Proteasome lid bridges mitochondrial stress with Cdc53/Cullin1 NEDDylation status.

Cycles of Cdc53/Cullin1 rubylation (a.k.a NEDDylation) protect ubiquitin-E3 SCF (Skp1-Cullin1-F-box protein) complexes from self-destruction and play an important role in mediating the ubiquitination of key protein substrates involved in cell cycle progression, development, and survival. Cul1 rubylation is balanced by the COP9 signalosome (CSN), a multi-subunit derubylase that shows 1:1 paralogy to the 26S proteasome lid. The turnover of SCF substrates and their relevance to various diseases is well studied, yet, the extent by which environmental perturbations influence Cul1 rubylation/derubylation cycles per se is still unclear. In this study, we show that the level of cellular oxidation serves as a molecular switch, determining Cullin1 rubylation/derubylation ratio. We describe a mutant of the proteasome lid subunit, Rpn11 that exhibits accumulated levels of Cullin1-Rub1 conjugates, a characteristic phenotype of csn mutants. By dissecting between distinct phenotypes of rpn11 mutants, proteasome and mitochondria dysfunction, we were able to recognize the high reactive oxygen species (ROS) production during the transition of cells into mitochondrial respiration, as a checkpoint of Cullin1 rubylation in a reversible manner. Thus, the study adds the rubylation cascade to the list of cellular pathways regulated by redox homeostasis.

Blood transcriptomic markers of Mycobacterium tuberculosis load in sputum.

BACKGROUND: Peripheral blood transcriptome signatures that distinguish active pulmonary tuberculosis (TB) from control groups have been reported, but correlations of these signatures with sputum mycobacterial load are incompletely defined. METHODS: We assessed the performance of published TB transcriptomic signatures in Haiti, and identified transcriptomic biomarkers of TB bacterial load in sputum as measured by Xpert® MTB/RIF molecular testing. People in Port au Prince, Haiti, with untreated pulmonary TB (n = 51) formed the study cohort: 19 people with low and 32 with high sputum Mycobacterium tuberculosis load. Peripheral whole blood transcriptomes were generated using RNA sequencing. RESULTS: Twenty of the differentially expressed transcripts in TB vs. no TB were differentially expressed in people with low vs. high sputum mycobacterial loads. The difference between low and high bacterial load groups was independent of radiographic severity. In a published data set of transcriptomic response to anti-tuberculosis treatment, this 20-gene subset was more treatment-responsive at 6 months than the full active TB signature. CONCLUSION: We identified genes whose transcript levels in the blood distinguish active TB with high vs. low M. tuberculosis loads in the sputum. These transcripts may reveal mechanisms of mycobacterial control of M. tuberculosis during active infection, as well as identifying potential biomarkers for bacterial response to anti-tuberculosis treatment.

Breast cancer survivors of different sexual orientations: which factors explain survivors' quality of life and adjustment?

BACKGROUND: Little is known about differences by sexual orientation in explanatory factors of breast cancer survivors' quality of life, anxiety, and depression. PATIENTS AND METHODS: Survivors were recruited from a cancer registry and additional survivors recruited through convenience methods. Data were collected via telephone survey from all 438 survivors, who were disease free and diagnosed with non-metastatic breast cancer an average of 5 years earlier. To explain quality of life, anxiety, and depression, we focused on sexual orientation as the primary independent factors, in addition, considering demographic, psychosocial, clinical, and functional factors as correlates. RESULTS: Sexual orientation had indirect associations with each of the outcomes, through disease-related and demographic factors as well as psychosocial and coping resources. The various explanatory models explain between 36% and 50% of the variance in outcomes and identified areas of strengths and vulnerabilities in sexual minority compared with heterosexual survivors. CONCLUSIONS: This study's findings of strengths among specific subgroups of sexual minority compared with heterosexual survivors require further explorations to identify the reasons for this finding. Most of the identified vulnerabilities among sexual minority compared with heterosexual survivors of breast cancer are amenable to change by interventions.

