Carnivorous mammals: nutrient digestibility and energy evaluation.
Estimating the energy content is the first step in diet formulation, as it determines the amount of food eaten and hence the concentration of nutrients required to meet the animal's requirements. Additionally, being able to estimate the energy content of a diet empirically known to maintain body condition in an animal will facilitate an estimation of maintenance energy requirements. We collated data on nutrient composition of diets fed to captive wild canids, felids, hyenids, mustelids, pinnipeds, and ursids and the digestibility coefficients from the literature (45 species, 74 publications) to test whether differences in protein and fat digestibility could be detected between species groups, and whether approaches suggested for the estimation of dietary metabolizable energy (ME) content in domestic carnivores (NRC [2006] Nutrient requirements of dogs and cats. Washington, DC: National Academy Press.) can be applied to wild carnivores as well. Regressions of digestible protein or fat content vs. the crude protein (CP) or fat content indicated no relevant differences in the digestive physiology between the carnivore groups. For diets based on raw meat, fish, or whole prey, applying the calculation of ME using "Atwater factors" (16.7 kJ/g CP; 16.7 kJ/g nitrogen-free extracts; 37.7 kJ/g crude fat) provided estimates that compared well to experimental results. This study suggests that ME estimation in such diets is feasible without additional digestion trials. For comparative nutrition research, the study implicates that highly digestible diets typically fed in zoos offer little potential to elucidate differences between species or carnivore groups, but research on diets with higher proportions of difficult-to-digest components (fiber, connective tissues) is lacking.


Steady state statistical correlations predict bistability in reaction motifs.
Various cellular decision making processes are regulated by bistable switches that take graded input signals and convert them to binary all-or-none responses. Traditionally, a bistable switch generated by a positive feedback loop is characterized either by a hysteretic signal response curve with two distinct signaling thresholds or by characterizing the bimodality of the response distribution in the bistable region. To identify the intrinsic bistability of a feedback regulated network, here we propose that bistability can be determined by correlating higher order moments and cumulants (≥2) of the joint steady state distributions of two components connected in a positive feedback loop. We performed stochastic simulations of four feedback regulated models with intrinsic bistability and we show that for a bistable switch with variation of the signal dose, the steady state variance vs. covariance adopts a signatory cusp-shaped curve. Further, we find that the (n + 1)th order cross-cumulant vs. nth order cross-cumulant adopts a closed loop structure for at least n = 3. We also propose that our method is capable of identifying systems without intrinsic bistability even though the system may show bimodality in the marginal response distribution. The proposed method can be used to analyze single cell protein data measured at steady state from experiments such as flow cytometry.


Contribution of E3-ubiquitin ligase activity to HIV-1 restriction by TRIM5alpha(rh): structure of the RING domain of TRIM5alpha.
TRIM5α(rh) is a cytosolic protein that potently restricts HIV-1 before reverse transcription. TRIM5α(rh) is composed of four different domains: RING, B-box 2, coiled coil, and B30.2(SPRY). The contribution of each of these domains to restriction has been extensively studied, with the exception of the RING domain. The RING domain of TRIM5α exhibits E3-ubiquitin ligase activity, but the contribution of this activity to the restriction of HIV-1 is not known. To test the hypothesis that the E3-ubiquitin ligase activity of the RING domain modulates TRIM5α(rh) restriction of HIV-1, we correlated the E3-ubiquitin ligase activity of a panel of TRIM5α(rh) RING domain variants with the ability of these mutant proteins to restrict HIV-1. For this purpose, we first solved the nuclear magnetic resonance structure of the RING domain of TRIM5α and defined potential functional regions of the RING domain by homology to other RING domains. With this structural information, we performed a systematic mutagenesis of the RING domain regions and tested the TRIM5α RING domain variants for the ability to undergo self-ubiquitylation. Several residues, particularly the ones on the E2-binding region of the RING domain, were defective in their self-ubiquitylation ability. To correlate HIV-1 restriction to self-ubiquitylation, we used RING domain mutant proteins that were defective in self-ubiquitylation but preserve important properties required for potent restriction by TRIM5α(rh), such as capsid binding and higher-order self-association. From these investigations, we found a set of residues that when mutated results in TRIM5α molecules that lost both the ability to potently restrict HIV-1 and their self-ubiquitylation activity. Remarkably, all of these changes were in residues located in the E2-binding region of the RING domain. Overall, these results demonstrate a role for TRIM5α self-ubiquitylation in the ability of TRIM5α to restrict HIV-1.


Transactivation of receptor tyrosine kinases by purinergic P2Y and adenosine receptors.
Transactivation of receptor tyrosine kinases (RTK) is a crosstalk mechanism exhibited by G-protein-coupled receptors (GPCR) to activate signaling pathways classically associated with growth factors. The discovery of RTK transactivation was a breakthrough in signal transduction that contributed to developing current concepts in intracellular signaling. RTK transactivation links GPCR signaling to important cellular processes, such as cell proliferation and differentiation, and explains the functional diversity of these receptors. Purinergic (P2Y and adenosine) receptors belong to class A of GPCR; in the present work, we systematically review the experimental evidence showing that purinergic receptors have the ability to transactivate RTK in multiple tissues and physiopathological conditions resulting in the modulation of cellular physiology. Of particular relevance, the crosstalk between purinergic receptors and epidermal growth factor receptor is a redundant pathway that participates in multiple pathophysiological processes. Specific and detailed knowledge of purinergic receptor-regulated pathways advances our understanding of the complexity of GPCR signal transduction and opens the way for pharmacologic intervention in the pathological context.


Effect of some monoamine oxidase inhibitors on the thiamin status of rabbits.
The monamine oxidase (MAO) inhibitors harmaline, tranylcypromine, deprenyl, clorgyline and iproniazid were injected intraperitoneally for five days to rabbits at doses that produced significant MAO inhibition. The first three inhibitors raised the concentrations of pyruvate and lactate in blood, decreased the activity of erythrocyte transketolase (TK) and increased the thiamin pyrophosphate (TPP) effect. The drugs also produced anorexia and loss of body weight. The changes were suggestive of an adverse effect on the thiamin status. Clorgyline and iproniazid, however, raised the blood concentrations of pyruvate and lactate but did not affect significantly erythrocyte TK activity or TPP effect. Treatment of rabbits for 14 days with the thiamin antagonists pyrithiamine (20 micrograms kg-1) or oxythiamine (0.8 mg kg-1) produced a significant drop in TK activity and increase in the lactate and pyruvate concentrations as well as an increase in TPP effect. Pyrithiamine (5, 10 or 20 micrograms kg-1, 14 days) lowered significantly the activity of MAO in the liver and brain of rabbits. Treatment with the other antagonist, oxythiamine, at doses of 0.2 or 0.4 mg kg-1 for 14 days had no significant effect on MAO activity. At a dose of 0.8 mg kg-1 a significant drop in MAO activity occurred. A pair-feeding trial indicated that the biochemical changes produced in animals treated with MAO inhibitors were attributable to the drugs per se, and not to the ensuing anorexia. Thiamin (100 micrograms kg-1, subcutaneously) when given concomitantly with pyrithiamine, oxythiamine, harmaline, deprenyl and tranylcypromine was effective in preventing the development of thiamin deficiency.


Intersexuality incidence, sex ratio fluctuations and intersex reproductive output as factors affecting the temporal variation of intersexed populations of the marine amphipod Echinogammarus marinus.
The occurrence of intersexuality in populations of the marine amphipod Echinogammarus marinus has been related to several causes, including anthropogenic contamination, microsporidian parasites and environmental sex determination (ESD). At the individual-level, the costs of intersexuality in E. marinus are reflected in reduced reproductive output of intersexes compared to normal specimens. The impacts of intersexuality and other reproductive disorders in nature will only be fully understood when individual-level effects are upscale to the population-level, which can be achieved through ecological models. The aim of the present work is to simulate the density variation of real populations of E. marinus with different intersexuality incidence, proportions of phenotypes and intersex female reproductive rates. To achieve that, we have incorporated empirical data obtained at the individual-level from three intersexed populations of E. marinus into a population dynamic model. The original model was calibrated and validated for a long-term stable population without intersexuality and sex ratio approximately 1. In the present version, the model includes an extra population group, intersex females, but the total sum of flow rates from juveniles to adults was kept the same as in the original model to respect the previous validation. In these circumstances, results indicate that the male-biased population is very sensitive to intersexuality, even at low incidence levels, and exhibits significant density decrease if conditions do not change. On the contrary, the two populations with a preponderance of female-biased sex ratios, low or moderate intersexuality incidence (up to an average value of 9% of the population) and with intersex reproductive rates corresponding to 80% the value of normal specimens, exhibit long-term persistence. The model is highly sensitive to adult female parameters (e.g. mortality), indicating that small variations on its values can significantly change population responses. Overall the present results suggest that, in spite the costs of intersexuality in E. marinus, the temporal variation of intersexed populations depend on intersex incidence level, sex ratio fluctuations and reproductive output of intersexes. Simulations also suggest that other processes (e.g. mortality rates of normal females) may play important roles in the maintenance of populations with intersexuality. In the future, it will be crucial to improve and validate this type of models with long-term observations and monitoring of intersexed populations.


