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Purpose: Magnetically controlled implant systems have been established to treat severe progressive spinal deformity in children. The purpose of this study was to evaluate (1) the ratio between achieved and expected distraction length, (2) the complication rate and its risk factors as well as (3) the correlation of the distraction length and the length of the spine., Methods: A total of 40 patients with an average follow-up of 34 (14 to 57) months were prospectively included in the study. Children underwent lengthening procedures every three months. The ratio between the distraction lengths was determined by comparing the measured distraction length of the rod on radiographs with the distraction length displayed on the external remote controller for the magnetically controlled growing rod (MCGR). Age, weight, height, and complications were repeatedly recorded., Results: The analysis of 746 procedures showed the actual distraction to be 94.4% of the expected one. No difference between implants on the concave and convex spinal side was observed. The overall complication rate was 4.6% mainly because of failure of the implant or lack of implant extension, which was directly related to an increased BMI. There was also a strong correlation between achieved implant distraction length and gain in spinal length., Conclusions: Our study demonstrates a high ratio (0.94) between achieved and expected distraction length of magnetically controlled spinal rods. The complication rate was low (4.6%) and correlated to a high BMI. The correlation between the achieved implant distraction length and spinal length indicates the efficiency of the MCGR therapy., Level of Evidence: Therapeutic Level IV.

Introduction: Perinatal mood and anxiety disorders (PMADs) are the most common complication of pregnancy and have been found to have long-term implications for both mother and child. In vulnerable patient populations such as those served at Denver Health, a federally qualified health center the prevalence of PMADs is nearly double the nationally reported rate of 15-20%. Nearly 17% of women will be diagnosed with major depression at some point in their lives and those numbers are twice as high in women who live in poverty. Women also appear to be at higher risk for depression in the child-bearing years. In order to better address these issues, an Integrated Perinatal Mental Health program was created to screen, assess, and treat PMADs in alignment with national recommendations to improve maternal-child health and wellness. This program was built upon a national model of Integrated Behavioral Health already in place at Denver Health., Methods: A multidisciplinary team of physicians, behavioral health providers, public health, and administrators was assembled at Denver Health, an integrated hospital and community health care system that serves as the safety net hospital to the city and county of Denver, CO. This team was brought together to create a universal screen-to-treat process for PMAD's in perinatal clinics and to adapt the existing Integrated Behavioral Health (IBH) model into a program better suited to the health system's obstetric population. Universal prenatal and postnatal depression screening was implemented at the obstetric intake visit, a third trimester prenatal care visit, and at the postpartum visit across the clinical system. At the same time, IBH services were implemented across our health system's perinatal care system in a stepwise fashion. This included our women's care clinics as well as the family medicine and pediatric clinics. These efforts occurred in tandem to support all patients and staff enabling a specially trained behavioral health provider (psychologists and L.C.S.W.'s) to respond immediately to any positive screen during or after pregnancy., Results: In August 2014 behavioral health providers were integrated into the women's care clinics. In January 2015 universal screening for PMADs was implemented throughout the perinatal care system. Screening has improved from 0% of women screened at the obstetric care intake visit in August 2014 to >75% of women screened in August 2016. IBH coverage by a licensed psychologist or licensed clinical social worker exists in 100% of perinatal clinics as of January 2016. As well, in order to gain sustainability, the ability to bill same day visits as well as to bill, and be reimbursed for screening and assessment visits, continues to improve and provide for a model that is self-sustaining for the future., Conclusion: Implementation of a universal screening process for PMADs alongside the development of an IBH model in perinatal care has led to the creation of a program that is feasible and has the capacity to serve as a national model for improving perinatal mental health in vulnerable populations.

The neutral silver(i) dithiocarboxylate complexes silver(i) dithioacetate, silver(i) pentafluorodithiobenzoate, silver(i) 2,4,6-trimethyldithiobenzoate, silver(i) 2,4,6-tri-iso-propyldithiobenzoate and silver(i) 2,6-dimesityl-dithiobenzoate were synthesized and characterized by IR spectroscopy, elemental analyses and single crystal X-ray diffraction. All complexes form primarily dinuclear units with eight-membered rings [Ag2S4C2], which in turn aggregate by argentophilic interactions and additional AgS contacts in their crystal structures. The molecular structures and shapes of aggregation in the solid state are presented and discussed. The layer-like and chain-like aggregates as well as the [Ag] clusters formed are compared with the (almost) analogous gold(i) complexes.

Neutral, homoleptic gold(i) dithiocarboxylate complexes with different-sized Me, iPr, Mes, Tip, Mes* and Dmp substituents at the dithiocarboxylate unit were synthesized by reacting the corresponding dithiocarboxylic acid with potassium dicyanoaurate(i). They were characterized by IR spectroscopy and elemental analyses and their structures were determined by single crystal X-ray diffraction. The dinuclear complexes with an intramolecular d10-d10 type aurophilic interaction aggregate through additional intermolecular aurophilic interactions in different binding motifs. The molecular structures and the patterns of aggregation in the solid state are presented and discussed with respect to the influence of the substituent size.

The unstable dithiocarboxylic acids dithioacetic acid, 2-methyl-dithiopropionic acid, 2,2-dimethyl-dithiopropionic acid and dithiobenzoic acid were synthesized and characterized by NMR spectroscopy and GC/MS. The stable dithiocarboxylic acids 2,4,6-trimethyl benzoic acid, 2,4,6-tri-iso-propylbenzoic acid and 2,6-dimesityl benzoic acid were synthesized, isolated and characterized by spectroscopic methods and in parts by mass spectrometry and X-ray crystallography. The new data were used to re-evaluate literature data on the synthesis, spectroscopy and structural data of dithiocarboxylic acids as a fundamental class of organic compounds in general., (© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The digitization of biocollections is a critical task with direct implications for the global community who use the data for research and education. Recent innovations to involve citizen scientists in digitization increase awareness of the value of biodiversity specimens; advance science, technology, engineering, and math literacy; and build sustainability for digitization. In support of these activities, we launched the first global citizen-science event focused on the digitization of biodiversity specimens: Worldwide Engagement for Digitizing Biocollections (WeDigBio). During the inaugural 2015 event, 21 sites hosted events where citizen scientists transcribed specimen labels via online platforms (DigiVol, Les Herbonautes, Notes from Nature, the Smithsonian Institution's Transcription Center, and Symbiota). Many citizen scientists also contributed off-site. In total, thousands of citizen scientists around the world completed over 50,000 transcription tasks. Here, we present the process of organizing an international citizen-science event, an analysis of the event's effectiveness, and future directions-content now foundational to the growing WeDigBio event.

Plant-released flavonoids induce the transcription of symbiotic genes in rhizobia and one of the first bacterial responses is the synthesis of so called Nod factors. They are responsible for the initial root hair curling during onset of root nodule development. This signal exchange is believed to be essential for initiating the plant symbiosis with rhizobia affiliated with the Alphaproteobacteria . Here, we provide evidence that in the broad host range strain Sinorhizobium fredii NGR234 the complete lack of quorum sensing molecules results in an elevated copy number of its symbiotic plasmid (pNGR234 a ). This in turn triggers the expression of symbiotic genes and the production of Nod factors in the absence of plant signals. Therefore, increasing the copy number of specific plasmids could be a widespread mechanism of specialized bacterial populations to bridge gaps in signaling cascades.