Legionella jordanis in hematopoietic SCT patients radiographically mimicking invasive mold infection.

Opportunistic pulmonary infections are a major cause of post-transplant morbidity and mortality. Among these infections, Aspergillus is a common cause of fatal pneumonia. Owing to the precarious clinical condition of many patients who acquire invasive mold infections, clinicians often treat them on the basis of radiographic findings, such as the halo sign. However, in patients who do not respond to treatment or who have uncommon presentations, bronchoscopy or lung biopsy looking for other pathogens should be considered. This study describes two cases in which the radiographic halo signs characteristic of Aspergillus were in fact due to Legionella jordanis, a pathogen that has been culture proven only in two patients previously (both of whom had underlying lung pathology) and diagnosed by serologic evidence in several other patients. In immunocompromised patients, Legionella can present as a cavitary lesion. Thus, presumptive treatment for this organism should be considered in post-transplant patients who do not have a classic presentation for invasive fungal infection and/or who fail to respond to conventional treatment. These cases illustrate the importance of obtaining tissue cultures to differentiate among the wide variety of pathogens present in this patient population.

Socioeconomic status and ischaemic heart disease mortality in 10 western European populations during the 1990s.

OBJECTIVE: To assess the association between socioeconomic status and ischaemic heart disease (IHD) mortality in 10 western European populations during the 1990s. DESIGN: Longitudinal study. SETTING: 10 European populations (95,009,822 person years). METHODS: Longitudinal data on IHD mortality by educational level were obtained from registries in Finland, Norway, Denmark, England/Wales, Belgium, Switzerland, Austria, Turin (Italy), Barcelona (Spain), and Madrid (Spain). Age standardised rates and rate ratios (RRs) of IHD mortality by educational level were calculated by using Poisson regression. RESULTS: IHD mortality was higher in those with a lower socioeconomic status than in those with a higher socioeconomic status among men aged 30-59 (RR 1.55, 95% confidence interval (CI) 1.51 to 1.60) and 60 years and over (RR 1.22, 95% CI 1.21 to 1.24), and among women aged 30-59 (RR 2.13, 95% CI 1.98 to 2.29) and 60 years and over (RR 1.36, 95% CI 1.33 to 1.38). Socioeconomic disparities in IHD mortality were larger in the Scandinavian countries and England/Wales, of moderate size in Belgium, Switzerland, and Austria, and smaller in southern European populations among men and younger women (p < 0.0001). For elderly women the north-south gradient was smaller and there was less variation between populations. No socioeconomic disparities in IHD mortality existed among elderly men in southern Europe. CONCLUSIONS: Socioeconomic disparities in IHD mortality were larger in northern than in southern European populations during the 1990s. This partly reflects the pattern of socioeconomic disparities in cardiovascular risk factors in Europe. Population wide strategies to reduce risk factor prevalence combined with interventions targeted at the lower socioeconomic groups can contribute to reduce IHD mortality in Europe.

Person, place or time? The effect of individual circumstances, area and changes over time on mortality in men, 1995-2001.

This article uses the ONS Longitudinal Study to explore, for a cohort of adult males who were aged 26 or over in 1971, the relative influence on mortality in 1995-2001 of their place of residence and individual socioeconomic circumstances, at three censuses over a 20-year period. Factors examined in this analysis include social class, neighbourhoo deprivation (at ward level), unemployment, residence in the South East region in 1971 or 1981 housing tenure, and change in social class and housing tenure between 1971 and 1991. The variation in mortality attributable to the local authority district of residence in 1991 was also investigated.

Education level inequalities and transportation injury mortality in the middle aged and elderly in European settings.

