BAI1 localizes AMPA receptors at the cochlear afferent post-synaptic density and is essential for hearing.

Type I spiral ganglion neurons (SGNs) convey sound information to the central auditory pathway by forming synapses with inner hair cells (IHCs) in the mammalian cochlea. The molecular mechanisms regulating the formation of the post-synaptic density (PSD) in the SGN afferent terminals are still unclear. Here, we demonstrate that brain-specific angiogenesis inhibitor 1 (BAI1) is required for the clustering of AMPA receptors GluR2-4 (glutamate receptors 2-4) at the PSD. Adult Bai1-deficient mice have functional IHCs but fail to transmit information to the SGNs, leading to highly raised hearing thresholds. Despite the almost complete absence of AMPA receptor subunits, the SGN fibers innervating the IHCs do not degenerate. Furthermore, we show that AMPA receptors are still expressed in the cochlea of Bai1-deficient mice, highlighting a role for BAI1 in trafficking or anchoring GluR2-4 to the PSDs. These findings identify molecular and functional mechanisms required for sound encoding at cochlear ribbon synapses.

Designing lightweight neutron absorbing composites using a comprehensive absorber areal density metric.

Efforts to lightweight neutron absorbing composites are limited by incomplete understandings of the interaction between absorbing particles and their matrices. In this study, analytical models and a more physically representative simulation evaluated the penalty to neutron absorbing performance due to neutron channeling between large absorbing particles. Models and simulation agreed that B4C particles smaller than 100µm and especially those smaller than 10µm did not cause excessive neutron channeling. A more comprehensive neutron absorbing composite design metric - boron-10 equivalent areal density, which considers the particle size penalty and the matrix contribution to absorptivity - was introduced and used to estimate lightweighting via matrix substitution. Calculations using this new metric showed that a non-absorbing Mg matrix reduced mass by up to 35% over Al, constrained by the difference in mass density, while an absorbing Mg-Li matrix reduced mass by up to 60%, exceeding the difference in mass densities alone. Measurement of apparent absorber areal density through two experimental techniques - foil activation and direct counting - validated estimated absorber areal density as a neutron absorbing composite design metric. This updated understanding of the particle size penalty, newly introduced design metric, and experimental validation demonstrate a path to lightweight neutron absorbing composites.

A universal stress protein upregulated by hypoxia has a role in Burkholderia cenocepacia intramacrophage survival: Implications for chronic infection in cystic fibrosis.

Universal stress proteins (USPs) are ubiquitously expressed in bacteria, archaea, and eukaryotes and play a lead role in adaptation to environmental conditions. They enable adaptation of bacterial pathogens to the conditions encountered in the human niche, including hypoxia, oxidative stress, osmotic stress, nutrient deficiency, or acid stress, thereby facilitating colonization. We previously reported that all six USP proteins encoded within a low-oxygen activated (lxa) locus in Burkholderia cenocepacia showed increased abundance during chronic colonization of the cystic fibrosis (CF) lung. However, the role of USPs in chronic cystic fibrosis infection is not well understood. Structural modeling identified surface arginines on one lxa-encoded USP, USP76, which suggested it mediated interactions with heparan sulfate. Using mutants derived from the B. cenocepacia strain, K56-2, we show that USP76 is involved in host cell attachment. Pretreatment of lung epithelial cells with heparanase reduced the binding of the wild-type and complement strains but not the Δusp76 mutant strain, indicating that USP76 is directly or indirectly involved in receptor recognition on the surface of epithelial cells. We also show that USP76 is required for growth and survival in many conditions associated with the CF lung, including acidic conditions and oxidative stress. Moreover, USP76 also has a role in survival in macrophages isolated from people with CF. Overall, while further elucidation of the exact mechanism(s) is required, we can conclude that USP76, which is upregulated during chronic infection, is involved in bacterial survival within CF macrophages, a hallmark of Burkholderia infection.

The involvement of the low-oxygen-activated locus of Burkholderia cenocepacia in adaptation during cystic fibrosis infection.

