A role for thioredoxin-interacting protein (Txnip) in cellular creatine homeostasis.
Creatine is important for energy metabolism, yet excitable cells such as cardiomyocytes do not synthesize creatine and rely on uptake via a specific membrane creatine transporter (CrT; SLC6A8). This process is tightly controlled with downregulation of CrT upon continued exposure to high creatine via mechanisms that are poorly understood. Our aim was to identify candidate endogenous CrT inhibitors. In 3T3 cells overexpressing the CrT, creatine uptake plateaued at 3 h in response to 5 mM creatine but peaked 33% higher (P < 0.01) in the presence of cycloheximide, suggesting CrT regulation depends on new protein synthesis. Global gene expression analysis identified thioredoxin-interacting protein (Txnip) as the only significantly upregulated gene (by 46%) under these conditions (P = 0.036), subsequently verified independently at mRNA and protein levels. There was no change in Txnip expression with exposure to 5 mM taurine, confirming a specific response to creatine rather than osmotic stress. Small-interfering RNA against Txnip prevented Txnip upregulation in response to high creatine, maintained normal levels of creatine uptake, and prevented downregulation of CrT mRNA. These findings were relevant to the in vivo heart since creatine-deficient mice showed 39.71% lower levels of Txnip mRNA, whereas mice overexpressing the CrT had 57.6% higher Txnip mRNA levels and 28.7% higher protein expression compared with wild types (mean myocardial creatine concentration 124 and 74 nmol/mg protein, respectively). In conclusion, we have identified Txnip as a novel negative regulator of creatine levels in vitro and in vivo, responsible for mediating substrate feedback inhibition and a potential target for modulating creatine homeostasis.


"No level up!": no effects of video game specialization and expertise on cognitive performance.
Previous research into the effects of action video gaming on cognition has suggested that long term exposure to this type of game might lead to an enhancement of cognitive skills that transfer to non-gaming cognitive tasks. However, these results have been controversial. The aim of the current study was to test the presence of positive cognitive transfer from action video games to two cognitive tasks. More specifically, this study investigated the effects that participants' expertise and genre specialization have on cognitive improvements in one task unrelated to video gaming (a flanker task) and one related task (change detection task with both control and genre-specific images). This study was unique in three ways. Firstly, it analyzed a continuum of expertise levels, which has yet to be investigated in research into the cognitive benefits of video gaming. Secondly, it explored genre-specific skill developments on these tasks by comparing Action and Strategy video game players (VGPs). Thirdly, it used a very tight experiment design, including the experimenter being blind to expertise level and genre specialization of the participant. Ninety-two university students aged between 18 and 30 (M = 21.25) were recruited through opportunistic sampling and were grouped by video game specialization and expertise level. While the results of the flanker task were consistent with previous research (i.e., effect of congruence), there was no effect of expertise, and the action gamers failed to outperform the strategy gamers. Additionally, contrary to expectation, there was no interaction between genre specialization and image type in the change detection task, again demonstrating no expertise effect. The lack of effects for game specialization and expertise goes against previous research on the positive effects of action video gaming on other cognitive tasks.


The Interplay of Relational and Non-relational Processes in Sentence Production: The Case of Relative Clause Planning in Japanese and Spanish.
Speech planning involves different steps in order to transform a conceptual message into speech. These include establishing structural relations among constituents (i.e., relational information), and selecting the appropriate lexical items to convey the intended message (non-relational elements). However, the precise way relational and non-relational information are computed when undertaking linguistic encoding is not clear. This paper explores how the pre-linguistic message undergoes linguistic encoding, and what kind of information (relational or non-relational) is prioritized in doing so. We analyze the production planning of Relative Clauses in Spanish (a head-initial language) and Japanese (a head-final language) by monolingual speakers, by means of the eye-tracking method while participants described colored pictures. Although in both Spanish and Japanese the structure under study is the same (with the same syntactic configuration), word order is entirely opposite between both languages. In Japanese, the head noun is not uttered until the end of the clause, thus making it possible to explore sentence planning in a structure where the syntactically most dominant element (the head noun, HN) is not the first element. Variables tested were type of relative clause, with either the agent or the patient as head noun, and the animacy of the agent and the patient of the event, the latter allowing the manipulation of the conceptual saliency of the elements involved. Results showed Japanese speakers focus extensively on the HN before directing their gazes to the element they are going to utter first, suggesting a speech planning process that prioritizes relational information, that is, structural scaffolding. Spanish monolinguals, in turn, showed a pattern in which both structural and linear information appear to be more closely related from the beginning. In both languages, the animacy of isolated elements had little effect on gaze patterns. Results point to a planning process that prioritizes structural relations over access to lexical elements in order in the planning of complex structures, with room for flexibility when the grammar of the language allows so.