Phenotypic heterogeneity describes the occurrence of "nonconformist" cells within an isogenic population. The nonconformists show an expression profile partially different from that of the remainder of the population. Phenotypic heterogeneity affects many aspects of the different bacterial lifestyles, and it is assumed that it increases bacterial fitness and the chances for survival of the whole population or smaller subpopulations in unfavorable environments. Well-known examples for phenotypic heterogeneity have been associated with antibiotic resistance and frequently occurring persister cells. Other examples include heterogeneous behavior within biofilms, DNA uptake and bacterial competence, motility (i.e., the synthesis of additional flagella), onset of spore formation, lysis of phages within a small subpopulation, and others. Interestingly, phenotypic heterogeneity was recently also observed with respect to quorum-sensing (QS)-dependent processes, and the expression of autoinducer (AI) synthase genes and other QS-dependent genes was found to be highly heterogeneous at a single-cell level. This phenomenon was observed in several Gram-negative bacteria affiliated with the genera Vibrio, Dinoroseobacter, Pseudomonas, Sinorhizobium, and Mesorhizobium. A similar observation was made for the Gram-positive bacterium Listeria monocytogenes. Since AI molecules have historically been thought to be the keys to homogeneous behavior within isogenic populations, the observation of heterogeneous expression is quite intriguing and adds a new level of complexity to the QS-dependent regulatory networks. All together, the many examples of phenotypic heterogeneity imply that we may have to partially revise the concept of homogeneous and coordinated gene expression in isogenic bacterial populations., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Populations of genetically identical Sinorhizobium fredii NGR234 cells differ significantly in their expression profiles of autoinducer (AI)-dependent and AI-independent genes. Promoter fusions of the NGR234 AI synthase genes traI and ngrI showed high levels of phenotypic heterogeneity during growth in TY medium on a single-cell level. However, adding very high concentrations of N-(3-oxooctanoyl-)-l-homoserine lactone resulted in a more homogeneous expression profile. Similarly, the lack of internally synthesized AIs in the background of the NGR234-ΔtraI or the NGR234-ΔngrI mutant resulted in a highly homogenous expression of the corresponding promoter fusions in the population. Expression studies with reporter fusions of the promoter regions of the quorum-quenching genes dlhR and qsdR1 and the type IV pilus gene cluster located on pNGR234b suggested that factors other than AI molecules affect NGR234 phenotypic heterogeneity. Further studies with root exudates and developing Arabidopsis thaliana seedlings provide the first evidence that plant root exudates have strong effects on the heterogeneity of AI synthase and quorum-quenching genes in NGR234. Therefore, plant-released octopine appears to play a key role in modulation of heterogeneous gene expression., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)

The alphaproteobacterium Sinorhizobium fredii NGR234 has an exceptionally wide host range, as it forms nitrogen-fixing nodules with more legumes than any other known microsymbiont. Within its 6.9-Mbp genome, it encodes two N-acyl-homoserine-lactone synthase genes (i.e., traI and ngrI) involved in the biosynthesis of two distinct autoinducer I-type molecules. Here, we report on the construction of an NGR234-ΔtraI and an NGR234-ΔngrI mutant and their genome-wide transcriptome analysis. A high-resolution RNA sequencing (RNA-seq) analysis of early-stationary-phase cultures in the NGR234-ΔtraI background suggested that up to 316 genes were differentially expressed in the NGR234-ΔtraI mutant versus the parent strain. Similarly, in the background of NGR234-ΔngrI 466 differentially regulated genes were identified. Accordingly, a common set of 186 genes was regulated by the TraI/R and NgrI/R regulon. Coregulated genes included 42 flagellar biosynthesis genes and 22 genes linked to exopolysaccharide (EPS) biosynthesis. Among the genes and open reading frames (ORFs) that were differentially regulated in NGR234-ΔtraI were those linked to replication of the pNGR234a symbiotic plasmid and cytochrome c oxidases. Biotin and pyrroloquinoline quinone biosynthesis genes were differentially expressed in the NGR234-ΔngrI mutant as well as the entire cluster of 21 genes linked to assembly of the NGR234 type III secretion system (T3SS-II). Further, we also discovered that genes responsible for rhizopine catabolism in NGR234 were strongly repressed in the presence of high levels of N-acyl-homoserine-lactones. Together with nodulation assays, the RNA-seq-based findings suggested that quorum sensing (QS)-dependent gene regulation appears to be of higher relevance during nonsymbiotic growth rather than for life within root nodules., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)

Ralstonia eutropha H16 is capable of utilizing 3,3'-thiodipropionic acid (TDP) and 3,3'-dithiodipropionic acid (DTDP) as precursor substrates for biosynthesis of a polythioester (PTE) heteropolymer consisting of 3-hydroxybutyric acid (3HB) and 3-mercaptopropionic acid (3MP). To elucidate the hitherto unknown catabolic pathways of TDP and DTDP in R. eutropha H16, 19 defined deletion mutants were generated based on extensive functional genome analyses. Deletions of two ABC-type transporter clusters (H16_A0357-0359, H16_A3658-3660) resulted in an alteration of poly(3HB-co-3MP) composition with TDP as precursor to only 10.2±1.9mol% 3MP in comparison to 15.1±5.5mol% in the wild type. A mutant strain of H16 lacking Bordetella uptake gene-like substrate binding proteins (H16_A2779, H16_A0337) incorporated only 7.4±3.8mol% 3MP into PTE heteropolymers with DTDP as precursor in comparison to 24.5±14.5mol% in the wild type. Therefore, both gene products are probably involved in transport processes of this compound into the cells. However, the most significant reduction in 3MP contents of the heteropolymers with DTDP as precursor occurred upon the deletion of a gene encoding the putative thiol-disulfide interchange protein DsbD (H16_A3455, 3.9±2.6mol% 3MP). DsbD is proposed to be involved in the reduction of DTDP into two molecules of 3MP, the common cleavage product of TDP and DTDP., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Strain HIMB11 is a planktonic marine bacterium isolated from coastal seawater in Kaneohe Bay, Oahu, Hawaii belonging to the ubiquitous and versatile Roseobacter clade of the alphaproteobacterial family Rhodobacteraceae. Here we describe the preliminary characteristics of strain HIMB11, including annotation of the draft genome sequence and comparative genomic analysis with other members of the Roseobacter lineage. The 3,098,747 bp draft genome is arranged in 34 contigs and contains 3,183 protein-coding genes and 54 RNA genes. Phylogenomic and 16S rRNA gene analyses indicate that HIMB11 represents a unique sublineage within the Roseobacter clade. Comparison with other publicly available genome sequences from members of the Roseobacter lineage reveals that strain HIMB11 has the genomic potential to utilize a wide variety of energy sources (e.g. organic matter, reduced inorganic sulfur, light, carbon monoxide), while possessing a reduced number of substrate transporters.