Chronic infection with opportunistic pathogens including Burkholderia cepacia complex (Bcc) is a hallmark of cystic fibrosis (CF). We investigated the adaptive mechanisms facilitating chronic lung infection in sequential Bcc isolates from two siblings with CF (P1 and P2), one of whom also experienced intermittent blood-stream infections (P2). We previously showed increased lung cell attachment with colonisation time in both P1 and P2. WGS analysis confirmed that the isolates are closely related. Twelve genes showed three or more mutations, suggesting these were genes under selection. Single nucleotide polymorphisms (SNVs) in 45 regulatory genes were also observed. Proteomic analysis showed that the abundance of 149 proteins increased over 61-months in sputum isolates, and both time- and source-related alterations in protein abundance between the second patient's isolates. A consistent time-dependent increase in abundance of 19 proteins encoded by a low-oxygen-activated (lxa) locus was observed in both sets of isolates. Attachment was dramatically reduced in a B. cenocepacia K56-2Δlxa-locus deletion mutant, further indicating that it encodes protein(s) involved in host-cell attachment. Time-related changes in virulence in Galleria mellonella or motility were not observed. We conclude that the lxa-locus, associated with anoxic persistence in vitro, plays a role in host-cell attachment and adaptation to chronic colonization in the hypoxic niche of the CF lung.

Quantum Dot Nanotoxicity Investigations Using Human Lung Cells and TOXOR Electrochemical Enzyme Assay Methodology.

Recent studies have suggested that certain nanomaterials can interfere with optically based cytotoxicity assays resulting in underestimations of nanomaterial toxicity. As a result there has been growing interest in the use of whole cell electrochemical biosensors for nanotoxicity applications. Herein we report application of an electrochemical cytotoxicity assay developed in house (TOXOR) in the evaluation of toxic effects of mercaptosuccinic acid capped cadmium telluride quantum dots (MSA capped CdTe QDs), toward mammalian cells. MSA capped CdTe QDs were synthesized, characterized, and their cytotoxicity toward A549 human lung epithelial cells investigated. The internalization of QDs within cells was scrutinized via confocal microscopy. The cytotoxicity assay is based on the measurement of changes in cellular enzyme acid phosphatase upon 24 h exposure to QDs. Acid phosphatase catalyzes dephosphorylation of 2-naphthyl phosphate to 2-naphthol (determined by chronocoulometry) and is indicative of metabolic activity in cells. The 24 h IC50 (concentration resulting in 50% reduction in acid phosphatase activity) value for MSA capped CdTe QDs was found to be 118 ± 49 µg/mL using the TOXOR assay and was in agreement with the MTT assay (157 ± 31 µg/mL). Potential uses of this electrochemical assay include the screening of nanomaterials, environmental toxins, in addition to applications in the pharmaceutical, food, and health sectors.

The Role of Universal Stress Proteins in Bacterial Infections.

Universal stress proteins are ubiquitously expressed in bacteria, archaea and plants and other eukaryotes. A general property of USPs is their role in adaptation of bacteria to oxidative stress, high temperature, low pH and/or hypoxia. There is increasing evidence that these proteins facilitate the adaption of bacterial pathogens to the human host environment, thereby facilitating colonisation and pathogenicity. USPs in Mycobacterium tuberculosis are well studied and may play a role in latency of tuberculosis. USP expressed by Acinetobacter baumannii, Listeria monocytogenes and Salmonella enterica serovar Typhimurium are involved in survival in vivo, while USPs expressed in Pseudomonas aeruginosa and Porphyromonas gingivalis are involved in biofilm formation. Burkholderia cepacia complex and Staphylococcus aureus express USPs that play roles in host cell or host protein adhesion. There is also increasing evidence that USPs also bind to antimicrobial agents and may be ideal candidates to target in the future design of new anti-virulence strategies.

Two fac-tricarbonylrhenium(I) azadipyrromethene (ADPM) complexes: ligand-substitution effect on crystal structure.

