Validation of an osteoporotic animal model for dental implant analyses: an in vivo densitometric study in rabbits.
The achievement of primary stability in porous and soft bone, where implants are more likely to fail, is one of the unresolved challenges of implant dentistry. Therefore, the aim of the study was to validate an osteoporotic animal model for analysis of poor-quality bone. Sixteen female New Zealand rabbits, each 6 months old and weighing 4 to 5 kg, were used in this study. The animals were anesthetized, and an in vivo densitometric analysis was performed by dual-energy x-ray absorptiometry (DEXA) to measure bone mineral density (BMD) in the calvaria, cervical spine, and tibia. Ovariectomy was then performed, and animals were fed a low-calcium diet that featured 0.07% calcium, rather than the 0.45% calcium of a standard diet, for 6 weeks. After this period, new densitometric measurements were carried out. Two-way analysis of variance was used for statistical evaluation. A P value of less than .05 was considered to be significant. Together, ovariectomy and a low-calcium diet were able to induce a quick decrease in BMD, as measured at 6 weeks by DEXA. This decrease was statistically significant in the calvaria (P < .001) and the cervical spine (P < .05) but not in the tibia. Based upon this study, ovariectomy and a low-calcium diet are able to induce experimental osteoporosis in rabbits in a short period of time.


Linear Free Energy Relationship Analysis of Transition State Mimicry by 3-Deoxy-d-arabino-heptulosonate-7-phosphate (DAHP) Oxime, a DAHP Synthase Inhibitor and Phosphate Mimic.
3-Deoxy-d-arabino-heptulosonate-7-phosphate (DAHP) synthase catalyzes an aldol-like reaction of phosphoenolpyruvate (PEP) with erythrose 4-phosphate (E4P) to form DAHP in the first step of the shikimate biosynthetic pathway. DAHP oxime, in which an oxime replaces the ketone, is a potent inhibitor, with Ki = 1.5 μM. Linear free energy relationship (LFER) analysis of DAHP oxime inhibition using DAHP synthase mutants revealed an excellent correlation between transition state stabilization and inhibition. The equations of LFER analysis were rederived to formalize the possibility of proportional, rather than equal, changes in the free energies of transition state stabilization and inhibitor binding, in accord with the fact that the majority of LFER analyses in the literature demonstrate nonunity slopes. A slope of unity, m = 1, indicates that catalysis and inhibitor binding are equally sensitive to perturbations such as mutations or modified inhibitor/substrate structures. Slopes <1 or >1 indicate that inhibitor binding is less sensitive or more sensitive, respectively, to perturbations than is catalysis. LFER analysis using the tetramolecular specificity constant, that is, plotting log(KM,MnKM,PEPKM,E4P/kcat) versus log(Ki), revealed a slope, m, of 0.34, with r2 = 0.93. This provides evidence that DAHP oxime is mimicking the first irreversible transition state of the DAHP synthase reaction, presumably phosphate departure from the tetrahedral intermediate. This is evidence that the oxime group can act as a functional, as well as structural, mimic of phosphate groups.


Detection and quantitation of human papillomavirus type 16, 18 and 52 DNA in the peripheral blood of cervical cancer patients.
To prospectively evaluate the feasibility of detecting human papillomavirus (HPV) type 16, 18 and 52 DNA in the peripheral blood of patients with cervical cancer using real-time polymerase chain reaction (PCR) and to determine its prognostic importance. Blood and cervical swab specimens from 135 consecutive patients with 60 invasive cervical cancers, 10 microinvasions, 20 cervical intraepithelial neoplasias (CIN) III, 10 CIN II, 10 CIN I and 25 controls were collected and examined for HPV type 16, 18 and 52 DNA using real-time PCR to investigate the prevalence and viral load of HPV DNA at the time of diagnosis and during follow-up in patients with positive blood samples. Of the 60 patients with invasive cervical cancer, 27% had positive test results for HPV DNA in blood samples in contrast to 0% of patients with microinvasions, CIN III, CIN II, CIN I and normal controls. The DNA detection rates of viral subtypes in blood samples of cervical cancer patients were 5% for HPV-16, 16.7% for HPV-18, 8.3% for HPV-52, 1.7% for both HPV-16 and HPV-18 and 1.7% for both HPV-18 and HPV-52, while the detection rates in cervical swab specimens were 36.2% for HPV-16, 15.5% for HPV-18 and 17.2% for HPV-52. During follow-up, 8 of 10 cervical cancer patients with viral DNA detected in blood within 3 months after treatment had recurrence, and a high percentage (87.5%, 7/8) of this recurrence involved distant metastases. In this study, real-time PCR detected HPV-16, -18 or -52 DNA in the peripheral blood of more than one-fourth of invasive cervical cancer patients. The association between risk of cancer recurrence and the amount of viral DNA detected in blood among cervical cancer patients after treatment is intriguing and deserves further investigation.


Manipulation of the vsg co-transposed region increases expression-site switching in Trypanosoma brucei.
Disruption of a region of DNA in Trypanosoma brucei immediately upstream of the expressed telomere-proximal variant surface glycoprotein gene (vsg), known as the co-transposed region (CTR), can cause a dramatic increase in the rate at which the active expression site (ES) is switched off and a new ES is switched on. Deletion of most of the CTR in two ESs caused a greater than 100-fold increase in the rate of ES switching, to about 1.3 x 10(-4) per generation. A more dramatic effect was observed when the entire CTR and the 5' coding region of the expressed vsg221 were deleted. In this case a new ES was activated within a few cell divisions. This switch also occurred in cell lines where a second vsg had been inserted into the ES, prior to CTR deletion. These cell lines, which stably co-expressed the inserted and endogenous Vsgs, in equal amounts, did not differ from the wild-type in growth rate or switching frequency, suggesting that simultaneous expression of two Vsgs has no intrinsic effect. CTR deletion did not disturb the inserted vsg117. We tentatively conclude that it was not the disruption of the vsg221 in itself that destabilized the ES. All of the observed switches occurred without additional detectable DNA rearrangements in the switched ES. Deletion of the 70-bp repeats and/or a vsg pseudogene upstream of the CTR did not affect ES stability. Several speculative interpretations of these observation are offered, the most intriguing of which is that the CTR plays some role in modulating chromatin conformation at an ES.


Cathepsin E-deficient mice show increased susceptibility to bacterial infection associated with the decreased expression of multiple cell surface Toll-like receptors.
Cathepsin E, an intracellular aspartic proteinase, is predominantly localized in the endosomal compartments of immune system cells. In the present study, we investigated the role of cathepsin E in immune defense systems against bacterial infection. Cathepsin E-deficient (CatE(-/-)) mice showed dramatically increased susceptibility to infection with both the Gram-positive bacterium Staphyrococcus aureus, and the Gram-negative bacterium Porphyromonas gingivalis when compared with syngeneic wild-type mice, most likely due to impaired regulation of bacterial elimination. Peritoneal macrophages from CatE(-/-) mice showed significantly impaired tumor necrosis factor-alpha and IL-6 production in response to S. aureus and decreased bactericidal activities toward this bacterium. Moreover, the cell surface levels of Toll-like receptor-2 (TLR2) and TLR4, which recognize specific components of Gram-positive and -negative bacteria, respectively, were decreased in CatE(-/-) macrophages, despite no significant difference in the total cellular expression levels of these receptors between the wild-type and CatE(-/-) macrophages, implying trafficking defects in these surface receptors in the latter. These results indicate an essential role of cathepsin E in immune defense against invading microorganisms, most probably due to regulation of the cell surface expression of TLR family members required for innate immune responses.


What does quality care mean to nurses in rural hospitals?
This paper is a report of a study conducted to answer the question: 'How do rural nurses and their chief nursing officers define quality care?' Established indicators of quality care were developed primarily in urban hospitals. Rural hospitals and their environments differ from urban settings, suggesting that there might be differences in how quality care is defined. This has measurement implications. Focus groups with staff nurses and interviews with chief nursing officers were conducted in 2006 at four rural hospitals in the South-Eastern United States of America. Data were analysed using conventional content analysis. The staff nurse and chief nursing officer data were analysed separately and then compared, exposing two major themes: 'Patients are what matter most' and 'Community connectedness is both a help and a hindrance'. Along with conveying that patients were the utmost priority and all care was patient-focused, the first theme included established indicators of quality such as falls, pressure ulcers, infection rates, readmission rates, and lengths of stay. A new discovery in this theme was a need for an indicator relevant for rural settings: transfer time to larger hospitals. The second theme, Community Connectedness, is unique to rural settings, exemplifying the rural culture. The community and hospital converge into a family of sorts, creating expectations for quality care by both patients and staff that are not typically found in urban settings and larger hospitals. Established quality indicators are appropriate for rural hospitals, but additional indicators need to be developed. These must include transfer times to larger facilities and the culture of the community.


Joint assembly and genetic mapping of the Atlantic horseshoe crab genome reveals ancient whole genome duplication.
Horseshoe crabs are marine arthropods with a fossil record extending back approximately 450 million years. They exhibit remarkable morphological stability over their long evolutionary history, retaining a number of ancestral arthropod traits, and are often cited as examples of "living fossils." As arthropods, they belong to the Ecdysozoa, an ancient super-phylum whose sequenced genomes (including insects and nematodes) have thus far shown more divergence from the ancestral pattern of eumetazoan genome organization than cnidarians, deuterostomes and lophotrochozoans. However, much of ecdysozoan diversity remains unrepresented in comparative genomic analyses. Here we apply a new strategy of combined de novo assembly and genetic mapping to examine the chromosome-scale genome organization of the Atlantic horseshoe crab, Limulus polyphemus. We constructed a genetic linkage map of this 2.7 Gbp genome by sequencing the nuclear DNA of 34 wild-collected, full-sibling embryos and their parents at a mean redundancy of 1.1x per sample. The map includes 84,307 sequence markers grouped into 1,876 distinct genetic intervals and 5,775 candidate conserved protein coding genes. Comparison with other metazoan genomes shows that the L. polyphemus genome preserves ancestral bilaterian linkage groups, and that a common ancestor of modern horseshoe crabs underwent one or more ancient whole genome duplications 300 million years ago, followed by extensive chromosome fusion. These results provide a counter-example to the often noted correlation between whole genome duplication and evolutionary radiations. The new, low-cost genetic mapping method for obtaining a chromosome-scale view of non-model organism genomes that we demonstrate here does not require laboratory culture, and is potentially applicable to a broad range of other species.