A psychro- and aerotolerant bacterium was isolated from the sulfidic water of a pelagic redox zone of the central Baltic Sea. The slightly curved rod- or spiral-shaped cells were motile by one polar flagellum or two bipolar flagella. Growth was chemolithoautotrophic, with nitrate or nitrite as electron acceptor and either a variety of sulfur species of different oxidation states or hydrogen as electron donor. Although the bacterium was able to utilize organic substances such as acetate, pyruvate, peptone and yeast extract for growth, these compounds yielded considerably lower cell numbers than obtained with reduced sulfur or hydrogen; in addition, bicarbonate supplementation was necessary. The cells also had an absolute requirement for NaCl. Optimal growth occurred at 15 °C and at pH 6.6-8.0. The predominant fatty acid of this organism was 16 : 1ω7c, with 3-OH 14 : 0, 16 : 0, 16 : 1ω5c+t and 18 : 1ω7c present in smaller amounts. The DNA G+C content was 33.6 mol%. As determined in 16S rRNA gene sequence phylogeny analysis, the isolate belongs to the genus Sulfurimonas, within the class Epsilonproteobacteria, with 93.7 to 94.2 % similarity to the other species of the genus Sulfurimonas, Sulfurimonas autotrophica, Sulfurimonas paralvinellae and Sulfurimonas denitrificans. However, the distinct physiological and genotypic differences from these previously described taxa support the description of a novel species, Sulfurimonas gotlandica sp. nov. The type strain is GD1(T) ( = DSM 19862(T) = JCM 16533(T)). Our results also justify an emended description of the genus Sulfurimonas.

Enhanced DC conductivity and photoconductivity of cationic carbazole tethered deoxyribonucleic acid (Cz-DNA) in film devices is achieved by incorporating mobility enhancers. An anthracene-based organic semiconductor (namely 4HPA-Ant) and the inorganic semiconductor cadmium sulfide (CdS) multipod nanocrystal (NC) were used as the mobility enhancers. Space charge limited current (SCLC) experiments show that hole mobility in CdS:Cz-DNA composite film is improved significantly, by about an order of magnitude, compared to the Cz-DNA film. Similarly, the DC conductivity of the composite film is slightly enhanced by 4HPA-Ant. The photoconductivity is also improved in the Cz-DNA composite, with both 4HPA-Ant and CdS multipod NCs. The enhancement in photocurrent is by more than an order of magnitude, as demonstrated by current-voltage (I-V) characterization using DNA composite photodetectors.

Despite its immense size, logistical and methodological constraints have largely limited microbiological investigations of the subseafloor basement biosphere. In this study, a unique sampling system was used to collect fluids from the subseafloor basaltic crust via a Circulation Obviation Retrofit Kit (CORK) observatory at Integrated Ocean Drilling Program borehole 1301A, located at a depth of 2667 m in the Pacific Ocean on the eastern flank of the Juan de Fuca Ridge. Here, a fluid delivery line directly accesses a 3.5 million years old basalt-hosted basement aquifer, overlaid by 262 m of sediment, which serves as a barrier to direct exchange with bottom seawater. At an average of 1.2 × 10(4) cells ml(-1), microorganisms in borehole fluids were nearly an order of magnitude less abundant than in surrounding bottom seawater. Ribosomal RNA genes were characterized from basement fluids, providing the first snapshots of microbial community structure using a high-integrity fluid delivery line. Interestingly, microbial communities retrieved from different CORKs (1026B and 1301A) nearly a decade apart shared major community members, consistent with hydrogeological connectivity. However, over three sampling years, the dominant gene clone lineage changed from relatives of Candidatus Desulforudis audaxviator within the bacterial phylum Firmicutes in 2008 to the Miscellaneous Crenarchaeotic Group in 2009 and a lineage within the JTB35 group of Gammaproteobacteria in 2010, and statistically significant variation in microbial community structure was observed. The enumeration of different phylogenetic groups of cells within borehole 1301A fluids supported our observation that the deep subsurface microbial community was temporally dynamic.

SAR11 is an ancient and diverse clade of heterotrophic bacteria that are abundant throughout the world's oceans, where they play a major role in the ocean carbon cycle. Correlations between the phylogenetic branching order and spatiotemporal patterns in cell distributions from planktonic ocean environments indicate that SAR11 has evolved into perhaps a dozen or more specialized ecotypes that span evolutionary distances equivalent to a bacterial order. We isolated and sequenced genomes from diverse SAR11 cultures that represent three major lineages and encompass the full breadth of the clade. The new data expand observations about genome evolution and gene content that previously had been restricted to the SAR11 Ia subclade, providing a much broader perspective on the clade's origins, evolution, and ecology. We found small genomes throughout the clade and a very high proportion of core genome genes (48 to 56%), indicating that small genome size is probably an ancestral characteristic. In their level of core genome conservation, the members of SAR11 are outliers, the most conserved free-living bacteria known. Shared features of the clade include low GC content, high gene synteny, a large hypervariable region bounded by rRNA genes, and low numbers of paralogs. Variation among the genomes included genes for phosphorus metabolism, glycolysis, and C1 metabolism, suggesting that adaptive specialization in nutrient resource utilization is important to niche partitioning and ecotype divergence within the clade. These data provide support for the conclusion that streamlining selection for efficient cell replication in the planktonic habitat has occurred throughout the evolution and diversification of this clade. IMPORTANCE The SAR11 clade is the most abundant group of marine microorganisms worldwide, making them key players in the global carbon cycle. Growing knowledge about their biochemistry and metabolism is leading to a more mechanistic understanding of organic carbon oxidation and sequestration in the oceans. The discovery of small genomes in SAR11 provided crucial support for the theory that streamlining selection can drive genome reduction in low-nutrient environments. Study of isolates in culture revealed atypical organic nutrient requirements that can be attributed to genome reduction, such as conditional auxotrophy for glycine and its precursors, a requirement for reduced sulfur compounds, and evidence for widespread cycling of C1 compounds in marine environments. However, understanding the genetic variation and distribution of such pathways and characteristics like streamlining throughout the group has required the isolation and genome sequencing of diverse SAR11 representatives, an analysis of which we provide here.