The effect of 5-fluorouracil on the long term survival and proliferation of cells in the rat hippocampus.
Chemotherapy has been reported to produce cognitive impairments in a significant number of cancer patients. These deficits frequently involve aspects of spatial or declarative memory which can persist for up to several years after completion of the treatment. We have recently shown that 5-fluorouracil (5-FU), a commonly used chemotherapy drug, induces cognitive impairment and a reduction in hippocampal neurogenesis using a rat model of chemotherapy (Elbeltagy et al. [17]). The present study examines the effects of two weeks of 5-FU treatment on cell proliferation in the sub granular zone (SGZ) of the dentate gyrus and the survival of newly dividing cells over a six week period after the end of treatment. Cell proliferation at each time point was quantified by staining for the cell proliferation marker Ki67 while the survival of cells, dividing at the start of treatment, was determined by quantification of BrdU positive cell numbers after pulse labelling with BrdU at the start of drug treatment. The results show that 2 weeks of 5-FU treatment did not significantly reduce cell proliferation in the SGZ immediately after treatment. However cell proliferation was significantly reduced, compared to saline treated controls, two weeks after the end of treatment and remained significantly reduced at 6 weeks. The survival of cells, dividing at the start of treatment, was significantly reduced when quantified immediately after the end of treatment and continued to decline compared with control animals over the following 2 weeks but no further change occurred at 6 weeks. Quantification of COX-2 positive cell numbers in the hippocampus did not correlate with the reduction in cell proliferation or survival suggesting that inflammation is not responsible for these changes. These results demonstrate that 5-FU has delayed and prolonged effects on hippocampal neurogenesis after the end of chemotherapy treatment. This correlates with patient reports of continued cognitive impairment after treatment and indicates that changes in neurogenesis may underlie these effects.


The surgical learning curve in aural atresia surgery.
The objective of this retrospective case review is to examine the effect of surgical learning on hearing outcomes and complications in congenital aural atresia surgery. Sixty-four consecutive ears (in 60 patients) operated on during the period of 1994 to 2004 at a tertiary referral center were studied. Intervention consisted of aural atresiaplasty through an anterior approach by the same surgeon (C.S.). Hearing outcomes and complication rates were compared between four temporally sequential groups of 16 ears. Acceptable hearing and complication rate outcomes were defined as results comparable to larger series in the literature. Hearing results, in the short term, comparable to larger series were achieved during the first group of ears (nos. 1-16). A plateau in the learning curve for short-term hearing outcomes was achieved after the first two groups, that is, after 32 ears. Hearing outcomes, in the long term (>1 year) comparable to larger series, were achieved in the second group of ears (nos. 16-32). The learning curve for long-term hearing demonstrated a significant improvement in outcomes in the final group of 16 ears compared with the first 48 ears. Long-term hearing results for the final group show closure of the postoperative air-bone gap to less than 30 dB in 94% of cases. Postoperative complication rates were equivalent to larger series in the first group of 16 ears and showed no statistically significant difference between the four groups. There was one patient with sensorineural hearing loss after surgery; there were no anacoustic ears and no facial palsies in the study group. A learning curve of at least 32 ears was required to achieve stable short-term hearing results. To achieve stable long-term hearing results required a learning curve of at least 48 patients in our series. Complication rates remained stable throughout the study period.