The crystal structures of fac-(acetonitrile-κN)(2-{[3,5-bis(4-methoxyphenyl)-2H-pyrrol-2-ylidene-κN(1)]amino}-3,5-bis(4-methoxyphenyl)-1H-pyrrol-1-ido-κN(1))tricarbonylrhenium(I)-hexane-acetonitrile (2/1/2), [Re(C36H30N3O4)(CH3CN)(CO)3]·0.5C6H14·CH3CN, (2), and fac-(2-{[3,5-bis(4-methoxyphenyl)-2H-pyrrol-2-ylidene-κN(1)]amino}-3,5-bis(4-methoxyphenyl)-1H-pyrrol-1-ido-κN(1))tricarbonyl(dimethyl sulfoxide-κO)rhenium(I), [Re(C36H30N3O4)(C2H6OS)(CO)3], (3), at 150 K are reported. Both complexes display a distorted octahedral geometry, with a fac-Re(CO)3 arrangement and one azadipyrromethene (ADPM) chelating ligand in the equatorial position. One solvent molecule completes the coordination sphere of the Re(I) centre in the remaining axial position. The ADPM ligand shows high flexibility upon coordination, while retaining its π-delocalized nature. Bond length and angle analyses indicate that the differences in the geometry around the Re(I) centre in (2) and (3), and those found in three reported fac-Re(CO)3-ADPM complexes, are dictated mainly by steric factors and crystal packing. Both structures display intramolecular C-H...N hydrogen bonding. Intermolecular interactions of the Csp(2)-H...π and Csp(2)-H...O(carbonyl) types link the discrete monomers into extended chains.

Hypertension awareness, treatment, and control in adults with CKD: results from the Chronic Renal Insufficiency Cohort (CRIC) Study.

BACKGROUND: A low rate of blood pressure control has been reported in patients with chronic kidney disease (CKD). These data were derived from population-based samples with a low rate of CKD awareness. STUDY DESIGN: Cross-sectional. SETTING & PARTICIPANTS: Data from the baseline visit of the Chronic Renal Insufficiency Cohort (CRIC) Study (n = 3,612) were analyzed. Participants with an estimated glomerular filtration rate of 20-70 mL/min/1.73 m(2) were identified from physician offices and review of laboratory databases. OUTCOMES: Prevalence and awareness of hypertension, treatment patterns, control rates, and factors associated with hypertension control. MEASUREMENTS: Following a standardized protocol, blood pressure was measured 3 times by trained staff, and hypertension was defined as systolic blood pressure > or =140 mm Hg and/or diastolic blood pressure > or =90 mm Hg and/or self-reported antihypertensive medication use. Patients' awareness and treatment of hypertension were defined using self-report, and 2 levels of hypertension control were evaluated: systolic/diastolic blood pressure <140/90 and <130/80 mm Hg. RESULTS: The prevalence of hypertension was 85.7%, and 98.9% of CRIC participants were aware of this diagnosis and 98.3% were treated with medications, whereas 67.1% and 46.1% had hypertension controlled to <140/90 and <130/80 mm Hg, respectively. Of CRIC participants with hypertension, 15%, 25%, 26%, and 32% were using 1, 2, 3, and > or =4 antihypertensive medications, respectively. After multivariable adjustment, older patients, blacks, and those with higher urinary albumin excretion were less likely, whereas participants using angiotensin-converting enzyme inhibitors and angiotensin receptor blockers were more likely to have controlled their hypertension to <140/90 and <130/80 mm Hg. LIMITATIONS: Data were derived from a single study visit. CONCLUSIONS: Despite almost universal hypertension awareness and treatment in this cohort of patients with CKD, rates of hypertension control were suboptimal.

The case for chronic kidney disease, diabetes mellitus, and myocardial infarction being equivalent risk factors for cardiovascular mortality in patients older than 65 years.