Angiotensin II enhances hyperpolarization-activated currents in rat aortic baroreceptor neurons: involvement of superoxide.
As an endogenous physiologically active peptide, angiotensin (ANG) II plays an important role in the maintenance of blood pressure. In the arterial baroreceptor reflex (a pivotal regulator of blood pressure), aortic baroreceptor (AB) neurons located in the nodose ganglia (NG) are a primary afferent limb of the baroreflex. Hyperpolarization-activated currents (I(h)) in the AB neurons contribute to the excitability of the AB neurons. Therefore, the present study was to measure the modulating effect of ANG II on the I(h) in the primary AB neurons isolated from rats. Data from immunofluorescent and Western blot analyses showed that protein of AT(1) and AT(2) receptors was expressed in the nodose neurons. In the whole cell patch-clamp recording, ANG II concentration dependently enhanced the I(h) density in the AB neurons (100 nM ANG II-induced 53.8 +/- 3.8% increase for A-type AB neurons and 30.4 +/- 7.7% increase for C-type AB neurons at test pulse -140 mV, P < 0.05). ANG II also decreased membrane excitability in the AB neurons. AT(1) receptor antagonist (1 muM losartan) but not AT(2) receptor antagonist (1 muM PD-123,319) totally abolished the effect of ANG II on the I(h) and neuronal excitability. In addition, NADPH oxidase inhibitor (100 muM apocynin) and superoxide scavenger (1 mM tempol) also significantly blunted the ANG II-induced increase of the I(h) and decrease of the membrane excitability in the AB neurons. Furthermore, losartan, apocynin, or tempol significantly attenuated the superoxide overproduction in the NG tissues induced by ANG II. These results suggest that ANG II-NADPH oxidase-superoxide signaling can activate the I(h) and subsequently decrease the membrane excitability of rat AB neurons.


Vibrationally resolved sum-frequency generation with broad-bandwidth infrared pulses.
We present a novel procedure for vibrationally resolved sum-frequency generation (SFG) in which a broad-bandwidth IR pulse is mixed with a narrow-bandwidth visible pulse. The resultant SFG spectrum is dispersed with a spectrograph and detected in parallel with a scientific-grade CCD detector, permitting rapid and high signal-to-noise ratio data acquisition over a 400-cm(-1) spectral region without scanning the IR frequency. Application to the study of a self-assembled monolayer of octadecanethiol is discussed.


Establishment of serum protein pattern for screening colorectal cancer using SELDI-TOF-MS.
The purpose of this study is to develop a proteomic pattern for distinguishing individuals with colorectal cancer from healthy controls and monitoring micrometastasis using SELDI-TOF-MS. A training set consisting of 63 patients with colorectal cancer, 20 patients with benign colorectal diseases and 26 healthy volunteers was used to develop a proteomic model that discriminated colorectal cancer effectively. The sensitivity and specificity of this model was validated by an independent test set. To explore serum proteins changed after operation, the protein profiles of 31 postoperative patients were compared with those of preoperative patients. We also analyzed protein profiles of patients with and without metastasis to monitor micrometastasis. Our study yielded a four-peak model (m/z: 3191.5, 3262.9, 3396.3 and 5334.4) that discriminated cancer from non-cancer samples with sensitivity of 90.3% and specificity of 95.7%. This model was validated in the test set with sensitivity of 87.5% and specificity of 93.8% which was significantly better than the combination use of CEA, CA199 and CA242 (sensitivity 62.4%) for early detection of colorectal cancer. Two peaks (m/z: 2753.8 and 4172.4) were found down-regulated in postoperative samples comparing with preoperative samples. We also detected two proteins (m/z: 9184.4 and 9340.9) that can discriminate patients with primary colorectal cancer from metastatic colorectal cancer. The four-peak model and two peaks (m/z: 2753.8 and 4172.4) detected in this study have the potential for assistance in diagnostics and therapeutic strategies in colorectal cancer and the two proteins (m/z: 9184.4 and 9340.9) were effective biomarkers for monitoring micrometastasis.


Relation Between Different Measures of Glycemic Exposure and Microvascular and Macrovascular Complications in Patients with Type 2 Diabetes Mellitus: An Observational Cohort Study.
This retrospective cohort study investigated the relation between different measures of glycemic exposure and micro- and macrovascular complications among patients with type 2 diabetes. The analysis included patients receiving oral antihyperglycemic agents between 1 January 2006 and 31 December 2014 from the General Practitioner Database from the PHARMO Database Network. All recorded HbA1c levels during follow-up were used to express glycemic exposure in four ways: index HbA1c, time-dependent HbA1c, exponential moving average (EMA) and glycemic burden. Association between glycemic exposure and micro-/macrovascular complications was analyzed by estimating hazard ratios and 95% confidence intervals using an adjusted (time-dependent) Cox proportional hazards model. The analysis included 32,725 patients (median age, 65 years; 47% female). Median follow-up was 5.4 years; median number of HbA1c measurements per patient was 18.0. From all measures, HbA1c at index showed the weakest relation between all micro-/macrovascular complications, with coronary artery disease (CAD) having the highest HR (95% CI): 1.18 (1.04-1.34) for HbA1c ≥64 mmol/mol (8%). The time-dependent HbA1c model showed a significant association only for microvascular complications, with retinopathy having the highest HR (95% CI): 1.55 (1.40-1.73) for HbA1c ≥64 mmol/mol (8%). EMA-defined exposure showed similar findings, although the effect of retinopathy was more pronounced [HR (95% CI): 1.81 (1.63-2.02) for HbA1c ≥64 mmol/mol (8%)] and was also predictive for CAD [HR (95% CI): 1.29 (1.10-1.50) for HbA1c ≥64 mmol/mol (8%)]. A statistically significant relation with glycemic burden was found for all selected micro-/macrovascular complications, with retinopathy having the highest HR (95%): 2.60 (2.19-3.07) for glycemic burden years >3. This study shows that greater and more prolonged exposure to hyperglycemia increases the risk of micro- and macrovascular complications. Janssen Pharmaceutica NV.


Five-Lumen Antibiotic-Impregnated Femoral Central Venous Catheters in Severely Burned Patients: An Investigation of Device Utility and Catheter-Related Bloodstream Infection Rates.
The objective of this study is to determine the catheter-related bloodstream infection (CRBSI) rate in a severely burned patient population, many of whom required prolonged use of central venous catheters (CVCs). Between January 2008 and June 2012, 151 patients underwent placement of 455 five-lumen minocycline/rifampin-impregnated CVCs. CRBSI was defined as at least one blood culture (>100,000 colonies) and one simultaneous roll-plate CVC tip culture (>15 colony forming units) positive for the same organism. Most patients had accidental burns (81.5%) with a mean TBSA of 50%. A mean of three catheters were inserted per patient (range, 1-25). CVCs were inserted in the femoral vein (91.2%), subclavian vein (5.3%), and internal jugular vein (3.3%). Mean overall catheter indwell time was 8 days (range, 0-39 days). The overall rate of CRBSI per 1000 catheter days was 11.2; patients with a TBSA >60% experienced significantly higher rates of CRBSI than patients with a TBSA ≤60% (16.2 vs 7.3, P = .01). CVCs placed through burned skin were four times more likely to be associated with CRBSI than CVCs placed through intact skin. The most common infectious organism was Acinetobacter baumannii. Deep venous thrombosis developed in eleven patients (7%). The overall rate of CRBSI was 11.2, consistent with published rates of CRBSI in burn patients. Thus, femoral placement of 5-lumen CVCs did not result in increased CRBSI rates. These data support the safety of femoral CVC placement in burn patients, contrary to the Centers for Disease Control recommendation to avoid femoral CVC insertion.


T2* mapping of articular cartilage: current status of research and first clinical applications.
T2* mapping is a relatively new method for the compositional assessment of the articular cartilage. Typically, a multigradient echo or an ultrashort echo time imaging technique with a range of short and very short echo times is used. In most studies, imaging is performed at a high field strength, that is, 3 and 7 T. Postprocessing includes exponential fitting of relaxation decay and manual region-of-interest-based measurements of T2* times on T2* maps. Detailed analyses of T2* times of articular cartilage have shown distinct T2* components with shorter and longer T2* times. Moreover, there is a zonal distribution with a significant depthwise gradient of T2*, with relatively short times near the osteochondral junction and relatively long times at the cartilage's surface. T2* times of normal articular cartilage at the knee are, when averaged over the whole cartilage thickness and using monoexponential fitting, approximately 20 milliseconds. The results of recent studies have shown a good test-retest as well as interreader and intrareader reliabilities for T2* mapping. This article provides a descriptive review of the current literature, briefly discusses the technique itself, and provides an outlook on future research questions and possible clinical applications.


Assessment of Minimal Residual Disease in Standard-Risk AML.
Despite the molecular heterogeneity of standard-risk acute myeloid leukemia (AML), treatment decisions are based on a limited number of molecular genetic markers and morphology-based assessment of remission. Sensitive detection of a leukemia-specific marker (e.g., a mutation in the gene encoding nucleophosmin [NPM1]) could improve prognostication by identifying submicroscopic disease during remission. We used a reverse-transcriptase quantitative polymerase-chain-reaction assay to detect minimal residual disease in 2569 samples obtained from 346 patients with NPM1-mutated AML who had undergone intensive treatment in the National Cancer Research Institute AML17 trial. We used a custom 51-gene panel to perform targeted sequencing of 223 samples obtained at the time of diagnosis and 49 samples obtained at the time of relapse. Mutations associated with preleukemic clones were tracked by means of digital polymerase chain reaction. Molecular profiling highlighted the complexity of NPM1-mutated AML, with segregation of patients into more than 150 subgroups, thus precluding reliable outcome prediction. The determination of minimal-residual-disease status was more informative. Persistence of NPM1-mutated transcripts in blood was present in 15% of the patients after the second chemotherapy cycle and was associated with a greater risk of relapse after 3 years of follow-up than was an absence of such transcripts (82% vs. 30%; hazard ratio, 4.80; 95% confidence interval [CI], 2.95 to 7.80; P<0.001) and a lower rate of survival (24% vs. 75%; hazard ratio for death, 4.38; 95% CI, 2.57 to 7.47; P<0.001). The presence of minimal residual disease was the only independent prognostic factor for death in multivariate analysis (hazard ratio, 4.84; 95% CI, 2.57 to 9.15; P<0.001). These results were validated in an independent cohort. On sequential monitoring of minimal residual disease, relapse was reliably predicted by a rising level of NPM1-mutated transcripts. Although mutations associated with preleukemic clones remained detectable during ongoing remission after chemotherapy, NPM1 mutations were detected in 69 of 70 patients at the time of relapse and provided a better marker of disease status. The presence of minimal residual disease, as determined by quantitation of NPM1-mutated transcripts, provided powerful prognostic information independent of other risk factors. (Funded by Bloodwise and the National Institute for Health Research; Current Controlled Trials number, ISRCTN55675535.).