Excess risk of chronic physical conditions associated with depression and anxiety.
Depression and anxiety have been reported to be associated with chronic physical conditions. We examined the excess risk of chronic physical conditions associated with depression and/or anxiety within a multivariate framework controlling for demographic and modifiable lifestyle risk factors. We used a retrospective cross-sectional study design. Study participants were adults aged 22-64 years from 2007 and 2009 Medical Expenditure Panel Survey. We defined presence of depression-anxiety based on self-reported depression and anxiety and classified adults into 4 groups: 1) depression only; 2) anxiety only; 3) comorbid depression and anxiety 4) no depression and no anxiety. We included presence/absence of arthritis, asthma, chronic obstructive pulmonary disorder, diabetes, heart disease, hypertension, and osteoporosis as dependent variables. Complementary log-log regressions were used to examine the excess risk associated with depression and/or anxiety for chronic physical conditions using a multivariate framework that controlled for demographic (gender, age, race/ethnicity) and modifiable lifestyle (obesity, lack of physical activity, smoking) risk factors. Bonferroni correction for multiple comparisons was applied and p ≤0.007 was considered statistically significant. Overall, 7% had only depression, 5.2% had only anxiety and 2.5% had comorbid depression and anxiety. Results from multivariable regressions indicated that compared to individuals with no depression and no anxiety, individuals with comorbid depression and anxiety, with depression only and with anxiety only, all had higher risk of all the chronic physical conditions. ARRs for comorbid depression and anxiety ranged from 2.47 (95% CI: 1.47, 4.15; P = 0.0007) for osteoporosis to 1.64 (95% CI: 1.33, 2.04; P < 0.0001) for diabetes. Presence of depression only was also found to be significantly associated with all chronic conditions except for osteoporosis. Individuals with anxiety only were found to have a higher risk for arthritis, COPD, heart disease and hypertension. Presence of depression and/or anxiety conferred an independent risk for having chronic physical conditions after adjusting for demographic and modifiable lifestyle risk factors.


Regulation of Wnt5a mRNA expression in human mammary epithelial cells by cell shape, confluence, and hepatocyte growth factor.
The Wnts are a family of genes with a role in cell fate and morphological development in numerous embryonic and adult tissues. In mouse mammary tissue a subset of the Wnts have a function in the normal development of the gland, and aberrant expression of Wnts normally silent in this tissue causes mammary carcinomas. We have previously shown that Wnt5a expression is elevated in the epithelial component of proliferative lesions of human breast and have therefore examined the regulation of Wnt5a mRNA expression in the human mammary epithelial cell line HB2, which has a luminal phenotype and thus represents the most commonly transformed cell type in human breast cancer. Wnt5a was up-regulated 30-fold at confluence. This up-regulation was induced specifically by confluence and not by the growth arrest that accompanied it. In addition, Wnt5a was down-regulated 3-fold by changes in cell shape associated with the transition from growth on a two-dimensional surface (flat cell morphology) to growth in three-dimensional gels (spherical cell morphology). Cytoskeletal disruption with non-toxic doses of colchicine also induced a spherical morphology and brought about a dose-dependent down-regulation of Wnt5a. Wnt5a was also down-regulated 10-fold during the hepatocyte growth factor-induced branching of HB2 cell aggregates in collagen gels. The down-regulation of Wnt5a preceded the branching process. A similar result was obtained with primary human breast epithelial populations and the breast cancer cell line MDA468. We conclude that regulation of Wnt5a expression is a down-stream effect of signaling by hepatocyte growth factor. These results are consistent with a role for Wnt5a in mammary epithelial cell motility and are in accord with Xwnt5a's function in embryonal cell migration. If Wnt5a's function in human mammary epithelial cells is similar to that of Xwnt5a, its up-regulation at confluence may be a mechanism for inhibition of cell migration beyond confluence.