OBJECTIVE: To study the differential distribution of transportation injury mortality by educational level in nine European settings, among people older than 30 years, during the 1990s. METHODS: Deaths of men and women older than 30 years from transportation injuries were studied. Rate differences and rate ratios (RR) between high and low educational level rates were obtained. RESULTS: Among men, those of low educational level had higher death rates in all settings, a pattern that was maintained in the different settings; no inequalities were found among women. Among men, in all the settings, the RR was higher in the 30-49 age group (RR 1.46, 95% CI 1.32 to 1.61) than in the age groups 50-69 and > or = 70 years, a pattern that was maintained in the different settings. For women for all the settings together, no differences were found among educational levels in the three age groups. In the different settings, only three had a high RR in the youngest age group, Finland (RR 1.33, 95% CI 1.01 to 1.74), Belgium (RR 1.38; 95% CI 1.13 to 1.67), and Austria (RR 1.49, 95% CI 0.75 to 2.96). CONCLUSION: This study provides new evidence on the importance of socioeconomic inequalities in transportation injury mortality across Europe. This applies to men, but not to women. Greater attention should be placed on opportunities to select intervention strategies tailored to tackle socioeconomic inequalities in transportation injuries.

Trends in socioeconomic disparities in stroke mortality in six european countries between 1981-1985 and 1991-1995.

This study assesses whether stroke mortality trends have been less favorable among lower than among higher socioeconomic groups. Longitudinal data on mortality by socioeconomic status were obtained for Finland, Norway, Denmark, Sweden, England/Wales, and Turin, Italy. Data covered the entire population or a representative sample. Stroke mortality rates were calculated for the period 1981-1995. Changes in stroke mortality rate ratios were analyzed using Poisson regression and compared with rate ratios in ischemic heat disease mortality. Trends in stroke mortality were generally as favorable among lower as among higher socioeconomic groups, such that socioeconomic disparities in stroke mortality persisted and remained of a similar magnitude in the 1990s as in the 1980s. In Norway, however, occupational disparities in stroke mortality significantly widened, and a nonsignificant increase was observed in some countries. In contrast, disparities in ischemic heart disease mortality widened throughout this period in most populations. Improvements in hypertension prevalence and treatment may have contributed to similar stroke mortality declines in all socioeconomic groups in most countries. Socioeconomic disparities in stroke mortality generally persisted and may have widened in some populations, which fact underlines the need to improve preventive and secondary care for stroke among the lower socioeconomic groups.

Keepers at the final gates: regulatory complexes and gating of the proteasome channel.

The proteolytic active sites of the 26S proteasome are sequestered within the central chamber of its 20S catalytic core particle. Access to this chamber is through a narrow channel defined by the outer alpha subunits. Free proteasome 20S core particles are found in an autoinhibited state in which the N-termini of neighboring alpha subunits are anchored by an intricate lattice of interactions blocking access to the channel. Entry of substrates into proteasomes can be enhanced by attachment of activators or regulatory particles. An important part of this activation is channel gating; regulatory particles rearrange the blocking residues to form an open pore and promote substrate entry into the proteolytic chamber. Interestingly, some substrates can open the entrance themselves and thus facilitate their own destruction. In this review, we will discuss the mechanisms proposed for channel gating and the interactions required to maintain stable closed and open conformations.

Is ultrasound more accurate than axial computed tomography for determination of maximal abdominal aortic aneurysm diameter?