The objective of the study was to determine whether chronic kidney disease (CKD) is as important a risk as either diabetes mellitus (DM) or previous myocardial infarction (MI). CKD and DM are important coronary artery disease risk factors. We hypothesized that the risk of cardiovascular mortality in elderly patients with CKD is equivalent to that for patients with either DM or previous MI. The CHS limited-access database was used to identify a cohort of patients with a baseline history of MI, DM, or CKD (estimated glomerular filtration rate <60 ml/min). Subjects were categorized in 1 of 3 groups as group 1, patients with DM (no CKD or MI); group 2, patients with previous MI (no DM or CKD); and group 3, patients with CKD (no DM or MI). Patients were followed up for a mean of 8.6 years, and rates of cardiovascular mortality were compared using proportional hazards regression. There were 789, 443, and 667 people in the MI, DM, and CKD groups, respectively. During follow-up, 124 patients (15.7%) died of cardiovascular causes in the MI group, and 69 (15.8%) and 87 (13%), in the DM and CKD groups, respectively. After adjusting for age, race, gender, smoking, hypertension, and total, high-density lipoprotein, and low-density lipoprotein cholesterol, the hazard ratio (HR) for cardiovascular mortality was similar between the DM (HR 1.0, 95% confidence interval 0.8 to 1.4)) and CKD cohorts (HR 0.8, 95% confidence interval 0.6 to 1.1) compared with the MI group. In conclusion, the risk of cardiovascular mortality in patients with moderate CKD was as high as that in patients with a history of MI or DM. Designation of CKD as a cardiovascular risk equivalent in patients >65 years of age appears justified.

The morbidity and cost implications of hemodialysis clinical performance measures.

Clinical performance measures, including dialysis dose, hemoglobin, albumin, and vascular access, are the focus of monitoring and quality improvement activities. However, little is known about the implications of clinical performance measures for hospital utilization and health care costs. We obtained clinical performance measures and hospitalization records for a national random sample of 10,650 hemodialysis patients and analyzed the relationship between changes in clinical performance measures and hospital utilization after adjustment for patient demographic and medical characteristics. Higher hemoglobin, higher albumin, and fistula or graft use were independently associated with fewer hospitalizations, fewer hospital days, and decreased Medicare inpatient reimbursement. For example, a 0.5 g/dL higher hemoglobin, a 0.25 g/dL higher albumin, fistula use, and graft use were associated with hospitalization rate ratios of 0.90 (95% confidence interval 0.85, 0.96), 0.64 (0.53, 0.77), 0.60 (0.52, 0.69), and 0.79 (0.71, 0.89), respectively. Moreover, there was a 2-3-fold variation in hospital utilization across end-stage renal disease networks that was still evident after adjustment for patient characteristics and clinical performance measures. Clinical performance measures, especially albumin and vascular access, are strongly associated with hospital utilization and health care costs. These results highlight the importance of targeting nutrition and vascular access in quality improvement efforts. The marked variation in hospital utilization across networks deserves further examination.

Changes in Medicare reimbursement and patient-nephrologist visits, quality of care, and health-related quality of life.

BACKGROUND: Medicare's reimbursement system was changed in January 2004 to encourage more frequent visits between dialysis patients and nephrologists. We sought to determine the impact of this policy change on patient-nephrologist visits, quality of care, and health-related quality of life. METHODS: We examined visits and outcomes for 2,043 patients at 12 hemodialysis facilities in northeast Ohio for 12 months before and 7 months after the reimbursement change. For comparison of outcomes, we used linear, logistic, or negative binomial regression models (for continuous, binary, and rate outcomes, respectively) to assess the significance of changes across the 2 periods. RESULTS: For patients seen before and after the reimbursement change for at least 6 months, the number of visits per patient-month increased from 1.52 before to 3.14 after (P < 0.001). The percentage of patients with no nephrologist visits per patient-month decreased from 16.6% before to 4.6% after (P < 0.001). However, there were no clinically important changes in Kt/V, albumin level, hemoglobin level, phosphorus level, calcium level, hemodialysis catheter use, ultrafiltration volume, shortened or skipped treatments, hospital admissions, hospitalization days, or health-related quality of life, including patient satisfaction. CONCLUSION: Despite a marked increase in visits between patients and nephrologists, there was no clinically important impact on parameters related to quality of care or health-related quality of life. Additional work is needed to determine effective payment strategies to improve dialysis patient outcomes.