[Prospective study on the relationship between physical fitness and all-cause mortality in Japanese men].
This study was conducted to examine the relationship between physical fitness and all-cause mortality in Japanese men. We evaluated the physical fitness and risk for all-cause mortality of 9,986 Japanese men who were given a submaximal exercise test and a medical examination between 1982 and 1984. Physical fitness was measured using a bicycle ergometer test, and maximal oxygen uptake was estimated. The average follow-up time was 14 years, for total of 139,836 person-years of observation. There were 247 deaths during the observation period. The relative risk and 95% confidence intervals (95% CI) for all-cause mortality were obtained using the Cox proportional hazards model. Following age adjustment, and using the lowest physical fitness (quintile I) group as a reference, the hazard ratios for quintiles II through V were, 0.54 (0.39-0.77), 0.66 (0.47-0.94), 0.58 (0.39-0.86), and 0.46 (0.27-0.78), respectively. After being adjusted for age, body mass index, hypertension, and urinary protein, the hazard ratios were, 0.52 (0.37-0.73), 0.60 (0.42-0.87), 0.50 (0.33-0.75), and 0.39 (0.22-0.67), respectively. The results presented here support the hypothesis that a low level of physical fitness in an important risk factor for all-cause mortality in Japanese men.


Scale-Up of Early Infant Male Circumcision Services for HIV Prevention in Lesotho: A Review of Facilitating Factors and Challenges.
The World Health Organization and the Joint United Nations Programme on HIV/AIDS recommend early infant male circumcision (EIMC) as a component of male circumcision programs in countries with high HIV prevalence and low circumcision rates. Lesotho began incorporating EIMC into routine maternal, newborn, and child health (MNCH) services in 2013 with funding from the United States Agency for International Development and United Nations Children's Fund. This presented unique challenges: Lesotho had no previous experience with EIMC and cultural traditions link removal of the foreskin to rites of passage. This process evaluation provides an overview of EIMC implementation. The Lesotho Ministry of Health and Jhpiego conducted a baseline assessment before service implementation. Baseline information from an initial assessment was used to develop and implement an EIMC program that had a pilot and a scale-up phase. Key program activities such as staff training, quality assurance, and demand creation were included at the program design phase. Facilitating factors and challenges were identified from a review of information collected during the baseline assessment as well as the pilot. Between September 2013 and March 2015, 592 infants were circumcised at 9 sites: 165 (28%) between 1 day and 6 days after birth; 196 (33%) between 7 and 30 days, and 231 (39%) between 31 and 60 days. Facilitating factors included strong support from the Ministry of Health, collaboration with stakeholders, and donor funding. Providers were enthusiastic about the opportunity to offer new services and receive training. Challenges included gaining consent from family members other than mothers, and parents' concern about pain and complications. The EIMC program also had to manage providers' expectations of compensation because overtime was paid to providers who took part in adult circumcision programming but not for EIMC. Limited human resources, including authorization only for doctors to perform EIMC, impeded provision of services. Despite communication, compensation, and task-shifting challenges, integrating EIMC services with MNCH services could be a sustainable model for EIMC service delivery in Lesotho.


Harnessing the active site triad: merging hemilability, proton responsivity, and ligand-based redox-activity.
Metalloenzymes catalyze many important reactions by managing the proton and electron flux at the enzyme active site. The motifs utilized to facilitate these transformations include hemilabile, redox-active, and so called proton responsive sites. Given the importance of incorporating and understanding these motifs in the area of coordination chemistry and catalysis, we highlight recent milestones in the field. Work incorporating the triad of hemilability, redox-activity, and proton responsivity into single ligand scaffolds will be described.