Wnt/β-catenin and FGF signalling direct the specification and maintenance of a neuromesodermal axial progenitor in ensembles of mouse embryonic stem cells.
The development of the central nervous system is known to result from two sequential events. First, an inductive event of the mesoderm on the overlying ectoderm that generates a neural plate that, after rolling into a neural tube, acts as the main source of neural progenitors. Second, the axial regionalization of the neural plate that will result in the specification of neurons with different anteroposterior identities. Although this description of the process applies with ease to amphibians and fish, it is more difficult to confirm in amniote embryos. Here, a specialized population of cells emerges at the end of gastrulation that, under the influence of Wnt and FGF signalling, expands and generates the spinal cord and the paraxial mesoderm. This population is known as the long-term neuromesodermal precursor (NMp). Here, we show that controlled increases of Wnt/β-catenin and FGF signalling during adherent culture differentiation of mouse embryonic stem cells (mESCs) generates a population with many of the properties of the NMp. A single-cell analysis of gene expression within this population reveals signatures that are characteristic of stem cell populations. Furthermore, when this activation is triggered in three-dimensional aggregates of mESCs, the population self-organizes macroscopically and undergoes growth and axial elongation that mimics some of the features of the embryonic spinal cord and paraxial mesoderm. We use both adherent and three-dimensional cultures of mESCs to probe the establishment and maintenance of NMps and their differentiation.


hTERT, BICD1 and chromosome 18 polymorphisms associated with telomere length affect kidney allograft function after transplantation.
It has been confirmed that telomere length (TL) correlates with chronological donor age and that telomere shortening is accelerated in allografts. The aim of this study was to analyse the associations between graft rs2735940 hTERT and rs2630578 BICD1 gene polymorphisms and rs7235755/rs2162440 chromosome 18 polymorphisms, relative TL and kidney function after transplantation. The study enrolled 119 Polish Caucasian kidney allograft recipients (64M/55F, mean age 47.3±14.0 years). The relative TL was assessed in biopsy specimens. To identify genotypes of the studied polymorphisms, real-time PCR was performed. The graft rs2735940 hTERT gene polymorphism TT genotype was associated with a significantly lower risk of delayed graft function (DGF) (TT vs. TC+CC; OR=0, p=0.009) and significantly shorter TL in the '0' biopsy (TT vs. CC: 207±153 vs. 400±161, p=0.036). The graft rs2630578 BICD1 gene polymorphism CC genotype was associated with lower creatinine concentrations in the first month (CC vs. GC: 1.11±0.06 vs. 2.0±1.25 mg/dL, p=0.03). The AA genotype of the graft rs7235755 chromosome 18 polymorphism was associated with longer relative TL in specimens collected 12 to 60 months after transplantation (AA vs. GG+GA p=0.04; AA vs. GG: 489±152 vs. 246±145, p=0.035) and the presence of A allele was associated with higher creatinine concentrations one month after transplantation (GA+AA vs. GG p=0.026; GA vs. GG: 2.18±1.59 vs. 1.76±0.88 mg/dL, p=0.02). It was found that shorter TL in the first six months was associated with higher creatinine concentrations 12 and 18 months after transplantation (Rs=-0.32; p=0.07 and Rs=-0.54; p=0.006, respectively). Graft rs2735940 hTERT and rs2630578 BICD1 gene polymorphisms and rs7235755/rs2162440 chromosome 18 polymorphisms, apart from the association with TL, affect early kidney function after transplantation. Relative TL correlated negatively with creatinine concentrations, allowing the use of TL as a predictor of long-term kidney function.


Primary structures and biological activities of substance-P-related peptides from the brain of the dogfish, Scyliorhinus canicula.
Two peptides with substance-P-like immunoreactivity were isolated in pure form from an extract of the brain of the elasmobranch fish, Scyliorhinus canicula (european common dogfish). One peptide was identical to scyliorhinin I, previously identified in dogfish intestine, and the second was the undecapeptide Lys-Pro-Arg-Pro-Gly-Gln-Phe-Phe-Gly-Leu-Met-CONH2 which is structurally similar to mammalian substance P. Scyliorhinin II or a peptide analogous to mammalian neurokinin A were not detected in the extract. Synthetic dogfish substance P ([Lys1, Arg3, Gly5]substance P) was approximately threefold more potent than mammalian substance P (Kd = 0.21 +/- 0.11 nM versus Kd = 0.74 +/- 0.17 nM; mean +/- SD; n = 6) in inhibiting the binding of 125I-labelled substance P to neurokinin (NK1) receptors in rat submandibular gland membranes. The vasodilator action of tachykinins in mammals is mediated primarily through interaction with NK1 receptors. Bolus intravenous injections of [Lys1, Arg3, Gly5]substance P (100 pmol) and scyliorhinin I (100 pmol) produced appreciable (> 4 kPa) decreases in arterial blood pressure in the rat whereas intravenous injections of up to 5 nmol of the peptides into conscious, unrestrained dogfish produced no change in arterial blood pressure, pulse amplitude or heart rate. Injections of greater amounts of the peptides (10-50 nmol) produced a slight increase (400-667 Pa) in blood pressure. The data indicate that mammalian-type NK1 tachykinin receptors are not involved in cardiovascular regulation in elasmobranch fish.


Modulation of Schistosoma japonicum pulmonary egg granulomas with monoclonal antibodies.
We have examined the ability of various Schistosoma japonicum egg antigens to sensitize mice for subsequent secondary pulmonary egg granuloma formation. Further, we produced monoclonal antibodies (Mabs) specific for egg antigens and evaluated their abilities to modulate pulmonary granuloma formation and inhibit soluble egg antigen (SEA)-stimulated lymphocyte blastogenesis. A homogeneous, 140 Kd egg glycoprotein, SJe; 140-GP was nearly as effective as intact eggs in sensitizing for egg-focused pulmonary granuloma formation, while SEA, or the heterogeneous immunoaffinity (IA)-purified C10 antigens were ineffective. The in vivo administration of chronic infection serum (CIS) or Mabs reactive with SJe; 140-GP, to egg-sensitized/egg-challenged mice, modulated pulmonary granuloma formation. Two of these modulatory SJe; 140-GP-specific Mabs (1 A1-1 or 14B3-8), an IgG1, and an IgG3, respectively, did not alter the responses of SEA-stimulated lymph node cells from mice with acute or chronic schistosomiasis japonica. In contrast, another SJe; 140-GP-specific IgG3 Mab (7A6-5), CIS, immunoaffinity purified anti-SEA from CIS, or the non-SEA-binding fraction of CIS, all resulted in dose-related inhibition of SEA-induced proliferation. These data confirm the antibody-driven nature of some immunoregulatory networks in murine schistosomiasis japonica, and extend these observations to include certain Mabs. The immunogenic nature of SJe; 140-GP, and the modulatory and inhibitory activities of Mabs specific for this glycoprotein, indicate it may play a central role in granuloma formation and modulation in this infection.


Spatial structure and dispersal dynamics in a house sparrow metapopulation.
The effects of spatial structure on metapopulation dynamics depend upon the interaction between local population dynamics and dispersal, and how this relationship is affected by the geographical isolation and spatial heterogeneity in habitat characteristics. Our aim is to examine how emigration and immigration of house sparrows Passer domesticus in a Norwegian archipelagic metapopulation are affected by key factors predicted by classic metapopulation models to affect dispersal-spatial and temporal variation in population size, inter-island distance, local demography and habitat characteristics. This metapopulation can be divided into two major habitat types: (a) islands closer to the mainland where sparrows breed in colonies on farms, and (b) islands without farms, situated farther away from the mainland where sparrows are exposed to harsher environmental conditions. Dispersal was spatially structured within the metapopulation; there was proportionally and numerically less emigration and immigration involving farm islands, as compared to non-farm islands. Furthermore, emigration and immigration occurred mostly between nearby islands. Moreover, emigration in response to spatial differences in mean population size differed between the habitat types, but populations with large mean received more immigrants in both habitat types. The number of emigrants and immigrants was negatively related to long-term recruit production, which was not the case in non-farm islands. The proportion and number of emigrants was positively related to temporal increases in recruit production on farm islands, however not on non-farm islands. Our results demonstrate that spatial heterogeneity in environmental conditions influences how spatial variation in long-term mean population size, and temporal and spatial variation in recruit production, affects dispersal dynamics. The spatial structure of this metapopulation is therefore best described by a spatially explicit model in which the exchange of individuals within each habitat type is strongly affected by the degree of geographical isolation, population size and recruit production. However, these relationships differed between the two habitat types; non-farm islands showing similarities to a mainland-island model type of structure, whereas farm islands showed features more associated with source-sink or balanced dispersal models. Such differential dispersal dynamics between habitat types are expected to have important consequences for the ecological and evolutionary dynamics within this metapopulation.


Crystal structure of a divalent metal ion transporter CorA at 2.9 angstrom resolution.
CorA family members are ubiquitously distributed transporters of divalent metal cations and are considered to be the primary Mg2+ transporter of Bacteria and Archaea. We have determined a 2.9 angstrom resolution structure of CorA from Thermotoga maritima that reveals a pentameric cone-shaped protein. Two potential regulatory metal binding sites are found in the N-terminal domain that bind both Mg2+ and Co2+. The structure of CorA supports an efflux system involving dehydration and rehydration of divalent metal ions potentially mediated by a ring of conserved aspartate residues at the cytoplasmic entrance and a carbonyl funnel at the periplasmic side of the pore.


Effectiveness of HIV prevention for youth in sub-Saharan Africa: systematic review and meta-analysis of randomized and nonrandomized trials.
Systematically assess the effectiveness of HIV-prevention interventions in changing sexual behaviour of young people (10-25 years) in sub-Saharan Africa. Three online databases were searched using prespecified terms. Additional articles were identified on websites of international organizations and by searching bibliographies. Randomized and nonrandomized trials of interventions aiming to reduce risk behaviour were included as well as single-arm studies reporting effects of differential exposure to an intervention. Data were extracted independently in duplicate using predefined data fields. Thirty-one studies on 28 interventions met the inclusion criteria, including 11 randomized trials. Difficulties with implementing planned activities were reportedly common and differential exposure to intervention was high. Two hundred and seventeen outcome measures were extracted: 88 early (within 1 year of intervention) and 129 late outcomes (more than 1 year after the end of the intervention). Sex education and condom promotion among youth did not increase sexual behaviour as well as risky sexual behaviour. No positive effects on sexual behaviour were detected either and condom use at last sex only increased among males [relative risk = 1.46; 95% confidence interval = 1.31-1.64]. One study reported a reduction of herpes simplex virus-2, but not HIV incidence. There remains a stark mismatch between the HIV burden in youth and the number of attempts to design and test prevention interventions - only two trials report biological outcomes. More effective interventions targeting youth are needed. Attention should go to studying implementation difficulties, sex differences in responses to interventions, determinants of exposure to interventions and perhaps inclusion of other factors apart from HIV/AIDS which influence sexual behaviour.


Reliability and validity of the Karaduman Chewing Performance Scale in paediatric neuromuscular diseases: A system for classification of chewing disorders.
Chewing disorders can be seen in children with neuromuscular diseases (NMDs), which may cause swallowing difficulties such as choking, retching and food stuck feeling in the throat. Due to these effects, it is important to determine chewing disorders early with appropriate assessment methods to plan appropriate therapies in NMDs. The aim of this study was to investigate reliability and validity of the Karaduman Chewing Performance Scale (KCPS) in children with NMDs. Age, sex and diagnosis were recorded. Children were asked to chew a standardised biscuit while video-recording. Two physical therapists scored each video according to the KCPS. The correlation between the KCPS scores of 2 therapists was analysed for interobserver reliability. One therapist rescored the recordings after an interval of 2 weeks for intra-observer reliability. The Pediatric Version of the Eating Assessment Tool (PEDI-EAT-10) was used for criterion-based validity. Sixty-eight children with a mean age of 8.34 ± 3.73 (min = 2.5, max = 14.5) years were included, of which 94.1% were male. Karaduman Chewing Performance Scale scores were found to be level 0 in 20 cases, level 1 in 35 cases, level 2 in 12 cases and level 3 in 1 case. A positive, very strong correlation was detected between 2 therapists (r = .93, P < .001) and between 2 examinations of 1 therapist (r = .83, P < .001). A good correlation between the KCPS and PEDI-EAT-10 was detected (r = .62, P < .001). Chewing function, especially food processing phase, deteriorated in children with NMDs. The KCPS could be used as a reliable and valid instrument in determining chewing performance level for children with NMDs.


