Fine mapping of 10q and 18q for familial Alzheimer's disease in Caribbean Hispanics.
Familial Alzheimer's disease (AD [MIM 104300]) has been a focus of intense investigation, primarily in Caucasian families from Europe and North America families. Although the late-onset form of familial AD, beginning after age 65 years, has been linked to regions on chromosomes 10q and 12p, the specific genetic variants have not yet been consistently identified. Using a unique cohort of families of Caribbean Hispanics ancestry, we screened the genome using 340 markers on 490 family members from 96 families with predominantly late-onset AD. We observed the strongest support for linkage on 18q (LOD=3.14). However, 17 additional markers (chromosomes 1-6, 8, 10, 12, and 14) exceeded a two-point LOD score of 1.0 under the affecteds-only autosomal dominant model or affected sibpair model. As we previously reported the fine-mapping effort on 12p showing modest evidence of linkage, we focused our fine-mapping efforts on two other candidate regions in the current report, namely 10q and 18q. We added 31 family members and eight additional Caribbean Hispanic families to fine map 10q and 18q. With additional microsatellite markers, the evidence for linkage for 18q strengthened near 112 cM, where the two-point LOD score for D18S541 was 3.37 and the highest NPL score in that region was 3.65 (P=0.000177). This narrow region contains a small number of genes expressed in the brain. However, at 10q (134-138 cM), the NPL score decreased from 3.15 (P=0.000486) to 2.1 (P=0.0218), but two broad peaks remained overlapping with previously reported peaks. Our results provide modest support for linkage on 10q and 12p in this cohort of Caribbean Hispanic families with familial Alzheimer's disease, and strong evidence for a new locus on 18q.


Regulation of mitochondrial iron accumulation by Yfh1p, a putative homolog of frataxin.
The gene responsible for Friedreich's ataxia, a disease characterized by neurodegeneration and cardiomyopathy, has recently been cloned and its product designated frataxin. A gene in Saccharomyces cerevisiae was characterized whose predicted protein product has high sequence similarity to the human frataxin protein. The yeast gene (yeast frataxin homolog, YFH1) encodes a mitochondrial protein involved in iron homeostasis and respiratory function. Human frataxin also was shown to be a mitochondrial protein. Characterizing the mechanism by which YFH1 regulates iron homeostasis in yeast may help to define the pathologic process leading to cell damage in Friedreich's ataxia.


Molecular Adsorption Changes the Quantum Structure of Oxide-Supported Gold Nanoparticles: Chemisorption versus Physisorption.
STM conductance spectroscopy and mapping has been used to analyze the impact of molecular adsorption on the quantized electronic structure of individual metal nanoparticles. For this purpose, isophorone and CO2, as prototype molecules for physisorptive and chemisorptive binding, were dosed onto monolayer Au islands grown on MgO thin films. The molecules attach exclusively to the metal-oxide boundary, while the interior of the islands remains pristine. The Au quantum well states are perturbed due to the adsorption process and increase their mutual energy spacing in the CO2 case but move together in isophorone-covered islands. The shifts disclose the nature of the molecule-Au interaction, which relies on electron exchange for the CO2 ligands but on dispersive forces for the organic species. Our experiments reveal how molecular adsorption affects individual quantum systems, a topic of utmost relevance for heterogeneous catalysis.


Longitudinal assessment of the neuroanatomical consequences of deep brain stimulation: Application of fornical DBS in an Alzheimer's mouse model.
Following its success in the treatment of refractory movement disorders, deep brain stimulation (DBS) is currently under clinical investigation as a possible treatment for several neuropsychiatric disorders, including Alzheimer's disease (AD). DBS's mechanism of action, delivery regimen, optimal brain target, and timeline of behavioural and neuroanatomical outcomes are all open fields of investigation. There is a critical need to develop methodologies that allow us to examine the time course of both behavioural changes and neuroanatomical remodelling in response to DBS. Here we present a proof-of-concept methodology for DBS experiments which incorporates both brain imaging and behaviour in a longitudinal fashion. We implanted triple transgenic AD mouse models (expressing mutations for both amyloid and tau) with custom magnetic resonance imaging (MRI)-compatible, carbon based electrodes. Mice received DBS or sham stimulation to the fornix (a critical white matter node in the brain's memory circuit) for 1 h (100 Hz, 100 μs pulses, 100 μA). Treatment was followed with an adapted Morris water maze (to test learning and memory; performed weekly) and structural MRI (to assess neuroanatomy; 3 days before and 3 days after DBS with a 6 week follow-up). The acute DBS treatment improved learning and long term memory in a delayed, sex specific, and transient manner relative to sham-stimulated controls. Significant, persistent, volumetric changes were seen in diverse brains structures, also heavily mediated by sex. We believe this methodology may be used as a template for DBS-related experimentation and will encourage preclinical studies to use longitudinal designs.


Functional coupling of adenine nucleotide translocase and mitochondrial creatine kinase is enhanced after exercise training in lung transplant skeletal muscle.
Mechanisms responsible for limitation of exercise capacity in lung transplant recipients (LR) and benefits gained by exercise training were studied. Mitochondrial respiration parameters, energy transfer, and cell structure were assessed in vastus lateralis biopsies using the permeabilized fiber technique with histochemical and morphometric measurements. Twelve male controls (C) and 12 LR performed exercise training over 12 wk. Before exercise training, there were strong correlations between exercise capacity (maximal O(2) consumption and endurance time at 70% maximal power output) and cellular events, as assessed by percentage of type I fibers and apparent K(m) for exogenous ADP. Anticalcineurins were not involved in LR exercise limitation, since there were no differences in maximal mitochondrial rate of respiration before exercise training and no abnormalities in respiratory chain complexes compared with C. Training resulted in a significant increase in physiological parameters both at the cellular (apparent K(m) for exogenous ADP and stimulating effect of creatine) and integrated (maximal O(2) consumption, power output at ventilatory threshold, maximal power output, and endurance time at 70% maximal power output) levels in LR and C. After the training period, improvements in maximal O(2) consumption and in maximal mitochondrial rate of respiration were noted, as well as changes in endurance time and percentage of type I fibers. Because there were no changes in diameters and fiber types, baseline alteration of apparent K(m) for exogenous ADP and its improvement after training might be related to changes within the intracellular energetic units. After the training period, intracellular energetic units exhibited a higher control of mitochondrial respiration by creatine linked to a more efficient functional coupling adenine nucleotide translocase-mitochondrial creatine kinase, resulting in better exercise performances in C and LR.


Development and psychometric properties of a scale assessing the needs of caregivers of children with disabilities.
Planning and evaluation of health care services for children with disabilities requires information on their caregivers' needs. This paper aims to present the development and psychometric properties of the Caregiver Needs Scale (CNS), a scale assessing the needs of caregivers of children with disabilities aged 0-12 years in Malaysia. Development of the scale went through a multistage process of literature review, modification of an existing instrument, input from experts and feedback from service users. Literature review identified content domains and response options. An exploratory factor analysis (EFA) was undertaken to identify subscales of caregiver needs. The internal consistency reliability, convergent validity and discriminant validity of the new scale were examined. 273 caregivers of children with disabilities completed the fielded questionnaire. EFA revealed 4 subscales of caregiver needs: need for 'Help getting information and services for the child,' 'Help coping with the child,' 'Help getting child care' and 'Help with finances.' Three items with factor loading <0.4 were dropped. Cronbach's alpha coefficients of the subscales ranged from 0.813 to 0.903. Total CNS score correlated with number of child's needs and unmet needs. The score was also higher in families with financial and employment problems. A new instrument was developed to assess the needs of caregivers of children with disabilities for use in the Malaysian population. The CNS showed satisfactory psychometric properties but further examination is warranted to confirm its validity.


Role of extracellular space in hyperosmotic suppression of potassium-induced electrographic seizures.
1. Focal electrographic seizures arose in the CA1 region of rat hippocampal slices bathed in elevated (8.5 mM) external potassium [( K+]o). High [K+]o also induced spontaneous interictal bursts that originated in area CA3 and propagated to CA1. To examine the contribution to electrographic seizure initiation of excitatory mechanisms that are influenced by extracellular volume, we studied the effect of hyperosmotic expansion of interstitial volume on seizure occurrence, interictal bursts, and excitatory synaptic transmission. The tissue electrical resistance was also measured leading up to and during seizures. 2. Media made 5-30 mosmol/kg hyperosmotic by addition of agents restricted to the extracellular space (mannitol, sucrose, raffinose, L-glucose, dextran) rapidly and reversibly abolished [K+]o-induced spontaneous CA1 seizures in 86% of slices tested. However, similar increases in osmolality effected by agents that access the intracellular compartment (D-glucose, glycerol) did not influence electrographic seizure occurrence. Hyperosmotic changes with plasma membrane impermeable compounds, but not permeable compounds, produced significant concentration-dependent decreases (1-10%) in the electrical resistance of CA1 stratum pyramidale. Because tissue resistance is proportional to extracellular volume, these results suggest that hyperosmotic suppression of electrographic seizures is associated with expansion of the extracellular space in hippocampal slices. 3. Measurement of electrical resistance of the CA1 stratum pyramidale during spreading depression and electrographic seizure revealed an increase in tissue resistance to 122% and 108% of control, respectively. Furthermore, a slight (approximately 2%) but significant increase in electrical resistance gradually occurred over the 20 s immediately preceding seizure generation. The observed increase in tissue resistance suggests extracellular space is decreased during these events. 4. Hyperosmolality did not alter CA3 interictal burst frequency. However, burst intensity, estimated from the total length of the burst waveform, was significantly reduced in both the CA3 (83% control) and CA1 region (67% control) when osmotic changes were imposed by plasma membrane impermeant compounds. Additionally, media made hypoosmotic by removal of 7.5 mM NaCl reversibly increased burst intensity. 5. High [K+]o potentiated excitatory synaptic transmission and excitatory postsynaptic potential (EPSP) spike coupling.(ABSTRACT TRUNCATED AT 400 WORDS)