Unexpectedly high prevalence of target-organ damage in newly diagnosed Nigerians with hypertension.
Hypertension remains a formidable cardiovascular problem in the entire world. Untreated or poorly controlled hypertension predisposes to target-organ damage and is often followed by serious cardiovascular events. Knowledge of the type and distribution of endorgan damage in newly diagnosed hypertensive patients would help to determine the interventions or preventative measures needed in such patients. To assess target-organ involvement in newly diagnosed, untreated hypertensive patients presenting at the University College Hospital, Ibadan, Nigeria. Newly diagnosed hypertensive men and women were recruited consecutively into the study. Target-organ damage was assessed using electroctrocardiography, echocardiography, renal function assessment (including tests for albuminuria) and fundoscopy. A total of 68 subjects were recruited, but only 54 (79.4%), with a mean age of 59.1 ( +/- 12.4) years, completed the study. There were 29 ( +/- 53.7%) men, mean age 63.5 ( +/- 12.6) years, and 25 (46.3%) women, mean age 54.1 ( +/- 10.4) years. Fifty per cent of the subjects had no symptoms. Some form of albuminuria was found in about 40.7%. Seventy-one per cent (71%) had between grade 1 and 2 hypertensive retinopathy. Only six (11.1%) had glomerular filtration rate greater than 90 ml/min. Systolic function was impaired in three (5.6%) of the subjects. Diastolic dysfunction was present in 36 (66.7%) subjects. Target-organ damage already exists in newly diagnosed hypertensive subjects before presentation. We suggest early screening for cardiovascular risk factors and possibly a reduction in the criteria for the definition of hypertension in black Africans.


Surgical treatment of peri-implantitis.
To review the literature on surgical treatment of peri-implantitis. A search of PubMed and as well as a hand search of articles were conducted. Publications and articles accepted for publication up to November 2007 were included. A total of 43 studies were selected for the review. Only 13 of these were studies in humans and only one study directly addressed disease resolution. Thus the available evidence for surgical treatment of peri-implantitis is extremely limited. Re-osseointegration can occur on previously contaminated surfaces. The surface characteristics are decisive for regeneration and re-osseointegration. No single surface decontamination method appears to be distinctly superior. Open debridement with surface decontamination can achieve resolution. Access surgery has been investigated in one study demonstrating that resolution occurred in 58% of the lesions. No single method of surface decontamination (chemical agents, air abrasives and lasers) was found to be superior. The use of regenerative procedures such as bone graft techniques with or without the use of barrier membranes has been reported with various degrees of success. However, it must be stressed that such techniques do not address disease resolution but rather merely attempt to fill the osseous defect.


Cooperative Chikungunya Virus Membrane Fusion and Its Substoichiometric Inhibition by CHK-152 Antibody.
Chikungunya virus (CHIKV) presents a major burden on healthcare systems worldwide, but specific treatment remains unavailable. Attachment and fusion of CHIKV to the host cell membrane is mediated by the E1/E2 protein spikes. We used an in vitro single-particle fusion assay to study the effect of the potent, neutralizing antibody CHK-152 on CHIKV binding and fusion. We find that CHK-152 shields the virions, inhibiting interaction with the target membrane and inhibiting fusion. The analysis of the ratio of bound antibodies to epitopes implied that CHIKV fusion is a highly cooperative process. Further, dissociation of the antibody at lower pH results in a finely balanced kinetic competition between inhibition and fusion, suggesting a window of opportunity for the spike proteins to act and mediate fusion, even in the presence of the antibody.


Recurrent Loss, Horizontal Transfer, and the Obscure Origins of Mitochondrial Introns in Diatoms (Bacillariophyta).
We sequenced mitochondrial genomes from five diverse diatoms (Toxarium undulatum, Psammoneis japonica, Eunotia naegelii, Cylindrotheca closterium, and Nitzschia sp.), chosen to fill important phylogenetic gaps and help us characterize broadscale patterns of mitochondrial genome evolution in diatoms. Although gene content was strongly conserved, intron content varied widely across species. The vast majority of introns were of group II type and were located in the cox1 or rnl genes. Although recurrent intron loss appears to be the principal underlying cause of the sporadic distributions of mitochondrial introns across diatoms, phylogenetic analyses showed that intron distributions superficially consistent with a recurrent-loss model were sometimes more complicated, implicating horizontal transfer as a likely mechanism of intron acquisition as well. It was not clear, however, whether diatoms were the donors or recipients of horizontally transferred introns, highlighting a general challenge in resolving the evolutionary histories of many diatom mitochondrial introns. Although some of these histories may become clearer as more genomes are sampled, high rates of intron loss suggest that the origins of many diatom mitochondrial introns are likely to remain unclear.