Standardization of the Food Composition Database Used in the Latin American Nutrition and Health Study (ELANS).
Between-country comparisons of estimated dietary intake are particularly prone to error when different food composition tables are used. The objective of this study was to describe our procedures and rationale for the selection and adaptation of available food composition to a single database to enable cross-country nutritional intake comparisons. Latin American Study of Nutrition and Health (ELANS) is a multicenter cross-sectional study of representative samples from eight Latin American countries. A standard study protocol was designed to investigate dietary intake of 9000 participants enrolled. Two 24-h recalls using the Multiple Pass Method were applied among the individuals of all countries. Data from 24-h dietary recalls were entered into the Nutrition Data System for Research (NDS-R) program after a harmonization process between countries to include local foods and appropriately adapt the NDS-R database. A food matching standardized procedure involving nutritional equivalency of local food reported by the study participants with foods available in the NDS-R database was strictly conducted by each country. Standardization of food and nutrient assessments has the potential to minimize systematic and random errors in nutrient intake estimations in the ELANS project. This study is expected to result in a unique dataset for Latin America, enabling cross-country comparisons of energy, macro- and micro-nutrient intake within this region.


Compression regulates mitotic spindle length by a mechanochemical switch at the poles.
Although the molecules involved in mitosis are becoming better characterized, we still lack an understanding of the emergent mechanical properties of the mitotic spindle. For example, we cannot explain how spindle length is determined. To gain insight into how forces are generated and responded to in the spindle, we developed a method to apply controlled mechanical compression to metaphase mitotic spindles in living mammalian cells while monitoring microtubules and kinetochores by fluorescence microscopy. Compression caused reversible spindle widening and lengthening to a new steady state. Widening was a passive mechanical response, and lengthening was an active mechanochemical process requiring microtubule polymerization but not kinesin-5 activity. Spindle morphology during lengthening and drug perturbations suggested that kinetochore fibers are pushed outward by pole-directed forces generated within the spindle. Lengthening of kinetochore fibers occurred by inhibition of microtubule depolymerization at poles, with no change in sliding velocity, interkinetochore stretching, or kinetochore dynamics. We propose that spindle length is controlled by a mechanochemical switch at the poles that regulates the depolymerization rate of kinetochore fibers in response to compression and discuss models for how this switch is controlled. Poleward force appears to be exerted along kinetochore fibers by some mechanism other than kinesin-5 activity, and we speculate that it may arise from polymerization pressure from growing plus ends of interpolar microtubules whose minus ends are anchored in the fiber. These insights provide a framework for conceptualizing mechanical integration within the spindle.


Rhabdoid tumor growth is inhibited by flavopiridol.
Rhabdoid tumors are aggressive and incurable pediatric malignancies. INI1/hSNF5, a tumor suppressor biallelically deleted/inactivated in rhabdoid tumors, directly represses cyclin D1. Rhabdoid tumors and cells are exquisitely dependent on cyclin D1 for genesis and survival, suggesting that targeting the cyclin/cyclin-dependent kinase (cdk) axis may be an effective therapeutic strategy for these tumors. Because cdk inhibitors have not been used for preclinical or clinical testing on rhabdoid tumors, we investigated the effect of flavopiridol, a pan-cdk inhibitor with promising clinical activity, on rhabdoid tumors. The effect of flavopiridol on rhabdoid cells was tested in vitro using survival, cell cycle, and apoptosis assays. Its effect was assessed in vivo using xenografted rhabdoid tumor models. Immunoblot and immunohistochemical analysis was used to assess the effect of flavopiridol on cyclin D1 and p21 expression in vitro and in vivo, respectively. Nanomolar concentrations of flavopiridol inhibited rhabdoid cell growth (IC(50) approximately 200 nmol/L), induced G(1) and G(2) arrest, and apoptosis in vitro in a concentration-dependent manner. These effects were correlated with the down-modulation of cyclin D1, up-regulation of p21, and induction of caspase 3/7 activities. Flavopiridol (at 7.5 mg/kg) significantly inhibited the growth of xenografted rhabdoid tumors, and its effect was correlated with the induction of p21 and down-modulation of cyclin D1. Flavopiridol is effective in inducing cell cycle arrest and cytotoxicity in rhabdoid tumors. Its effects are correlated with the down-regulation of cyclin D1 and the up-regulation of p21. Flavopiridol is potentially a novel chemotherapeutic agent for rhabdoid tumors.


A haplotype inference algorithm for trios based on deterministic sampling.
In genome-wide association studies, thousands of individuals are genotyped in hundreds of thousands of single nucleotide polymorphisms (SNPs). Statistical power can be increased when haplotypes, rather than three-valued genotypes, are used in analysis, so the problem of haplotype phase inference (phasing) is particularly relevant. Several phasing algorithms have been developed for data from unrelated individuals, based on different models, some of which have been extended to father-mother-child "trio" data. We introduce a technique for phasing trio datasets using a tree-based deterministic sampling scheme. We have compared our method with publicly available algorithms PHASE v2.1, BEAGLE v3.0.2 and 2SNP v1.7 on datasets of varying number of markers and trios. We have found that the computational complexity of PHASE makes it prohibitive for routine use; on the other hand 2SNP, though the fastest method for small datasets, was significantly inaccurate. We have shown that our method outperforms BEAGLE in terms of speed and accuracy for small to intermediate dataset sizes in terms of number of trios for all marker sizes examined. Our method is implemented in the "Tree-Based Deterministic Sampling" (TDS) package, available for download at http://www.ee.columbia.edu/~anastas/tds Using a Tree-Based Deterministic sampling technique, we present an intuitive and conceptually simple phasing algorithm for trio data. The trade off between speed and accuracy achieved by our algorithm makes it a strong candidate for routine use on trio datasets.


A yeast artificial chromosome contig containing the complete Duchenne muscular dystrophy gene.
A contig of 36 overlapping yeast artificial chromosome (YAC) clones has been constructed for the complete Duchenne muscular dystrophy (DMD) gene in Xp21. The YACs were isolated from a human 48,XXXX YAC library using the DMD cDNA and brain promoter fragments as hybridization probes. The YAC clones were characterized for exon content using HindIII or EcoRI digests, hybridization of individual DMD cDNA probes, and polymerase chain reaction (PCR) amplification of specific exons near the 5' end of the gene. For comparison to the known long-range restriction map of the DMD gene, YAC clones were digested with SfiI and hybridized with DMD cDNA probes. The combined analysis of the exon content and the SfiI map allowed an approximately 3.2-Mb YAC contig to be constructed. The complete 2.4-Mb DMD gene could be represented in a minimum set of 7 overlapping YAC clones.


Extensional flow-based assessment of red blood cell deformability using hyperbolic converging microchannel.
The deformability of the red blood cell (RBC), is known to be closely related to microcirculation and diagnosis of specific diseases such as malaria, arterial sclerosis, sepsis, and so on. From the viewpoint of the flow type, conventional methods to measure the cell deformability have exploited simple shear or complex flow field with little focus on extensional flow field. In this paper, we present a new approach to assess cell deformability under the extensional flow field. For this purpose, a hyperbolic converging microchannel was designed, and the cell deformation in the extensional flow region was continuously monitored. It overcomes the limitation of conventional methods by reducing experiment time. As quantified by the degree of deformation, the extensional flow (Deformation Index = 0.51 at 3.0 Pa) was found to be more efficient in inducing cell deformation compared to the shear flow (Deformation Index = 0.29 at 3.0 Pa). This indicates the insufficiency of the existing models that predict the blood damage in artificial organs, which only consider shear flow. Also, this method could detect the heat-induced difference in deformability of RBCs. It provides a new platform to study the clinical effect of RBC deformability under extensional flow, and is expected to contribute the association of several diseases and deformability of RBCs.


Physical access barriers to care for diagnosis and treatment of breast cancer among women with mobility impairments.
To explore the perceptions of patients with breast cancer with mobility impairments of the physical accessibility of healthcare facilities and equipment. Individual audiotaped interviews lasting one to two hours. Interviews in homes or workplaces or by telephone. 20 women with chronic mobility impairments who developed early-stage breast cancer prior to age 60. Three were recruited from oncologist panels and 17 from informal social networks of disabled women nationwide. Qualitative analyses of interview transcripts to identify common themes. Extent and nature of mobility impairments and concerns raised by patients about barriers to care. The 20 participants identified issues with inaccessible equipment, including mammography machines, examining tables, and weight scales. The patients sometimes needed to insist on being transferred to an examining table when physicians preferred to examine them seated in their wheelchairs. When staff would transfer them, patients feared injury or felt badly when clinical personnel were injured during transfers. Other issues included difficulties with positioning and handling patients' uncontrollable movements. Even when clinical sites had accessible equipment, this equipment was sometimes unavailable for the appointment. Women with major mobility issues who developed breast cancer confronted numerous physical barriers during the course of their breast cancer diagnosis and treatment. With the aging of the baby boomer generation, an increasing number of people with mobility impairments will be seeking healthcare services. Healthcare providers should be proactive in planning to accommodate these patients by considering accessibility whenever they acquire new equipment, renovate older structures, or build new facilities. They also should establish policies and procedures to ensure that equipment is available during appointments of patients with mobility issues and that staff are trained in safe transferring procedures. Ensuring accommodations and accessibility will benefit patients with impaired mobility and clinical staff.


Gene expression profiling as a tool for positional cloning of genes-shortcut or the longest way round.
The identification of quantitative trait loci, QTL, in arthritis animal models is a straight forward process. However, to identify the underlying genes is a great challenge. One strategy frequently used, is to combine QTL analysis with genomic/proteomic screens. This has resulted in a number of publications where carefully performed genomic analyses present likely candidate genes for their respective QTL s. However, seldom the findings are reconnected to the QTL controlled phenotypes. In this review, we use our own data as an illustrative example that "very likely candidate genes" identified by genomic/proteomics is not necessarily the same as true QTL underlying genes.