Molecular cloning and characterization of an L-epinephrine transporter from sympathetic ganglia of the bullfrog, Rana catesbiana.
Chemical signaling by dopamine (DA) and L-norepinephrine (L-NE) at synapses is terminated by uptake via specialized presynaptic transport proteins encoded by the DA transporter (DAT) and L-NE transporter (NET) genes, respectively. In some vertebrate neurons, particularly the sympathetic neurons of amphibians, L-NE is converted to L-epinephrine (L-Epi, adrenaline) and released as the primary neurotransmitter. Although evidence exists for a molecularly distinct L-Epi transporter (ET) in the vertebrate brain and peripheral nervous system, a transporter specialized for extracellular L-Epi clearance has yet to be identified. To pursue this issue, we cloned transporter cDNAs from bullfrog (Rana catesbiana) paravertebral sympathetic ganglia and characterized functional properties via heterologous expression in non-neuronal cells. A cDNA of 2514 bp (fET) was identified for which the cognate 3.1 kb mRNA is highly enriched in frog sympathetic ganglia. Sequence analysis of the fET cDNA reveals an open reading frame coding for a protein of 630 amino acids. Inferred fET protein sequence bears 75, 66, and 48% amino acid identity with human NET, DAT, and the 5-hydroxytryptamine transporter (SERT), respectively. Transfection of fET confers Na+- and Cl--dependent catecholamine uptake in HeLa cells. Uptake of [3H]-L-NE by fET is inhibited by catecholamines in a stereospecific manner. L-Epi and DA inhibit fET-mediated [3H]-L-NE uptake more potently than they inhibit [3H]-L-NE uptake by human NET (hNET), whereas L-NE exhibits equivalent potency between the two carriers. Moreover, fET exhibits a greater maximal velocity (Vmax) for the terminal products of catecholamine biosynthesis (L-Epi > L-NE >> DA), unlike hNET, in which a Vmax rank order of L-NE > DA > L-Epi is observed. fET-mediated transport of catecholamines is sensitive to cocaine and tricyclic antidepressants, with antagonist potencies significantly correlated with hNET inhibitor sensitivity. Amino acid conservation and divergence of fET with mammalian catecholamine transporters help define residues likely to be involved in catecholamine recognition and translocation as well as blockade by selective reuptake inhibitors.


Passivation of PbS Quantum Dot Surface with l-Glutathione in Solid-State Quantum-Dot-Sensitized Solar Cells.
Surface oxidation of quantum dots (QDs) is one of the biggest challenges in quantum dot-sensitized solar cells (QDSCs), because it introduces surface states that enhance electron-hole recombination and degrade device performance. Protection of QDs from surface oxidation by passivating the surface with organic or inorganic layers can be one way to overcome this issue. In this study, solid-state QDSCs with a PbS QD absorber layer were prepared from thin mesoporous TiO2 layers by the successive ionic layer adsorption/reaction (SILAR) method. Spiro-OMeTAD was used as the organic p-type hole transporting material (HTM). The effects on the solar cell performance of passivating the surface of the PbS QDs with the tripeptide l-glutathione (GSH) were investigated. Current-voltage characteristics and external quantum efficiency measurements of the solar cell devices showed that GSH-treatment of the QD-sensitized TiO2 electrodes more than doubled the short circuit current and conversion efficiency. Impedance spectroscopy, intensity-modulated photovoltage and photocurrent spectroscopy analysis of the devices revealed that the enhancement in solar cell performance of the GSH-treated cells originates from improved charge injection from PbS QDs into the conduction band of TiO2. Time-resolved photoluminescence decay measurements show that passivation of the surface of QDs with GSH ligands increases the exciton lifetime in the QDs.


Stress-induced stimulation of choline transport in cultured choroid plexus epithelium exposed to low concentrations of cadmium.
The choroid plexus epithelium forms the blood-cerebrospinal fluid barrier and accumulates essential minerals and heavy metals. Choroid plexus is cited as being a "sink" for heavy metals and excess minerals, serving to minimize accumulation of these potentially toxic agents in the brain. An understanding of how low doses of contaminant metals might alter transport of other solutes in the choroid plexus is limited. Using primary cultures of epithelial cells isolated from neonatal rat choroid plexus, our objective was to characterize modulation of apical uptake of the model organic cation choline elicited by low concentrations of the contaminant metal cadmium (CdCl₂). At 50-1,000 nM, cadmium did not directly decrease or increase 30-min apical uptake of 10 μM [(3)H]choline. However, extended exposure to 250-500 nM cadmium increased [(3)H]choline uptake by as much as 75% without marked cytotoxicity. In addition, cadmium induced heat shock protein 70 and heme oxygenase-1 protein expression and markedly induced metallothionein gene expression. The antioxidant N-acetylcysteine attenuated stimulation of choline uptake and induction of stress proteins. Conversely, an inhibitor of glutathione synthesis l-buthionine-sulfoximine (BSO) enhanced stimulation of choline uptake and induction of stress proteins. Cadmium also activated ERK1/2 MAP kinase. The MEK1 inhibitor PD98059 diminished ERK1/2 activation and attenuated stimulation of choline uptake. Furthermore, inhibition of ERK1/2 activation abated stimulation of choline uptake in cells exposed to cadmium with BSO. These data indicate that in the choroid plexus, exposure to low concentrations of cadmium may induce oxidative stress and consequently stimulate apical choline transport through activation of ERK1/2 MAP kinase.


Unique catalytic activities and scaffolding of p21 activated kinase-1 in cardiovascular signaling.
P21 activated kinase-1 (Pak1) has diverse functions in mammalian cells. Although a large number of phosphoproteins have been designated as Pak1 substrates from in vitro studies, emerging evidence has indicated that Pak1 may function as a signaling molecule through a unique molecular mechanism - scaffolding. By scaffolding, Pak1 delivers signals through an auto-phosphorylation-induced conformational change without transfer of a phosphate group to its immediate downstream effector(s). Here we review evidence for this regulatory mechanism based on structural and functional studies of Pak1 in different cell types and research models as well as in vitro biochemical assays. We also discuss the implications of Pak1 scaffolding in disease-related signaling processes and the potential in cardiovascular drug development.


Biomechanical evaluation of lateral lumbar interbody fusion with secondary augmentation.
OBJECTIVE Lateral lumbar interbody fusion (LLIF) has emerged as a popular method for lumbar fusion. In this study the authors aimed to quantify the biomechanical stability of an interbody implant inserted using the LLIF approach with and without various supplemental fixation methods, including an interspinous plate (IP). METHODS Seven human cadaveric L2-5 specimens were tested intact and in 6 instrumented conditions. The interbody implant was intended to be used with supplemental fixation. In this study, however, the interbody was also tested without supplemental fixation for a relative comparison of these conditions. The instrumented conditions were as follows: 1) interbody implant without supplemental fixation (LLIF construct); and interbody implant with supplemental fixation performed using 2) unilateral pedicle screws (UPS) and rod (LLIF + UPS construct); 3) bilateral pedicle screws (BPS) and rods (LLIF + BPS construct); 4) lateral screws and lateral plate (LP) (LLIF + LP construct); 5) interbody LP and IP (LLIF + LP + IP construct); and 6) IP (LLIF + IP construct). Nondestructive, nonconstraining torque (7.5 Nm maximum) induced flexion, extension, lateral bending, and axial rotation, whereas 3D specimen range of motion (ROM) was determined optoelectronically. RESULTS The LLIF construct reduced ROM by 67% in flexion, 52% in extension, 51% in lateral bending, and 44% in axial rotation relative to intact specimens (p < 0.001). Adding BPS to the LLIF construct caused ROM to decrease by 91% in flexion, 82% in extension and lateral bending, and 74% in axial rotation compared with intact specimens (p < 0.001), providing the greatest stability among the constructs. Adding UPS to the LLIF construct imparted approximately one-half the stability provided by LLIF + BPS constructs, demonstrating significantly smaller ROM than the LLIF construct in all directions (flexion, p = 0.037; extension, p < 0.001; lateral bending, p = 0.012) except axial rotation (p = 0.07). Compared with the LLIF construct, the LLIF + LP had a significant reduction in lateral bending (p = 0.012), a moderate reduction in axial rotation (p = 0.18), and almost no benefit to stability in flexion-extension (p = 0.86). The LLIF + LP + IP construct provided stability comparable to that of the LLIF + BPS. The LLIF + IP construct provided a significant decrease in ROM compared with that of the LLIF construct alone in flexion and extension (p = 0.002), but not in lateral bending (p = 0.80) and axial rotation (p = 0.24). No significant difference was seen in flexion, extension, or axial rotation between LLIF + BPS and LLIF + IP constructs. CONCLUSIONS The LLIF construct that was tested significantly decreased ROM in all directions of loading, which indicated a measure of inherent stability. The LP significantly improved the stability of the LLIF construct in lateral bending only. Adding an IP device to the LLIF construct significantly improves stability in sagittal plane rotation. The LLIF + LP + IP construct demonstrated stability comparable to that of the gold standard 360° fixation (LLIF + BPS).


Cell type of origin as well as genetic alterations contribute to breast cancer phenotypes.
Breast cancer is classified into different subtypes that are associated with different patient survival outcomes, underscoring the importance of understanding the role of precursor cell and genetic alterations in determining tumor subtypes. In this study, we evaluated the oncogenic phenotype of two distinct mammary stem/progenitor cell types designated as K5+/K19- or K5+/K19+ upon introduction of identical combinations of oncogenes-mutant H-Ras (mRas) and mutant p53 (mp53), together with either wild-type ErbB2(wtErbB2) or wild-type EGFR (wtEGFR). We examined their tumor forming and metastasis potential, using both in-vitro and in-vivo assays. Both the combinations efficiently transformed K5+/K19- or K5+/K19+ cells. Xenograft tumors formed by these cells were histologically heterogeneous, with variable proportions of luminal, basal-like and claudin-low type components depending on the cell types and oncogene combinations. Notably, K5+/K19- cells transformed with mRas/mp53/wtEGFR combination had a significantly longer latency for primary tumor development than other cell lines but more lung metastasis incidence than same cells expressing mRas/mp53/wtErbB2. K5+/K19+ cells exhibit shorter overall tumor latency, and high metastatic potential than K5+/K19- cells, suggesting that these K19+ progenitors are more susceptible to oncogenesis and metastasis. Our results suggest that both genetic alterations and cell type of origin contribute to oncogenic phenotype of breast tumors.