Effects of carbamazepine on serum parathormone, 25- hydroxyvitamin D, bone specific alkaline phosphatase, C-telopeptide, and osteocalcin levels in healthy rats.
It is still not completely clear whether carbamazepine causes alterations in vitamin D status and in bone metabolism. The objective of this study was to investigate the effects of carbamazepine on serum levels of 25-hydroxyvitamin D and on biomarkers of bone formation and resorption in healthy rats. Levels of calcium, 25- hydroxyvitamin D, parathormone, C-telopeptide, bone specific alkaline phosphatase and osteocalcin were measured in 3 groups of rats consisting of controls (n=10), isotonic saline solution group (n=10) and carbamazepine group (n=10). Mean calcium levels were found to be significantly lower in healthy controls in comparison to isotonic saline solution and carbamazepine groups (10.0±0.24, 10.81±0.16, 10.93±0.22 mg/dL, respectively, p<0.05). Mean levels of 25- hydroxyvitamin D, were found to be significantly higher in control group compared to isotonic saline solution group (25- hydroxyvitamin D; 25.91±1.12, 19.99±0.99 ng/mL, respectively, p<0.01). Mean levels of parathormone and osteocalcin were found to be significantly higher in control group compared to isotonic saline solution group and carbamazepine group. Parathormone levels were measured as 3.46±0.83, 1.08±0.08, 0.94±0.02 pg/mL, respectively (p<0.01). Osteocalcine levels were measured as 1.66±0.001, 1.32±0.002, 1.32±0.001 ng/mL, respectively (p<0.001). A significant difference in terms of mean serum bone specific alkaline phosphatase and C-telopeptide levels among groups was not observed. The main outcome of this prospective study in healthy rats showed no change in biochemical parameters of bone turnover during treatment with carbamazepine.


Mining the secretome of the root-knot nematode Meloidogyne chitwoodi for candidate parasitism genes.
Parasite proteins secreted at the interface of nematode and host are believed to play an essential role in parasitism. Here, we present an efficient pipeline of bio-informatic algorithms and laboratory experiments to identify candidate parasitism genes within nematode secretomes, i.e. the repertoire of secreted proteins in an organism. We performed our approach on 12 218 expressed sequence tags (ESTs) originating from three life stages of the plant parasitic nematode Meloidogyne chitwoodi--a molecularly unexplored root-knot nematode species. The ESTs from M. chitwoodi were assembled into 5880 contigs and open reading frames translated from the consensus sequences were searched for features of putative signal peptides for protein secretion and trans-membrane regions, resulting in the identification of 398 secretome members. The products of parasitism genes are secreted by a range of organs, including the oesophageal, amphidial and rectal glands, the intestine, and the hypodermis. To localize the site of expression in M. chitwoodi, we subjected the most abundant secretome members to in situ hybridization microscopy. We found hybridization of one tag in the dorsal oesophageal gland, seven in the two subventral oesophageal glands, two in the intestine and one tag hybridized to the tail tip in the proximity of the phasmids. Four sequences showed similarity to putative parasitism genes from other nematode species, whereas seven represented pioneering sequences. Our approach presents an efficient method to identify candidate parasitism genes, which does not require sophisticated cDNA isolation and selection protocols, and can therefore be used as a powerful starting point for the molecular investigation of parasites.


Differential item functioning analysis of the 2003-04 NHANES physical activity questionnaire.
Using differential item functioning (DIF) analyses, this study examined whether there were any DF items in the National Health and Nutrition Examination Survey (NHANES) physical activity (PA) questionnaire. A subset of adult data from the 2003-04 NHANES study (n = 3,083) was used. PA items related to respondents' occupational, transportation, domestic, leisure-time, strength-related, and sedentary activities were analyzed for DIF using Mantel-Haenszel, SIBTEST, and analysis of variance procedures. Some items were identified as DF items, with the majority of those items favoring advantaged social groups. For example, items about domestic activity and moderate-intensity leisure-time activity were DF items favoring non-Hispanic Whites, and persons with higher levels of education (e.g., some college or more) or income (e.g, annual family income [AFI] > $34,999). The vigorous activity item was a DIF item favoring younger adults (ages 20-59 years) and persons with higher levels of education or income. Occupational PA presented DF favoring Hispanics and persons with lower levels of education (e.g., high school or less), and the transportation-related PA item presented DF favoring persons with lower incomes (e.g., AFI $34,999). These findings highlight the importance of conducting DF analysis in PA survey construction and emphasize the need to introduce DIF concepts and methods to PA researchers.


Seasonal variation in haematological and biochemical variables in free-ranging subadult brown bears (Ursus arctos) in Sweden.
Free-ranging brown bears exhibit highly contrasting physiological states throughout the year. They hibernate 6 months of the year, experiencing a decrease in body temperature, heart rate, respiratory rate and metabolism. An increase in food consumption and the resulting weight gain (mostly through fat storage) prior to hibernation are also part of the brown bear's annual cycle. Due to these physiological changes, haematological and biochemical variables vary dramatically throughout the year. Seasonal changes in 12 haematological and 34 biochemical variables were evaluated in blood samples collected from 40 free-ranging subadult brown bears (22 females, 18 males) immobilised in Sweden in winter (February-March), spring (April-May), and summer (June). Higher levels of haemoglobin, haematocrit and red blood cell count, and a lower white blood cell count and mean cell volume was found during hibernation than in spring and summer. Lower values of the enzymes; aspartate aminotransferase (AST), alanine transaminase (ALT), alkaline phosphatase (AP), γ-glutamyl transpeptidase (GGT), glutamate dehydrogenase (GD) and amylase, and increased values of β-hydroxybutyrate (β-HBA) and blood lipids; triglycerides, cholesterol and free fatty acids, were present during hibernation compared to spring and summer. This study documents significant shifts in haematological and biochemical variables in samples collected from brown bears anaesthetised in winter (February-March) compared to in spring and summer (April-June), reflecting the lowered metabolic, renal and hepatic activity during hibernation. Lower values of enzymes and higher values of blood lipids during hibernation, likely reflect a lipid-based metabolism.


Updated phylogenetic description of the Mycoplasma hominis cluster (Weisburg et al. 1989) based on 16S rDNA sequences.
The fastidious nature of the mollicutes (mycoplasmas), their lack of a classic bacterial cell wall, and their very small genome, make phylogenetic placements of new species in this enlarging group of prokaryotes an important and valuable aid in their classification. In this report we have determined the phylogeny of the Mycoplasma hominis cluster of the hominis group. The 16S rDNA sequences from several previously described Mycoplasma species were determined and ten species were found to belong to the M. hominis cluster. With almost complete sequences available, the phylogenetic analysis revealed that the M. hominis cluster currently comprises 19 species, forming a distinct clade as judged from branch lengths, bootstrap percentage values, nucleotide signature analysis, and structural elements in the 16S rRNA molecule. The 16S rRNA gene sequences of species in the M. hominis cluster were found to be > or = 94% similar and the range within which similarities can be used in the classification of new species is discussed. Members of the M. hominis cluster all share a major biochemical property of M. hominis, in that they hydrolyse arginine and are incapable of fermenting glucose. This consistency in phenotypic pattern has not been found in any of the other phylogenetic clusters of the hominis group. Two species, the non-cultivable agent of Grey Lung disease in rodents (tentatively named 'Candidatus Mycoplasma ravipulmonis') and the avian species Mycoplasma gypis strain B1/T1T, were regarded as close relatives to the M. hominis cluster, but are clearly separated from the species of this cluster. Both species formed early branches of the M. hominis cluster and should be regarded as individual lines containing one species.


Two independently regulated cytochromes P-450 in a Rhodococcus rhodochrous strain that degrades 2-ethoxyphenol and 4-methoxybenzoate.
A red-pigmented coryneform bacterium, identified as Rhodococcus rhodochrous strain 116, that grew on 2-ethoxyphenol and 4-methoxybenzoate as sole carbon and energy sources was isolated. Phylogenetic analysis based on the 16S rDNA sequences indicates that the strain clusters more closely to other rhodococci than to other gram-positive organisms with a high G + C content. Each of the abovementioned growth substrates was shown to induce a distinct cytochrome P-450: cytochrome P-450RR1 was induced by 2-ethoxyphenol, and cytochrome P-450RR2 was induced by 4-methoxybenzoate. A type I difference spectrum typical of substrate binding was induced in cytochrome P-450RR1 by both 2-ethoxyphenol (KS = 4.2 +/- 0.3 microM) and 2-methoxyphenol (KS = 2.0 +/- 0.1 microM), but not 4-methoxybenzoate or 4-ethoxybenzoate. Similarly, a type I difference spectrum was induced in cytochrome P-450RR2 by both 4-methoxybenzoate (KS = 2.1 +/- 0.1 microM) and 4-ethoxybenzoate (KS = 1.6 +/- 0.1 microM), but not 2-methoxyphenol or 2-ethoxyphenol. A purified polyclonal antiserum prepared against cytochrome P-450RR1 did not cross-react with cytochrome P-450RR2, indicating that the proteins are immunologically distinct. The cytochromes appear to catalyze the O-dealkylation of their respective substrates. The respective products of the O-dealkylation are further metabolized via ortho cleavage enzymes, whose expression is also regulated by the respective aromatic ethers.


A cluster of cholinergic premotor interneurons modulates mouse locomotor activity.
Mammalian motor programs are controlled by networks of spinal interneurons that set the rhythm and intensity of motor neuron firing. Motor neurons have long been known to receive prominent "C bouton" cholinergic inputs from spinal interneurons, but the source and function of these synaptic inputs have remained obscure. We show here that the transcription factor Pitx2 marks a small cluster of spinal cholinergic interneurons, V0(C) neurons, that represents the sole source of C bouton inputs to motor neurons. The activity of these cholinergic interneurons is tightly phase locked with motor neuron bursting during fictive locomotor activity, suggesting a role in the modulation of motor neuron firing frequency. Genetic inactivation of the output of these neurons impairs a locomotor task-dependent increase in motor neuron firing and muscle activation. Thus, V0(C) interneurons represent a defined class of spinal cholinergic interneurons with an intrinsic neuromodulatory role in the control of locomotor behavior.