Eutrophication and global climate change lead to expansion of hypoxia in the ocean, often accompanied by the production of hydrogen sulfide, which is toxic to higher organisms. Chemoautotrophic bacteria are thought to buffer against increased sulfide concentrations by oxidizing hydrogen sulfide before its diffusion to oxygenated surface waters. Model organisms from such environments have not been readily available, which has contributed to a poor understanding of these microbes. We present here a detailed study of "Sulfurimonas gotlandica" str. GD1, an Epsilonproteobacterium isolated from the Baltic Sea oxic-anoxic interface, where it plays a key role in nitrogen and sulfur cycling. Whole-genome analysis and laboratory experiments revealed a high metabolic flexibility, suggesting a considerable capacity for adaptation to variable redox conditions. S. gotlandica str. GD1 was shown to grow chemolithoautotrophically by coupling denitrification with oxidation of reduced sulfur compounds and dark CO(2) fixation. Metabolic versatility was further suggested by the use of a range of different electron donors and acceptors and organic carbon sources. The number of genes involved in signal transduction and metabolic pathways exceeds those of other Epsilonproteobacteria. Oxygen tolerance and environmental-sensing systems combined with chemotactic responses enable this organism to thrive successfully in marine oxygen-depletion zones. We propose that S. gotlandica str. GD1 will serve as a model organism in investigations that will lead to a better understanding how members of the Epsilonproteobacteria are able to cope with water column anoxia and the role these microorganisms play in the detoxification of sulfidic waters.

Strain HIMB100 is a planktonic marine bacterium in the class Alphaproteobacteria. This strain is of interest because it is one of the first known isolates from a globally ubiquitous clade of marine bacteria known as SAR116 within the family Rhodospirillaceae. Here we describe preliminary features of the organism, together with the draft genome sequence and annotation. This is the second genome sequence of a member of the SAR116 clade. The 2,458,945 bp genome contains 2,334 protein-coding and 42 RNA genes.

The sucCD gene of Advenella mimigardefordensis strain DPN7(T) encodes a succinyl coenzyme A (succinyl-CoA) synthetase homologue (EC 6.2.1.4 or EC 6.2.1.5) that recognizes, in addition to succinate, the structural analogues 3-sulfinopropionate (3SP) and itaconate as substrates. Accumulation of 3SP during 3,3'-dithiodipropionic acid (DTDP) degradation was observed in Tn5::mob-induced mutants of A. mimigardefordensis strain DPN7(T) disrupted in sucCD and in the defined deletion mutant A. mimigardefordensis ΔsucCD. These mutants were impaired in growth with DTDP and 3SP as the sole carbon source. Hence, it was proposed that the succinyl-CoA synthetase homologue in A. mimigardefordensis strain DPN7(T) activates 3SP to the corresponding CoA-thioester (3SP-CoA). The putative genes coding for A. mimigardefordensis succinyl-CoA synthetase (SucCD(Am)) were cloned and heterologously expressed in Escherichia coli BL21(DE3)/pLysS. Purification and characterization of the enzyme confirmed its involvement during degradation of DTDP. 3SP, the cleavage product of DTDP, was converted into 3SP-CoA by the purified enzyme, as demonstrated by in vitro enzyme assays. The structure of 3SP-CoA was verified by using liquid chromatography-electrospray ionization-mass spectrometry. SucCD(Am) is Mg²⁺ or Mn²⁺ dependent and unspecific regarding ATP or GTP. In kinetic studies the enzyme showed highest enzyme activity and substrate affinity with succinate (V(max) = 9.85 ± 0.14 μmol min⁻¹ mg⁻¹, K(m) = 0.143 ± 0.001 mM). In comparison to succinate, activity with 3SP was only ca. 1.2% (V(max) = 0.12 ± 0.01 μmol min⁻¹ mg⁻¹) and the affinity was 6-fold lower (K(m) = 0.818 ± 0.046 mM). Based on the present results, we conclude that SucCD(Am) is physiologically associated with the citric acid cycle but is mandatory for the catabolic pathway of DTDP and its degradation intermediate 3SP.

Background: Current unitage for the calciferols suggests that equimolar quantities of vitamins D(2) (D2) and D(3) (D3) are biologically equivalent. Published studies yield mixed results., Objective: The aim of the study was to compare the potencies of D2 and D3., Design: The trial used a single-blind, randomized design in 33 healthy adults. Calciferols were dosed at 50,000 IU/wk for 12 wk. Principal outcome variables were area under the curve for incremental total 25-hydroxyvitamin D [25(OH)D] and change in calciferol content of sc fat., Results: Incremental mean (sd) 25(OH)D area under the curve at 12 wk was 1366 ng · d/ml (516) for the D2-treated group and 2136 (606) for the D3 (P < 0.001). Mean (sd) steady-state 25(OH)D increments showed similar differences: 24 ng/ml for D2 (10.3) and 45 ng/ml (16.2) for D3 (P <0.001). Subcutaneous fat content of D2 rose by 50 μg/kg in the D2-treated group, and D3 content rose by 104 μg/kg in the D3-treated group. Total calciferol in fat rose by only 33 ng/kg in the D2-treated, whereas it rose by 104 μg/kg in the D3-treated group. Extrapolating to total body fat D3, storage amounted to just 17% of the administered dose., Conclusion: D3 is approximately 87% more potent in raising and maintaining serum 25(OH)D concentrations and produces 2- to 3-fold greater storage of vitamin D than does equimolar D2. For neither was there evidence of sequestration in fat, as had been postulated for doses in this range. Given its greater potency and lower cost, D3 should be the preferred treatment option when correcting vitamin D deficiency.

The Evaluation of the US Department of Housing and Urban Development Lead-Based Paint Hazard Control Grant Program studied the effectiveness of the housing intervention performed in reducing the blood lead of children at four post-intervention times (6-months, 1-year, 2-years, and 3-years). A repeat measures analysis showed that blood lead levels declined up to three-years post-intervention. The results at each successive collection time were significantly lower than at the previous post-intervention time except for the difference between the levels at two and three years. At two-years post-intervention, geometric mean blood lead levels were approximately 37% lower than at pre-intervention. Children with pre-intervention blood lead levels as low as 10 μg/dL experienced substantial declines in blood lead levels. Previous studies have found substantial improvements only if a child's pre-intervention blood lead level was above 20 μg/dL. Individual interior lead hazard control treatments as grouped by Interior Strategy were not a significant predictor of post-intervention blood lead levels. However, children living in dwellings where exterior lead hazard control interventions were done had lower blood lead levels at one-year post-intervention than those living in dwellings without the exterior interventions (all other factors being equal), but those differences were only significant when the mean exterior paint lead loading at pre-intervention was about the 90th percentile (7.0mg/cm(2)). This observation suggests that exterior lead hazard control can be an important component of a lead hazard control plan. Children who were six to eleven months of age at pre-intervention had a significant increase in blood lead at one-year post-intervention, probably due to other exposures., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Mitochondria share a common ancestor with the Alphaproteobacteria, but determining their precise origins is challenging due to inherent difficulties in phylogenetically reconstructing ancient evolutionary events. Nonetheless, phylogenetic accuracy improves with more refined tools and expanded taxon sampling. We investigated mitochondrial origins with the benefit of new, deeply branching genome sequences from the ancient and prolific SAR11 clade of Alphaproteobacteria and publicly available alphaproteobacterial and mitochondrial genome sequences. Using the automated phylogenomic pipeline Hal, we systematically studied the effect of taxon sampling and missing data to accommodate small mitochondrial genomes. The evidence supports a common origin of mitochondria and SAR11 as a sister group to the Rickettsiales. The simplest explanation of these data is that mitochondria evolved from a planktonic marine alphaproteobacterial lineage that participated in multiple inter-specific cell colonization events, in some cases yielding parasitic relationships, but in at least one case producing a symbiosis that characterizes modern eukaryotic life.