The indirect immunobead test for seminal antisperm antibodies and fertilization rates at in-vitro fertilization.
A series of 183 patients with positive indirect immunobead tests on semen was studied to determine the correlation in semen between specific antibody types, binding sites, antibody concentration, and fertilizing ability. IgM was present in only 44 ejaculates and was present in sufficient quantity to cause significant binding to immunobeads (i.e. >20% of motile donor spermatozoa) in only three of them. There was no correlation between the percentages of motile donor spermatozoa that bound IgA and IgG immunobeads but the two classes of beads generally bound to the same region of the spermatozoa. A total of 63 couples went on to attempt in-vitro fertilization (IVF) treatment, all with mature eggs recovered. Of these mature eggs, 44% were fertilized and cleaved normally in comparison to 68% in a group of patients with tubal disease. Fertilization rates in individuals followed a bimodal distribution with a substantial number of couples experiencing zero or very poor rates (0-20%), the mode for the remainder lying between 60 and 80%. The fertilization rate tended to decrease as the amount of antibody increased. The percentage of donor spermatozoa that bound to immunobeads, taken as the greater of IgA and IgG, was selected by logistic regression as a significant predictor of poor fertilization (rate <=25%). The predictive power of the equation was improved by including the motile normal sperm concentration but the equation could only account for a small proportion of the total variation in fertilization rate. The presence of antibodies to the sperm head was highly correlated with the antibody concentration but was not selected as a predictor of fertilization. We conclude that the nature of the antigen against which the seminal antisperm antibody is directed may be as important as the antibody concentration in affecting sperm function. There seems to be little practical value in measuring IgM in seminal plasma.


Early detection of liver cancer based on bioluminescence tomography.
As a new modality of molecular imaging, bioluminescence imaging has been widely used in tumor detection and drug evaluation. However, BLI cannot present the depth of information for internal diseases such as a liver tumor in situ or a lung tumor in situ. In this paper, we describe a bioluminescence tomography (BLT) method based on the bioluminescent intensity attenuation calibration and applied it to the early detection of liver cancer in situ. In comparison with BLT without calibration, this method could improve the reconstruction accuracy by more than 10%. In comparison with micro-computed tomography and other traditional imaging modalities, this method can detect a liver tumor at a very early stage and provide reliable location information.


Heart disease and dementia: a population-based study.
There are conflicting reports on the possible positive association between coronary disease and dementia. The objectives of this study were to examine the association between coronary disease, as measured by myocardial infarction and cardiac death, and dementia in a population-based study. By use of the record-linkage system of the Rochester Epidemiology Project, 916 cases of dementia and 916 age (+/-1 year)- and sex-matched controls were identified in Rochester, Minnesota, between 1985 and 1994. From the same population, the authors identified all subjects who experienced a myocardial infarction (defined using standardized criteria) during the period 1979-1998. For myocardial infarction occurring prior to the index year of dementia, the authors used conditional logistic regression (case-control analysis), while for myocardial infarction and death occurring after the index year, they used competing risk survival analysis to account for informative censoring (cohort analysis). Before the index year, the odds ratio for myocardial infarction among cases with dementia compared with controls was 1.00 (95% confidence interval (CI): 0.62, 1.62; p = 1.00). After the index year, patients with dementia had a 46% decreased risk of subsequent myocardial infarction (hazard ratio = 0.54, 95% CI: 0.36, 0.82; p = 0.004) and an 18% decreased risk of cardiac death (hazard ratio = 0.82, 95% CI: 0.70, 0.95; p = 0.010). There was no evidence of a positive association between dementia and preceding myocardial infarction, while there was a decreased risk of myocardial infarction and cardiac death following dementia.


Pollination, seed set and seed predation on a landscape scale.
We analysed the combined effects of pollination and seed predation on seed set of Centaurea jacea in 15 landscapes differing in structural complexity. In the centre of each landscape, a patch of Centaurea plants was established for standardized measurements of flower visitation, seed predation and seed set. Both the number of flower-visiting bees and the proportion of flower heads damaged by seed predators increased with landscape complexity, which was measured as the proportion of semi-natural habitats. The mean number of seeds per flower head did not increase with the proportion of semi-natural habitats, presumably because of the counterbalancing effects of pollination and seed predation. For a subset of undamaged flower heads, the number of seeds per flower head was positively correlated with the number of flower visits. Further reasons for the unexpected failure to detect a correlation between landscape complexity and seed set appeared to be changes in flower-visitor behaviour and the contrasting responses of honeybees and wild bees to habitat context. Landscape analyses at eight spatial scales (radius of landscape sectors, 250-3000 m) showed that different groups perceived the landscape at different spatial scales. Changes in pollinator numbers could be explained only at small scales (up to 1000 m), while the seed predators also responded to large scales (up to 2500 m).


Demystifying excessively volatile human learning: A Bayesian persistent prior and a neural approximation.
Understanding how humans and animals learn about statistical regularities in stable and volatile environments, and utilize these regularities to make predictions and decisions, is an important problem in neuroscience and psychology. Using a Bayesian modeling framework, specifically the Dynamic Belief Model (DBM), it has previously been shown that humans tend to make the default assumption that environmental statistics undergo abrupt, unsignaled changes, even when environmental statistics are actually stable. Because exact Bayesian inference in this setting, an example of switching state space models, is computationally intensive, a number of approximately Bayesian and heuristic algorithms have been proposed to account for learning/prediction in the brain. Here, we examine a neurally plausible algorithm, a special case of leaky integration dynamics we denote as EXP (for exponential filtering), that is significantly simpler than all previously suggested algorithms except for the delta-learning rule, and which far outperforms the delta rule in approximating Bayesian prediction performance. We derive the theoretical relationship between DBM and EXP, and show that EXP gains computational efficiency by foregoing the representation of inferential uncertainty (as does the delta rule), but that it nevertheless achieves near-Bayesian performance due to its ability to incorporate a "persistent prior" influence unique to DBM and absent from the other algorithms. Furthermore, we show that EXP is comparable to DBM but better than all other models in reproducing human behavior in a visual search task, suggesting that human learning and prediction also incorporates an element of persistent prior. More broadly, our work demonstrates that when observations are information-poor, detecting changes or modulating the learning rate is both difficult and thus unnecessary for making Bayes-optimal predictions.


Genome sequence of the phage-gene rich marine Phaeobacter arcticus type strain DSM 23566(T.).
Phaeobacter arcticus Zhang et al. 2008 belongs to the marine Roseobacter clade whose members are phylogenetically and physiologically diverse. In contrast to the type species of this genus, Phaeobacter gallaeciensis, which is well characterized, relatively little is known about the characteristics of P. arcticus. Here, we describe the features of this organism including the annotated high-quality draft genome sequence and highlight some particular traits. The 5,049,232 bp long genome with its 4,828 protein-coding and 81 RNA genes consists of one chromosome and five extrachromosomal elements. Prophage sequences identified via PHAST constitute nearly 5% of the bacterial chromosome and included a potential Mu-like phage as well as a gene-transfer agent (GTA). In addition, the genome of strain DSM 23566(T) encodes all of the genes necessary for assimilatory nitrate reduction. Phylogenetic analysis and intergenomic distances indicate that the classification of the species might need to be reconsidered.


Effect of calcium chloride, zinc chloride, and water infusion on metmyoglobin reducing activity and fresh lamb color.
Calcium chloride (CaCl2), zinc chloride (ZnCl2), or water infusions were used to investigate the biochemical factors that affect fresh lamb color, and to examine the role of metmyoglobin-reducing activity in regulating this important quality attribute. Immediately after exsanguination, lamb carcasses (n = 6 per treatment) were infused (10% of BW) with 0.3 M CaCl2, 0.05 M ZnCl2, or water via a catheter inserted into the left carotid artery. The right LM was excised at 24-h postmortem and divided into two halves. The caudal portion was cut into 2.5-cm-thick chops and displayed for 6 d under 1,076 lx of white fluorescent lighting at 2 degrees C, whereas the cranial half was vacuum-packaged and stored at 2 degrees C for 3 wk before retail display. Objective color measurements and samples for biochemical analysis were taken at 0, 1, 3, and 6 d of display. In infused carcasses, pH decline was more rapid (P < 0.05) than in untreated controls, and it was greatest for CaCl2-infused carcasses. Calcium chloride-infused carcasses had lower (P < 0.01) NAD and higher (P < 0.001) NADPH concentrations than water- and ZnCl2-infused or untreated control carcasses. The negative effects of calcium infusion on fresh lamb color, higher (P < 0.01) metmyoglobin accumulation rate, and lower (P < 0.01) L*, a*, and b* color measurements could be explained by the lower amounts of unbound water (P < 0.01), shorter sarcomere length (P < 0.01), lower NAD concentrations (P < 0.01), and higher lipid peroxidation (P < 0.01). Zinc and water-infusions produced less (P < 0.01) lipid oxidation and improved the color and color stability of fresh lamb (P < 0.001). Rate of lipid oxidation in LM chops was greater (P < 0.01) after 3 wk of vacuum-packaged storage than 24-h postmortem. Metmyoglobin-reducing activities (sarcoplasmic and myofibrillar) were decreased in response to infusion treatments (P < 0.001), and ZnCl2 infusion resulted in the lowest metmyoglobin-reducing activities (P < 0.001). A significant association between the myofibrillar metmyoglobin-reducing activity and lipid peroxidation was observed, but metmyoglobin-reducing activities were not associated with any improvement in lamb color. Strategies to increase the antioxidant levels in lamb are very important to improve lamb quality, especially during vacuum-packaging storage.


Evaluating RNAlater® as a preservative for using near-infrared spectroscopy to predict Anopheles gambiae age and species.
Mosquito age and species identification is a crucial determinant of the efficacy of vector control programmes. Near-infrared spectroscopy (NIRS) has previously been applied successfully to rapidly, non-destructively, and simultaneously determine the age and species of freshly anesthetized African malaria vectors from the Anopheles gambiae s.l. species complex: An. gambiae s. s. and Anopheles arabiensis. However, this has only been achieved on freshly-collected specimens and future applications will require samples to be preserved between field collections and scanning by NIRS. In this study, a sample preservation method (RNAlater(®)) was evaluated for mosquito age and species identification by NIRS against scans of fresh samples. Two strains of An. gambiae s.s. (CDC and G3) and two strains of An. arabiensis (Dongola, KGB) were reared in the laboratory while the third strain of An. arabiensis (Ifakara) was reared in a semi-field system. All mosquitoes were scanned when fresh and rescanned after preservation in RNAlater(®) for several weeks. Age and species identification was determined using a cross-validation. The mean accuracy obtained for predicting the age of young (<7 days) or old (≥ 7 days) of all fresh (n = 633) and all preserved (n = 691) mosquito samples using the cross-validation technique was 83% and 90%, respectively. For species identification, accuracies were 82% for fresh against 80% for RNAlater(®) preserved. For both analyses, preserving mosquitoes in RNAlater(®) was associated with a highly significant reduction in the likelihood of a misclassification of mosquitoes as young or old using NIRS. Important to note is that the costs for preserving mosquito specimens with RNAlater(®) ranges from 3-13 cents per insect depending on the size of the tube used and the number of specimens pooled in one tube. RNAlater(®) can be used to preserve mosquitoes for subsequent scanning and analysis by NIRS to determine their age and species with minimal costs and with accuracy similar to that achieved from fresh insects. Cold storage availability allows samples to be stored longer than a week after field collection. Further study to develop robust calibrations applicable to other strains from diverse ecological settings is recommended.