OBJECTIVE(S): Clinical assessment of maximal abdominal aortic aneurysm (AAA) diameter assumes clinical equivalency between ultrasound (US) and axial computed tomography (CT). Three-dimensional (3D) CT reconstruction allows for the assessment of AAA in the orthogonal plane and avoids oblique cuts due to AAA angulation. This study was undertaken to compare maximal AAA diameter by US, axial CT, and orthogonal CT, and to assess the effect that AAA angulation has on each measurement. METHODS: Maximal AAA diameter by US (US(max)), axial CT (axial(max)), and orthogonal CT (orthogonal(max)) along with aortic angulation and minor axis diameters were measured prospectively. Spiral CT data was processed by Medical Media Systems (West Lebanon, NH) to produce computerized axial CT and reformatted orthogonal CT images. The US technologists were blinded to all CT results and vice versa. RESULTS: Thirty-eight patients were analyzed. Mean axial(max) (58.0 mm) was significantly larger (P<0.05) than US(max) (53.9 mm) or orthogonal(max) (54.7 mm). The difference between US(max) and orthogonal(max) (0.8 mm) was insignificant (P>0.05). When aortic angulation was <==25 degrees, axial(max) (55.3 mm), US(max) (54.3 mm), and orthogonal(max) (54.1 mm) were similar (P>0.05); however, when aortic angulation was >25 degrees, axial(max) (60.1 mm) was significantly larger (P<0.001) than US(max) (53.8 mm) and orthogonal(max) (55.0 mm). The limits of agreement (LOA) between axial(max) and both US(max) and orthogonal(max) was poor and exceeded clinical acceptability (+/-5 mm). The variation between US(max) and orthogonal(max) was minimal with an acceptable LOA of -2.7 to 4.5 mm. CONCLUSION: Compared to axial CT, US is a better approximation of true perpendicular AAA diameter as determined by orthogonal CT. When aortic angulation is greater than 25 degrees axial CT becomes unreliable. However, US measurements are not affected by angulation and agree strongly with orthogonal CT measurements.

Challenges of hemodialysis access for high risk patients: Impact of mesenteric vein bioprosthetic graft.

OBJECTIVE: The purpose of this study is to compare in a prospective fashion the performance of a new bioprosthesis, the mesenteric vein bioprosthesis (MVB), in patients who have had multiple failed ePTFE grafts. Performance measures include primary patency rates, assisted-primary patency rates, secondary patency rates, complications, and the number of interventions required to maintain graft patency. STUDY: From October 1999 to February 2002, 276 hemodialysis access grafts were implanted in a multicenter study. Of those grafts, 74 were placed in patients with a prior history of 3 failed prosthetic grafts (mean = 3.5 grafts, range = 3-6 grafts). Fifty-nine grafts were constructed with MVB, and 15 grafts with ePTFE as a concomitant control. Mean follow-up was 11.5 months. In the MVB group, 79.7% were African-Americans, 61% were females, and 23.7% were hypercoagulable. Of the ePTFE group, 86.7% were African-Americans, 46.7% were female, and 13.2% were hypercoagulable. Results : Per Kaplan-Meier curves, the primary patency rate of the MVB group at 12 months was 33% vs the ePTFE group of 18% (p=0.120); the assisted-primary patency rates at 12 months were 45% MVB vs 18% ePTFE (p=0.011). The secondary patency rates at 12 and 24 months for the MVB group were 67% and 59%, respectively, vs 45% and 15% for the ePTFE group (p=0.006). During the follow-up time period, 80% of the ePTFE grafts were abandoned compared to 34% of the MVB group. Infection and thrombosis rates in the MVB group were lower than the ePTFE group. The infection rate for the MVB group requiring intervention was 0.07 events/graft year (gt/y) compared to 0.30 events/gt-y for ePTFE (p=0.04). A thrombosis rate of 0.69 events/gt-y occurred in the MVB group whereas 2.50 events/gt-y presented in the ePTFE group (p<0.01). CONCLUSION: In this study, high-risk patients (defined as those having multiple failed prosthetic grafts for hemodialysis) in whom the MVB conduit for hemoaccess was implanted, showed significant improvement in assisted-primary and secondary patency rates compared to the ePTFE cohort. The MVB group, however, did not have a statistically better primary patency rate compared to the ePTFE group. The MVB patient also had fewer thrombotic and infectious events and an overall reduction in the number of interventions while maintaining a permanent access site. This new bioprosthesis should be the conduit of choice in the complex group of patients as it offers assisted-primary and secondary patency rates similar to those commonly experienced by patients without a history of multiple graft failures.

Regulating the 26S proteasome.