Dot-blot is a versatile and simple analysis to perform. We adapted this method as a simple identity test for monoclonal antibodies to a number of small compounds: three transplant drugs, an anticonvulsant, a steroid, an anticancer drug, and an antibiotic. Immunology-based identity tests using low-molecular-mass organic compounds have historically been a challenge to develop. We modified the traditional dot-blot assay to serve as an identity test for monoclonal antibodies to carbamazepine, sirolimus, tacrolimus, cyclosporine, cortisol, methotrexate, and gentamicin. The primary obstacle was the immobilization of these organic compounds on nitrocellulose as nitrocellulose is also soluble in most of the organic solvents in which the compounds are soluble. We evaluated different membranes, solvents, and chemical forms of these organic compounds to overcome this challenge. A number of incubation and washing solutions were also investigated. By varying the chemical form, concentration, and incubation conditions, a set of effective and reproducible identity tests were developed for these monoclonal antibodies., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Objective: We investigated the mechanism by which the ARCHITECT cyclosporine (CsA) chemiluminescent microparticle immunoassay (CMIA) eliminates cross-reactivity to CsA metabolites AM1 and AM9, despite its use of a monoclonal antibody which shows cross-reactivity in fluorescence polarization immunoassays., Design and Methods: The CMIA was accomplished by incubating an extracted blood sample with magnetic microparticles coated with a very low amount of anti-CsA antibody. After a wash step the microparticles were incubated with a chemiluminescent CsA tracer, followed by a second wash step and measurement of chemiluminescence. The reagent concentrations of salt and detergent were optimized to maximize CsA binding and minimize metabolite interference., Results: Elimination of CsA metabolite cross-reactivity was shown using purified metabolites and blood samples containing native CsA metabolites. The CMIA demonstrated precision and sensitivity acceptable for use in a clinical setting., Conclusion: We conclude that it is possible to eliminate CsA metabolite immuno-cross-reactivity by careful assay design., (Copyright (c) 2010 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.)

Owing to their narrow bright emission band, broad size-tunable emission wavelength, superior photostability, and excellent flexible-substrate compatibility, light-emitting diodes based on quantum dots (QD-LEDs) are currently under intensive research and development for multiple consumer applications including flat-panel displays and flat lighting. However, their commercialization is still precluded by the slow development to date of efficient QD-LEDs as even the highest reported efficiency of 2.0% cannot favorably compete with their organic counterparts. Here, we report QD-LEDs with a record high efficiency (approximately 4%), high brightness (approximately 6580 cd/m(2)), low turn-on voltage (approximately 2.6 V), and significantly improved color purity by simply using deoxyribonucleic acid (DNA) complexed with cetyltrimetylammonium (CTMA) (DNA-CTMA) as a combined hole transporting and electron-blocking layer (HTL/EBL). This, together with controlled thermal decomposition of ligand molecules from the QD shell, represents a novel combined, but simple and very effective, approach toward the development of highly efficient QD-LEDs with a high color purity.

Recent studies have indicated that chemoautotrophic Epsilonproteobacteria might play an important role, especially as anaerobic or microaerophilic dark CO(2)-fixing organisms, in marine pelagic redoxclines. However, knowledge of their distribution and abundance as actively CO(2)-fixing microorganisms in pelagic redoxclines is still deficient. We determined the contribution of Epsilonproteobacteria to dark CO(2) fixation in the sulfidic areas of central Baltic Sea and Black Sea redoxclines by combining catalyzed reporter deposition-fluorescence in situ hybridization with microautoradiography using [(14)C]bicarbonate and compared it to the total prokaryotic chemoautotrophic activity. In absolute numbers, up to 3 x 10(5) (14)CO(2)-fixing prokaryotic cells ml(-1) were enumerated in the redoxcline of the central Baltic Sea and up to 9 x 10(4) (14)CO(2)-fixing cells ml(-1) were enumerated in the Black Sea redoxcline, corresponding to 29% and 12%, respectively, of total cell abundance. (14)CO(2)-incorporating cells belonged exclusively to the domain Bacteria. Among these, members of the Epsilonproteobacteria were approximately 70% of the cells in the central Baltic Sea and up to 100% in the Black Sea. For the Baltic Sea, the Sulfurimonas subgroup GD17, previously assumed to be involved in autotrophic denitrification, was the most dominant CO(2)-fixing group. In conclusion, Epsilonproteobacteria were found to be mainly responsible for chemoautotrophic activity in the dark CO(2) fixation maxima of the Black Sea and central Baltic Sea redoxclines. These Epsilonproteobacteria might be relevant in similar habitats of the world's oceans, where high dark CO(2) fixation rates have been measured.

Members of the class Epsilonproteobacteria are known to be of major importance in biogeochemical processes at oxic-anoxic interfaces. In pelagic redoxclines of the central Baltic Sea, an uncultured epsilonproteobacterium related to Sulfurimonas denitrificans was proposed to play a key role in chemolithotrophic denitrification (I. Brettar, M. Labrenz, S. Flavier, J. Bötel, H. Kuosa, R. Christen, and M. G. Höfle, Appl. Environ. Microbiol. 72:1364-1372, 2006). In order to determine the abundance, activity, and vertical distribution of this bacterium in high-resolution profiles, 16S rRNA cloning and catalyzed reporter deposition and fluorescence in situ hybridization (CARD-FISH) and quantitative PCR measurements were carried out. The results showed that 21% of the derived clone sequences, which in the present study were grouped together under the name GD17, had >99% similarity to the uncultured epsilonproteobacterium. A specific gene probe against GD17 (S-*-Sul-0090-a-A-18) was developed and used for enumeration by CARD-FISH. In different pelagic redoxclines sampled during August 2003, May 2005, and February 2006, GD17 cells were always detected from the lower oxic area to the sulfidic area. Maximal abundance was detected around the chemocline, where sulfide and nitrate concentrations were close to the detection limit. The highest GD17 numbers (2 x 10(5) cells ml(-1)), representing up to 15% of the total bacteria, were comparable to those reported for Epsilonproteobacteria in pelagic redoxclines of the Black Sea and the Cariaco Trench (X. Lin, S. G. Wakeham, I. F. Putnam, Y. M. Astor, M. I. Scranton, A. Y. Chistoserdov, and G. T. Taylor, Appl. Environ. Microbiol. 72:2679-2690, 2006). However, in the Baltic Sea redoxclines, Epsilonproteobacteria consisted nearly entirely of cells belonging to the distinct GD17 group. This suggested that GD17 was the best-adapted epsilonproteobacterium within this ecological niche.