Antibiotics for preterm rupture of membranes.
Premature birth carries substantial neonatal morbidity and mortality. Subclinical infection is associated with preterm rupture of membranes (PROM). Prophylactic maternal antibiotic therapy might lessen infectious morbidity and delay labour, but could suppress labour without treating underlying infection. To evaluate the immediate and long-term effects of administering antibiotics to women with PROM before 37 weeks, on maternal infectious morbidity, neonatal morbidity and mortality, and longer-term childhood development. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (30 September 2013). Randomised controlled trials comparing antibiotic administration with placebo that reported clinically relevant outcomes were included as were trials of different antibiotics. Trials in which no placebo was used were included for the outcome of perinatal death alone. We extracted data from each report without blinding of either the results or the treatments that women received. We sought unpublished data from a number of authors. We included 22 trials, involving 6872 women and babies.The use of antibiotics following PROM is associated with statistically significant reductions in chorioamnionitis (average risk ratio (RR) 0.66, 95% confidence interval (CI) 0.46 to 0.96, and a reduction in the numbers of babies born within 48 hours (average RR 0.71, 95% CI 0.58 to 0.87) and seven days of randomisation (average RR 0.79, 95% CI 0.71 to 0.89). The following markers of neonatal morbidity were reduced: neonatal infection (RR 0.67, 95% CI 0.52 to 0.85), use of surfactant (RR 0.83, 95% CI 0.72 to 0.96), oxygen therapy (RR 0.88, 95% CI 0.81 to 0.96), and abnormal cerebral ultrasound scan prior to discharge from hospital (RR 0.81, 95% CI 0.68 to 0.98). Co-amoxiclav was associated with an increased risk of neonatal necrotising enterocolitis (RR 4.72, 95% CI 1.57 to 14.23).One study evaluated the children's health at seven years of age (ORACLE Children Study) and found antibiotics seemed to have little effect on the health of children. Routine prescription of antibiotics for women with preterm rupture of the membranes is associated with prolongation of pregnancy and improvements in a number of short-term neonatal morbidities, but no significant reduction in perinatal mortality. Despite lack of evidence of longer-term benefit in childhood, the advantages on short-term morbidities are such that we would recommend antibiotics are routinely prescribed. The antibiotic of choice is not clear but co-amoxiclav should be avoided in women due to increased risk of neonatal necrotising enterocolitis.


Supernumerary teeth--an overview of classification, diagnosis and management.
Most supernumerary teeth are located in the anterior maxillary region. They are classified according to their form and location. Their presence may give rise to a variety of clinical problems. Detection of supernumerary teeth is best achieved by thorough clinical and radiographic examination. Their management should form part of a comprehensive treatment plan. This article presents an overview of the clinical problems associated with supernumerary teeth and includes a discussion of the classification, diagnosis and management of this difficult clinical entity.


Natural Products as Sources of New Drugs over the Nearly Four Decades from 01/1981 to 09/2019.
This review is an updated and expanded version of the five prior reviews that were published in this journal in 1997, 2003, 2007, 2012, and 2016. For all approved therapeutic agents, the time frame has been extended to cover the almost 39 years from the first of January 1981 to the 30th of September 2019 for all diseases worldwide and from ∼1946 (earliest so far identified) to the 30th of September 2019 for all approved antitumor drugs worldwide. As in earlier reviews, only the first approval of any drug is counted, irrespective of how many "biosimilars" or added approvals were subsequently identified. As in the 2012 and 2016 reviews, we have continued to utilize our secondary subdivision of a "natural product mimic", or "NM", to join the original primary divisions, and the designation "natural product botanical", or "NB", to cover those botanical "defined mixtures" now recognized as drug entities by the FDA (and similar organizations). From the data presented in this review, the utilization of natural products and/or synthetic variations using their novel structures, in order to discover and develop the final drug entity, is still alive and well. For example, in the area of cancer, over the time frame from 1946 to 1980, of the 75 small molecules, 40, or 53.3%, are N or ND. In the 1981 to date time frame the equivalent figures for the N* compounds of the 185 small molecules are 62, or 33.5%, though to these can be added the 58 S* and S*/NMs, bringing the figure to 64.9%. In other areas, the influence of natural product structures is quite marked with, as expected from prior information, the anti-infective area being dependent on natural products and their structures, though as can be seen in the review there are still disease areas (shown in Table 2) for which there are no drugs derived from natural products. Although combinatorial chemistry techniques have succeeded as methods of optimizing structures and have been used very successfully in the optimization of many recently approved agents, we are still able to identify only two de novo combinatorial compounds (one of which is a little speculative) approved as drugs in this 39-year time frame, though there is also one drug that was developed using the "fragment-binding methodology" and approved in 2012. We have also added a discussion of candidate drug entities currently in clinical trials as "warheads" and some very interesting preliminary reports on sources of novel antibiotics from Nature due to the absolute requirement for new agents to combat plasmid-borne resistance genes now in the general populace. We continue to draw the attention of readers to the recognition that a significant number of natural product drugs/leads are actually produced by microbes and/or microbial interactions with the "host from whence it was isolated"; thus we consider that this area of natural product research should be expanded significantly.


Interrogating Genomic-Scale Data for Squamata (Lizards, Snakes, and Amphisbaenians) Shows no Support for Key Traditional Morphological Relationships.
Genomics is narrowing uncertainty in the phylogenetic structure for many amniote groups. For one of the most diverse and species-rich groups, the squamate reptiles (lizards, snakes, and amphisbaenians), an inverse correlation between the number of taxa and loci sampled still persists across all publications using DNA sequence data and reaching a consensus on the relationships among them has been highly problematic. In this study, we use high-throughput sequence data from 289 samples covering 75 families of squamates to address phylogenetic affinities, estimate divergence times, and characterize residual topological uncertainty in the presence of genome-scale data. Importantly, we address genomic support for the traditional taxonomic groupings Scleroglossa and Macrostomata using novel machine-learning techniques. We interrogate genes using various metrics inherent to these loci, including parsimony-informative sites (PIS), phylogenetic informativeness, length, gaps, number of substitutions, and site concordance to understand why certain loci fail to find previously well-supported molecular clades and how they fail to support species-tree estimates. We show that both incomplete lineage sorting and poor gene-tree estimation (due to a few undesirable gene properties, such as an insufficient number of PIS), may account for most gene and species-tree discordance. We find overwhelming signal for Toxicofera, and also show that none of the loci included in this study supports Scleroglossa or Macrostomata. We comment on the origins and diversification of Squamata throughout the Mesozoic and underscore remaining uncertainties that persist in both deeper parts of the tree (e.g., relationships between Dibamia, Gekkota, and remaining squamates; among the three toxicoferan clades Iguania, Serpentes, and Anguiformes) and within specific clades (e.g., affinities among gekkotan, pleurodont iguanians, and colubroid families).


Cleaning Ability of Irrigants and Orange Oil Solvent Combination in the Removal of Root Canal Filling Materials.
This study investigated the influence of different irrigants and the use of orange oil solvent in the removal of filling materials during root canal retreatment. Forty maxillary premolars were shaped using the ProTaper System up to file F3 (size 30, 0.09 taper) and filled by Tagger's hybrid technique using the AH Plus. Samples were randomly assigned to four groups (n=10) according to the irrigating protocol during endodontic retreatment with the ProTaper Universal Re-treatment System: G1, 2% chlorhexidine (CHX) gel; G2, 2% CHX gel with an orange oil solvent; G3, 5% sodium hypochlorite (NaOCl); and G4, 5% NaOCl with an orange oil solvent. Afterwards, the samples were longitu-dinally split into two halves, and the root wall images were prepared by scanning electron microscopy. Two pre-calibrated evaluators analyzed the images using a filling materials remnants score system. Data were statistically analyzed using the Kruskal-Wallis and Dunn's tests (p<0.05). All samples had residual filling materials in the root canal walls after instrumentation. According to the presence of the filling material remnants in the total area of samples, the groups were ranked in the follow-ing order: G2=G4>G1=G3. No statistical differences were found when the CHX and NaOCl were used (p>0.05). Groups in which a solvent was used showed a less effective cleaning ability (p<0.05). The use of NaOCl with solvent presented the highest amounts of filling materials remnants in the critical apical area (p<0.05). The use of orange oil with NaOCl or CHX does not improve the removal of residual root canal filling materials.


Do flame retardant chemicals increase the risk for thyroid dysregulation and cancer?
Flame retardant chemicals are added to consumer products to reduce fire incidence and severity; approximately 1.5 million tons of these chemicals are used annually. However, their widespread use has led to their ubiquitous presence in the environment and chronic accumulation in human tissues. We summarize current trends in human flame retardant chemical exposure, and review recent data highlighting concerns for thyroid dysregulation and cancer risk in human populations. Polybrominated diphenyl ethers were once commonly used as flame retardant chemicals, but recently were phased out. Exposure is associated with thyroid dysregulation (mainly T4 reductions) in animals, with new work focusing on specific mechanisms of action. Polybrominated diphenyl ethers also impact human thyroid regulation and are related to clinical thyroid disease, but associations appear both dose and life-stage dependent. Emerging data suggest that common alternate flame retardant chemicals may be more potent thyroid disruptors than their predecessors, which is particularly concerning given increasing levels of exposure. Potential health impacts of flame retardant chemicals are only beginning to be understood for 'legacy flame retardant chemicals' (i.e., polybrominated diphenyl ethers), and are largely unevaluated for newer-use chemicals. Cumulatively, current data suggest impact on thyroid regulation is likely, potentially implicating flame retardant chemicals in thyroid disease and cancers for which thyroid dysregulation impacts risk or prognosis.