Renal phenotype is exacerbated in Os and lpr double mutant mice.

BACKGROUND: ROP-Os/+ mice are born with oligosyndactyly and oligonephronia and develop renal dysfunction, which includes renal tubular epithelial cell (RTC) Fas-dependent apoptosis and tubular atrophy. MRL/lpr mice harbor a Fas-inactivating mutation and develop glomerulonephritis, whereas mice expressing lpr on a C3H background demonstrate no renal phenotype. We hypothesized that crossing ROP-Os/+ with CH3-lpr/lpr mice would rescue the Os/+ renal phenotype by reducing Fas-dependent RTC apoptosis. METHODS: ROP-Os/+ mice were intercrossed with C3H-lpr/lpr mice and F(2) generation animals were phenotyped by kidney weight, serum creatinine, and albuminuria. Kidney sections were scored for histopathology and apoptosis. Univariate and multivariate analyses were used to examine additive effects of Os and lpr on renal phenotype. RESULTS: By 16 weeks, F(2)Os/+ lpr/lpr mice developed significantly more albuminuria, glomerulosclerosis, and interstitial inflammation compared to Os/++/+ mice. Glomerular cell apoptosis was increased in Os/+ lpr/lpr compared to Os/++/+ mice, with no significant difference in RTC apoptosis. A statistically significant Os-lpr effect on renal phenotype was demonstrated by multivariate analysis, which exceeded the combined independent effects if Os and lpr, indicating a biologic interaction exists between Os and lpr. CONCLUSION: Os/+ mice with a superimposed lpr mutation displayed a more severe renal phenotype, rather than phenotype rescue, suggesting that Fas pathway activation is necessary to delete cells resulting from Os-dependent injury. We further propose that an Os-lpr gene interaction and/or mixed ROP-C3H genetic background regulated the renal phenotype, consistent with the concept that chronic renal disease pathogenesis reflects effects of multiple nephropathy susceptibility alleles.

The relative predictive ability of four different measures of hemodialysis dose.

BACKGROUND: The amount of hemodialysis that patients receive is an independent predictor of mortality. However, the relative predictive ability of four common measures of dialysis dose (urea reduction ratio, single-pool Kt/V, double-pool Kt/V, and urea index) is unclear. METHODS: Using The Renal Network Data System, we identified 14,810 incident hemodialysis patients in Indiana, Kentucky, Ohio, and Illinois from 1997 to 2000. We calculated each measure of hemodialysis dose during the first 6 months of treatment, then prospectively followed up patients for an additional 6 months. For each measure of dialysis dose, we developed a logistic regression model to examine the relationship between dose and patient mortality after adjustment for age, race, sex, cause of renal failure, comorbid conditions, and albumin level. We compared the predictive ability of the four models using the c statistic, a measure of how frequently survivors have a lower predicted probability of death compared with nonsurvivors. C statistics can vary from 0.50 (no predictive ability) to 1.00 (perfect predictive ability). RESULTS: Of all patients, 11.3% died during follow-up. Mortality was independently associated with low dialysis dose, advanced age, white race, female sex, specific comorbid conditions, and low albumin level. All four predictive models had virtually identical c statistics (range, 0.69 to 0.70). CONCLUSION: Models including hemodialysis dose and patient characteristics have a modest ability to predict mortality. Moreover, all four measures of dialysis dose have an equivalent predictive ability. Decisions to use a specific measure should be based on other considerations, such as ease of use, need to troubleshoot inadequate dialysis delivery, or research on urea kinetics or nutritional factors.