Background: S100 proteins, a multigenic family of non-ubiquitous cytoplasmic Ca2+-binding proteins, have been linked to human pathologies in recent years. Dysregulated expression of S100 proteins, including S100A9, has been reported in the epidermis as a response to stress and in association with neoplastic disorders. Recently, we characterized a regulatory element within the S100A9 promotor, referred to as MRE that drives the S100A9 gene expression in a cell type-specific, activation- and differentiation-dependent manner (Kerkhoff et al. (2002) J. Biol. Chem. 277, 41879-41887)., Results: In the present study, we investigated transcription factors that bind to MRE. Using the MRE motif for a pull-down assay, poly(ADP-ribose)polymerase-1 (PARP-1) and the heterodimeric complex Ku70/Ku80 were identified by mass spectrometry and confirmed by chromatin immunoprecipitation. Furthermore, TPA-induced S100A9 gene expression in HaCaT keratinocytes was blocked after the pharmacologic inhibition of PARP-1 with 1,5-isoquinolinediol (DiQ)., Conclusion: The candidates, poly(ADP-ribose)polymerase-1 (PARP-1) and the heterodimeric complex Ku70/Ku80, are known to participate in inflammatory disorders as well as tumorgenesis. The latter may indicate a possible link between S100 and inflammation-associated cancer.

Background: Cardioversion for atrial fibrillation (AF) is associated with impairment of left atrial mechanical function and increased risk of thrombus formation with subsequent embolisation. Measuring atrial mechanical function is of interest to determine the individual risk of thromboembolism and the risk of recurrent AF., Methods: We included 112 consecutive patients with AF and planned cardioversion. Serial echocardiographic measurements of left atrial size and Doppler measurement of mitral valve peak A wave velocities were obtained at days 0, 1, 2, 3, and 28 following cardioversion. These measurements and clinical parameters were related to clinical events and recurrent AF within 4 weeks following cardioversion. Cardioversion was achieved in 100 patients., Results: AF-recurrence within 4 weeks was 23.9% and 45.8% for patients with < or = and > 6 weeks AF-duration, respectively (p=0.04). Peak A wave velocities were significantly lower up to 2 days following cardioversion in patients with AF-recurrence. A peak A wave velocity < 52 cm/s at day 1 resulted in an odds ratio of 5.0 (95% CI: 1.4-18.6) for recurrence of AF. In multiple logistic regression analysis, peak A wave velocity at day 1 remained the only independent predictor of recurrent AF. Left atrial diameter did not correlate with recurrence of AF., Conclusions: A single measurement of mitral peak A wave velocity 1 day following cardioversion is predictive of AF recurrence. This method is feasible for risk estimation with potential therapeutic implications.

Occupational exposure to perchloroethylene (PERC) in dry cleaning is analyzed by calculating airborne concentrations for a large set of possible exposure scenarios. Different generations of dry-cleaning machines ranging from vented machines used until the 1980s to nonvented closed-loop machines used since the 1990s are investigated; the geographical scope of the study is Germany. Concentrations are calculated for diffuse emissions, release of contaminated air during loading and unloading, and residual solvent releases from cleaned garments with a dynamic two-box model for near field and far field. The concentration estimates for near field and far field agree well with measured concentrations. The airborne concentrations are compared with maximum workplace concentrations (MAK values). The full set of scenarios shows for which historical situations the workplace concentrations were near the MAK value and that the transition to newer dry-cleaning machines reduced the occupational exposure by more than one order of magnitude. In addition, numbers of exposed workers are estimated for different years. Whereas more than 25,700 workers in the near field were exposed to PERC in 1975, the number has been below 5900 since 2001, which is due mainly to a reduced amount of garments being dry cleaned and, to a lesser extent, to replacement of PERC by nonchlorinated solvents. The results from the scenario-based analysis are consistent with exposure ranges obtained from a Monte Carlo calculation using frequency distributions of key input parameters, such as emission factors, batch times, and room volumes.

The US Department of Housing and Urban Development (HUD) undertook an evaluation of its Lead Hazard Control Grant Program between 1994 and 1999. The Evaluation is the largest study ever done on the effectiveness of lead hazard controls implemented in residential dwellings. The Evaluation had several major objectives: determining the effectiveness of various lead hazard controls in reducing residential dust lead levels and children's blood lead levels, establishing the costs of doing lead hazard control work and factors that influence those costs, determining the rate of clearance testing failures and their causes, and identifying possible negative effects of lead hazard control work on children's blood lead levels. This paper reports the overall research design and data collection methods of the Evaluation. The large number of dwelling units enrolled in the Evaluation was possible only by the innovative partnership among HUD, the Evaluators, and the grantees. HUD and the Evaluators relied on the grantees for essentially all of the data collection. The 14 participating HUD Lead Hazard Control Grantees were responsible for implementing the lead hazard control programs in their communities and collecting the study data. This paper describes the methods for recruiting and enrolling dwellings and families, collecting environmental and housing data, interviewing participating families, and collecting data on lead hazard control work performed and its costs. The paper also describes the basic quality control and quality assurance procedures used. The principal outcome measures were lead in dust collected using wipes from floors, window sills, and window troughs and lead in blood collected from children who were 6 years old or younger at enrollment. Data collection was conducted before intervention, immediately postintervention, and 6 and 12 months postintervention. For a subset of dwellings undergoing an extended follow-up data were also collected at 24 and 36 months postintervention. This paper provides the context for subsequent reports that will describe such findings as the influence of lead hazard control work on serial dust lead levels, the influence of lead hazard control work on serial blood lead levels in children, the nature and costs of the lead hazard control work done at the dwellings, and the experience of the grantees in meeting clearance testing requirements.

A strategy for combining surface plasmon resonance (SPR) biomolecular interaction analysis, and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is reported. Both techniques are highly complementary but need separate optimization to improve their individual specificity and sensitivity. Sensor surfaces that are optimal for kinetic analysis are not well suited for MALDI-MS and vice versa. In addition, the transfer of analyte from SPR to MS is crucial and often accompanied by sample loss. To address both of these points, a bifunctional SPR fluid cell was constructed where optimized surfaces can be used for binding studies and MS simultaneously with regard to the special need of each technique. The setup guarantees that the SPR and the loading experiment for MS are performed at identical conditions. A removable pin carries the affinity-surface-bound analyte to the mass spectrometer so that handling is minimized, avoiding analyte elution. Functionalized transfer pins can also be used independently of SPR for microaffinity capture-MS.