High- and low-risk profiles for the development of multiple sclerosis within 10 years after optic neuritis: experience of the optic neuritis treatment trial.
To identify factors associated with a high and low risk of developing multiple sclerosis after an initial episode of optic neuritis. Three hundred eighty-eight patients who experienced acute optic neuritis between July 1, 1988, and June 30, 1991, were followed up prospectively for the development of multiple sclerosis. Consenting patients were reassessed after 10 to 13 years. The 10-year risk of multiple sclerosis was 38% (95% confidence interval, 33%-43%). Patients (160) who had 1 or more typical lesions on the baseline magnetic resonance imaging (MRI) scan of the brain had a 56% risk; those with no lesions (191) had a 22% risk (P<.001, log rank test). Among the patients who had no lesions on MRI, male gender and optic disc swelling were associated with a lower risk of multiple sclerosis, as was the presence of the following atypical features for optic neuritis: no light perception vision; absence of pain; and ophthalmoscopic findings of severe optic disc edema, peripapillary hemorrhages, or retinal exudates. The 10-year risk of multiple sclerosis following an initial episode of acute optic neuritis is significantly higher if there is a single brain MRI lesion; higher numbers of lesions do not appreciably increase that risk. However, even when brain lesions are seen on MRI, more than 40% of the patients will not develop clinical multiple sclerosis after 10 years. In the absence of MRI lesions, certain demographic and clinical features seem to predict a very low likelihood of developing multiple sclerosis. This natural history information is a critical input for estimating a patient's 10-year multiple sclerosis risk and for weighing the benefit of initiating prophylactic treatment at the time of optic neuritis or other initial demyelinating events in the central nervous system.


Actinoplanes teichomyceticus ATCC 31121 as a cell factory for producing teicoplanin.
Teicoplanin is a glycopeptide antibiotic used clinically in Europe and in Japan for the treatment of multi-resistant Gram-positive infections. It is produced by fermenting Actinoplanes teichomyceticus. The pharmaceutically active principle is teicoplanin A2, a complex of compounds designated T-A2-1-A2-5 differing in the length and branching of the fatty acid moiety linked to the glucosamine residue on the heptapeptide scaffold. According to European and Japanese Pharmacopoeia, components of the drug must be reproduced in fixed amounts to be authorized for clinical use. We report our studies on optimizing the fermentation process to produce teicoplanin A2 in A. teichomyceticus ATCC 31121. Robustness of the process was assessed on scales from a miniaturized deep-well microtiter system to flasks and 3-L bioreactor fermenters. The production of individual factors T-A2-1-A2-5 was modulated by adding suitable precursors to the cultivation medium. Specific production of T-A2-1, characterized by a linear C10:1 acyl moiety, is enhanced by adding methyl linoleate, trilinoleate, and crude oils such as corn and cottonseed oils. Accumulation of T-A2-3, characterized by a linear C10:0 acyl chain, is stimulated by adding methyl oleate, trioleate, and oils such as olive and lard oils. Percentages of T-A2-2, T-A2-4, and, T-A2-5 bearing the iso-C10:0, anteiso-C11:0, and iso-C11:0 acyl moieties, respectively, are significantly increased by adding precursor amino acids L-valine, L-isoleucine, and L-leucine. Along with the stimulatory effect on specific complex components, fatty acid esters, oils, and amino acids (with the exception of L-valine) inhibit total antibiotic productivity overall. By adding industrial oils to medium containing L-valine the total production is comparable, giving unusual complex compositions. Since the cost and the quality of teicoplanin production depend mainly on the fermentation process, we developed a robust and scalable fermentation process by using an industrial medium in which a complex composition can be modulated by the combined addition of suitable precursors. This work was performed in the wild-type strain ATCC 31121, which has a clear genetic background. This is important for starting a rational improvement program and also helps to better control teicoplanin production during process and strain development.


The Rubber Hand Illusion: Top-down attention modulates embodiment.
The Rubber Hand Illusion (RHI) creates distortions of body ownership through multimodal integration of somatosensory and visual inputs. This illusion largely rests on bottom-up (automatic multisensory and perceptual integration) mechanisms. However, the relative contribution from top-down factors, such as controlled processes involving attentional regulation, remains unclear. Following previous work that highlights the putative influence of higher-order cognition in the RHI, we aimed to further examine how modulations of working memory load and task instructions-two conditions engaging top-down cognitive processes-influence the experience of the RHI, as indexed by a number of psychometric dimensions. Relying on exploratory factor analysis for assessing this phenomenology within the RHI, our results confirm the influence of higher-order, top-down mental processes. Whereas task instruction strongly modulated embodiment of the rubber hand, cognitive load altered the affective dimension of the RHI. Our findings corroborate that top-down processes shape the phenomenology of the RHI and herald new ways to improve experimental control over the RHI.


The impact of reference gene selection in quantification of gene expression levels in guinea pig cervical tissues and cells.
Accurate measurements of mRNA expression levels in tissues or cells are crucially dependent on the use of relevant reference genes for normalization of data. In this study we used quantitative real-time PCR and two Excel-based applets (geNorm and BestKeeper) to determine the best reference genes for quantification of target gene mRNA in a complex tissue organ such as the guinea pig cervix. Gene expression studies were conducted in cervical epithelium and stroma during pregnancy and parturition and in cultures of primary cells from this tissue. Among 15 reference gene candidates examined, both geNorm and BestKeeper found CLF1 and CLTC to be the most stable in cervical stroma and cervical epithelium, ACTB and PPIB in primary stroma cells, and CLTC and PPIB in primary epithelial cells. The order of stability among the remaining candidate genes was not in such an agreement. Commonly used reference such as GAPDH and B2M demonstrated lower stability. Determination of pairwise variation values for reference gene combinations using geNorm revealed that the geometric mean of the two most stable genes provides sufficient normalization in most cases. However, for cervical stroma tissue in which many reference gene candidates displayed low stability, inclusion of three reference genes in the geometric mean may improve accuracy of target gene expression level analyses. Using the top ranked reference genes we examined the expression levels of target gene PTGS2 in cervical tissue and cultured cervical cells. We compared the results with PTGS2 expression normalized to the least stable gene and found significant differences in gene expression, up to 10-fold in some samples, emphasizing the importance of appropriately selecting reference genes. We recommend using the geometric mean of CFL1 and CLTC for normalization of qPCR studies in guinea pig cervical tissue studies, ACTB and PPIB in primary stroma cells and CLTC and PPIB in primary epithelial cells from guinea pig.


Systems analysis of biological networks in skeletal muscle function.
Skeletal muscle function depends on the efficient coordination among subcellular systems. These systems are composed of proteins encoded by a subset of genes, all of which are tightly regulated. In the cases where regulation is altered because of disease or injury, dysfunction occurs. To enable objective analysis of muscle gene expression profiles, we have defined nine biological networks whose coordination is critical to muscle function. We begin by describing the expression of proteins necessary for optimal neuromuscular junction function that results in the muscle cell action potential. That action potential is transmitted to proteins involved in excitation-contraction coupling enabling Ca(2+) release. Ca(2+) then activates contractile proteins supporting actin and myosin cross-bridge cycling. Force generated by cross-bridges is transmitted via cytoskeletal proteins through the sarcolemma and out to critical proteins that support the muscle extracellular matrix. Muscle contraction is fueled through many proteins that regulate energy metabolism. Inflammation is a common response to injury that can result in alteration of many pathways within muscle. Muscle also has multiple pathways that regulate size through atrophy or hypertrophy. Finally, the isoforms associated with fast muscle fibers and their corresponding isoforms in slow muscle fibers are delineated. These nine networks represent important biological systems that affect skeletal muscle function. Combining high-throughput systems analysis with advanced networking software will allow researchers to use these networks to objectively study skeletal muscle systems.


Cost-effectiveness of HIV testing referral strategies among tuberculosis patients in India.
Indian guidelines recommend routine referral for HIV testing of all tuberculosis (TB) patients in the nine states with the highest HIV prevalence, and selective referral for testing elsewhere. We assessed the clinical impact and cost-effectiveness of alternative HIV testing referral strategies among TB patients in India. We utilized a computer model of HIV and TB disease to project outcomes for patients with active TB in India. We compared life expectancy, cost, and cost-effectiveness for three HIV testing referral strategies: 1) selective referral for HIV testing of those with increased HIV risk, 2) routine referral of patients in the nine highest HIV prevalence states with selective referral elsewhere (current standard), and 3) routine referral of all patients for HIV testing. TB-related data were from the World Health Organization. HIV prevalence among TB patients was 9.0% in the highest prevalence states, 2.9% in the other states, and 4.9% overall. The selective referral strategy, beginning from age 33.50 years, had a projected discounted life expectancy of 16.88 years and a mean lifetime HIV/TB treatment cost of US$100. The current standard increased mean life expectancy to 16.90 years with additional per-person cost of US$10; the incremental cost-effectiveness ratio was US$650/year of life saved (YLS) compared to selective referral. Routine referral of all patients for HIV testing increased life expectancy to 16.91 years, with an incremental cost-effectiveness ratio of US$730/YLS compared to the current standard. For HIV-infected patients cured of TB, receiving antiretroviral therapy increased survival from 4.71 to 13.87 years. Results were most sensitive to the HIV prevalence and the cost of second-line antiretroviral therapy. Referral of all patients with active TB in India for HIV testing will be both effective and cost-effective. While effective implementation of this strategy would require investment, routine, voluntary HIV testing of TB patients in India should be recommended.


Misidentification of OLGA-PH-J/92, believed to be the only crustacean cell line.
Continuous cell lines from aquatic invertebrate species are few and the development of crustacean cell lines remains an elusive goal. Although a crayfish cell line derived from neural ganglia of Orconectes limosus was reported in 2000, this cell line OLGA-PH-J/92 failed to be authenticated as such. In this report, we describe our attempts to identify the taxonomic identity of the cell line through immunological and molecular techniques. Immunohistochemical screening for the expression of a suite of invertebrate neuropeptides gave negative results, precluding an invertebrate neural origin. PCR amplification and DNA sequencing for the mitochondrial cytochrome c oxydase I, and 18S ribosomal RNA genes that had been widely used to confirm species identity, could not confirm the OLGA-PH-J/92 cells as originating from crayfish. Subsequent attempts to identify the cells provided moderate homology (82%) to Gephyramoeba sp. (AF293897) following PCR amplification of an 18S rDNA fragment after a BLAST search. A literature search provided morphological evidence of the similarity of OLGA-PH-J/92 to the Gephyramoeba distributed by the American Type Culture Collection as ATCC 50654, which also had been misidentified and was renamed Acramoeba dendroida (Smirnov et al., Eur J Protistol 44:35-44, 2008). The morphology of the OLGA-PH-J/92 cells which remains identical to the original report (Neumann et al., In Vivo 14:691-698, 2000) and matched corresponding micrographs that were available from the ATCC before the cell line was dropped from their catalog (ATCC CRL 1494) is very similar to A. dendroida and could thus belong to the Acramoebidae. These results unequivocally indicate that the OLGA-PH-J/92 cell line is not derived from the crayfish O. limosus, and the search for an immortal crustacean cell line continues.