Despite the fact that the composition of proteasomes purified from different species is almost identical, and the basic components of the proteasome are remarkably conserved among all eukaryotes, there are quite a few additional proteins that show up in certain purifications or in certain screens. There is increasing evidence that the proteasome is in fact a dynamic structure forming multiple interactions with transiently associated subunits and cellular factors that are necessary for functions such as cellular localization, presentation of substrates, substrate-specific interactions, or generation of varied products. Harnessing the eukaryotic proteasome to its defined regulatory roles has been achieved by a number of means: (a) increasing the complexity of the proteasome by gene duplication, and differentiation of members within each gene family (namely the CP and RPT subunits); (b) addition of regulatory particles, complexes, and factors that influence both what enters and what exits the proteasome; and (c) signal-dependent alterations in subunit composition (for example, the CP beta to beta i exchange). It is not be surprising that the proteasome plays diverse roles, and that its specific functions can be fine-tuned depending on biological context or need.

Subunit interaction maps for the regulatory particle of the 26S proteasome and the COP9 signalosome.

The 26S proteasome plays a major role in eukaryotic protein breakdown, especially for ubiquitin-tagged proteins. Substrate specificity is conferred by the regulatory particle (RP), which can dissociate into stable lid and base subcomplexes. To help define the molecular organization of the RP, we tested all possible paired interactions among subunits from Saccharomyces cerevisiae by yeast two-hybrid analysis. Within the base, a Rpt4/5/3/6 interaction cluster was evident. Within the lid, a structural cluster formed around Rpn5/11/9/8. Interactions were detected among synonymous subunits (Csn4/5/7/6) from the evolutionarily related COP9 signalosome (CSN) from Arabidopsis, implying a similar quaternary arrangement. No paired interactions were detected between lid, base or core particle subcomplexes, suggesting that stable contacts between them require prior assembly. Mutational analysis defined the ATPase, coiled-coil, PCI and MPN domains as important for RP assembly. A single residue in the vWA domain of Rpn10 is essential for amino acid analog resistance, for degrading a ubiquitin fusion degradation substrate and for stabilizing lid-base association. Comprehensive subunit interaction maps for the 26S proteasome and CSN support the ancestral relationship of these two complexes.

Multicenter evaluation of a polyurethaneurea vascular access graft as compared with the expanded polytetrafluoroethylene vascular access graft in hemodialysis applications.

OBJECTIVES: The purpose of this study was to compare in a randomized, prospective, and controlled study, the performance of a multilayered, self-sealing polyurethane vascular access graft (PVAG) and expanded polytetrafluoroethylene (ePTFE) vascular access grafts in hemodialysis applications. Performance measures included graft survival, complications, time to early cannulation, and hemostasis times after cannulation. STUDY DESIGN: A total of 142 patients were randomized equally to receive one of the two grafts after meeting all eligibility requirements. All patients were followed up prospectively to 12 months or to the end of secondary patency. Specifically, this study documented the performance of the PVAG and ePTFE grafts by determining the patencies and complications for both grafts. RESULTS: Patient characteristics between the two groups were similar with respect to risk factors and demographic characteristics (P >.05). Life-table patencies from the date of first dialysis were primary patency: PVAG 55% versus ePTFE 47% (6 months) and PVAG 44% versus ePTFE 36% (12 months) and secondary patency: PVAG 87% versus ePTFE 90% (6 months) and PVAG 78% versus ePTFE 80% (12 months). None of these differences were significant (P >.05). Both primary and secondary patencies were also not significantly different when the date of implantation was the starting point. Adverse events and complications were similar for the two groups, except the PVAG group had a higher incidence of technical complications manifested by graft kinking when compared with the control cohort (P <.05). Additionally, there was no significant difference in complication rates between these two groups with regard to infection and bleeding. When the time to hemostasis after cannulation was compared at 5minutes or less, there were more PVAG cannulation sites that achieved hemostasis compared with ePTFE sites, and this difference was significant (P <.0001). When time to first dialysis access was compared between the two grafts, 53.9% of all PVAG grafts were cannulated before 9 days versus none with the ePTFE grafts (P <.001). However, long-term graft survival was not significantly different when PVAG patients were stratified into early (< 9 days) and the late access (9 >/= days) groups (P =.29). CONCLUSIONS: The PVAG graft allows for early access without compromising long-term performance. Both PVAG and standard ePTFE grafts have similar long-term outcomes, despite early access with the PVAG vascular access grafts.