CVD-grown monolayer MoS2 in bioabsorbable electronics and biosensors.
Transient electronics represents an emerging technology whose defining feature is an ability to dissolve, disintegrate or otherwise physically disappear in a controlled manner. Envisioned applications include resorbable/degradable biomedical implants, hardware-secure memory devices, and zero-impact environmental sensors. 2D materials may have essential roles in these systems due to their unique mechanical, thermal, electrical, and optical properties. Here, we study the bioabsorption of CVD-grown monolayer MoS2, including long-term cytotoxicity and immunological biocompatibility evaluations in biofluids and tissues of live animal models. The results show that MoS2 undergoes hydrolysis slowly in aqueous solutions without adverse biological effects. We also present a class of MoS2-based bioabsorbable and multi-functional sensor for intracranial monitoring of pressure, temperature, strain, and motion in animal models. Such technology offers specific, clinically relevant roles in diagnostic/therapeutic functions during recovery from traumatic brain injury. Our findings support the broader use of 2D materials in transient electronics and qualitatively expand the design options in other areas.


A small-scale serum-free liquid cell culture model of erythropoiesis to assess the effects of exogenous factors.
Anaemia is an important global health problem. Therefore, it is crucial to understand its pathophysiology in various genetic or infectious diseases where dyserythropoiesis is a key pathological feature. To this effect, reproducible and reliable models of erythropoiesis in vitro are much needed as investigative tools. We have developed a serum-free liquid culture model of erythropoiesis using human umbilical cord blood CD34(+) cells cultured in the cytokine combination, interleukin-3 (IL-3), IL-6, stem cell factor (SCF) and erythropoietin (Epo), over 14 days. We found that these culture conditions favored erythroid differentiation over the expansion of the more primitive erythroid precursors. With an initiating culture density of 5x10(4) cells per ml, the nucleated cell fold expansion increased from 7.9+/-3.9 (range 4.5 to 11.1) after 4 days to 2990.2+/-1936.1 (range 626.6 to 6912.0) after 14 days in culture. Day-14 burst-forming unit-erythroid (BFU-E) frequencies peaked at day 4 (24.0+/-8.9%), with a marked decrease in BFU-E burst size as the cultures progressed. Time-course immunophenotypical profiles were characteristically erythroid with a decrease in CD34 expression (from 96.8+/-3.0% at day 0 to 0.8+/-0.8% at day 14), and a concomitant increase in the expression of erythroid-specific markers, CD36, glycophorin A (GpA) and CD71 (from 14.8+/-5.0%, 1.7+/-1.0% and 37.9+/-18.0% to 93.0+/-7.0%, 82.1+/-14.0% and 95.7+/-3.0%, respectively). Morphological studies revealed the presence of normoblasts with the complete absence of reticulocytes and mature erythrocytes after 14 days in culture. Once the culture conditions were optimized, we scaled down our culture model from 24-well plate (large-scale) to 96-well plate cultures (small-scale). We found that the small-scale cultures compared favorably with their large-scale counterpart in terms of erythroid progenitor cell proliferation and differentiation, particularly at low CD34(+) initiating cell doses. By using tumor necrosis factor-alpha (TNF-alpha), a known inhibitor of erythropoiesis, we validated our model system and showed a dose-dependent inhibition of erythroid differentiation with TNF-alpha in our cultures. Therefore, our results demonstrate a small-scale serum-free liquid culture model of erythropoiesis that is comparable with and complements our well-defined large-scale model. Our system would prove useful for screening the effects of exogenous factors on erythropoiesis in vitro.


Toxicity of Turmeric Extracts to the Termite Reticulitermes flavipes (Blattodea: Rhinotermitidae).
Turmeric is an important spice crop with documented human health benefits associated with chemicals called curcuminoids. In this study, the termite Reticulitermes flavipes (Kollar) was exposed to different solvent extracts of turmeric to investigate potential termiticidal properties. Treating termites with hexane extracts of purified lab-grade curcuminoids had no effect on termites. However, in continuous exposure assays, the LC(50) for hexane extracts of crude turmeric powder was 9.6 mg, or 1.0 mg starting material per square centimeter of filter paper substrate. These active components were soluble in a range of polar and apolar solvents, but only hexane could selectively fractionate active components away from the inactive curcuminoids. The active constituents of turmeric separated by thin layer chromatography (TLC) fluoresced in short-wave UV light but were not visible in long-wave UV light. By re-extracting TLC-separated bands in hexane and performing bioassays and gas chromatography-mass spectrometry, we demonstrated that termiticidal components of turmeric are extractable as a blend containing mainly ar-turmerone, turmerone, and curlone. This determination is consistent with findings of preceding work by other researchers that investigated insecticidal properties of turmeric in other pest insects.


Mold allergens in respiratory allergy: from structure to therapy.
Allergic reactions to fungi were described 300 years ago, but the importance of allergy to fungi has been underestimated for a long time. Allergens from fungi mainly cause respiratory and skin symptoms in sensitized patients. In this review, we will focus on fungi and fungal allergens involved in respiratory forms of allergy, such as allergic rhinitis and asthma. Fungi can act as indoor and outdoor respiratory allergen sources, and depending on climate conditions, the rates of sensitization in individuals attending allergy clinics range from 5% to 20%. Due to the poor quality of natural fungal allergen extracts, diagnosis of fungal allergy is hampered, and allergen-specific immunotherapy is rarely given. Several factors are responsible for the poor quality of natural fungal extracts, among which the influence of culture conditions on allergen contents. However, molecular cloning techniques have allowed us to isolate DNAs coding for fungal allergens and to produce a continuously growing panel of recombinant allergens for the diagnosis of fungal allergy. Moreover, technologies are now available for the preparation of recombinant and synthetic fungal allergen derivatives which can be used to develop safe vaccines for the treatment of fungal allergy.


Slower reaction times and impaired learning in young adults with birth weight <1500 g.
Children with very low birth weight (VLBW; <1500 g) perform worse on cognitive tests than do children who are born at term. Whether this difference persists into adulthood has been little studied. We assessed core neurocognitive abilities (processing speed, working memory, attention, and learning capacity) in young adults with VLBW and in term-born control subjects. In conjunction with the Helsinki Study of Very Low Birth Weight Adults, 147 VLBW and 171 control subjects who were aged 18 to 27 years and did not have neurosensory impairments performed a computerized test battery (CogState Ltd, Melbourne, Australia). T tests and linear regression models were used. Cohen's d was used to express effect size (ES). VLBW adults had slower reaction times than did control subjects on all 5 tasks: simple reaction time (mean difference: 4.0% [95% confidence interval (CI): 1.1%-7.0%]; ES: 0.30), choice reaction time (mean difference: 3.2% [95% CI: 0.3%-6.2%]; ES: 0.24), working memory (mean difference: 8.4% [95% CI: 3.7%-13.4%]; ES: 0.40), divided attention (mean difference: 7.2% [95% CI: 2.7%-11.9%]; ES: 0.36), and associated learning reaction time (mean difference: 6.4% [95% CI: 1.3%-11.9%]; ES: 0.28). In addition, VLBW adults showed impaired learning abilities on the associated learning task (percentage of correct responses: 85.7 vs 80.2; P < .001; ES: 0.64). The results were little affected by adjustment for confounders. Nonimpaired VLBW individuals exhibited slower psychomotor speed and lower accuracy on the associated learning task. These results indicate that very preterm birth, even when obvious neurosensory deficits are absent, may have long-term consequences on core neurocognitive abilities.


Severe nitrofurantoin lung disease resolving without the use of steroids.
We report a case of an elderly woman who developed a severe, chronic pulmonary reaction to nitrofurantoin therapy that she had taken continuously for three years to prevent urinary tract infections. The patient was taking no other drug known to cause lung disease but the diagnosis was delayed by failure to recognize the association between nitrofurantoin and adverse drug reactions affecting the lung. When originally seen, the patient was unable to care for herself due to dyspnea. Bronchoscopy with biopsy ruled out other causes of her pulmonary disease. Immediate withdrawal of nitrofurantoin led to substantial, sustained improvement and disappearance of symptoms over several months without administration of corticosteroids. Nitrofurantoin toxicity should always be considered in any person taking that drug who develops bilateral infiltrates.


Evaluating the Effect of Cognitive Dysfunction on Mental Imagery in Patients with Stroke Using Temporal Congruence and the Imagined 'Timed Up and Go' Test (iTUG).
Motor imagery (MI) capacity may be altered following stroke. MI is evaluated by measuring temporal congruence between the timed performance of an imagined and an executed task. Temporal congruence between imagined and physical gait-related activities has not been evaluated following stroke. Moreover, the effect of cognitive dysfunction on temporal congruence is not known. To assess temporal congruence between the Timed Up and Go test (TUG) and the imagined TUG (iTUG) tests in patients with stroke and to investigate the role played by cognitive dysfunctions in changes in temporal congruence. TUG and iTUG performance were recorded and compared in twenty patients with chronic stroke and 20 controls. Cognitive function was measured using the Montreal Cognitive Assessment (MOCA), the Frontal Assessment Battery at Bedside (FAB) and the Bells Test. The temporal congruence of the patients with stroke was significantly altered compared to the controls, indicating a loss of MI capacity (respectively 45.11 ±35.11 vs 24.36 ±17.91, p = 0.02). Furthermore, iTUG test results were positively correlated with pathological scores on the Bells Test (r = 0.085, p = 0.013), likely suggesting that impairment of attention was a contributing factor. These results highlight the importance of evaluating potential attention disorder in patients with stroke to optimise the use of MI for rehabilitation and recovery. However further study is needed to determine how MI should be used in the case of cognitive dysfunction.


Quantitative trait loci for yield and grain plumpness relative to maturity in three populations of barley (Hordeum vulgare L.) grown in a low rain-fall environment.
Identifying yield and grain plumpness QTL that are independent of developmental variation or phenology is of paramount importance for developing widely adapted and stable varieties through the application of marker assisted selection. The current study was designed to dissect the genetic basis of yield performance and grain plumpness in southern Australia using three doubled haploid (DH) populations developed from crosses between adapted parents that are similar in maturity and overall plant development. Three interconnected genetic populations, Commander x Fleet (CF), Commander x WI4304 (CW), and Fleet x WI4304 (FW) developed from crossing of Australian elite barley genotypes, were used to map QTL controlling yield and grain plumpness. QTL for grain plumpness and yield were analysed using genetic linkage maps made of genotyping-by-sequencing markers and major phenology genes, and field trials at three drought prone environments for two growing seasons. Seventeen QTL were detected for grain plumpness. Eighteen yield QTL explaining from 1.2% to 25.0% of the phenotypic variation were found across populations and environments. Significant QTL x environment interaction was observed for all grain plumpness and yield QTL, except QPlum.FW-4H.1 and QYld.FW-2H.1. Unlike previous yield QTL studies in barley, none of the major developmental genes, including Ppd-H1, Vrn-H1, Vrn-H2 and Vrn-H3, that drive barley adaption significantly affected grain plumpness and yield here. Twenty-two QTL controlled yield or grain plumpness independently of known maturity QTL or genes. Adjustment for maturity effects through co-variance analysis had no major effect on these yield QTL indicating that they control yield per se.