Fitness Costs and the Rapid Spread of kelch13-C580Y Substitutions Conferring Artemisinin Resistance.
Fitness costs are key determinants of whether drug resistance alleles establish and how fast they spread within populations. More than 125 different kelch13 alleles, each containing a different amino acid substitution, have arisen in Southeast Asian malaria parasite (Plasmodium falciparum) populations under artemisinin selection over the past 15 years in a dramatic example of a soft selective event. However, just one of these alleles (C580Y) is now outcompeting other alleles in multiple different countries and is spreading toward fixation. Here we examine the fitness consequences of C580Y, relative to another less successful kelch13 mutation (R561H), to try to explain the distinctive dynamics of C580Y. We hypothesized that C580Y will show lower fitness costs than other kelch13 substitutions in the absence of artemisinin treatment. We used CRISPR/Cas9 methods to introduce single mutations (C580Y or R561H) or synonymous control edits into a wild-type parasite isolated on the Thailand-Myanmar border, conducted replicated head-to-head competition assays, and determined the outcome of competition using deep sequencing of kelch13 amplicons. Contrary to our predictions, these experiments reveal that C580Y carries higher fitness costs (s [selection coefficient] = 0.15 ± 0.008 [1 standard error {SE}]) than R561H (s = 0.084 ± 0.005). Furthermore, R561H outcompetes C580Y in direct competition (s = 0.065 ± 0.004). We conclude that fitness costs of C580Y in isolation are unlikely to explain the rapid spread of this substitution.


Socioeconomic inequalities in very preterm birth rates.
To investigate the extent of socioeconomic inequalities in the incidence of very preterm birth over the past decade. Ecological study of all 549 618 births in the former Trent health region, UK, from 1 January 1994 to 31 December 2003. All singleton births of 22(+0) to 32(+6) weeks gestation (7 185 births) were identified from population surveys of neonatal services and stillbirths. Poisson regression was used to calculate incidence of very preterm birth (22-32 weeks) and extremely preterm birth (22-28 weeks) by year of birth and decile of deprivation (child poverty section of the Index of Multiple Deprivation). Incidence of very preterm singleton birth rose from 11.9 per 1000 births in 1994 to 13.7 per 1000 births in 2003. Those from the most deprived decile were at nearly twice the risk of very preterm birth compared with those from the least deprived decile, with 16.4 per 1000 births in the most deprived decile compared with 8.5 per 1000 births in the least deprived decile (incidence rate ratio 1.94; 95% CI (1.73 to 2.17)). This deprivation gap remained unchanged throughout the 10-year period. The magnitude of socio-economic inequalities was the same for extremely preterm births (22-28 weeks incidence rate ratio 1.94; 95% CI (1.62 to 2.32)). This large, unique dataset of very preterm births shows wide socio-economic inequalities that persist over time. These findings are likely to have consequences on the burden of long-term morbidity. Our research can assist future healthcare planning, the monitoring of socio-economic inequalities and the targeting of interventions in order to reduce this persistent deprivation gap.


Assessing Oxidative Stress in Tumors by Measuring the Rate of Hyperpolarized [1-13C]Dehydroascorbic Acid Reduction Using 13C Magnetic Resonance Spectroscopy.
Rapid cancer cell proliferation promotes the production of reducing equivalents, which counteract the effects of relatively high levels of reactive oxygen species. Reactive oxygen species levels increase in response to chemotherapy and cell death, whereas an increase in antioxidant capacity can confer resistance to chemotherapy and is associated with an aggressive tumor phenotype. The pentose phosphate pathway is a major site of NADPH production in the cell, which is used to maintain the main intracellular antioxidant, glutathione, in its reduced state. Previous studies have shown that the rate of hyperpolarized [1-13C]dehydroascorbic acid (DHA) reduction, which can be measured in vivo using non-invasive 13C magnetic resonance spectroscopic imaging, is increased in tumors and that this is correlated with the levels of reduced glutathione. We show here that the rate of hyperpolarized [1-13C]DHA reduction is increased in tumors that have been oxidatively prestressed by depleting the glutathione pool by buthionine sulfoximine treatment. This increase was associated with a corresponding increase in pentose phosphate pathway flux, assessed using 13C-labeled glucose, and an increase in glutaredoxin activity, which catalyzes the glutathione-dependent reduction of DHA. These results show that the rate of DHA reduction depends not only on the level of reduced glutathione, but also on the rate of NADPH production, contradicting the conclusions of some previous studies. Hyperpolarized [1-13C]DHA can be used, therefore, to assess the capacity of tumor cells to resist oxidative stress in vivo However, DHA administration resulted in transient respiratory arrest and cardiac depression, which may prevent translation to the clinic.


An epidemiologic model of the incidence of acute illness in Plasmodium falciparum malaria.
We propose a stochastic model for simulating malaria tolerance. The model relates the probability of a clinical attack of malaria to the peripheral parasite densities via a pyrogenic threshold that itself responds dynamically to the parasite load. The parameters of the model have been estimated by fitting it to the relationship between incidence of clinical episodes and the entomologic inoculation rate, using age-specific incidence data from two villages in Senegal and one village in Tanzania. The model reproduces the shifts in age distribution of clinical episodes associated with variation in transmission intensity, and in keeping with the data, predicts a slightly higher lifetime number of episodes in the mesoendemic village of Ndiop than in the holoendemic village of Dielmo. This model provides a parsimonious explanation of counter-intuitive relationships between the overall incidence of clinical malaria and transmission intensity. In contrast to the theory of endemic stability, recently proposed to apply to P. falciparum, it does not assume any intrinsic age dependence in the outcome of infection. This model can be used to explore the consequences for predictions of the effects of different anti-malarial interventions on the incidence of clinical malaria.


Left and right ventricular structure and function in subclinical hypothyroidism: the effects of one-year levothyroxine treatment.
The aim of this study was to investigate left ventricular (LV) and right ventricular (RV) structure, function, and mechanics in patients with subclinical hypothyroidism (SHT), and to evaluate the effect of a 1-year levothyroxine treatment. We compared 45 untreated women with subclinical hypothyroidism and 35 healthy control women matched by age. All the subjects underwent laboratory analyses, which included a thyroid hormone levels (free T3, free T4, and TSH) test, and a complete 2-dimensional echocardiographic study. All the SHT patients received levothyroxine therapy and were followed for a year after euthyroid state was achieved. The LV mass index in the SHT participants before and after replacement therapy was significantly higher than in controls. In the SHT patients before the treatment, LV diastolic function and global function estimated by the Tei index were significantly impaired, whereas the LV systolic function was decreased. The results show that LV mechanics was significantly impaired in the SHT patients at baseline. Additionally, the SHT participants before levothyroxine substitution had increased RV wall thickness and significantly impaired RV diastolic and global function in comparison with the controls or the SHT subjects after the treatment. Furthermore, RV mechanics was also significantly deteriorated in the SHT patients before the treatment. Subclinical hypothyroidism significantly affected LV and RV structure, systolic, diastolic and global function, and LV and RV mechanics. Levothyroxine replacement therapy significantly improved cardiac structure, function, and mechanics in the SHT patients.


6-Gingerol alleviates exaggerated vasoconstriction in diabetic rat aorta through direct vasodilation and nitric oxide generation.
The aim of the present study is to investigate the effect and potential mechanism of action of 6-gingerol on alterations of vascular reactivity in the isolated aorta from diabetic rats. Male Wistar rats were divided into two experimental groups, control and diabetics. Diabetes was induced by a single intraperitoneal injection of streptozotocin (50 mg kg(-1)), and the rats were left for 10 weeks to develop vascular complications. The effect of in vitro incubation with 6-gingerol (0.3-3 μM) on the vasoconstrictor response of the isolated diabetic aortae to phenylephrine and the vasodilator response to acetylcholine was examined. Effect of 6-gingerol was also examined on aortae incubated with methylglyoxal as an advanced glycation end product (AGE). To investigate the mechanism of action of 6-gingerol, the nitric oxide synthase inhibitor Nω-nitro-l-arginine methyl ester hydrochloride (100 μM), guanylate cyclase inhibitor methylene blue (5 μM), calcium-activated potassium channel blocker tetraethylammonium chloride (10 mM), and cyclooxygenase inhibitor indomethacin (5 μM) were added 30 minutes before assessing the direct vasorelaxant effect of 6-gingerol. Moreover, in vitro effects of 6-gingerol on NO release and the effect of 6-gingerol on AGE production were examined. Results showed that incubation of aortae with 6-gingerol (0.3-10 μM) alleviated the exaggerated vasoconstriction of diabetic aortae to phenylephrine in a concentration-dependent manner with no significant effect on the impaired relaxatory response to acetylcholine. Similar results were seen in the aortae exposed to methylglyoxal. In addition, 6-gingerol induced a direct vasodilation effect that was significantly inhibited by Nω-nitro-l-arginine methyl ester hydrochloride and methylene blue. Furthermore, 6-gingerol stimulated aortic NO generation but had no effect on AGE formation. In conclusion, 6-gingerol ameliorates enhanced vascular contraction in diabetic aortae, which may be partially attributed to its ability to increase the production of NO and stimulation of cyclic guanosine monophosphate.


Positive personal changes in the aftermath of head and neck cancer diagnosis: a qualitative study in patients and their spouses.
There is increasing evidence that serious and life-threatening illnesses such as malignant tumors not only can lead to increased psychological stress, but also can lead to positive changes. In this qualitative study, 31 patients with head and neck cancer and 25 women partners were interviewed with regard to positive personal changes (posttraumatic growth). The audiotaped responses were analyzed using qualitative context analysis. A total of 25 patients (81%) and their women partners (84%) reported positive changes. Qualitative content analysis revealed 3 different categories of growth: attitudes toward life, personal strength, and relationships. Partners reported significantly more positive changes in relationships, especially, within the partnership. The total amount of positive change was almost equal. Positive personal changes are frequently reported by patients and by significant others. The total amount of positive change is almost equal in patients and their women partners, but wives experience more positive changes within the partnership.