Purpose: Patients with advanced hepatocellular carcinoma (HCC) have shown to benefit from tamoxifen treatment. The mechanisms of tamoxifen action in HCC, however, are not yet clearly understood. Results from studies on the human hepatoblastoma cell line HepG2 provide evidence that estrogen-receptor-alpha-independent antiproliferative actions of tamoxifen in HCC are mediated by the suppression of telomerase activity [5]., Materials and Methods: We investigate the pathway of the tamoxifen-induced down-regulation of telomerase activity, using HepG2 cells incubated over 24 h or 48 h in the presence of 20 microM tamoxifen., Results: The transcriptional levels of the three telomerase core components-human telomerase RNA (hTR), human telomerase reverse transcriptase (hTERT) (all variants), and telomerase-associated protein (TP1)-did not change during tamoxifen treatment, as revealed by RT-PCR analysis. Furthermore, the hTERT splice pattern was not shifted from the active full-length variant (+alpha/+beta) to the inactive deletion variants (-alpha; -beta; -alpha/-beta) and the level of the 120 kDa hTERT full-length protein remained constant, as shown by Western blot analysis. Protein kinase C (PKC) activity has been suggested to be crucial for post-translational up-regulation of telomerase activity. In HepG2 cells, we observed a tamoxifen-induced suppression of the total protein kinase C (PKC) activity (cytosolic and membrane-bound). Inhibition of PKC with bisindolylmaleimide I resulted in a reduction of telomerase activity, as revealed by TRAP-assay. Alpha-tocopherol (vitamin E) diminished the effects of tamoxifen on PKC-activity as well as on telomerase activity., Conclusions: We conclude that the tamoxifen-induced decrease of telomerase activity in HepG2 cells is mediated post-translationally via suppression of PKC-activity.

This study is an examination of the effect of lead hazard control strategies on children's blood lead levels immediately after an intervention was conducted as part of the US Department of Housing and Urban Development's Lead-Based Paint Hazard Control Grant Program. Fourteen state and local government grantees participated in the evaluation. The findings indicated an overall average reduction in the blood lead levels of 869 children soon after the implementation of lead hazard controls. However, 9.3% of these children (n = 81) had blood lead increases of 5 microg/dL or more. Data routinely collected as part of the evaluation, as well as additional information supplied by the individual programs, were used to determine potential reasons for these observed increases in blood lead. A logistic regression analysis indicated that three principal factors were associated with the blood lead increases: the number of exterior deteriorations present in the child's home (prior to intervention), the educational level of the female parent or guardian of the child, and the child's age. The statistical analysis did not find evidence that children living in households that either did not relocate or relocated for less than the full work period were significantly more likely to have a blood lead increase equal to or greater than 5 microg/dL than children living in households that fully relocated. Statistical analyses also did not reveal any single interior strategy to be more or less likely than others to be associated with a blood lead increase of 5 microg/dL or more.

Many studies of otitis media with effusion (OME) have been published, most of them dealing with risk factors. The literature correlates this condition with various patient characteristics and socio-economic factors, but none of these have been identified as the sole causative factor. A review of the literature suggests that otitis media with effusion is a response to pathogenic bacteria and thus a normal protective reaction of the body. Therefore, the decision on whether or not treatment is indicated should take the natural course of that response into account. In light of the literature reviewed here, we conclude that there is usually no need to treat middle ear effusion in young children.

Background/aims: Antiproliferative action of tamoxifen in the estrogen receptor-alpha-negative human hepatoblastoma cell line HepG2 was investigated., Methods: HepG2 cells, seeded at different densities (4000-36 000 cells/cm(2)), were incubated with tamoxifen (1, 10, or 20 microM) or the telomerase inhibitor 3'-azido-3'-deoxythymidine (AZT) (0.6-3.0 mM) up to 72 h. Cell viability was assessed (MTT-test), flow cytometric analysis was performed, and telomerase activity was measured (telomeric repeat amplification protocol assay)., Results: Ten or 20 microM tamoxifen induced a reduction of cell viability. Basically reduction of viability was related to an increase in the fraction of G0/1-phase. When tamoxifen was present at higher concentration (20 microM) or at low cell density (4000/cm(2)) an additional increase of the rate of apoptotic cells occurred with a delay, aggravating the effect of tamoxifen on cell viability substantially. When apoptosis was induced a significant suppression of telomerase activity preceded regularly. Direct inhibition of telomerase activity with AZT resulted in a decrease of cell viability and apoptosis., Conclusion: The tamoxifen-induced reduction of cell viability in HepG2 cells depends on drug concentration and cell density and is due to cytostatic and cytocide effects. The latter may be mediated by a down-regulation of telomerase activity.

Occupational exposure to trichloroethylene (TRIC) and perchloroethylene (PERC) in metal degreasing is analyzed by calculating airborne concentrations for a large set of possible exposure scenarios (Scenario-Based Risk Assessment, SceBRA). Different types of degreasing machines ranging from open-top machines used until the 1980s to closed-loop nonvented machines used since the 1990s are investigated; the scope of the study is Germany. Concentrations are calculated for different kinds of releases (emissions from open baths, leakage, release of contaminated air during loading and unloading) with a dynamic two-box model for the near-field and the far-field. The concentration estimates are in good agreement with measured data. The airborne concentrations are compared to maximum workplace concentrations (MAK values). The full set of scenarios shows for which situations MAK values were exceeded and how the transition to newer degreasing machines reduced the occupational exposure by more than one order of magnitude. In addition, numbers of exposed workers are estimated for different years. While more than 25,000 workers in the near-field were exposed to TRIC and PERC in 1985, the number is below 3000 since 1996, which is mainly due to technology changes, rationalization, automatization, and replacement of TRIC and PERC by nonchlorinated solvents.

Reaction of a rhodamine 2'-ester with an excess of alkyldiamines provides amino-functionalized rhodamine spirolactams, which when subsequently conjugated with carboxyfluorescein, provides probes which are fluorescent at acidic, neutral, and basic pH ranges.

The impact of teaching interventions and the availability of additional dispensers for alcoholic hand disinfection was measured during three study periods. A nursing care system of relevant nursing interventions was found to be a useful tool for the assessment of compliance with hand hygiene requirements if combined with an analysis of hand disinfectant use. This study shows that compliance with hand disinfection can be improved through targeted teaching and supportive structural improvements, and that this positive impact may persist even after targeted teaching activities have ended., (Copyright 2002 The Hospital Infection Society.)

The risk through chemical exposure is commonly characterized by ratios of exposure concentrations and effect levels (risk quotients). For chemicals with many different applications such as solvents, however, in addition to the risk quotients of different exposure situations it is useful to determine the corresponding numbers of exposed individuals, that is, not only the magnitude but also the extent of the risk. To this end, the Scenario-Based Risk Assessment (SceBRA) method has been developed that makes use of a large set of scenarios, each of which describes a typical situation regarding handling a solvent or solvent-containing product. The scenarios cover the life-cycle steps of production, distribution, and use of solvents. For each scenario, SceBRA provides the risk quotient, r, and the number of exposed individuals, N. This study investigated seven solvents that are used in large amounts in Switzerland. For each solvent, characteristic distributions of r and N values were calculated, making it possible to compare different solvents with respect to their risk profile. Graphical representations of the r, N data provide an informative way for analyzing and communicating the results of SceBRA.