The ionic basis of the hypo-osmotic depolarization in neurons from the opisthobranch mollusc Elysia chlorotica.
The resting potential of identified cells (Parker cells) in the abdominal ganglion of Elysia chlorotica (Gould) depolarizes by about 30 mV in response to a 50% reduction in osmolality and returns to the original potential in 20 min. Cell volume recovery requires approximately 2 h. Thus, recovery of the resting potential is not dependent on recovery of cell volume. The hypo-osmotic depolarization persists following inhibition of the electrogenic Na+/K(+)-ATPase with ouabain, and the levels of extracellular K+ and Cl- have little effect on the magnitude of the depolarization, while decreasing extracellular Na+ concentration produces a depolarization of only 10 mV. This suggests that the hypo-osmotic depolarization in Parker cells results mostly from increased relative permeability to Na+. Following transfer from 920 to 460 mosmol kg-1, Na+, Cl- and proline betaine leave the cells while intracellular K+ is conserved. Loss of intracellular Na+ and conservation of intracellular K+ are dependent on active transport by the Na+/K(+)-ATPase. Na+ and proline betaine leave the cells with a time course that is much longer than that of the hypo-osmotic depolarization. Unlike the other solutes, most of the reduction in intracellular Cl- concentration occurs coincidentally with the hypo-osmotic depolarization. However, unlike the hypo-osmotic depolarization, bulk loss of Cl- does not require the reduction in osmolality, only the reduction in extracellular ion concentrations. There is no apparent relationship between membrane depolarization and the regulation of intracellular osmolytes in Elysia neurons following hypo-osmotic stress.


Effect of intermittent preventive treatment of malaria on the outcome of pregnancy among women attending antenatal clinic of a new Nigerian teaching hospital, Ado-Ekiti.
Malaria is a public health problem globally especially in the Sub-Saharan Africa and among the under five children and pregnant women and is associated with a lot of maternal and foetal complications. The study was on the effect of intermittent preventive treatment of malaria in pregnancy on the prevalence of malaria in pregnancy and the outcome of pregnancy. In a descriptive cross-sectional study, a semi-structured questionnaire was administered to women admitted in Ekiti State University Teaching Hospital labour ward, Ado-Ekiti. About 4,200 women participated in the study and the inclusion criteria were women who were booked in the hospital, attended at least four antenatal clinic visits, and consented to the study while the exclusion criteria were those who didn't book in the hospital and failed to give their consent. The study revealed that about 75% of the pregnant women studied had access to intermittent preventive treatment of malaria. Among the women attending the antenatal clinic that received sulphadoxine-pyrimethamine (SP), about 78% of them took two doses of SP. The prevalence of clinical malaria was statistically higher in women who did not receive intermittent preventive treatment with SP during pregnancy (44.7% vs. 31.3%, P = 0.0001) and among women who had one dose of the drug instead of two doses (40.0% vs. 28.7%, P = 0.0001). There was no statistical significant difference in the mean age in years (31.53 ± 5.238 vs. 31.07 ± 4.751, P = 0.09 and the gestational age at delivery (38.76 ± 1.784 vs. 38.85 ± 1.459, P = 0.122) between the women who did not receive SP and those who had it. There was a statistical significant difference in the outcome of pregnancy among women who had Intermittent Preventive Treatment in pregnancy (IPTp) and those who did not viz.-a-viz. in the duration of labor (8.6 ± 1.491 vs. 8.7 ± 1.634, P = 0.011) and the birth weight of the babies (3.138 ± 0.402 vs. 3.263 ± 0.398, P = 0.0001). SP is an effective malarial prophylaxis in pregnancy.


IMPACT OF FATHERS' ALCOHOLISM AND ASSOCIATED RISK FACTORS ON PARENT-INFANT ATTACHMENT STABILITY FROM 12 TO 18 MONTHS.
This study examined short-term attachment stability and sought to identify predictors of stability and change within a sample characterized by fathers' alcoholism. Results suggest moderate stability of attachment classifications (60% for mothers, 53% for fathers) from 12 to 18 months. Higher paternal and maternal alcohol symptoms, maternal depression, and maternal antisocial behavior were found in families with stable insecure mother-infant attachment compared to those who were stable secure. Mother-infant stable insecurity was associated with higher levels of maternal negative affect expression during play. Father-infant stable insecurity was associated with lower levels of paternal positive affect expression and decreased sensitivity during play. Stable insecure children also had higher levels of negative affect during parent-infant interactions and higher negative emotionality during other episodes compared to stable secure children. Results indicate that infants who were insecure at both time points had the highest constellation of family risk characteristics.


Family Caregiving for Older Adults.
Family members are the primary source of support for older adults with chronic illness and disability. Thousands of published empirical studies and dozens of reviews have documented the psychological and physical health effects of caregiving, identified caregivers at risk for adverse outcomes, and evaluated a wide range of intervention strategies to support caregivers. Caregiving as chronic stress exposure is the conceptual driver for much of this research. We review and synthesize the literature on the impact of caregiving and intervention strategies for supporting caregivers. The impact of caregiving is highly variable, driven largely by the intensity of care provided and the suffering of the care recipient. The intervention literature is littered with many failures and some successes. Successful interventions address both the pragmatics of care and the emotional toll of caregiving. We conclude with both research and policy recommendations that address a national agenda for caregiving.


Centile charts of cervical length between 18 and 32 weeks of gestation.
To establish a centile chart of cervical length between 18 and 32 weeks of gestation in a low-risk population of women. A prospective longitudinal cohort study of women with a low risk, singleton pregnancy using public healthcare facilities in Cape Town, South Africa. Transvaginal measurement of cervical length was performed between 16 and 32 weeks of gestation and used to construct centile charts. The distribution of cervical length was determined for gestational ages and was used to establish estimates of longitudinal percentiles. Centile charts were constructed for nulliparous and multiparous women together and separately. Centile estimation was based on data from 344 women. Percentiles showed progressive cervical shortening with increasing gestational age. Averaged over the entire follow-up period, mean cervical length was 1.5 mm shorter in nulliparous women compared with multiparous women (95% CI, 0.4-2.6). Establishment of longitudinal reference values of cervical length in a low-risk population will contribute toward a better understanding of cervical length in women at risk for preterm labor.


An evaluation of the potential to use tumor-associated antigens as targets for antitumor T cell therapy using transgenic mice expressing a retroviral tumor antigen in normal lymphoid tissues.
A major obstacle to the development of T cell therapy for the treatment of human tumors has been the difficulty generating T cells specifically reactive with the tumor. Most of the characterized human tumor antigens have been classified as tumor associated, because of demonstrable expression at low levels in some normal cells, and thus have not been extensively studied as potential targets of a therapeutic immune response. However, the quantitative difference in expression of such antigens between the tumor and normal cells might permit the generation of antigen-specific T cells capable of selective antitumor and not autoimmune activity. To address this issue, transgenic (TG) mice were generated that expressed low levels of Friend murine leukemia virus (FMuLV) envelope protein in lymphoid cells under the control of an immunoglobulin promoter. This protein is expressed at high levels by a Friend virus-induced erythroleukemia of C57BL/6 (B6) origin, FBL, and has been shown to serve as an efficient tumor-specific rejection antigen in B6 mice. The env-TG mice were tolerant to envelope, as reflected by the failure to detect an envelope-specific response after in vivo priming and in vitro stimulation with preparations of FMuLV envelope. However, adoptively transferred envelope-specific T cells from immunized non-TG B6 mice mediated complete eradication of FBL tumor cells in TG mice, and did not induce detectable autoimmune damage to TG lymphoid tissues. The transferred immune cells were not permanently inactivated in the TG mice, since donor T cells responded to envelope after removal from the TG mice. The lack of autoimmune injury did not reflect inadequate expression of envelope by TG lymphocytes for recognition by T cells, since TG lymphocytes functioned effectively in vitro as stimulators for envelope-specific T cells. The results suggest that this and analogous strains of TG mice may prove useful for elucidating principles for the generation and therapeutic use of tumor-reactive T cells specific for tumor-associated antigens.


Analysis of interactions between mismatch repair initiation factors and the replication processivity factor PCNA.
In eukaryotes, the DNA replication factor PCNA is loaded onto primer-template junctions to act as a processivity factor for DNA polymerases. Genetic and biochemical studies suggest that PCNA also functions in early steps in mismatch repair (MMR) to facilitate the repair of misincorporation errors generated during DNA replication. These studies have shown that PCNA interacts directly with several MMR components, including MSH3, MSH6, MLH1, and EXO1. At present, little is known about how these interactions contribute to the mismatch repair mechanism. The interaction between MLH1 and PCNA is of particular interest because MLH1-PMS1 is thought to act as a matchmaker to signal mismatch recognition to downstream repair events; in addition, PCNA has been hypothesized to act in strand discrimination steps in MMR. Here, we utilized both genetic and surface plasmon resonance techniques to characterize the MLH1-PMS1-PCNA interaction. These analyses enabled us to determine the stability of the complex (K(D) = 300 nM) and to identify residues (572-579) in MLH1 and PCNA (126,128) that appear important to maintain this stability. We favor a model in which PCNA acts as a scaffold for consecutive protein-protein interactions that allow for the coordination of MMR steps.


TLR Agonists as Vaccine Adjuvants Targeting Cancer and Infectious Diseases.
Modern vaccines have largely shifted from using whole, killed or attenuated pathogens to being based on subunit components. Since this diminishes immunogenicity, vaccine adjuvants that enhance the immune response to purified antigens are critically needed. Further advantages of adjuvants include dose sparing, increased vaccine efficacy in immunocompromised individuals and the potential to protect against highly variable pathogens by broadening the immune response. Due to their ability to link the innate with the adaptive immune response, Toll-like receptor (TLR) agonists are highly promising as adjuvants in vaccines against life-threatening and complex diseases such as cancer, AIDS and malaria. TLRs are transmembrane receptors, which are predominantly expressed by innate immune cells. They can be classified into cell surface (TLR1, TLR2, TLR4, TLR5, TLR6) and intracellular TLRs (TLR3, TLR7, TLR8, TLR9), expressed on endosomal membranes. Besides a transmembrane domain, each TLR possesses a leucine-rich repeat (LRR) segment that mediates PAMP/DAMP recognition and a TIR domain that delivers the downstream signal transduction and initiates an inflammatory response. Thus, TLRs are excellent targets for adjuvants to provide a "danger" signal to induce an effective immune response that leads to long-lasting protection. The present review will elaborate on applications of TLR ligands as vaccine adjuvants and immunotherapeutic agents, with a focus on clinically relevant adjuvants.