Chitosan-stabilized platinum nanoparticles as effective oxidase mimics for colorimetric detection of acid phosphatase.
Capping molecules on the surface of nanomaterials not only enhance the dispersion and stability of nanomaterials but also greatly facilitate their surface modification and biological applications. However, most capping molecules can severely block the active sites of the catalytic core, thereby decreasing the enzymatic activity of nanomaterial-based enzyme mimics. This work demonstrates the superiority of chitosan (Ch) as a capping molecule for synthesizing catalytic platinum nanoparticles (PtNPs). The experimental results show that Ch simultaneously exhibits an excellent stabilizing effect and enhances the oxidase-like activity of PtNPs. Kinetic studies indicate that Ch-PtNPs have a higher affinity for 3,3',5,5'-tetramethylbenzidine (TMB) than other kinds of oxidase mimics. Furthermore, the TMB chromogenic reaction catalyzed by Ch-PtNPs is found to be much faster in an acidic medium, thus adapting well to the optimal pH for acid phosphatase (ACP). Therefore, a novel colorimetric approach for ACP determination is developed for the first time, which is based on the Ch-PtNP-catalyzed oxidation of TMB, the inhibitory effect of ascorbic acid (AA) on the oxidase-like activity of Ch-PtNPs, and the ACP-catalyzed hydrolysis of AA 2-phosphate (AAP) into AA. The linear range for ACP is 0.25-2.5 U L-1 and the limit of detection is measured to be 0.016 U L-1. This new colorimetric method is utilized to detect ACP in real biological samples and to screen ACP inhibitors. We believe that these new PtNPs, which exhibit high colloidal stability, excellent catalytic performance, good biocompatibility, simple preparation, and easy modification, can be promising candidates for a broad range of applications in optical sensing, environmental monitoring, clinical diagnosis, and drug discovery.


Klebsiella pneumoniae bloodstream infections among neonates in a high-risk nursery in Cali, Colombia.
To determine the cause of an outbreak of Klebsiella pneumoniae bloodstream infections (BSIs) among neonates in a high-risk nursery and to institute control measures. During the on-site investigation, a cohort study to identify risk factors for K. pneumoniae BSI, a point-prevalence study to assess K. pneumoniae colonization, a maternal cohort study to determine maternal K. pneumoniae colonization, and an observational study to evaluate healthcare worker (HCW) compliance with infection control practices were conducted. Neonates in a 40-bed high-risk nursery in a 700-bed university hospital in Cali, Colombia. Cohorting of neonates colonized with K. pneumoniae. The overall K. pneumoniae BSI attack rate was 10 of 105 (9.5%). In the retrospective cohort study, the number of blood transfusions (OR, 3.1 per transfusion; P = .02; CI95, 1.4-9.7) and intravenous injections (OR, 1.2 per injection; P = .04; CI95, 1.0-1.5) were independently associated with K. pneumoniae BSI. The overall prevalence of K. pneumoniae colonization was 61% among neonates and 7% among mothers. During the HCW assessment, suboptimal intravenous therapy practices were observed. A cohorting intervention resulted in a significant reduction in K. pneumoniae colonization (12% vs 61%; RR, 0.19; P < .001). During the intervention period, no K. pneumoniae BSIs occurred. This investigation suggested that the outbreak probably occurred due to widespread colonization and suboptimal infection control and intravenous therapy practices. Cohorting successfully reduced the overall prevalence of K. pneumoniae colonization and, along with improved infection control practices, probably prevented K. pneumoniae BSIs


Meta-analysis of diagnostic accuracy studies with multiple thresholds: Comparison of different approaches.
Methods for standard meta-analysis of diagnostic test accuracy studies are well established and understood. For the more complex case in which studies report test accuracy across multiple thresholds, several approaches have recently been proposed. These are based on similar ideas, but make different assumptions. In this article, we apply four different approaches to data from a recent systematic review in the area of nephrology and compare the results. The four approaches use: a linear mixed effects model, a Bayesian multinomial random effects model, a time-to-event model and a nonparametric model, respectively. In the case study data, the accuracy of neutrophil gelatinase-associated lipocalin for the diagnosis of acute kidney injury was assessed in different scenarios, with sensitivity and specificity estimates available for three thresholds in each primary study. All approaches led to plausible and mostly similar summary results. However, we found considerable differences in results for some scenarios, for example, differences in the area under the receiver operating characteristic curve (AUC) of up to 0.13. The Bayesian approach tended to lead to the highest values of the AUC, and the nonparametric approach tended to produce the lowest values across the different scenarios. Though we recommend using these approaches, our findings motivate the need for a simulation study to explore optimal choice of method in various scenarios.


Use of reference chemicals to determine passive uptake rates of common indoor air VOCs by collocation deployment of active and passive samplers.
Passive samplers have become more popular in their application in the measurement of airborne chemicals. For volatile organic compounds, the rate of a chemical's diffusivity is a determining factor in the quantity of the chemical being collected for a given passive sampler. While uptake rate of a chemical in the passive sampler can be determined either by collocation deployment of both active and passive samplers or use of controlled facilities such as environmental chambers, a new approach without a need for accurate active flow rate in the collocation experiment was demonstrated in this study. This approach uses chemicals of known uptake rates as references to calculate the actual flow rate of the active sampling in the collocation experiment. The active sampling rate in turn can be used in the determination of the uptake rates of all other chemicals present in the passive samplers. The advantage of such approach is the elimination of the errors in actual active sampling rate associated with low flow employed in the collocation experiment. Using this approach, passive uptake rates of more than 80 volatile organic compounds commonly present in indoor air were determined. These experimentally determined uptake rates correlate well with air diffusivity of the chemicals, indicating the regression equation describing such correlation might be useful in predicting the uptake rates of other volatile organic chemicals in indoor air based on their air diffusivity.


In vitro potency and stability of fortified ophthalmic antibiotics.
This study evaluates the in vitro potency, stability and contamination of the fortified ophthalmic antibiotic preparations of cefazolin, vancomycin, gentamicin and tobramycin stored for 4 weeks. The effects of the different solvents and storage temperatures on the antimicrobial potency and stability were also examined. The fortified stock solution of cefazolin and vancomycin were prepared by reconstituting with 0.9% sodium chloride and with artificial tears. Gentamicin and tobramycin were prepared by adding parenteral forms into their commercial ophthalmic solutions. The antimicrobial potency was measured by the minimum bactericidal concentration for cefazolin (33.3 mg/mL) and vancomycin (31 mg/mL) against Staphylococcus aureus, and for tobramycin (13.5 mg/mL) and gentamicin (13.5 mg/mL) against Pseudomonas aeruginosa. The stability of solution was evaluated by measuring absorbance spectra and pH. During the study period the levels of contamination of the stock solutions were examined by culturing on blood and on Sabouraud-dextrose media. There were no differences in the antimicrobial potency of the cefazolin, vancomycin and tobramycin within the 4-week period; however, the potency of gentamicin was decreased at both temperatures after 21 days. It was found that different solvents or storage temperatures had no effect on the potency. Throughout the 4-week period, no change was found in the absorbance spectra of gentamicin and tobramycin, whereas the absorbance spectra of cefazolin and vancomycin stored at 24 degrees C increased in both of the solvents used (P<0.05). In all of the antibiotics, pH changed at 24 degrees C after 7-10 days; however, at 4 degrees C, only the pH of tobramycin exhibited changes after 14 days (P<0.05). No contamination was detected in the stock solutions during the study period. Topical fortified antibiotic solutions used for longer than 7 days should be stored at < or =4 degrees C, those stored at 24 degrees C should be discarded after 7 days.


Endometrial lymphomyeloid cells in abnormal uterine bleeding due to levonorgestrel (Norplant).
Endometrial lymphomyeloid cell subsets were evaluated in samples from normal women and from women with abnormal uterine bleeding due to subcutaneous levonorgestrel implants (Norplant) or an intrauterine device (IUD). The frequency of CD3(+), CD68(+), CD43(+) and endometrial granulated lymphoid cells was evaluated by immunohistochemical or phloxine-tartrazine staining of formalin-fixed paraffin-embedded samples. In normal women, cyclic variation in lymphomyeloid subsets was seen. In women using Norplant for contraception, the frequency of CD3(+), CD68(+) and CD43(+) cells was dramatically decreased, compatible with endometrial atrophy. When Norplant users with abnormal bleeding were compared to women without bleeding, however, the number of CD68(+) cells was significantly increased and the number of CD3(+) and CD43(+) cells was preserved, contrary to the hypothesis that this group would show a greater degree of atrophy and hence, tissue fragility. A similar pattern was seen in a preliminary study of women with bleeding associated with use of copper-only IUD contraception, and in samples taken from late secretory and menstrual biopsies from normal cycling women. Whether these changes in endometrial lymphomyeloid cells represent a result of bleeding arising from a common mechanism or rather cause the uterine bleeding is discussed.


Drought tolerance strategies highlighted by two Sorghum bicolor races in a dry-down experiment.
Drought stress is the major environmental stress that affects more and more frequently plant growth and productivity due to the current climate change scenario. Unravelling the physiological mechanism underlying the response of plants to water stress and discover traits related to drought tolerance provide new and powerful tools for the selection in breeding programmes. Four genotypes of Sorghum bicolor (L.) Moench were screened in a dry-down experiment using different approaches to discover physiological and molecular indicators of drought tolerance. Different strategies were identified in response to drought among the four genotypes and the two Sorghum race allowing to state the tolerance of durra race compared to the caudatum one and, within the durra race, the drought tolerance of the genotype IS22330. It retained high biomass production and high tolerance index, it had a low threshold of fraction of transpirable soil water and high capacity to recover leaf apparatus after drought stress. Furthermore in this study, the expression levels of four genes highlighted that they could be used as proxy for drought tolerance. Dehdrine (DHN) could be used for screening drought tolerance both in durra and in caudatum races. NADP-Malic Enzyme, Carbonic Anhydrase (CA) and Plasma membrane Intrinsic Protein (PIP2-5), being up-regulated by drought stress only in durra race, have a more limited, though nonetheless useful application. In the tolerant durra genotype IS22330 in particular, the regulation of stomatal openings was strongly related to NADP-Malic Enzyme expression.