Otitis media with effusion (OME) is characterized by the presence of fluid in the middle ear without signs or symptoms of acute infection and by persistent changes in the middle ear mucosa. These are mainly induced by gram-negative bacterial infection and dysfunction of the eustachian tube (ET). Gram-negative bacteria (GNB) contain lipopolysaccharide (LPS) in their outer membrane that is responsible for inflammatory reactions in the middle ear. In this study we investigated the therapeutic effect of a recombinant LPS-binding protein, bactericidal/permeability-increasing protein (rBPI21), on the repair of mucosal damage in rats with experimentally induced OME. OME was induced by obstruction of the eustachian tube in combination with LPS injection. Twelve weeks after OME induction, secretory cells in the tympanic orifice of the middle ear were increased from an average of 14 +/- 2 to 31 +/- 5, ciliated cells were decreased from 24 +/- 4 to 6 +/- 4, and the number of macrophages in the subepithelial layer increased from 13 +/- 4 to 27 +/- 3. A single dose of rBPI21 was administered directly into the middle ear cavity 2 weeks after the induction of OME. Histologic examination of the middle ear mucosa at 4 and 12 weeks after OME induction showed that mucosal changes were restored by rBPI21 treatment. These results demonstrate that the middle ear mucosa recovers from inflammatory changes associated with OME after treatment with rBPI21. This suggests that rBPI21 may be useful in the treatment of OME and of mucosal infections of the respiratory tract.

This is a randomized, double-blind, controlled study of the effects of aprotinin (Trasylol) during primary total hip arthroplasty. Sixty patients were randomized to receive either 1.5 x 10(6) KIU of aprotinin or a similar volume of normal saline as a bolus preoperatively. Blood loss was measured from the femoral canal at the time of surgery. An estimate of the total blood loss during the operation was made, and the transfusion requirement was recorded. There was no significant difference between the groups in terms of total blood loss, postoperative hemoglobin, or transfusion requirement. In the group that received aprotinin, there was a trend toward reduced blood loss from the femoral canal, but this was not statistically significant. The results of this study do not support the routine use of aprotinin in primary total hip arthroplasty.

Hypothesis/background: Endotoxin can induce morphologic changes to middle ear epithelium, which can disturb the mucociliary clearance system (MCS) and lead to otitis media with effusion (OME). The bactericidal/permeability-increasing (BPI) protein is a major component of neutrophil granules and binds with high affinity to endotoxin. In this study, the capacity to inhibit the effects of endotoxin by rBPI21, a recombinant amino-terminal analog derived from BPI, was investigated on cultured human middle ear epithelium using light microscopy and scanning- and transmission electron microscopy., Methods: Human middle ear epithelium was air-exposed cultured on a collagenous underlayer with different additions of endotoxin and rBPI21 to the culture medium. The tissue specimens were inspected after 4 weeks for the number of ciliated and secretory cells, thickness of the mucosal layer, and cell size., Results: The morphologic changes induced by endotoxin were increased thickness of the mucosal layer and increased number of secretory cells. These changes were significantly diminished or even absent when endotoxin was added with rBPI21 to the culture medium., Conclusion: rBPI21 can inhibit morphologic changes in the middle ear epithelium due to endotoxin. Hence, the authors believe that rBPI21 can be a new therapeutic agent in the treatment of OME.

O-(Acridinium)hydroxylamine (AHA) reacted with a representative sample of oxo-steroids (6-oxoestradiol, estrone, norethindrone, cortisol, progesterone, digoxin dialdehyde, and digitoxin dialdehyde) to produce chemiluminescent acridinium oxime (AO) conjugates in a single step in 37-68% yield after preparative HPLC.

Conventional pure-tone thresholds were collected as determined at ages between 4 and 8 years from a group of 163 infants, tested by auditory brainstem response (ABR) in the age range between 1 and 3 years old for objective hearing assessment. The subjects suffered from a variety of degrees and types of sensorineural hearing impairment. The prognostic value of the ABR peak V thresholds in response to 0.1 ms clicks with respect to the behavioural thresholds at octave frequencies from 125 to 8,000 Hz obtained later is evaluated. Correlation between ABR and behavioural thresholds is largest in the 1,000- to 8,000-Hz frequency range. Predicted pure-tone audiograms (mean and SD) were determined for each 10-dB class of ABR thresholds. SDs are in the order of 15 to 18 dB in the 500- to 4,000-Hz range and slightly higher at adjacent frequencies (i.e., somewhat larger than in comparable adult studies). Mean pure-tone thresholds in the 1,000- to 8,000-Hz frequency range are up to 20 dB worse than ABR thresholds, which is opposite to findings in normally-hearing subjects. Thus, with an increasing degree of sensorineural hearing impairment, pure-tone thresholds increase at a significantly higher rate than ABR thresholds. The observation is explained in terms of reduced temporal integration in cochlear hearing loss. ABR thresholds worse than 80 dB nHL are demonstrated to have very limited predictive value with respect to the amount of residual hearing, not only in the low- but also in the high-frequency range. The presence of otitis media during ABR testing is shown to make estimation errors increase to more than 25 dB (SD).

Objectives/hypothesis: This study was performed to elucidate the role of endotoxin and tumor necrosis factor-alpha (TNF-alpha) in the middle ear effusions (MEEs) of children with otitis media with effusion (OME) in relation to the chronicity of the disease and the presence of upper respiratory tract infection (URTI)., Study Design: In a retrospective study 140 MEEs were collected from 101 children between 2 and 12 years of age, and evaluated for the cytokine TNF-alpha and the lipopolysaccharide endotoxin. The amounts were quantified and correlated with the type of MEE, OME duration, and the presence of URTI., Methods: Endotoxin levels were measured using a limulus amebocyte lysate assay and TNF-alpha levels were measured with an enzyme-linked immunosorbent assay (ELISA). Means of the different variables were compared using the one-way ANOVA least significance difference test with P<.05., Results: In MEEs classified as mucopurulent (22.8%) both endotoxin and TNF-alpha levels (11.9+/-3 ng/mg total protein and 61.1+/-21 pg/mg total protein, respectively) were significantly higher compared with serous- (23.6%) or mucoid- (53.6%) type effusions. Fifty-five percent of the children who were classified as having chronic OME also had significantly higher amounts of endotoxin and TNF-alpha. The majority of the children (61%) had no URTI, although children with URTI (36%) did also have significantly higher levels of endotoxin and TNF-alpha in their middle ears., Conclusions: These results indicate that there is a strong correlation between the endotoxin and the TNF-alpha concentration in the middle ear and the type of MEE, the presence of URTI, and the chronicity of the disease.

The effect of endotoxin was investigated on air-exposed cultured human middle ear epithelium. Concentrations of 0, 1 and 100 ng/ml endotoxin were used. Complete differentiation of the cells was not reached at 12 days. After 21 days, endotoxin had induced an increased proliferation of the epithelial layer. Furthermore, an increase in the number of secretory cells and in the amount and length of microvilli was observed at this time. There were no significant morphological differences between the high and the low endotoxin concentrations, which supports our hypothesis that endotoxin induces an all-or-nothing reaction. These findings are in agreement with our previous results on serially submerged cultured rat middle ear epithelium. From these results we conclude that endotoxin is an important factor in the disturbance of the morphology of the middle ear epithelium, which may lead to chronic otitis media with effusion. In addition, our tissue culture method proved to be a good model for further studies on human middle ear mucosa.