Development of a master health facility list in Nigeria.
Abstract. Routine Health Information Systems (RHIS) are increasingly transitioning to electronic platforms in several developing countries. Establishment of a Master Facility List (MFL) to standardize the allocation of unique identifiers for health facilities can overcome identification issues and support health facility management. The Nigerian Federal Ministry of Health (FMOH) recently developed a MFL, and we present the process and outcome. The MFL was developed from the ground up, and includes a state code, a local government area (LGA) code, health facility ownership (public or private), the level of care, and an exclusive LGA level health facility serial number, as part of the unique identifier system in Nigeria. To develop the MFL, the LGAs sent the list of all health facilities in their jurisdiction to the state, which in turn collated for all LGAs under them before sending to the FMOH. At the FMOH, a group of RHIS experts verified the list and identifiers for each state. The national MFL consists of 34,423 health facilities uniquely identified. The list has been published and is available for worldwide access; it is currently used for planning and management of health services in Nigeria. Unique identifiers are a basic component of any information system. However, poor planning and execution of implementing this key standard can diminish the success of the RHIS. Development and adherence to standards is the hallmark for a national health information infrastructure. Explicit processes and multi-level stakeholder engagement is necessary to ensuring the success of the effort.


Long-term administration of Aspalathus linearis infusion affects spatial memory of adult Sprague-Dawley male rats as well as increases their striatal dopamine content.
Everyday use of the herbal tea rooibos, produced from Aspalathus linearis (Brum.f) Dahlg. (Fabaceae) is customary in South Africa, a continuation of its historical use by indigenous people. Although evidence of its traditional indications is anecdotal, rooibos tea is regarded as a general health tea. Available contemporary research indicates to broad cell protective activity of rooibos focusing on its antioxidative, anti-inflammatory, anti-hyperglycaemic and antithrombotic features affecting metabolic syndrome, cardiovascular risk and neuroprotection. Nevertheless little is known about its impact on brain functions. The present experiment aimed to evaluate the possible behavioural and neurochemical effects of long-term oral administration of "fermented"" rooibos herbal tea (FRHT) infusions to adult male Sprague-Dawley rats. Infusions, prepared using 1, 2 and 4 g of "fermented"" (oxidised) A. linearis leaves for 100 ml of hot water, were characterised in terms of flavonoid content by ultra-high and high performance liquid chromatography (UHPLC-qTOF-MS, HPLC-DAD) and administered to rats as sole drinking fluid for 12 weeks. Spatial memory behaviour was assessed in a modified version of the Morris water maze. Dopamine, noradrenaline, serotonin and their metabolite levels (DOPAC, 3-MT, HVA, MHPG, 5-HIAA) were quantified in prefrontal cortex, hippocampus and striatum by HPLC-ECD. Body weight and blood glucose level were additionally estimated. All FRHT-treated rats showed improvement of long-term spatial memory defined as increased number of crossings over the previous platform position in SE quadrant of the water maze. It was not accompanied by excessive motor activity. Striatal dopamine and its metabolite 3-MT (3-methoxytyramine) levels were increased in treated rats. There were no differences in body weight gain between control and treated animals but blood glucose level was significantly lower in the latter ones. The improvement of long-term memory in FRHT-treated rats and stimulating impact of FRHT on their dopaminergic striatal transmission support the wellness enhancing effect of rooibos tea, contributing to a better understanding of the neurological background of traditional habitual consumption of this herbal tea.


A Universal Probe Set for Targeted Sequencing of 353 Nuclear Genes from Any Flowering Plant Designed Using k-Medoids Clustering.
Sequencing of target-enriched libraries is an efficient and cost-effective method for obtaining DNA sequence data from hundreds of nuclear loci for phylogeny reconstruction. Much of the cost of developing targeted sequencing approaches is associated with the generation of preliminary data needed for the identification of orthologous loci for probe design. In plants, identifying orthologous loci has proven difficult due to a large number of whole-genome duplication events, especially in the angiosperms (flowering plants). We used multiple sequence alignments from over 600 angiosperms for 353 putatively single-copy protein-coding genes identified by the One Thousand Plant Transcriptomes Initiative to design a set of targeted sequencing probes for phylogenetic studies of any angiosperm group. To maximize the phylogenetic potential of the probes, while minimizing the cost of production, we introduce a k-medoids clustering approach to identify the minimum number of sequences necessary to represent each coding sequence in the final probe set. Using this method, 5-15 representative sequences were selected per orthologous locus, representing the sequence diversity of angiosperms more efficiently than if probes were designed using available sequenced genomes alone. To test our approximately 80,000 probes, we hybridized libraries from 42 species spanning all higher-order groups of angiosperms, with a focus on taxa not present in the sequence alignments used to design the probes. Out of a possible 353 coding sequences, we recovered an average of 283 per species and at least 100 in all species. Differences among taxa in sequence recovery could not be explained by relatedness to the representative taxa selected for probe design, suggesting that there is no phylogenetic bias in the probe set. Our probe set, which targeted 260 kbp of coding sequence, achieved a median recovery of 137 kbp per taxon in coding regions, a maximum recovery of 250 kbp, and an additional median of 212 kbp per taxon in flanking non-coding regions across all species. These results suggest that the Angiosperms353 probe set described here is effective for any group of flowering plants and would be useful for phylogenetic studies from the species level to higher-order groups, including the entire angiosperm clade itself.


Ultrafast electron-induced desorption of water from nanometer amorphous solid water films.
Laser-induced desorption of water molecules from nanometer amorphous solid water films supported on a single-crystal platinum substrate is reported. A femtosecond laser pulse creates hot substrate electrons, which are injected into the water layer, resulting in significant desorption at the water-vacuum interface. The dependence of the desorption yield on film thickness and results for isotopic spacer and capping layers reveal that the desorbing water originates from relatively deep down into the water layer, i.e., from several nanometers below the surface. This is proposed to be the result of cooperative electronic effects resulting from the high electron densities in the thin water film, which cause a transient destabilization of the water H-bonded network. Motion of excited water molecules through the layer is enabled by mixing within the layer on ultrafast timescales during the desorption process.


Sound recognition and localization in man: specialized cortical networks and effects of acute circumscribed lesions.
Functional imaging studies have shown that information relevant to sound recognition and sound localization are processed in anatomically distinct cortical networks. We have investigated the functional organization of these specialized networks by evaluating acute effects of circumscribed hemispheric lesions. Thirty patients with a primary unilateral hemispheric lesion, 15 with right-hemispheric damage (RHD) and 15 with left-hemispheric damage (LHD), were evaluated for their capacity to recognise environmental sounds, to localize sounds in space and to perceive sound motion. One patient with RHD and 2 with LHD had a selective deficit in sound recognition; 3 with RHD a selective deficit in sound localization; 2 with LHD a selective deficit in sound motion perception; 4 with RHD and 3 with LHD a combined deficit of sound localization and motion perception; 2 with RHD and 1 with LHD a combined deficit of sound recognition and motion perception; and 1 with LHD a combined deficit of sound recognition, localization and motion perception. Five patients with RHD and 6 with LHD had normal performance in all three domains. Deficient performance in sound recognition, sound localization and/or sound motion perception was always associated with a lesion that involved the shared auditory structures and the specialized What and/or Where networks, while normal performance was associated with lesions within or outside these territories. Thus, damage to regions known to be involved in auditory processing in normal subjects is necessary, but not sufficient for a deficit to occur. Lesions of a specialized network was not always associated with the corresponding deficit. Conversely, specific deficits tended not be associated predominantly with lesions of the corresponding network; e.g. deficits in auditory spatial tasks were observed in patients whose lesions involved to a larger extent the shared auditory structures and the specialized What network than the specialized Where network, and deficits in sound recognition in patients whose lesions involved mostly the shared auditory structures and to a varying degree the specialized What network. The human auditory cortex consists of functionally defined auditory areas, whose intrinsic organization is currently not understood. In particular, areas involved in the What and Where pathways can be conceived as: (1) specialized regions, in which lesions cause dysfunction limited to the damaged part; observed deficits should be then related to the specialization of the damaged region and their magnitude to the extent of the damage; or (2) specialized networks, in which lesions cause dysfunction that may spread over the two specialized networks; observed deficits may then not be related to the damaged region and their magnitude not proportional to the extent of the damage. Our results support strongly the network hypothesis.


The phytoalexin camalexin induces fundamental changes in the proteome of Alternaria brassicicola different from those caused by brassinin.
Camalexin is the major phytoalexin produced by Alternaria thaliana, but is absent in Brassica species that usually produce phytoalexin blends containing brassinin and derivatives. The protein profiles of A. brassicicola treated with camalexin were evaluated using proteomics and metabolic analyses and compared with those treated with brassinin. Conidial germination and mycelial growth of A. brassicicola in liquid media amended with camalexin and brassinin showed that fungal growth was substantially slower in presence of camalexin than brassinin; chemical analyses revealed that A. brassicicola detoxified camalexin at much slower rate than brassinin. Two-dimensional gel electrophoresis (2-DE) followed by tryptic digestion and capillary liquid chromatography-mass spectrometric analyses identified 158 different proteins, of which 45 were up-regulated and 113 were down-regulated relative to controls. Venn diagram analyses of differentially expressed proteins in cultures of A. brassicicola incubated with camalexin and brassinin indicated clear differences in the effect of each phytoalexin, with camalexin causing down-regulation of a larger number of proteins than brassinin. Overall, results of this work suggest that each phytoalexin has several different targets in the cells of A. brassicicola, and that camalexin appears to have greater potential to protect cultivated Brassica species against A. brassicicola than brassinin.