ACC claims for sports-related dental trauma from 1999 to 2008: a retrospective analysis.
To describe trends in sports-related dental injuries reported to ACC in the past 10 years. Retrospective case series. De-identified data on orofacial injuries were obtained from ACC for the financial years 1999-2008, and new claims were identified for each year (with recurring claims omitted from the analysis). Patterns in new-claim data were identified by age, sex, region and the sport involved. Trends in claims for the sport categories were also identified The annual number of claims ranged from 24,998 to 31,257; overall, 38.7% of claims were made by females and 61.3% by males, and these proportions remained largely unchanged during the observation period. For sports-related claims, those aged 11-20 years had the highest percentage of claims (with between 41.7% and 44.4%, depending on year) while those older than 60 had the smallest percentage of claims, with 0.5% to 1.1%. Sport was involved in 20.6% to 26.2% of new claims. The highest percentage of injuries was attributable to rugby (between 22.2% and 33.1%, depending on the year). Water sports contributed to between 14.2% and 20.8% of claims. Cycling increased from 1.5% in 1999 to 15.3% in 2006, and then decreased to 10.6% in 2008. Hockey, basketball, soccer, cricket and netball had 4.4%, 4.8%, 6.9%, 4.7% and 3.9% respectively. Although its share has decreased in recent years, rugby remained the greatest contributor to sport-related orofacial injuries, with water sports consistently second (cycling had the largest change, with a rapid increase in the past 3 years). There is a need to re-examine mouthguard (and other injury prevention) policies for particular sports.


Context-dependent competition in a model gut bacterial community.
Understanding the ecological processes that generate complex community structures may provide insight into the establishment and maintenance of a normal microbial community in the human gastrointestinal tract, yet very little is known about how biotic interactions influence community dynamics in this system. Here, we use natural strains of Escherichia coli and a simplified model microbiota to demonstrate that the colonization process on the strain level can be context dependent, in the sense that the outcome of intra-specific competition may be determined by the composition of the background community. These results are consistent with previous models for competition between organisms where one competitor has adapted to low resource environments whereas the other is optimized for rapid reproduction when resources are abundant. The genomic profiles of E. coli strains representing these differing ecological strategies provide clues for deciphering the genetic underpinnings of niche adaptation within a single species. Our findings extend the role of ecological theory in understanding microbial systems and the conceptual toolbox for describing microbial community dynamics. There are few, if any, concrete examples of context-dependent competition on a single trophic level. However, this phenomenon can have potentially dramatic effects on which bacteria will successfully establish and persist in the gastrointestinal system, and the principle should be equally applicable to other microbial ecosystems.


Nitrogen utilization, nutrient digestibility, and excretion of purine derivatives in dairy cattle consuming rations containing corn milling co-products.
The objectives of this experiment were to determine the effects of feeding a combination of modified wet distillers grains with solubles (WDGS) and wet corn gluten feed (WCGF) on nutrient digestion, purine derivative excretion, and N utilization. Multiparous (n=20) and primiparous (n=20) cows were arranged in a replicated 5 x 5 Latin square with 21-d periods. Animals were fed one of 5 treatment diets during each period: 1) 0% co-products (control); 2) 15% WDGS (15WDGS); 3) 15% WCGF (15WCGF); 4) 7.5% WDGS and 7.5% WCGF (15MIX); and 5) 15% WDGS and 15% WCGF (30MIX; dry matter basis). A portion of forages, corn, and soy-based protein was replaced with WDGS, or WCGF, or both. Dry matter intake was greater for 15WDGS (25.1 kg/d) and 30MIX (25.5 kg/d) than for control (22.4 kg/d), 15WCGF (23.2 kg/d), or 15MIX (23.5 kg/d). Dry matter digestibility was greatest for 15WCGF and 30MIX (63.6 and 64.1%, respectively) and least for 15WDGS (59.8%), and neutral detergent fiber and N digestibility were greatest for 30MIX (50.7 and 68.6%, respectively) and lowest for 15WDGS (41.3 and 61.5%, respectively). Excretion of purine derivatives in urine was greater for co-product treatment diets than for control. Fecal N was greatest for 15WDGS compared with other treatment diets (311.0 vs. 263.3 g/d), whereas urinary N was greatest for 30MIX (330.0 g/d), intermediate for 15WCGF and 15MIX (319.3 and 320.5 g/d, respectively), and lowest for control and 15WDGS (308.5 and 312.2 g/d, respectively). Manure N (fecal+urinary N) was greatest for 15WDGS, intermediate for 15MIX and 30MIX, and lowest for control and 15WCGF. Treatment diets did not differ in 4% fat-corrected milk production. Compared with the ration containing WDGS, the ration with a 30% mixture of WDGS and WCGF improved nutrient digestibility and N utilization with reduced manure N excretion and increased N retention. Thus, it appears feeding WDGS and WCGF in combination reduces some of the negative effects of feeding WDGS alone.


Diethylstilbestrol regulates expression of avian apolipoprotein D during regression and recrudescence of the oviduct and epithelial-derived ovarian carcinogenesis.
Apolipoprotein D (APOD) is a glycoprotein which is widely expressed in mammalian tissues. It is structurally and functionally similar to the lipocalins which are multiple lipid-binding proteins that transport hydrophobic ligands and other small hydrophobic molecules, including cholesterol and several steroid hormones. Although multiple functions for APOD in various tissues have been reported, its expression, biological function, and hormonal regulation in the female reproductive system are not known. Thus, in this study, we focused on correlations between APOD and estrogen during development, differentiation, regression, and regeneration of the oviduct in chickens and in the development of ovarian carcinogenesis in laying hens. Results of the present study indicated that APOD messenger RNA (mRNA) expression increased (P < 0.001) in the luminal and glandular (GE) epithelia of the chicken oviduct in response to diethylstilbestrol (a nonsteroidal synthetic estrogen). In addition, the expression of APOD mRNA and protein decreased (P < 0.001) as the oviduct regressed during induced molting, and gradually increased (P < 0.001) with abundant expression in GE of the oviduct during recrudescence after molting. Furthermore, APOD mRNA and protein were predominantly localized in GE of cancerous, but not normal ovaries from laying hens. Collectively, results of the present study suggest that APOD is a novel estrogen-stimulated gene in the chicken oviduct which likely regulates growth, differentiation, and remodeling of the oviduct during oviposition cycles. Moreover, up-regulated expression of APOD in epithelial cell-derived ovarian cancerous tissue suggests that it could be a candidate biomarker for early detection and treatment of ovarian cancer in laying hens and in women.


Pre-procedural glucose levels and the risk for contrast-induced acute kidney injury in patients undergoing coronary angiography.
We sought to evaluate whether pre-procedural glucose levels are associated with contrast-induced acute kidney injury (CI-AKI) after coronary angiography. Although diabetes is a known risk factor for CI-AKI in patients undergoing coronary angiography, whether elevated pre-procedural glucose levels (regardless of pre-existing diabetes) are associated with higher risk for CI-AKI is unknown. We evaluated 6,358 patients with acute myocardial infarctions undergoing coronary angiography. Patients were stratified into 5 pre-procedural glucose groups: <110 mg/dl, 110 to <140 mg/dl, 140 to <170 mg/dl, 170 to <200 mg/dl, and >or=200 mg/dl. Logistic regression models were used to evaluate the relationship between glucose levels and risk for CI-AKI, first in the entire cohort and then in patients with and without established diabetes. The primary outcome was CI-AKI (>or=0.3 mg/dl absolute or >or=50% relative serum creatinine increase during 48 h after the procedure). The relationship between pre-procedural glucose and CI-AKI varied markedly in patients with and without diabetes. There was a strong association between glucose and CI-AKI risk in patients without diabetes (CI-AKI rates across the 5 glucose groups from lowest to highest: 8.2%, 9.9%, 12.4%, 14.9%, and 24.3%; p<0.001), but not in patients with diabetes (20.9%, 16.1%, 16.3%, 14.8%, and 19.2%, respectively; p=0.24; p for glucose x diabetes interaction<0.001). After adjusting for confounders (including baseline glomerular filtration rate), the relationship between higher glucose and greater CI-AKI risk persisted in patients without diabetes (odds ratios [95% confidence intervals] for glucose groups of 110 to <140 mg/dl, 140 to <170, mg/dl 170 to <200 mg/dl, and >or=200 mg/dl: 1.31 [1.00 to 1.71], 1.51 [1.11 to 2.10], 1.58 [1.03 to 2.43], and 2.14 [1.46 to 3.14] vs. glucose<110 mg/dl, respectively), but this relationship was not seen in patients with established diabetes. Elevated pre-procedural glucose is associated with greater risk for CI-AKI in patients without known diabetes who undergo coronary angiography in the setting of acute myocardial infarction. Measures used to prevent CI-AKI should be considered in these patients.


Mathematical modelling of metabolic regulation in aging.
The underlying cellular mechanisms that characterize aging are complex and multifaceted. However, it is emerging that aging could be regulated by two distinct metabolic hubs. These hubs are the pathway defined by the mammalian target of rapamycin (mTOR) and that defined by the NAD+-dependent deacetylase enzyme, SIRT1. Recent experimental evidence suggests that there is crosstalk between these two important pathways; however, the mechanisms underpinning their interaction(s) remains poorly understood. In this review, we propose using computational modelling in tandem with experimentation to delineate the mechanism(s). We briefly discuss the main modelling frameworks that could be used to disentangle this relationship and present a reduced reaction pathway that could be modelled. We conclude by outlining the limitations of computational modelling and by discussing opportunities for future progress in this area.


Evidence for independent genetic influences on fat mass and body mass index in a pediatric twin sample.
Insight into genetic and environmental influences on fat mass, independent of body mass index (BMI; kg/m2), is expected to enhance methods for treating pediatric obesity. However, few studies have estimated the heritability of fat mass in pediatric samples, and those conducted have relied primarily on BMI measurements. PRESENT STUDY: Using bioimpedance analysis, the present study tested a series of hypotheses predicting significant genetic and environmental influences on percent body fat (PBF) above and beyond BMI. Subjects were 66 pairs of twins, including 41 monozygotic and 25 dizygotic pairs, from 3 to 17 years of age. Structural equation modeling tested hypotheses, adjusting for demographic variables. Analyses indicated significant genetic influences on PBF, with genes estimated to account for 75% to 80% of the phenotypic variation. The remaining variation was attributable to nonshared environmental influences. Multivariate analyses revealed sizable genetic correlations and environmental correlations between BMI and PBF (rg =.74 and re =.67, respectively), suggesting that some genes and environmental experiences influence both phenotypes. However, analyses confirmed genetic and environmental influences on PBF above and beyond BMI. For example, 62.5% of the total genetic variation in PBF was attributable to genes that influenced PBF but not BMI. There seems to be a substantial genetic contribution to fat mass distinct from BMI in a sample of children and adolescents. Studies testing putative genetic or environmental determinants of pediatric obesity might be strengthened further by including research-based body composition methods.