Donkin psychosis.
Donkin psychoses are eclamptic psychoses without seizures. As symptomatic psychoses resulting from cerebral endothelial damage, they may explain the lucid intervals that sometimes occur between eclampsia and the eruption of psychosis. They have the same features as eclamptic psychoses, with onset during pregnancy or the early puerperium, especially in first-time mothers, a short duration and full recovery in most. The clinical picture is usually delirium, but mania is also seen, and some patients have retrograde amnesia or other cognitive defects. Donkin psychosis should be considered in the differential diagnosis of childbearing psychoses, and collaborative research is needed to clarify their differences.


Determining standard criteria for endotracheal suctioning in the paediatric intensive care patient: an exploratory study.
This four-phase mixed method study developed an evidence based "Endotracheal Suction Assessment Tool" (ESAT) as a guide for nurses undertaking "endotracheal tube" (ETT) suction within "Paediatric Intensive Care" (PIC). Phase 1 involved a comprehensive literature review to determine the most commonly used criteria for assessing the need for ETT suction. In Phase 2 an "Endotracheal Suction Questionnaire" (ESQ) was developed to survey experienced PIC nurses in Australia and New Zealand regarding their ETT suction decision making process and validity testing of the ESQ. In Phase 3, the ESQ was administered to target group (n=104). In Phase 4, the empirical evidence generated from this study, based upon the criteria rated by nurses in this study as being most clinically important and essential during the decision making process, determined the ESAT design. Analyses of quantitative results showed a positive correlation between the perceived frequency of use of a criterion and the appropriateness of the assessment. Where a criterion was used less frequently as a clinical indicator for the requirement for ETT suction, participants had a lower regard for this when rating the criterion as a specific single indicator to perform suction. Findings from qualitative data identified six criteria not previously documented within the literature. Further testing and validation of the tool within the PIC setting will determine the clinical viability of the ESAT.


A novel, dynamic pattern-based analysis of NF-κB binding during the priming phase of liver regeneration reveals switch-like functional regulation of target genes.
Following partial hepatectomy, a coordinated series of molecular events occurs to regulate hepatocyte entry into the cell cycle to recover lost mass. In rats during the first 6 h following resection, hepatocytes are primed by a tightly controlled cytokine response to prepare hepatocytes to begin replication. Although it appears to be a critical element driving regeneration, the cytokine response to resection has not yet been fully characterized. Specifically, the role of one of the key response elements to cytokine signaling (NF-κB) remains incompletely characterized. In this study, we present a novel, genome-wide, pattern-based analysis characterizing NF-κB binding during the priming phase of liver regeneration. We interrogated the dynamic regulation of priming by NF-κB through categorizing NF-κB binding in different temporal profiles: immediate sustained response, early transient response, and delayed response to partial hepatectomy. We then identified functional regulation of NF-κB binding by relating the temporal response profile to differential gene expression. We found that NF-κB bound genes govern negative regulation of cell growth and inflammatory response immediately following hepatectomy. NF-κB also transiently regulates genes responsible for lipid biosynthesis and transport as well as induction of apoptosis following hepatectomy. By the end of the priming phase, NF-κB regulation of genes involved in inflammatory response, negative regulation of cell death, and extracellular structure organization became prominent. These results suggest that NF-κB regulates target genes through binding and unbinding in immediate, transient, and delayed patterns. Such dynamic switch-like patterns of NF-κB binding may govern different functional transitions that drive the onset of regeneration.


Apical localization of ITPK1 enhances its ability to be a modifier gene product in a murine tracheal cell model of cystic fibrosis.
A new aspect of research into the pathogenesis of cystic fibrosis (CF) is a genetics-based search for ;modifier genes' that may affect the severity of CF lung disease. Using an alternative, cell biological approach, we show that ITPK1 should be considered a modifier gene. ITPK1 synthesizes an intracellular signal, inositol (3,4,5,6)-tetrakisphosphate [Ins(3,4,5,6)P4]. A bio-activatable, cell-permeable analogue of Ins(3,4,5,6)P4 inhibited Ca2+-dependent secretion of Cl- from polarized monolayers of immortalized mouse tracheal epithelial cells (MTEs). Analysis by high-pressure liquid chromatography showed endogenous Ins(3,4,5,6)P4 levels in CF MTEs were approximately 60% below those in wild-type MTEs (P<0.03). This adaptation, which improves purinergic activation of Ca2+-dependent Cl- secretion in CF MTEs, was exceptionally specific; there was no effect upon the cellular levels of all the other inositol phosphate signals. Real-time PCR provided the explanation: the level of ITPK1 expression in wild-type MTEs was twice as high as that in CF MTEs (P<0.002). The biological impact of this differential gene expression is amplified by ITPK1 being concentrated at the apical membrane of MTEs, which we discovered following confocal immunofluorescence microscopy. Compartmentalization of Ins(3,4,5,6)P4 synthesis adjacent to its site of action will enhance its regulatory capacity.


Exploration of in-fiber nanostructures from capillary instability.
A new class of multi-material fiber that incorporates micrometer-thickness concentric-cylindrical sheets of glass into polymer matrix has emerged. The ultimate lower limit of feature size and recent observation of interesting instability phenomenon in fiber system motivate us to examine fluid instabilities during the complicated thermal drawing fabrication processing. In this paper, from the perspective of a single instability mechanism, classical Plateau-Rayleigh instabilities in the form of radial fluctuation, we explore the stability of various microstructures (such as shells and filaments) in our composite fibers. The attained uniform structures are consistent with theoretical analysis. Furthermore, a viscous materials map is established from calculations and agrees well with various identified materials. These results not only shed insights into other forms of fluid instabilities, but also provide guidance to achieve more diverse nanostructures (such as filaments, wires, and particles) in the microstructured fibers.


Contents of mRNAs encoding endosome/lysosome components in normal human aorta and in stage II of atherogenesis: a hidden regulation.
Contents of mRNAs encoding endosome/lysosome components EEA1, Rab5a, Lamp1, Lamp2, p62 (SQSTM1), and CD63 were measured by quantitative PCR and compared in intact fragments of human aorta and in aorta fragments with atherosclerotic lesions of stage II (fatty streaks) of the same donors. During atherogenesis an increase was detected only in the level of p62 mRNA but not in other mRNAs. Nevertheless, correlation analysis revealed a profound rearrangement of inter-gene correlations: only 30% of correlations found in the fatty streaks coincided with the correlations in normal fragments. Thus, new constellations were formed in fatty streaks concurrently with disappearance of correlations between mRNAs under study and mRNAs encoding factors of lipid accumulation, reverse cholesterol transfer, and some lipid sensors/transcription regulators of lipid metabolism.


Minimal NMR distance information for rigidity of protein graphs.
Nuclear Magnetic Resonance (NMR) experiments provide distances between nearby atoms of a protein molecule. The corresponding structure determination problem is to determine the 3D protein structure by exploiting such distances. We present a new order on the atoms of the protein, based on information from the chemistry of proteins and NMR experiments, which allows us to formulate the problem as a combinatorial search. Additionally, this order tells us what kind of NMR distance information is crucial to understand the cardinality of the solution set of the problem and its computational complexity.


Trabecular Meshwork Response to Pressure Elevation in the Living Human Eye.
The mechanical characteristics of the trabecular meshwork (TM) are linked to outflow resistance and intraocular pressure (IOP) regulation. The rationale behind this technique is the direct observation of the mechanical response of the TM to acute IOP elevation. Prior to scanning, IOP is measured at baseline and during IOP elevation. The limbus is scanned by spectral-domain optical coherence tomography at baseline and during IOP elevation (ophthalmodynamometer (ODM) applied at 30 g force). Scans are processed to enhance visualization of the aqueous humor outflow pathway using ImageJ. Vascular landmarks are used to identify corresponding locations in baseline and IOP elevation scan volumes. Schlemm canal (SC) cross-sectional area (SC-CSA) and SC length from anterior to posterior along its long axis are measured manually at 10 locations within a 1 mm segment of SC. Mean inner to outer wall distance (short axis length) is calculated as the area of SC divided by its long axis length. To examine the contribution of adjacent tissues to the effect IOP elevations, measurements are repeated without and with smooth muscle relaxation with instillation of tropicamide. TM migration into SC is resisted by TM stiffness, but is enhanced by the support of its attachment to adjacent smooth muscle within the ciliary body. This technique is the first to measure the living human TM response to pressure elevation in situ under physiological conditions within the human eye.


An integrated malaria control program with community participation on the Pacific Coast of Colombia.
The study focuses on integrated malaria control in 23 communities on the Pacific Coast of Colombia, with several elements of an ecosystem approach to human health, including malaria-related sociopolitical, ecological, and economic factors. The program fostered community participation. The program presented here had 2 components: implementation and research. The first was conducted in 23 communities, 21 of which lacked adequate health services in terms of education, community participation, prompt diagnosis and complete treatment, and vector control. Research focused on specific vector control measures and the current national health services decentralization process. The project: 1) created a malaria prevention culture in the community; 2) avoided deaths from malaria (no fatal cases in the 3-year period, compared to 5-8 deaths a year previously); 3) avoided cases of cerebral malaria (no cases, as compared to 90-110 per year previously); 4) reduced malaria incidence by 45.36%; 5) decreased length of sick leave from 7.52 to 3.7 days; 6) established a permanent network of microscope technicians and 2-way radio communications; 7) integrated work by local, regional, and outside institutions; 8) demonstrated efficacy of insecticide-impregnated bednets to reduce malaria transmission.


Inactive Matrix Gla-Protein Is Associated With Arterial Stiffness in an Adult Population-Based Study.
Increased pulse wave velocity (PWV) is a marker of aortic stiffness and an independent predictor of mortality. Matrix Gla-protein (MGP) is a vascular calcification inhibitor that needs vitamin K to be activated. Inactive MGP, known as desphospho-uncarboxylated MGP (dp-ucMGP), can be measured in plasma and has been associated with various cardiovascular markers, cardiovascular outcomes, and mortality. In this study, we hypothesized that high levels of dp-ucMGP are associated with increased PWV. We recruited participants via a multicenter family-based cross-sectional study in Switzerland. Dp-ucMGP was quantified in plasma by sandwich ELISA. Aortic PWV was determined by applanation tonometry using carotid and femoral pulse waveforms. Multiple regression analysis was performed to estimate associations between PWV and dp-ucMGP adjusting for age, renal function, and other cardiovascular risk factors. We included 1001 participants in our analyses (475 men and 526 women). Mean values were 7.87±2.10 m/s for PWV and 0.43±0.20 nmol/L for dp-ucMGP. PWV was positively associated with dp-ucMGP both before and after adjustment for sex, age, body mass index, height, systolic and diastolic blood pressure (BP), heart rate, renal function, low- and high-density lipoprotein, glucose, smoking status, diabetes mellitus, BP and cholesterol lowering drugs, and history of cardiovascular disease (P≤0.01). In conclusion, high levels of dp-ucMGP are independently and positively associated with arterial stiffness after adjustment for common cardiovascular risk factors, renal function, and age. Experimental studies are needed to determine whether vitamin K supplementation slows arterial stiffening by increasing MGP carboxylation.