Parkinson's disease is positively associated with periodontal inflammation.
Parkinson's disease (PA) affects 1% of the global population above 60 years. PA pathogenesis involves severe neuroinflammation that impacts systemic and local inflammatory changes. We tested the hypothesis that PA is associated with periodontal tissue inflammation promoting a greater systemic inflammatory burden. We recruited 60 patients with Stage III, Grade B periodontitis (P) with and without PA (n = 20 for each). We also included systemically and periodontally healthy individuals as controls (n = 20). Clinical periodontal parameters were recorded. Serum, saliva, and gingival crevicular fluid (GCF) samples were collected to measure the inflammatory and neurodegenerative targets (YKL-40, fractalkine, S100B, alpha-synuclein, tau, vascular cell adhesion protein-1 (VCAM-1), brain-derived neurotrophic factor (BDNF), neurofilament light chain (NfL). Parkinson's patients in this study had mild to moderate motor dysfunctions, which did not prevent them from performing optimal oral hygiene control. Periodontal parameters and GCF volume were significantly higher in the P and P+PA groups than in the control group. PA was associated with significantly increased bleeding on probing (BOP) compared to P-alone (p < 0.05), while other clinical parameters were similar between P and P+PA groups. In saliva and serum, YKL-40 levels were higher in the P+PA group than in P and C groups (p < 0.001). GCF NfL levels from shallow sites were significantly higher in the P+PA group compared to the C group (p = 0.0462). GCF S100B levels from deep sites were higher in the P+PA group than in healthy individuals (p = 0.0194). The data suggested that PA is highly associated with increased periodontal inflammatory burden-bleeding upon probing and inflammatory markers-in parallel with PA-related neuroinflammation.


What Factors Influence the Biomechanical Properties of Allograft Tissue for ACL Reconstruction? A Systematic Review.
Allograft tissue is used in 22% to 42% of anterior cruciate ligament (ACL) reconstructions. Clinical outcomes have been inconsistent with allograft tissue, with some series reporting no differences in outcomes and others reporting increased risk of failure. There are numerous variations in processing and preparation that may influence the eventual performance of allograft tissue in ACL reconstruction. We sought to perform a systematic review to summarize the factors that affect the biomechanical properties of allograft tissue for use in ACL reconstruction. Many factors might impact the biomechanical properties of allograft tissue, and these should be understood when considering using allograft tissue or when reporting outcomes from allograft reconstruction. What factors affect the biomechanical properties of allograft tissue used for ACL reconstruction? We performed a systematic review to identify studies on factors that influence the biomechanical properties of allograft tissue through PubMed and SCOPUS databases. We included cadaveric and animal studies that reported on results of biomechanical testing, whereas studies on fixation, histologic evaluation, and clinical outcomes were excluded. There were 319 unique publications identified through the search with 48 identified as relevant to answering the study question. For each study, we recorded the type of tissue tested, parameters investigated, and the effects on biomechanical behavior, including load to failure and stiffness. Primary factors identified to influence allograft tissue properties were graft tissue type, sterilization methods (irradiation and chemical processing), graft preparation, donor parameters, and biologic adjuncts. Load to failure and graft stiffness varied across different tissue types, with nonlooped tibialis grafts exhibiting the lowest values. Studies on low-dose irradiation showed variable effects, whereas high-dose irradiation consistently produced decreased load to failure and stiffness values. Various chemical sterilization measures were also associated with negative effects on biomechanical properties. Prolonged freezing decreased load to failure, ultimate stress, and ultimate strain. Up to eight freeze-thaw cycles did not lead to differences in biomechanical properties of cadaveric grafts. Regional differences were noted in patellar tendon grafts, with the central third showing the highest load to failure and stiffness. Graft diameter strongly contributed to load-to-failure measurements. Age older than 40 years, and especially older than 65 years, negatively impacted biomechanical properties, whereas gender had minimal effect on the properties of allograft tissue. Biologic adjuncts show potential for improving in vivo properties of allograft tissue. Future clinical studies on allograft ACL reconstruction should investigate in vivo graft performance with standardized allograft processing and preparation methods that limit the negative effects on the biomechanical properties of tissue. Additionally, biologic adjuncts may improve the biomechanical properties of allograft tissue, although future preclinical and clinical studies are necessary to clarify the role of these treatments. Based on the findings of this systematic review that emphasize biomechanical properties of ACL allografts, surgeons should favor the use of central third patellar tendon or looped soft tissue grafts, maximize graft cross-sectional area, and favor grafts from donors younger than 40 years of age while avoiding grafts subjected to radiation doses > 20 kGy, chemical processing, or greater than eight freeze-thaw cycles.


Silibinin suppresses NPM-ALK, potently induces apoptosis and enhances chemosensitivity in ALK-positive anaplastic large cell lymphoma.
Nucleophosmin-anaplastic lymphoma kinase (NPM-ALK), an oncogenic fusion protein carrying constitutively active tyrosine kinase, is known to be central to the pathogenesis of ALK-positive anaplastic large cell lymphoma (ALK+ALCL). Here, it is reported that silibinin, a non-toxic naturally-occurring compound, potently suppressed NPM-ALK and effectively inhibited the growth and soft agar colony formation of ALK+ALCL cells. By western blots, it was found that silibinin efficiently suppressed the phosphorylation/activation of NPM-ALK and its key substrates/downstream mediators (including STAT3, MEK/ERK and Akt) in a time- and dose-dependent manner. Correlating with these observations, silibinin suppressed the expression of Bcl-2, survivin and JunB, all of which are found to be upregulated by NPM-ALK and pathogenetically important in ALK+ALCL. Lastly, silibinin augmented the chemosensitivity of ALK+ALCL cells to doxorubicin, particularly the small cell sub-set expressing the transcriptional activity of Sox2, an embryonic stem cell marker. To conclude, the findings suggest that silibinin might be useful in treating ALK+ALCL.


Indoxyl sulfate down-regulates SLCO4C1 transporter through up-regulation of GATA3.
The accumulated uremic toxins inhibit the expression of various renal transporters and this inhibition may further reduce renal function and subsequently cause the accumulation of uremic toxins. However, the precise mechanism of the nephrotoxicity of uremic toxins on renal transport has been poorly understood. Here we report that indoxyl sulfate, one of the potent uremic toxins, directly suppresses the renal-specific organic anion transporter SLCO4C1 expression through a transcription factor GATA3. The promoter region of SLCO4C1 gene has several GATA motifs, and indoxyl sulfate up-regulated GATA3 mRNA and subsequently down-regulated SLCO4C1 mRNA. Overexpression of GATA3 significantly reduced SLCO4C1 expression, and silencing of GATA3 increased SLCO4C1 expression vice versa. Administration of indoxyl sulfate in rats reduced renal expression of slco4c1 and under this condition, plasma level of guanidinosuccinate, one of the preferable substrates of slco4c1, was significantly increased without changing plasma creatinine. Furthermore, in 5/6 nephrectomized rats, treatment with oral adsorbent AST-120 significantly decreased plasma indoxyl sulfate level and conversely increased the expression of slco4c1, following the reduction of plasma level of guanidinosuccinate. These data suggest that the removal of indoxyl sulfate and blocking its signal pathway may help to restore the SLCO4C1-mediated renal excretion of uremic toxins in CKD.


Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria.
Endothelial cells were isolated from freshly obtained human umbilical cords by collagenase digestion of the interior of the umbilical vein. The cells were grown in tissue culture as a homogeneous population for periods up to 5 mo and some lines were subcultured for 10 serial passages. During the logarithmic phase of cell growth, cell-doubling time was 92 h. Light, phase contrast, and scanning electron microscopy demonstrated that cultured human endothelial cells grew as monolayers of closely opposed, polygonal large cells whereas both cultured human fibroblasts and human smooth muscle cells grew as overlapping layers of parallel arrays of slender, spindle-shaped cells. By transmission electron microscopy, cultured endothelial cells were seen to contain cytoplasmic inclusions (Weibel-Palade bodies) characteristic of in situ endothelial cells. These inclusions were also found in endothelial cells lining umbilical veins but were not seen in smooth muscle cells or fibroblasts in culture or in situ. Cultured endothelial cells contained abundant quantities of smooth muscle actomyosin. Cultured endothelial cells also contained ABH antigens appropriate to the tissue donor's blood type; these antigens were not detectable on cultured smooth muscle cells or fibroblasts. These studies demonstrate that it is possible to culture morphologically and immunologically identifiable human endothelial cells for periods up to 5 mo.


Expression of the human germinal center-associated lymphoma (HGAL) protein, a new marker of germinal center B-cell derivation.
We identified the human germinal center-associated lymphoma (HGAL) in gene-expression profiling studies of diffuse large B-cell lymphoma (DLBCL). The expression of HGAL correlated with survival in patients with DLBCL. The HGAL gene is the human homolog of M17, a mouse gene expressed specifically in normal germinal center (GC) B cells. We generated a monoclonal antibody against the HGAL protein and show that HGAL is expressed in the cytoplasm of GC lymphocytes and in lymphomas of GC derivation. Among 727 lymphomas tested by immunohistochemistry on tissue microarrays, HGAL staining was found in follicular lymphomas (103 of 107), Burkitt lymphomas (40 of 40), mediastinal large B lymphomas (7 of 8), and in DLBCLs (103 of 151). Most marginal zone lymphomas lacked HGAL staining. Lymphocyte-predominant Hodgkin lymphomas (12 of 17) and, surprisingly, classical Hodgkin lymphomas (78 of 107) were found to be positive. Hierarchical clustering of comparative immunohistologic results in DLBCLs demonstrates that the expression of HGAL is similar to 2 other GC-associated proteins, BCL6 and CD10, but different from 2 markers associated with a non-GC phenotype, MUM1/IRF4 and BCL2. The restricted expression and GC specificity of HGAL protein suggest that it may have an important role in the diagnosis of specific lymphomas, and, potentially in the identification of subtypes associated with different prognoses.


Alternative mechanisms for delivery of medication in South Africa: A scoping review.
The number of people in South Africa with chronic conditions is a challenge to the health system. In response to the coronavirus infection, health services in Cape Town introduced home delivery of medication by community health workers. In planning for the future, they requested a scoping review of alternative mechanisms for delivery of medication to patients in primary health care in South Africa. Databases were systematically searched using a comprehensive search strategy to identify studies from the last 10 years. A methodological guideline for conducting scoping reviews was followed. A standardised template was used to extract data and compare study characteristics and findings. Data was analysed both quantitatively and qualitatively. A total of 4253 publications were identified and 26 included. Most publications were from the last 5 years (n = 21), research (n = 24), Western Cape (n = 15) and focused on adherence clubs (n = 17), alternative pick-up-points (n = 14), home delivery (n = 5) and HIV (n = 17). The majority of alternative mechanisms were supported by a centralised dispensing and packaging system. New technology such as smart lockers and automated pharmacy dispensing units have been piloted. Patients benefited from these alternatives and had improved adherence. Available evidence suggests alternative mechanisms were cheaper and more beneficial than attending the facility to collect medication. A mix of options tailored to the local context and patient choice that can be adequately managed by the system would be ideal. More economic evaluations are required of the alternatives, particularly before going to scale and for newer technology.


