inhibit neutrophil elastase in a fashion parallel to that of the M1 and Z proteins. Quantification of ELF alpha 1AT levels and antineutrophil elastase capacity demonstrated that the SS ELF parameters were, as in plasma, intermediate between M1 homozygotes and Z homozygotes. Using the association rate constant together with the quantification of ELF alpha 1AT levels, the "in vivo lung inhibition time" was estimated, yielding an assessment of the relative antineutrophil elastase screen of the PISS lower respiratory tract.(ABSTRACT TRUNCATED AT 250 WORDS)     

General 2.5 power law of metallic glasses

Metallic glass (MG) is an important new category of materials, but very few rigorous laws are currently known for defining its “disordered” structure. Recently we found that under compression, the volume (V) of an MG changes precisely to the 2.5 power of its principal diffraction peak position (1/q₁). In the present study, we find that this 2.5 power law holds even through the first-order polyamorphic transition of a Ce₆₈Al₁₀Cu₂₀Co₂ MG. This transition is, in effect, the equivalent of a continuous “composition” change of 4f-localized “big Ce” to 4f-itinerant “small Ce,” indicating the 2.5 power law is general for tuning with composition. The exactness and universality imply that the 2.5 power law may be a general rule defining the structure of MGs.Metallic glasses (MGs) possess many unique and superior properties, such as extremely high strength, hardness, and corrosion resistance, etc., making them promising metallic materials with widespread applications (1, 2). Thousands of MGs with a wide range of compositions and properties have been synthesized over the past decades. However, so far the development of MGs is mainly based on tedious composition mapping in multicomponent space to pinpoint the combination of elements with optimized glass-forming ability (GFA). This method for development of MGs is a time- and resource-intensive strategy of trial and error which highlights the need for the guidance of a general theory (2, 3). Intensive research effort has been devoted to finding general rules in various MGs to understand the fundamentals and to guide the development of new MGs (4, 5). Quantitative correlations between their properties have been observed. For instance, compressive yield strength and elastic moduli of MGs are found to be intimately connected with their glass transition temperature T_g (6–10), and the ductility, fragility (11, 12), and Poisson’s ratio of MGs are closely related (13–16). The extensive correlations in properties suggest that the disordered MGs may share general rules in their structure. To clarify this scenario, detailed and accurate structural information spanning short range to long range is required. However, the current experimental probes and theories are limited to local structure in MGs (17). Therefore, understanding how the atoms efficiently fill up the 3D space and how this controls the bulk properties of MGs remains a long-standing theoretical challenge (18–23). To date, few general and exact rules regarding structure–property relationships have been established in MGs (23).Encouraging progress on understanding structure–property relationships in MGs has recently been made through the discoveries of the noncubic (2.3 or 2.5) power laws that correlate the principal diffraction peak (PDP) position q₁ with the bulk density ρ or average atomic volume, V_a, i.e., ρ∝(q₁)^D or V_a∝(1/q₁)^D, where D equals ∼2.3 with varying the composition of MGs at ambient pressure (19) or ∼2.5 for tuning the density of MGs with pressure (22, 24). Whereas composition and pressure show similar exponents in the power laws in MGs, composition and pressure are two independent variables for controlling the density (volume) of materials; they usually have dramatically different effects on MGs. For example, pressure is thought to cause only elastic densification in MGs without obvious structural change because of their already densely packed structure; the structure and properties of MGs are very sensitive to even minor compositional variations (25, 26). In addition, to achieve composition change, different samples usually have to be synthesized. And, many other variables are thought to be inevitably involved, making the compositional change complex (23). Therefore, some basic questions have been perplexing to the glass community: Why do “complex” compositional and “simple” pressure power laws show similar exponents? Is there any connection between them? These questions remain unanswered and have been the major obstacle in understanding the nature of these noncubic power laws.To address these questions, a systematic study in the 2D pressure-composition space seems to be required. However, the consistency of the data in this kind of study will be questionable. Alternatively, in the present study, we choose the polyamorphous Ce₆₈Al₁₀Cu₂₀Co₂ MG as a model system. It is well known that Ce-based MG systems show a polyamorphic transition between ∼2 GPa and ∼5 GPa caused by the pressure-induced 4f electron localized-to-itinerant transition (27, 28). During this polyamorphic transition, both the atomic size and the electronegativity of Ce are significantly changed (29). Composition tuning in MGs mainly means the variation of atomic size and electronegativity of components, which controls the formation of MGs (30). Therefore, although nothing changes in the nucleus, for MGs this pressure-induced polyamorphic transition is equivalent to a continuous “composition” change with the 4f-localized “big Ce” gradually substituted by 4f-itinerant “small Ce.” As a result, we are able to vary both pressure and composition of a MG in a well-controlled way for the first time, to our knowledge.     

Health-related quality of life in patients with human immunodeficiency virus infection in the United States: results from the HIV Cost and Services Utilization Study

PURPOSE: To measure health-related quality of life among adult patients with human immunodeficiency virus (HIV) disease; to compare the health-related quality of life of adults with HIV with that of the general population and with patients with other chronic conditions; and to determine the associations of demographic variables and disease severity with health-related quality of life. SUBJECTS AND METHODS: We studied 2,864 HIV-infected adults participating in the HIV Cost and Services Utilization Study, a probability sample of adults with HIV receiving health care in the contiguous United States (excluding military hospitals, prisons, or emergency rooms). A battery of 28 items covering eight domains of health (physical functioning, emotional well-being, role functioning, pain, general health perceptions, social functioning, energy, disability days) was administered. The eight domains were combined into physical and mental health summary scores. SF-36 physical functioning and emotional well-being scales were compared with the US general population and patients with other chronic diseases on a 0 to 100 scale. RESULTS: Physical functioning was about the same for adults with asymptomatic HIV disease as for the US population [mean (+/- SD) of 92+/-16 versus 90+/-17) but was much worse for those with symptomatic HIV disease (76+/-28) or who met criteria for the acquired immunodeficiency syndrome (AIDS; 58+/-31). Patients with AIDS had worse physical functioning than those with other chronic diseases (epilepsy, gastroesophageal reflux disease, clinically localized prostate cancer, clinical depression, diabetes) for which comparable data were available. Emotional well-being was comparable among patients with various stages of HIV disease (asymptomatic, 62+/-9; symptomatic, 59+/-11; AIDS, 59+/-11), but was significantly worse than the general population and patients with other chronic diseases except depression. In multivariate analyses, HIV-related symptoms were strongly associated with physical and mental health, whereas race, sex, health insurance status, disease stage, and CD4 count were at most weakly associated with physical and mental health. CONCLUSIONS: There is substantial morbidity associated with HIV disease in adults. The variability in health-related quality of life according to disease progression is relevant for health policy and allocation of resources, and merits the attention of clinicians who treat patients with HIV disease.