Transient full-field vibration measurement using spectroscopical stereo photogrammetry.
Contrasted with other vibration measurement methods, a novel spectroscopical photogrammetric approach is proposed. Two colored light filters and a CCD color camera are used to achieve the function of two traditional cameras. Then a new calibration method is presented. It focuses on the vibrating object rather than the camera and has the advantage of more accuracy than traditional camera calibration. The test results have shown an accuracy of 0.02 mm.


Direct observation of incommensurate magnetism in Hubbard chains.
The interplay between magnetism and doping is at the origin of exotic strongly correlated electronic phases and can lead to novel forms of magnetic ordering. One example is the emergence of incommensurate spin-density waves, which have wavevectors that do not belong to the reciprocal lattice. In one dimension this effect is a hallmark of Luttinger liquid theory, which also describes the low-energy physics of the Hubbard model1. Here we use a quantum simulator that uses ultracold fermions in an optical lattice2-8 to directly observe such incommensurate spin correlations in doped and spin-imbalanced Hubbard chains using fully spin- and density-resolved quantum gas microscopy. Doping is found to induce a linear change in the spin-density wavevector, in excellent agreement with predictions from Luttinger theory. For non-zero polarization we observe a reduction in the wavevector with magnetization, as expected from the antiferromagnetic Heisenberg model in a magnetic field. We trace the microscopic-scale origin of these incommensurate correlations to holes, doublons (double occupancies) and excess spins, which act as delocalized domain walls for the antiferromagnetic order. In addition, by inducing interchain coupling we observe fundamentally different spin correlations around doublons and suppression of incommensurate magnetism at finite (low) temperature in the two-dimensional regime9. Our results demonstrate how access to the full counting statistics of all local degrees of freedom can be used to study fundamental phenomena in strongly correlated many-body physics.


The potential of exosomes in diagnosis and treatment of inborn errors of metabolism.
Extracellular vesicles, in particular exosomes, have gained much attention as potent mediators of intercellular signaling. Exosomes are 50-130 nm intraluminal vesicles of multivesicular bodies (MVB) that are secreted into the extracellular environment upon fusion of MVB with the plasma membrane. Current research on exosomes focuses on their biogenesis, including specific sorting mechanisms, their potential to transfer proteins and RNA from their cells of origin to target cells, specific methods of vesicle isolation, and their possible application as diagnostic and therapeutic devices. Exosomes are vesicles of endocytic origin that contain a portion of the cytoplasm. Their molecular components represent the composition and thereby the physiological state of the cells from which they originate. In this review, we recapitulate the discovery of exosomes and the subsequent expansion of exosome research into a variety of different areas of interest, with a specific focus on how exosomes could prove to be invaluable for both diagnostic and therapeutic applications within the research field of inborn errors of metabolism.


Clinics in diagnostic imaging (102).
A 19-year-old man presented with cough and haemoptysis of ten days duration. He also had mild right hypochondrial pain. Chest radiograph and computed tomography (CT) showed a rounded soft tissue density opacity with an air crescent sign. CT showed multiple cystic lesions in the liver with a daughter cyst in its lateral wall. Diagnosis of hydatid disease of lung and liver was made. The contents of the liver cyst were aspirated, hypertonic saline instilled, re-aspirated, and absolute alcohol injected. Hydatid disease is endemic in certain parts of the world. Although the lungs and liver are most frequently affected, the disease can arise in any part of the body and should be kept in differential diagnosis whenever a cystic lesion is encountered. Hydatid cysts typically demonstrate characteristic imaging findings, however, the appearances may become complicated due to cyst rupture or secondary infection. Ultrasonography is the imaging modality of choice particularly in hepatic disease. CT best demonstrates cyst wall calcification and cyst infection.


Stigmatization as a social control mechanism for persons living with HIV and AIDS.
Stigmatization contributes to inequity by marginalizing persons living with HIV and AIDS (PHAs). In this study we examined the stigmatizing practices in health care settings from the perspectives of PHAs and health care providers (HCPs). A qualitative design, using a participatory action research approach, was used. Interviews and focus groups were completed with 16 aboriginal and 17 nonaboriginal individuals living with HIV (APHAs and PHAs) and 27 HCPs in Ottawa and Edmonton, Canada. We present findings to support the premise that stigmatization can be used as a social control mechanism with PHAs. Participants described both active and passive social control mechanisms: shunning and ostracizing, labeling, and disempowering health care practices. Forgiving behavior, balancing disclosure, practicing universal precautions, bending the rules, shifting services, and reducing labeling were strategies to manage, resist, and mitigate social control. The findings illustrate the urgent need for multilevel interventions to manage, resist, and mitigate stigma.


A High-Resolution Comparative Chromosome Map of Cricetus cricetus and Peromyscus eremicus Reveals the Involvement of Constitutive Heterochromatin in Breakpoint Regions.
Compared to humans and other mammals, rodent genomes, specifically Muroidea species, underwent intense chromosome reshuffling in which many complex structural rearrangements occurred. This fact makes them preferential animal models for studying the process of karyotype evolution. Here, we present the first combined chromosome comparative maps between 2 Cricetidae species, Cricetus cricetus and Peromyscus eremicus, and the index species Mus musculus and Rattus norvegicus. Comparative chromosome painting was done using mouse and rat paint probes together with in silico analysis from the Ensembl genome browser database. Hereby, evolutionary events (inter- and intrachromosomal rearrangements) that occurred in C. cricetus and P. eremicus since the putative ancestral Muroidea genome could be inferred, and evolutionary breakpoint regions could be detected. A colocalization of constitutive heterochromatin and evolutionary breakpoint regions in each genome was observed. Our results suggest the involvement of constitutive heterochromatin in karyotype restructuring of these species, despite the different levels of conservation of the C. cricetus (derivative) and P. eremicus (conserved) genomes.


Decreased bronchodilating effect of salbutamol in relieving methacholine induced moderate to severe bronchoconstriction during high dose treatment with long acting beta2 agonists.
In vitro the long acting beta2 agonist salmeterol can, in contrast to formoterol, behave as a partial agonist and become a partial antagonist to other beta2 agonists. To study this in vivo, the bronchodilating effect of salbutamol was measured during methacholine induced moderate to severe bronchoconstriction in patients receiving maintenance treatment with high dose long acting beta2 agonists. A randomised double blind crossover study was performed in 19 asthmatic patients with mean forced expiratory volume in one second (FEV1) of 88.4% predicted and median concentration of methacholine provoking a fall in FEV1 of 20% or more (PC(20)) of 0.62 mg/ml at entry. One hour after the last dose of 2 weeks of treatment with formoterol (24 microg twice daily by Turbuhaler), salmeterol (100 microg twice daily by Diskhaler), or placebo a methacholine provocation test was performed and continued until there was at least a 30% decrease in FEV1. Salbutamol (50 microg) was administered immediately thereafter, followed by ipratropium bromide (40 microg) after a further 30 minutes. Lung function was monitored for 1 hour after provocation. There was a significant bronchodilating and bronchoprotective effect after 2 weeks of active treatment. The dose of methacholine needed to provoke a fall in FEV1 of > or = 30% was higher after pretreatment with formoterol (2.48 mg) than with salmeterol (1.58 mg) or placebo (0.74 mg). The difference between formoterol and salmeterol was statistically significant: 0.7 doubling dose steps (95% CI 0.1 to 1.2, p=0.016). The immediate bronchodilating effect of subsequently administered salbutamol was significantly impaired after pretreatment with both drugs (p<0.0003 for both). Three minutes after inhaling salbutamol the increase in FEV1 relative to the pre-methacholine baseline was 15.8%, 7.3%, and 5.5% for placebo, formoterol and salmeterol, respectively (equivalent to increases of 26%, 14%, and 12%, respectively, from the lowest FEV1 after methacholine). At 30 minutes significant differences remained, but 1 hour after completing the methacholine challenge FEV1 had returned to baseline values in all three treatment groups. Formoterol has a greater intrinsic activity than salmeterol as a bronchoprotective agent, indicating that salmeterol is a partial agonist compared with formoterol in contracted human airways in vivo. Irrespective of this, prior long term treatment with both long acting beta2 agonists reduced the bronchodilating effect of an additional single dose of salbutamol equally, indicating that the development of tolerance or high receptor occupancy overshadowed any possible partial antagonistic activity of salmeterol. Patients on regular treatment with long acting beta2 agonists should be made aware that an additional single dose of a short acting beta2 agonist may become less effective.


A direct role for cohesin in gene regulation and ecdysone response in Drosophila salivary glands.
Developmental abnormalities observed in Cornelia de Lange syndrome have been genetically linked to mutations in the cohesin machinery. These and other recent experimental findings have led to the suggestion that cohesin, in addition to its canonical function of mediating sister chromatid cohesion, might also be involved in regulating gene expression. We report that cleavage of cohesin's kleisin subunit in postmitotic Drosophila salivary glands induces major changes in the transcript levels of many genes. Kinetic analyses of changes in transcript levels upon cohesin cleavage reveal that a subset of genes responds to cohesin cleavage within a few hours. In addition, cohesin binds to most of these loci, suggesting that cohesin is directly regulating their expression. Among these genes are several that are regulated by the steroid hormone ecdysone. Cytological visualization of transcription at selected ecdysone-responsive genes reveals that puffing at Eip74EF ceases within an hour or two of cohesin cleavage, long before any decline in ecdysone receptor could be detected at this locus. We conclude that cohesin regulates expression of a distinct set of genes, including those mediating the ecdysone response.