Physical activity promotion in Latin American populations: a systematic review on issues of internal and external validity.
The purpose of this review was to determine the degree to which physical activity interventions for Latin American populations reported on internal and external validity factors using the RE-AIM framework (reach & representativeness, effectiveness, adoption, implementation, maintenance). We systematically identified English (PubMed; EbscoHost) and Spanish (SCIELO; Biblioteca Virtual en Salud) language studies published between 2001 and 2012 that tested physical activity, exercise, or fitness promotion interventions in Latin American populations. Cross-sectional/descriptive studies, conducted in Brazil or Spain, published in Portuguese, not including a physical activity/fitness/exercise outcome, and with one time point assessment were excluded. We reviewed 192 abstracts and identified 46 studies that met the eligibility criteria (34 in English, 12 in Spanish). A validated 21-item RE-AIM abstraction tool was used to determine the quality of reporting across studies (0-7 = low, 8-14 = moderate, and 15-21 = high). The number of indicators reported ranged from 3-14 (mean = 8.1 ± 2.6), with the majority of studies falling in the moderate quality reporting category. English and Spanish language articles did not differ on the number of indicators reported (8.1 vs. 8.3, respectively). However, Spanish articles reported more across reach indicators (62% vs. 43% of indicators), while English articles reported more across effectiveness indicators (69% vs 62%). Across RE-AIM dimensions, indicators for reach (48%), efficacy/effectiveness (67%), and implementation (41%) were reported more often than indicators of adoption (25%) and maintenance (10%). Few studies reported on the representativeness of participants, staff that delivered interventions, or the settings where interventions were adopted. Only 13% of the studies reported on quality of life and/or potential negative outcomes, 20% reported on intervention fidelity, and 11% on cost of implementation. Outcomes measured after six months of intervention, information on continued delivery and institutionalization of interventions, were also seldom reported. Regardless of language of publication, physical activity intervention research for Latin Americans should increase attention to and measurement of external validity and cost factors that are critical in the decision making process in practice settings and can increase the likelihood of translation into community or clinical practice.


Unravelling the Structural and Molecular Basis Responsible for the Anti-Biofilm Activity of Zosteric Acid.
The natural compound zosteric acid, or p-(sulfoxy)cinnamic acid (ZA), is proposed as an alternative biocide-free agent suitable for preventive or integrative anti-biofilm approaches. Despite its potential, the lack of information concerning the structural and molecular mechanism of action involved in its anti-biofilm activity has limited efforts to generate more potent anti-biofilm strategies. In this study a 43-member library of small molecules based on ZA scaffold diversity was designed and screened against Escherichia coli to understand the structural requirements necessary for biofilm inhibition at sub-lethal concentrations. Considerations concerning the relationship between structure and anti-biofilm activity revealed that i) the para-sulfoxy ester group is not needed to exploit the anti-biofilm activity of the molecule, it is the cinnamic acid scaffold that is responsible for anti-biofilm performance; ii) the anti-biofilm activity of ZA derivatives depends on the presence of a carboxylate anion and, consequently, on its hydrogen-donating ability; iii) the conjugated aromatic system is instrumental to the anti-biofilm activities of ZA and its analogues. Using a protein pull-down approach, combined with mass spectrometry, the herein-defined active structure of ZA was matrix-immobilized, and was proved to interact with the E. coli NADH:quinone reductase, WrbA, suggesting a possible role of this protein in the biofilm formation process.


Bacillus sphaericus asporogenous mutants: morphology, protein pattern and larvicidal activity.
Asporogenous mutants from Bacillus sphaericus strains 2297 and 1593-4, blocked at different stages of the sporulation process, were isolated. Two mutants (2297 Aspo30A and 2297 Aspo34) which were blocked early in sporulation did not possess any crystalline inclusions and were poorly toxic to Culex pipiens mosquito larvae. Other mutants (2297 Aspo115, 2297 Aspo24 and 1593-4 Aspo12) which were blocked at later stages synthesized crystal-like inclusions next to the forespores, and were highly toxic to mosquito larvae. Electrophoretic protein analysis of alkali extracts from mutants and wild type strains confirmed the absence of toxic crystal-related proteins in early-blocked mutants and their presence in later ones. Western blots with antisera directed against the crystal proteins confirmed those observations.


Variation in Use of Prostate Biopsy Following Changes in Prostate Cancer Screening Guidelines.
Prostate biopsy rates have paralleled decreasing prostate specific antigen screening rates since 2012. We hypothesized that biopsy rates and the change in rates since 2012 would vary considerably across hospital referral regions. Using Medicare data from 2012 through 2014 we identified prostate biopsies performed by physicians who performed 11 or more biopsies annually. We calculated annual biopsy rates and changes in rates from 2012 to 2014 across 306 hospital referral regions. We performed multivariable regression adjusting for factors associated with annual biopsy rates (eg percent of patients older than 75 who were screened with prostate specific antigen and percent of the population that was African American). We also estimated adjusted prostate biopsy rates and changes with time across regions. We identified 395,993 biopsies. The overall rates decreased from 11.68 biopsies per 1,000 men in 2012 to 10.23 per 1,000 in 2014 (-12.4%, p = 0.11). Biopsy rates were higher in regions in which a greater percentage of the population was African American (β = 0.810, 95% CI 0.235-1.384, p = 0.006), ambulatory surgical centers were available where biopsy could be performed (β = 0.892, 95% CI 0.108-1.676, p = 0.026) and prostate specific antigen testing occurred more frequently (β = 2.462, 95% CI 1.153-3.771, p <0.001). There was marked geographic variation in the adjusted average biopsy rate (median adjusted rate 9.08 biopsies per 1,000 men, IQR 7.65-10.76) and in the change in biopsy rates with time (median adjusted rate change -1.49 biopsies per 1,000 men, IQR -1.94--1.22 per 1,000). Since 2012 there has been considerable geographic variation in the performance of prostate biopsies as well as changes with time after prostate specific antigen recommendations changed. Characterizing the role of unmeasured patient and physician level factors is crucial to optimize the use and minimize the harms of prostate biopsy.


Effect of finishing method on the microleakage of Class V resin composite restorations.
To evaluate the effect of three finishing methods on the microleakage of Class V hybrid resin composite. Facial and lingual Class V cavities were prepared at the cemento-enamel junction of bovine incisors. Restorations of the two materials (Z100; Prodigy) were placed, using the supplied dentin-adhesives (Scotchbond Multipurpose; OptiBond FL, respectively). Finishing methods were carbide (C) and diamond (D) finishing instruments operated at high-speed, and finishing disks (S). After thermal cycling, the teeth were immersed in methylene blue dye, then sections of the restored teeth (n = 12) were visually assessed for leakage. No leakage was observed at the resin-enamel interface in any restoration. At gingival margins, the incidence of leakage (slight/severe) was: Z100 D = 4/3, C = 5/3, S = 2/0; Prodigy D = 3/0, C = 2/4, S = 3/2. The greatest incidence of leakage was observed in restorations finished with carbide finishing burs. A two-way ANOVA of leakage data rankings showed no significant difference among the three methods (P = 0.07) or between the two materials (P = 0.55), and no significant interaction of material and finishing method (P = 0.07).


Tissue Dimensionality Influences the Functional Response of Cytotoxic T Lymphocyte-Mediated Killing of Targets.
Cytotoxic T lymphocyte (CTL)-mediated killing of virus infections and tumors occurs over a wide range of conditions. The spatial environments in which CTLs encounter target cells vary from narrow vessels, to two-dimensional epithelial tissues, to densely populated 3-dimensional (3D) T cell areas within lymphoid tissues. How such spatial environments alter the functional response of CTL-mediated killing, i.e., how the killing efficiency depends on cell densities, is unclear. In this study, we perform cellular Potts model simulations in different spatial configurations to investigate how the dimensionality of the space affects the functional response of CTL-mediated killing. Irrespective of the spatial configuration, the function with separate saturation constants for CTL and for target cell densities that we previously proposed can in all cases describe the response, demonstrating its generality. However, the tissue dimensionality determines at which cell densities the killing rate starts to saturate. We show that saturation in a fully 3D environment is stronger than in a "flat" 3D environment, which is largely due to accompanying differences in the CTL-target encounter rates.


Hyperentanglement source by intersubband two-photon emission from semiconductor quantum wells.
We propose an efficient hyperentanglement source emitting photon pairs entangled in both energy and polarization. The compact electrically driven room-temperature source, based on intersubband two-photon emission from semiconductor quantum wells (QWs) exhibits pair generation rates several orders of magnitude higher than alternative conventional schemes. A theoretical formalism is derived for the calculation of photon pair generation spectra and rates. The results are presented for superlattice structures similar to quantum cascade lasers of GaAs/AlGaAs QWs emitting in the mid-IR and far-IR and for InN/AlN QW structures suitable for telecommunication wavelengths.


The K65R mutant reverse transcriptase of HIV-1 cross-resistant to 2', 3'-dideoxycytidine, 2',3'-dideoxy-3'-thiacytidine, and 2',3'-dideoxyinosine shows reduced sensitivity to specific dideoxynucleoside triphosphate inhibitors in vitro.
The K65R mutation in human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) encodes cross-resistance to 2',3'-dideoxycytidine (ddC), 2',3'-dideoxy-3'-thiacytidine (3TC), and 2',3'-dideoxyinosine (ddI). We characterized the in vitro sensitivities of recombinant wild type (wt) and K65R mutant RT to dideoxynucleoside triphosphate (ddNTP) inhibitors, using a variety of primer-templates. With poly(rA)-oligo(dT), the K65R mutant showed slight increases in Ki for ddTTP and 3'-azido, 3'-deoxythymidine triphosphate (AZTTP) compared to wt RT, but neither wt nor K65R RT was inhibited by ddCTP or ddATP. With poly(rI)-oligo(dC), the K65R mutant showed a 2-fold increase in Km for dCTP and a 20-fold increase in Ki for ddCTP compared to wt, whereas ddATP, ddTTP, and AZTTP failed to inhibit either enzyme. With a heteropolymeric primer-template, the K65R mutant showed 10-fold reduced sensitivities to ddCTP, 3TCTP, and ddATP, and 4-fold reduced sensitivity to AZTTP, compared to wt. In contrast, both enzymes were equally inhibited by ddTTP and ddGTP. HIV-1 cross-resistance to ddC/3TC/ddI resulting from the K65R mutation may therefore involve selective alterations in substrate/inhibitor recognition. Additionally, competitive inhibition by ddNTPs noncomplementary to the template base appears to be unimportant in the mechanism of inhibition of HIV-1 RT by dideoxynucleoside analogs.