The substrate translocation channel of the proteasome.

The core particle (CP) of the yeast proteasome is composed of four heptameric rings of subunits arranged in a hollow, barrel-like structure. We have found that the CP is autoinhibited by the N-terminal tails of the outer (alpha) ring subunits. Crystallographic analysis showed that deletion of the tail of the alpha3 subunit opens a channel into the proteolytically active interior chamber of the CP, thus derepressing peptide hydrolysis. In the latent state of the particle, the tails prevent substrate entry by imposing topological closure on the CP. Inhibition by the alpha subunit tails is relieved upon binding of the regulatory particle to the CP to form the proteasome holoenzyme. Opening of the CP channel by assembly of the holoenzyme is regulated by the ATPase domain of Rpt2, one of 17 subunits in the RP. Thus, open-channel mutations in CP subunits suppress the closed-channel phenotype of an rpt2 mutant. These results identify a specific mechanism for allosteric regulation of the CP by the RP.

The Mycobacterium tuberculosis cmaA2 gene encodes a mycolic acid trans-cyclopropane synthetase.

Infection with Mycobacterium tuberculosis remains a major global health emergency. Although detailed understanding of the molecular events of M. tuberculosis pathogenesis is still limited, recent genetic analyses have implicated specific lipids of the cell envelope as important effectors in M. tuberculosis pathogenesis. We have shown that pcaA, a novel member of a family of M. tuberculosis S-adenosyl methionine (SAM)-dependent methyl transferases, is required for alpha-mycolic acid cyclopropanation and lethal chronic persistent M. tuberculosis infection. To examine the apparent redundancy between pcaA and cmaA2, another cyclopropane synthetase of M. tuberculosis thought to be involved in alpha-mycolate synthesis, we have disrupted the cmaA2 gene in virulent M. tuberculosis by specialized transduction. Inactivation of cmaA2 causes accumulation of unsaturated derivatives of both the methoxy- and ketomycolates. Analysis by proton NMR indicates that the mycolic acids of the cmaA2 mutant lack trans-cyclopropane rings but are otherwise intact with respect to cyclopropane and methyl branch content. Thus, cmaA2 is required for the synthesis of the trans cyclopropane rings of both the methoxymycolates and ketomycolates. These results define cmaA2 as a trans-cyclopropane synthetase and expand our knowledge of the substrate specificity of a large family of highly homologous mycolic acid methyl transferases recently shown to be critical to M. tuberculosis pathogenesis.

A gated channel into the proteasome core particle.

The core particle (CP) of the yeast proteasome is composed of four heptameric rings of subunits arranged in a hollow, barrel-like structure. We report that the CP is autoinhibited by the N-terminal tails of the outer (alpha) ring subunits. Crystallographic analysis showed that deletion of the tail of the alpha 3-subunit opens a channel into the proteolytically active interior chamber of the CP, thus derepressing peptide hydrolysis. In the latent state of the particle, the tails prevent substrate entry by imposing topological closure on the CP. Inhibition by the alpha-subunit tails is relieved upon binding of the regulatory particle to the CP to form the proteasome holoenzyme.

Cognitive-behavioral therapy for anxiety disorders: why and how it works.

Cognitive-behavioral therapy (CBT) has been found to be effective in the treatment of anxiety disorders such as obsessive-compulsive disorder (OCD) and posttraumatic stress disorder (PTSD). There is a gap, however, between reports of controlled clinical trials and actual clinical use of the methods of CBT in general medical and psychiatric settings. While psychiatric medications are commonly the first line of treatment, pharmacotherapy may not completely eradicate symptomatology or may have a delayed effectiveness, during which time patients continue to suffer. Cognitive and behavioral interventions can complement, if not replace, pharmacotherapy for relief of symptoms of OCD and PTSD. This article explains how CBT works in the treatment of OCD and PTSD. Basic instructions for implementing these treatment methods are provided.