Distribution of hepatitis B virus genotypes in two different pediatric populations from Argentina.
Differences in pathogenesis and the probability of becoming a chronic carrier depend on the age at which hepatitis B virus (HBV) infection is acquired, ranging from 82% in infants less than 6 months of age to 15 to 30% in older children. HBV genotypes from 22 pediatric patients from two areas that differ in prevalence have been determined. Phylogenetic analysis shows a clear difference between the genotype distribution in Buenos Aires, a low-prevalence area, and that found in Gualeguay, Entre Ríos, a high-prevalence area. While the analysis allocated the sequences in the Buenos Aires group to genotypes A (36%), D (9%), and F (55%), the Gualeguay group presented exclusively genotype A isolates with very low nucleotide divergence, which suggests a strong founder viral population. The high prevalence of genotype F in the Buenos Aires group and its high intragroup heterogeneity agree with the American origin of this genotype.


Technical and policy approaches to balancing patient privacy and data sharing in clinical and translational research.
Clinical researchers need to share data to support scientific validation and information reuse and to comply with a host of regulations and directives from funders. Various organizations are constructing informatics resources in the form of centralized databases to ensure reuse of data derived from sponsored research. The widespread use of such open databases is contingent on the protection of patient privacy. We review privacy-related problems associated with data sharing for clinical research from technical and policy perspectives. We investigate existing policies for secondary data sharing and privacy requirements in the context of data derived from research and clinical settings. In particular, we focus on policies specified by the US National Institutes of Health and the Health Insurance Portability and Accountability Act and touch on how these policies are related to current and future use of data stored in public database archives. We address aspects of data privacy and identifiability from a technical, although approachable, perspective and summarize how biomedical databanks can be exploited and seemingly anonymous records can be reidentified using various resources without hacking into secure computer systems. We highlight which clinical and translational data features, specified in emerging research models, are potentially vulnerable or exploitable. In the process, we recount a recent privacy-related concern associated with the publication of aggregate statistics from pooled genome-wide association studies that have had a significant impact on the data sharing policies of National Institutes of Health-sponsored databanks. Based on our analysis and observations we provide a list of recommendations that cover various technical, legal, and policy mechanisms that open clinical databases can adopt to strengthen data privacy protection as they move toward wider deployment and adoption.


Fundamental performance comparison of a Hartmann and a shearing interferometer wave-front sensor.
The performance of ground-based optical imaging systems is severely degraded from the diffraction limit by the random effects of the atmosphere. Adaptive-optics techniques have been used to compensate for atmospheric-turbulence effects. A critical component in the adaptive-optics system is the wave-front sensor. At present, two types of sensors are common: the Hartmann-Shack wave-front sensor and the shearing interferometer wave-front sensor. In this paper we make a direct performance comparison of these two sensors. The performance calculations are restricted to common configurations of these two sensors and the fundamental limits imposed by shot noise and atmospheric effects. These two effects encompass the effects of extended reference beacons and sensor subaperture spacings larger than the Fried parameter r(0). Our results indicate comparable performance for good seeing conditions and small beacons. However, for poor seeing conditions and extended beacons, the Hartmann sensor has lower error levels than the shearing interferometer.


No clinical utility of KRAS variant rs61764370 for ovarian or breast cancer.
Clinical genetic testing is commercially available for rs61764370, an inherited variant residing in a KRAS 3' UTR microRNA binding site, based on suggested associations with increased ovarian and breast cancer risk as well as with survival time. However, prior studies, emphasizing particular subgroups, were relatively small. Therefore, we comprehensively evaluated ovarian and breast cancer risks as well as clinical outcome associated with rs61764370. Centralized genotyping and analysis were performed for 140,012 women enrolled in the Ovarian Cancer Association Consortium (15,357 ovarian cancer patients; 30,816 controls), the Breast Cancer Association Consortium (33,530 breast cancer patients; 37,640 controls), and the Consortium of Modifiers of BRCA1 and BRCA2 (14,765 BRCA1 and 7904 BRCA2 mutation carriers). We found no association with risk of ovarian cancer (OR=0.99, 95% CI 0.94-1.04, p=0.74) or breast cancer (OR=0.98, 95% CI 0.94-1.01, p=0.19) and results were consistent among mutation carriers (BRCA1, ovarian cancer HR=1.09, 95% CI 0.97-1.23, p=0.14, breast cancer HR=1.04, 95% CI 0.97-1.12, p=0.27; BRCA2, ovarian cancer HR=0.89, 95% CI 0.71-1.13, p=0.34, breast cancer HR=1.06, 95% CI 0.94-1.19, p=0.35). Null results were also obtained for associations with overall survival following ovarian cancer (HR=0.94, 95% CI 0.83-1.07, p=0.38), breast cancer (HR=0.96, 95% CI 0.87-1.06, p=0.38), and all other previously-reported associations. rs61764370 is not associated with risk of ovarian or breast cancer nor with clinical outcome for patients with these cancers. Therefore, genotyping this variant has no clinical utility related to the prediction or management of these cancers.


Chronic pain treatment and scrambler therapy: a multicenter retrospective analysis.
Scrambler Therapy is a novel neuromodulation that works by electrocutaneous stimulation in a non-invasive manner through C fibers surface receptors. It substitutes pain information with synthetic "non pain" information. The primary aim of this study was to analyze the efficacy and safety of Scrambler Therapy after ten sessions related to different usage conditions and different learning curves that occur in a multi-center study. 201 patients have been treated with Scrambler Therapy. All the patients were suffering from chronic pain with a mean pain NRS of 7.41 (SD 2.06) before treatment. Main causes of chronic pain: post herpetic neuralgia 18.40 %, chronic low back pain (LBP) 37.31%, polyneuropathy 10.94%, and peripheral neuropathy 14.42 %. The remaining 18.93 % included chronic pain due to other causes. The difference between pre-treatment NRS 7.41 (SD 2.06) and post-treatment 1.60 (SD 2.22) was statistically significant (P< 0.0001). The mean number of sessions per patient was 10, but 39 had complete absence of pain sooner and used fewer sessions. Only 7 patients stopped treatment due to lack of results, and 2 for personal reasons not ascribable to the treatment. Stimulation pain score of 0 during treatment, and not just pain reduction, predicts long term effectiveness, so this must be pursued by optimizing electrode positioning and correct fine-tuning of stimulation intensity. Scrambler Therapy is an efficient and safe alternative for several different types of refractory chronic neuropathic pain, with a very rare possibility of adverse events.


Recommendations for Preclinical Renal MRI: A Comprehensive Open-Access Protocol Collection to Improve Training, Reproducibility, and Comparability of Studies.
Renal MRI holds incredible promise for making a quantum leap in improving diagnosis and care of patients with a multitude of diseases, by moving beyond the limitations and restrictions of current routine clinical practice. Clinical and preclinical renal MRI is advancing with ever increasing rapidity, and yet, aside from a few examples of renal MRI in routine use, it is still not good enough. Several roadblocks are still delaying the pace of progress, particularly inefficient education of renal MR researchers, and lack of harmonization of approaches that limits the sharing of results among multiple research groups.Here we aim to address these limitations for preclinical renal MRI (predominantly in small animals), by providing a comprehensive collection of more than 40 publications that will serve as a foundational resource for preclinical renal MRI studies. This includes chapters describing the fundamental principles underlying a variety of renal MRI methods, step-by-step protocols for executing renal MRI studies, and detailed guides for data analysis. This collection will serve as a crucial part of a roadmap toward conducting renal MRI studies in a robust and reproducible way, that will promote the standardization and sharing of data.This chapter is based upon work from the COST Action PARENCHIMA, a community-driven network funded by the European Cooperation in Science and Technology (COST) program of the European Union, which aims to improve the reproducibility and standardization of renal MRI biomarkers.


Development of potential methods for testing congestion control algorithm implemented in vehicle-to-vehicle communications.
A channel congestion problem might occur when the traffic density increases because the number of basic safety messages carried on the communication channel also increases in vehicle-to-vehicle communications. A remedy algorithm proposed in SAE J2945/1 is designed to address the channel congestion issue by decreasing transmission frequency and radiated power. This study is to develop potential test procedures for evaluating or validating the congestion control algorithm. Simulations of a reference unit transmitting at a higher frequency are implemented to emulate a number of onboard equipment (OBE) transmitting at the normal interval of 100 ms (10 Hz). When the transmitting interval is reduced to 1.25 ms (800 Hz), the reference unit emulates 80 vehicles transmitting at 10 Hz. By increasing the number of reference units transmitting at 800 Hz in the simulations, the corresponding channel busy percentages are obtained. An algorithm for Global Positioning System (GPS) data generation of virtual vehicles is developed for facilitating the validation of transmission intervals in the congestion control algorithm. Channel busy percentage is the channel busy time over a specified period of time. Three or 4 reference units are needed to generate channel busy percentages between 50 and 80%, and 5 reference units can generate channel busy percentages above 80%. The proposed test procedures can verify the operation of congestion control algorithm when channel busy percentages are between 50 and 80% and above 80%. By using a GPS data generation algorithm, the test procedures can also verify the transmission intervals when traffic densities are 80 and 200 vehicles in a radius of 100 m. A suite of test tools with functional requirements is also proposed for facilitating the implementation of test procedures. The potential test procedures for a congestion control algorithm are developed based on the simulation results of channel busy percentage and the GPS data generation algorithm. The test tools can examine the requirement compliance automatically and objectively. The required number of reference units should be further validated using real OBEs before implementing these potential test procedures.


Localization and expression of clarin-1, the Clrn1 gene product, in auditory hair cells and photoreceptors.
The Usher syndrome 3A (CLRN1) gene encodes clarin-1, which is a member of the tetraspanin family of transmembrane proteins. Although identified more than 6 years ago, little is known about its localization or function in the eye and ear. We developed a polyclonal antibody that react with all clarin-1 isoforms and used it to characterize protein expression in cochlea and retina. In the cochlea, we observe clarin-1expression in the stereocilia of P0 mice, and in synaptic terminals present at the base of the auditory hair cells from E18 to P6. In the retina, clarin-1 localizes to the connecting cilia, inner segment of photoreceptors and to the ribbon synapses. RT-PCR from P0 cochlea and P28 retina show mRNAs encoding only isoforms 2 and 3. Western blots show that only isoform 2 is present in protein extracts from these same tissues. We examined clarin-1 expression in the immortomouse-derived hair cell line UB/OC-1. Only isoform 2 is expressed in UB/OC-1 at both mRNA and protein levels, suggesting this isoform is biologically relevant to hair cell function. The protein co-localizes with microtubules and post-transgolgi vesicles. The subcellular localization of clarin-1 in hair cells and photoreceptors suggests it functions at both the basal and apical poles of neurosensoriepithelia.