Animal model estimation of genetic parameters and response to selection for litter size and weight, growth, and backfat in closed seedstock populations of large white and Landrace swine.
Records from 2,495 litters and 14,605 Landrace and Large White pigs from two farms, but established from the same base population and run as replicated selection lines, were analyzed. Selection within herd was on estimated breeding values weighted by economic values. Animal models and REML procedures were used to estimate genetic, phenotypic, and environmental parameters for the number of pigs born alive (NBA), litter weight at 21 d (LW), average daily gain form approximately 30 to 104 kg (ADG), and backfat thickness adjusted to 104 kg (BF). Random animal genetic effects (o), permanent (NBA and LW) or litter (ADG and BF) environmental effects, maternal genetic effects (m), and the covariance between o and m were sequentially added to the model. Estimates of total heritability calculated from all data (ht2 = sigma o2 + 1/2 sigma m2 + 3/2 sigma om) ranged from .01 to .14 for NBA, from .18 to .22 for LW, from .23 to .34 for ADG, and from .40 to .50 for BF. Maternal genetic variance was from 2.4 to 3.8% of phenotypic variance in NBA, from 1.2 to 3.6% in LW, from .5 to 1.5% in ADG, and from 1.9 to 3.4% in BF. The correlation between o and m was -.07 for NBA, -.25 for LW, -.34 for ADG, and -.26 for BF. Permanent environmental effects explained from 16 to 17% of total phenotypic variation for NBA and from 1.6 to 5.3% for LW. Approximately 7% of the variation in ADG and 5% in BF was due to litter environmental effects.(ABSTRACT TRUNCATED AT 250 WORDS)


Fatty liver is associated with reduced SIRT3 activity and mitochondrial protein hyperacetylation.
Acetylation has recently emerged as an important mechanism for controlling a broad array of proteins mediating cellular adaptation to metabolic fuels. Acetylation is governed, in part, by SIRTs (sirtuins), class III NAD(+)-dependent deacetylases that regulate lipid and glucose metabolism in liver during fasting and aging. However, the role of acetylation or SIRTs in pathogenic hepatic fuel metabolism under nutrient excess is unknown. In the present study, we isolated acetylated proteins from total liver proteome and observed 193 preferentially acetylated proteins in mice fed on an HFD (high-fat diet) compared with controls, including 11 proteins not previously identified in acetylation studies. Exposure to the HFD led to hyperacetylation of proteins involved in gluconeogenesis, mitochondrial oxidative metabolism, methionine metabolism, liver injury and the ER (endoplasmic reticulum) stress response. Livers of mice fed on the HFD had reduced SIRT3 activity, a 3-fold decrease in hepatic NAD(+) levels and increased mitochondrial protein oxidation. In contrast, neither SIRT1 nor histone acetyltransferase activities were altered, implicating SIRT3 as a dominant factor contributing to the observed phenotype. In Sirt3⁻(/)⁻ mice, exposure to the HFD further increased the acetylation status of liver proteins and reduced the activity of respiratory complexes III and IV. This is the first study to identify acetylation patterns in liver proteins of HFD-fed mice. Our results suggest that SIRT3 is an integral regulator of mitochondrial function and its depletion results in hyperacetylation of critical mitochondrial proteins that protect against hepatic lipotoxicity under conditions of nutrient excess.


Anatomical segregation of representations of personally familiar and famous people in the temporal and parietal cortices.
Person recognition has been assumed to entail many types of person-specific cognitive responses, including retrieval of knowledge, episodic recollection, and emotional responses. To demonstrate the cortical correlates of this modular structure of multimodal person representation, we investigated neural responses preferential to personally familiar people and responses dependent on familiarity with famous people in the temporal and parietal cortices. During functional magnetic resonance imaging (fMRI) measurements, normal subjects recognized personally familiar names (personal) or famous names with high or low degrees of familiarity (high or low, respectively). Effects of familiarity with famous people (i.e., high-low) were identified in the bilateral angular gyri, the left supramarginal gyrus, the middle part of the bilateral posterior cingulate cortices, and the left precuneus. Activation preferentially relevant to personally familiar people (i.e., personal-high) was identified in the bilateral temporo-parietal junctions, the right anterolateral temporal cortices, posterior middle temporal gyrus, posterior cingulate cortex (with a peak in the posterodorsal part), and the left precuneus; these activation foci exhibited varying degrees of activation for high and low names. An equivalent extent of activation was observed for all familiar names in the bilateral temporal poles, the left orbito-insular junction, the middle temporal gyrus, and the anterior part of the posterior cingulate cortex. The results demonstrated that distinct cortical areas supported different types of cognitive responses, induced to different degrees during recognition of famous and personally familiar people, providing neuroscientific evidence for the modularity of multimodal person representation.


Policies, activities, and structures supporting research mentoring: a national survey of academic health centers with clinical and translational science awards.
To document the frequency of policies and activities in support of mentoring practices at institutions receiving a U.S. National Institutes of Health's Clinical and Translational Science Award (CTSA). The study consisted of a 69-item survey with questions about the inclusion (formal or informal) of policies, activities, and structures supporting mentoring within CTSA-sponsored research (i.e., KL2 programs) and, more broadly, in the CTSA's home institution. The survey, conducted from November 2010 through January 2011, was sent to the 55 institutions awarded CTSAs at the time of the survey. Follow-up phone interviews were conducted to clarify responses as needed. Fifty-one of 55 (92%) institutions completed the survey for institutional programs and 53 of 55 (96%) for KL2 programs. Responses regarding policies and activities involving mentor criteria, mentor-mentee relationship, incentives, and evaluative mechanisms revealed considerable variability between KL2 and institutional programs in some areas, such as having mentor qualification criteria and processes to evaluate mentors. The survey also identified areas, such as training and women and minority mentoring programs, where there was frequent sharing of activities between the institutional and KL2 programs. KL2 programs and institutional programs tend to have different preferences for policies versus activities to optimize qualification of mentors, the mentor-mentee relationship, incentives, and evaluation mechanisms. Frequently, these elements are informal. Individuals in charge of implementing and maintaining mentoring initiatives can use the results of the study to consider their current mentoring policies, structures, and activities by comparing them with national patterns within CTSA institutions.


Quasimonochromatic white light fringe interferometer.
Quasimonochromatic light sources, such as laser diodes and high power LEDs, are investigated to determine their suitability for zero path difference determination using white light fringes in a Michelson interferometer. Fringe visibility curves are theoretically determined for various combinations of light sources and compared with experimental results when used in a Michelson interferometer with a 25-m path length. A resolution of 2-3 microm was obtained for a pair of multimode laser diodes and also for a single multimode laser diode operated as an LED. This is more than adequate for the calibration of survey baselines.


A Novel Resolution of Diabetes: C-C Chemokine Motif Ligand 4 Is a Common Target in Different Types of Diabetes by Protecting Pancreatic Islet Cell and Modulating Inflammation.
Systemic inflammation is related to hyperglycemia in diabetes mellitus (DM). C-C chemokine motif ligand (CCL) 4 is upregulated in type 1 & type 2 DM patients. This study aimed to investigate if CCL4 could be a potential target to improve blood sugar control in different experimental DM models. Streptozotocin-induced diabetic mice, Leprdb /JNarl diabetic mice, and C57BL/6 mice fed a high fat diet were used as the type 1 DM, type 2 DM, and metabolic syndrome model individually. Mice were randomly assigned to receive an anti-CCL4 neutralizing monoclonal antibody. The pancreatic β-cells were treated with streptozotocin for in vitro experiments. In streptozotocin-induced diabetic mice, inhibition of CCL4 controlled blood sugar, increased serum insulin levels, increased islet cell proliferation and decreased pancreatic interleukin (IL)-6 expression. In the type 2 diabetes and metabolic syndrome models, CCL4 inhibition retarded the progression of hyperglycemia, reduced serum tumor necrosis factor (TNF)-α and IL-6 levels, and improved insulin resistance via reducing the phosphorylation of insulin receptor substrate-1 in skeletal muscle and liver tissues. CCL4 inhibition directly protected pancreatic β-cells from streptozotocin stimulation. Furthermore, CCL4-induced IL-6 and TNF-α expressions could be abolished by siRNA of CCR2/CCR5. In summary, direct inhibition of CCL4 protected pancreatic islet cells, improved insulin resistance and retarded the progression of hyperglycemia in different experimental models, suggesting the critical role of CCL4-related inflammation in the progression of DM. Future experiments may investigate if CCL4 could be a potential target for blood sugar control in clinical DM.


Impact of antibiotic treatments on the expression of the R plasmid tra genes and on the host innate immune activity during pRAS1 bearing Aeromonas hydrophila infection in zebrafish (Danio rerio).
The transfer of R plasmids between bacteria has been well studied under laboratory conditions and the transfer frequency has been found to vary between plasmids and under various physical conditions. For the first time, we here study the expression of the selected plasmid mobility genes traD, virB11 and virD4 in the 45 kb IncU plasmid, pRAS1, conferring resistance to tetracycline, trimethoprim and sulphonamide, using an in vivo zebrafish infection- treatment model. Three days after oral infection of adult zebrafish with Aeromonas hydrophila harboring pRAS1, elevated expression of pro-inflammatory cytokine (TNF α, IL-1β and IL-8) and complement C3 genes in the intestine coincided with disease symptoms. Tetracycline, trimethoprim and an ineffective concentration of flumequine given 48 h prior to sampling, strongly increased expression of plasmid mobility genes, whereas an effective dosage of flumequine resulted in lower levels of mRNA copies of these genes relative to placebo treatment. Following effective treatment with flumequine, and ineffective treatments with a low concentration of flumequine, with trimethoprim or with sulphonamide, the intestinal expression of immune genes was strongly induced compared to placebo treated control fish. Treatment of zebrafish infected with an antibiotic resistant (TcR, TmR, SuR) A. hydrophila with ineffective concentrations of flumequine or the ineffective antimicrobials tetracycline and trimethoprim strongly induced expression of genes mediating conjugative transfer of the R-plasmid pRAS1. Simultaneously, there was a strong induction of selected inflammatory and immune response genes, which was again evident in fish subjected to ineffective treatment protocols. Our findings point to the essential role of therapeutic practices in escalation or control of antibiotic resistance transfer, and suggest that antibiotic substances, even in sub-inhibitory concentrations, may stimulate innate defenses against bacterial infections.