Efficacy of interventions to increase the uptake of chlamydia screening in primary care: a systematic review.
As most genital chlamydia infections are asymptomatic, screening is the main way to detect and cases for treatment. We undertook a systematic review of studies assessing the efficacy of interventions for increasing the uptake of chlamydia screening in primary care. We reviewed studies which compared chlamydia screening in the presence and the absence of an intervention. The primary endpoints were screening rate or total tests. We identified 16 intervention strategies; 11 were randomised controlled trials and five observational studies, 10 targeted females only, five both males and females, and one males only. Of the 15 interventions among females, six were associated with significant increases in screening rates at the 0.05 level including a multifaceted quality improvement program that involved provision of a urine jar to patients at registration (44% in intervention clinics vs. 16% in the control clinic); linking screening to routine Pap smears (6.9% vs. 4.5%), computer alerts for doctors (12.2% vs. 10.6%); education workshops for clinic staff; internet-based continuing medical education (15.5% vs. 12.4%); and free sexual health consultations (16.8% vs. 13.2%). Of the six interventions targeting males, two found significant increases including the multifaceted quality improvement program in which urine jars were provided to patients at registration (45% vs. 15%); and the offering by doctors of a test to all presenting young male clients, prior to consultation (29 vs. 4%). Interventions that promoted the universal offer of a chlamydia test in young people had the greatest impact on increasing screening in primary care.


Evidence for Asthma in the Lungs of Mice Inoculated with Different Doses of Toxocara canis.
Toxocara canis, a common roundworm that mainly causes toxocariasis, is a zoonotic parasite found worldwide. Humans, an accidental host, can acquire T. canis infection through accidental ingestion of T. canis-embryonated egg-contaminated food, water, and soil, and by encapsulated larvae in a paratenic host's viscera or meat. Long-term residence of T. canis larvae in a paratenic host's lungs may induce pulmonary inflammation that contributes to lung injury, airway inflammatory hyperresponsiveness, and collagen deposition in mice and clinical patients. This study intended to investigate the relationship between T. canis infection and allergic asthma in BALB/c mice inoculated with high, moderate, and low doses of T. canis eggs for a 13-week investigation. The airway hyperresponsiveness (AHR) to methacholine, collagen deposition, cytokine levels, and pathological changes in lung tissues was assessed in infected mice at weeks 1, 5, and 13 postinfection. The cell composition in bronchoalveolar lavage fluid of infected mice was assessed at weeks 5 and 13 postinfection. Compared with uninfected control mice, all groups of T. canis-infected mice exhibited significant AHR, a dose-dependent increase in eosinophilic infiltration leading to multifocal interstitial and alveolar inflammation with abundant mucus secretion, and collagen deposition in which the lesion size increased with the infective dose. Infected mice groups also showed significant expressions of eotaxin and type 2 T-helper-dominant cytokines such as interleukin (IL)-4, IL-5, and IL-13. Overall, these results suggest that T. canis larval invasion of the lungs may potentially cause pulmonary inflammatory injury and could subsequently contribute to the development of allergic manifestations such as asthma.


The Role of Spirituality Among Military En Route Care Nurses: Source of Strength or Moral Injury?
Military nurses provide care to seriously injured service members in flight, on the ground, or at sea during transport from the point of injury to a facility capable of providing higher levels of care. From this experience nurses are at increased risk of developing negative behavioral health symptoms. Spirituality, a belief in someone or something greater than oneself, could provide behavioral health support for military nurses who serve in this role. To determine the impact of spirituality on the behavioral health of nurses who provided en route care while deployed to Iraq or Afghanistan. This exploratory mixed-methods study used 5 instruments to determine levels of anxiety, depression, posttraumatic stress, posttraumatic growth, and resilience among 119 military nurses. Interviews provided rich data about the experiences of these nurses and extended quantitative outcomes. Posttraumatic Growth Inventory findings showed no significant change in spirituality based on deployment experiences (mean, 3.07; SD, 3.26). However, interviews revealed that spirituality served as a buffer against developing behavioral health issues. Many relied on spirituality to get them through difficult experiences. There was also a sense of moral injury as a few expressed regrets for things they witnessed or experienced. Spirituality can insulate military nurses from negative behavioral health symptoms. Nurses included in the study relied on their spirituality to stay mentally fit. For nurses who experienced moral injury, supervisory recognition of this and appropriate referral may decrease the long-term effects of deployment on their behavioral health.