[Docosahexaenoic acid in the treatment of idiopathic male infertility].
docosahexaenoic acid is one of the most common fatty acids in the cell membranes of sperm. This substance is a structural component of cell membranes, and is responsible for such properties as plasticity and fluidity which are necessary for the implementation of the process of capacitation and acrosome reaction. In addition, docosahexaenoic acid has antioxidant properties. Aim of our study was to assess the effect of dietary supplement docosahexaenoic acid (Brudi PLUS) on such markers of male fertility as the sperm DNA integrity, standard indicators of semen analysis and cryotolerance in infertile men. a randomized, multicenter, double-blind, placebo-controlled study was conducted. A total of 109 infertile men were recruited to participate in the study. All patients underwent different analyzes, including semen analysis, sperm DNA fragmentation, MAR-test, test of cryotolerance and electron microscopy analysis of spermatozoa. In active treatment group patients were prescribed to drug containing 350 mg of docosahexaenoic acid 3 times a day for 90 days. In the control group, patients were administered to a placebo with a similar scheme. All analyzes were repeatedly performed after 3-months of therapy. An increase in the progressive mobility and sperm viability was determined after the test of cryotolerance in the active treatment group. According to electron microscopy analysis of spermatozoa, positive changes were observed in the number of heads of the normal form with the normal structure of chromatin and acrosome, as well as a decrease in the number of spermatozoa with insufficiently condensed chromatin. the therapy with drug Brudi PLUS in patients with male infertility allows to increase in cryotolerance of sperms as well as decrease in number of sperm ultrastructure defects.


Proposed factor structure of the Geriatric Depression Scale.
The Geriatric Depression Scale (GDS) is commonly used to measure depression in the elderly. However, there have been no reports of the underlying structure of the GDS. To this end, the GDS was administered to 326 community-dwelling elderly subjects, and the data were subjected to a factor analysis. A five-factor solution was selected and, after a varimax rotation, the factors that emerged could be described as: (1) sad mood, (2) lack of energy, (3) positive mood, (4) agitation, and (5) social withdrawal. This solution accounted for 42.9% of the variance. Knowledge of the factor structure should aid both clinicians and researchers in the interpretation of responses on the GDS.


Molecular characterisation and expression analysis of SEREX-defined antigen NUCB2 in gastric epithelium, gastritis and gastric cancer.
NUCB2 is an EF-hand Ca2+ binding protein that has been implicated in various physiological processes like calcium homeostasis, hypothalamic regulation of feeding and TNF receptor shedding. In our previous study we identified NUCB2 as a potential tumour antigen eliciting autoantibody responses in 5.4% of gastric cancer patients but not in the healthy individuals. The current study aimed to elucidate the molecular mechanism underlying NUCB2 immunogenicity and to gain an insight into the physiological functions of NUCB2 in the stomach. mRNA expression analysis demonstrated that NUCB2 is ubiquitously expressed in normal tissues, including lymphoid tissues, and downregulated in gastric tumours when compared with the adjacent relatively normal stomach tissues. The search for molecular alterations resulted in the identification of novel mRNA variants transcribed from an alternative promoter and expressed predominantly in gastric cancers. Western blot analysis demonstrated that the protein levels correspond to mRNA levels and revealed that NUCB2 is phosphorylated in gastric mucosa. Furthermore, a 55 kDa isoform, generated presumably by yet an unidentified post-translational modification was detected in gastric tumours and AGS gastric cancer cells but was absent in the relatively normal gastric mucosa and thereby might have served as a trigger for the immune response against NUCB2. Staining of stomach tissue microarray with anti-NUCB2 antibody revealed that it is expressed in the secretory granules of chief cells and in the cytoplasm of parietal cells in the functioning gastric glands which are lost in atrophic glands and tumour cells. Hence we propose that NUCB2 may be implicated in gastric secretion by establishing an agonist-releasable Ca2+ store in ER or Golgi apparatus, signalling via heterotrimeric Gα proteins and/or mediating the exocytosis of the secretory granules.


The Oxidosqualene Cyclase from the Oomycete Saprolegnia parasitica Synthesizes Lanosterol as a Single Product.
The first committed step of sterol biosynthesis is the cyclisation of 2,3-oxidosqualene to form either lanosterol (LA) or cycloartenol (CA). This is catalyzed by an oxidosqualene cyclase (OSC). LA and CA are subsequently converted into various sterols by a series of enzyme reactions. The specificity of the OSC therefore determines the final composition of the end sterols of an organism. Despite the functional importance of OSCs, the determinants of their specificity are not well understood. In sterol-synthesizing oomycetes, recent bioinformatics, and metabolite analysis suggest that LA is produced. However, this catalytic activity has never been experimentally demonstrated. Here, we show that the OSC of the oomycete Saprolegnia parasitica, a severe pathogen of salmonid fish, has an uncommon sequence in a conserved motif important for specificity. We present phylogenetic analysis revealing that this sequence is common to sterol-synthesizing oomycetes, as well as some plants, and hypothesize as to the evolutionary origin of some microbial sequences. We also demonstrate for the first time that a recombinant form of the OSC from S. parasitica produces LA exclusively. Our data pave the way for a detailed structural characterization of the protein and the possible development of specific inhibitors of oomycete OSCs for disease control in aquaculture.


Fast image reconstruction with L2-regularization.
We introduce L2-regularized reconstruction algorithms with closed-form solutions that achieve dramatic computational speed-up relative to state of the art L1- and L2-based iterative algorithms while maintaining similar image quality for various applications in MRI reconstruction. We compare fast L2-based methods to state of the art algorithms employing iterative L1- and L2-regularization in numerical phantom and in vivo data in three applications; (i) Fast Quantitative Susceptibility Mapping (QSM), (ii) Lipid artifact suppression in Magnetic Resonance Spectroscopic Imaging (MRSI), and (iii) Diffusion Spectrum Imaging (DSI). In all cases, proposed L2-based methods are compared with the state of the art algorithms, and two to three orders of magnitude speed up is demonstrated with similar reconstruction quality. The closed-form solution developed for regularized QSM allows processing of a three-dimensional volume under 5 s, the proposed lipid suppression algorithm takes under 1 s to reconstruct single-slice MRSI data, while the PCA based DSI algorithm estimates diffusion propagators from undersampled q-space for a single slice under 30 s, all running in Matlab using a standard workstation. For the applications considered herein, closed-form L2-regularization can be a faster alternative to its iterative counterpart or L1-based iterative algorithms, without compromising image quality.


alpha 1-adrenoceptors in normal and hyperalgesic human skin.
1. Evidence of an adrenergic component of cutaneous hyperalgesia has recently been obtained in animal models of painful peripheral neuropathy. These findings have prompted speculation that an increased density or sensitivity of peripheral alpha-adrenoceptors contributes to sensory abnormalities and chronic neuropathic pain in conditions such as reflex sympathetic dystrophy. However, it is not known whether alpha-adrenoceptors are present at the site of nociception, either in hyperalgesic or normal skin. 2. We used the selective radioligand 125I-hydroxyphenyl-ethyl-aminomethyl-tetralone (HEAT) to label alpha 1-adrenoceptors, and quantitative autoradiography to assess the relative density of these receptors in skin samples from seven normal individuals and from the hyperalgesic and pain-free limbs of five patients with reflex sympathetic dystrophy. The distribution of autoradiographic grains over the epidermis and dermis was investigated in 10 microns serial transverse sections. 3. alpha 1-Adrenoceptors were identified in the epidermis and dermal papillae of normal individuals, and in the hyperalgesic and pain-free skin of patients with reflex sympathetic dystrophy. The density of alpha 1-adrenoceptors was greater in the epidermis and dermal papillae than further down in the dermis. 4. The mean density of alpha 1-adrenoceptors was significantly greater in the hyperalgesic skin of patients than in the skin of normal individuals (35.4 grains/1000 microns2 compared with 15.5 grains/ 1000 microns2, P < 0.01). The mean density of alpha 1-adrenoceptors in the pain-free skin of patients (26.9 grains/1000 microns2) fell midway between receptor density in hyperalgesic skin and in the skin of normal individuals, and did not differ significantly from either. 5. Our findings indicate that alpha 1-adrenoceptors are present in the epidermis, and suggest that their numbers may be increased in the hyperalgesic skin of patients with reflex sympathetic dystrophy. Further studies need to identify the dermal and epidermal cell types that express high densities of alpha 1-adrenoceptors, and to investigate their normal function and role in neuropathic pain.


Test-retest reliability study of the Pennsylvania Bi-Manual Worksample.
Test-retest reliability of a vocational assessment battery called the Pennsylvania Bi-Manual Worksample (PBWS) (Roberts, 1969) was examined in this study. The PBWS was administered to 30 adults with developmental disabilities at two different times, with an interval of 7 to 10 days between administrations. The subjects, who had mild to moderate degrees of mental retardation and ranged in age from 25 to 51 years, were chosen randomly from a group of 95 employees of a sheltered workshop for adults with developmental disabilities in western New York. When the scores from the two test administrations were compared to determine test-retest reliability, a high intraclass correlation value of .98 was found, which indicates that the PBWS provides consistent information about the vocational skills of adults with developmental disabilities.


Serum leptin, C-reactive protein, and cancer mortality in the NHANES III.
Adipokines, such as leptin, may affect cancer through its link with inflammation and obesity. We investigated the association between leptin, C-reactive protein, and risk of cancer death while accounting general and abdominal obesity. From the Third National Health and Examination Survey (NHANES III), we selected 5957 adult men and women with baseline measurements of serum leptin and CRP. Multivariable Cox regression was used to assess leptin and CRP levels (low, moderate, high) in relation to risk of cancer death. Stratification analyses were performed for obesity as defined by body mass index (BMI) and waist circumference. Fine and Gray regression was performed to account for death from cardiovascular disease and other causes as competing events. A total of 385 participants died of cancer during a mean follow-up of 18 years. After adjusting for BMI and waist circumference, an inverse association with log-transformed leptin was found for women, with a hazard ratio (HR) of 0.81 (95% confidence interval [CI]: 0.51-1.30) and 0.40 (95% CI: 0.24-0.68) for moderate and high compared to low levels of leptin, respectively; P(trend) = 0.0007). No association for leptin was observed in men, but higher CRP corresponded to increased risk of dying from cancer (HR: 2.98; 95% CI: 1.57-5.64 for the highest vs. lowest categories of CRP). Similar associations were observed with competing risk analysis also adjusted for BMI and waist circumference. Contrasting associations of serum leptin and CRP with cancer mortality may indicate sex-specific biological or environmental pathways linking obesity and cancer in men and women which warrant mechanistic investigations.