Safety and efficacy of a feed additive consisting on the bacteriophages PCM F/00069, PCM F/00070, PCM F/00071 and PCM F/00097 infecting Salmonella Gallinarum B/00111 (Bafasal®) for all avian species (Proteon Pharmaceuticals S.A.).
Following a request from the European Commission, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the feed additive consisting of four bacteriophages infecting Salmonella Gallinarum B/00111 (PCM F/00069, PCM F/00070, PCM F/00071 and PCM F/00097, trade name: Bafasal®) when used as a zootechnical additive in water for drinking and liquid complementary feed for all avian species. The effects sought are the reduction of the Salmonella spp. carriage in chickens for fattening, the improvement of their performance, or both. The host strain harbours an acquired antimicrobial resistance gene. No viable cells or DNA from the host organism were found in the additive. The four phages proved to be strictly lytic and to have a machinery allowing to package a unit-length of the viral genome. The manufacturing process excludes the presence of remnants from the propagation process in the final additive. Consequently, no concerns are expected from the nature and manufacture of the product. Considering this and the results of the tolerance study with chickens for fattening, the Panel concluded that Bafasal® is safe for all avian species. Considering the nature and manufacturing process of the additive, Bafasal® is not expected to pose a risk for consumers. The results of the subchronic oral toxicity study and genotoxicity studies provided support this conclusion. Exposure of users via inhalation is expected to be low, but Bafasal® should be considered a respiratory sensitiser. No conclusions were drawn on the irritancy of Bafasal® to skin and eyes or on its dermal sensitisation potential due to lack of data. Considering the nature and manufacturing process of the additive, Bafasal® is safe for the environment. The Panel was not in the position to conclude on the efficacy of Bafasal® for any avian species due to insufficient data.


Chemical modification of halloysite nanotubes for controlled loading and release.
Clay minerals have been used for medical purposes from ancient times. Among them, the halloysite nanotube, an aluminosilicate of the kaolin group, is an emerging nanomaterial which possesses peculiar chemical characteristics. By means of suitable modifications, such as supramolecular functionalization or covalent modifications, it is possible to obtain novel nanomaterials with tunable properties for several applications. In this context the covalent grafting of suitable organic moieties on the external surface or in the halloysite lumen has been exploited to improve the loading and release of several biologically active molecules. The resulting hybrid nanomaterials have been applied as drug carrier and delivery systems, as fillers for hydrogels, in tissue regeneration and in the gene delivery field. Furthermore the loading and release of specific molecules have been also investigated for environmental purposes. This review summarizes the main developments in the halloysite modifications in the last 20 years with a particular attention to the development in the past two years.


Neurochemical and histologic characterization of striatal excitotoxic lesions produced by the mitochondrial toxin 3-nitropropionic acid.
An impairment of energy metabolism may underlie slow excitotoxic neuronal death in neurodegenerative diseases. We therefore examined the effects of intrastriatal, subacute systemic, or chronic systemic administration of the mitochondrial toxin 3-nitropropionic acid (3-NP) in rats. Following intrastriatal injection 3-NP produced dose-dependent striatal lesions. Neurochemical and histologic evaluation showed that markers of both spiny projection neurons (GABA, substance P, calbindin) and aspiny interneurons (somatostatin, neuropeptide Y, NADPH-diaphorase) were equally affected. Subacute systemic administration of 3-NP produced age-dependent bilateral striatal lesions with a similar neurochemical profile. However, in contrast to the intrastriatal injections, striatal dopaminergic afferent projections were spared. Both freeze-clamp measurements and chemical shift magnetic resonance spectroscopy showed that 3-NP impairs energy metabolism in the striatum in vivo. Microdialysis showed no increase in extracellular glutamate concentrations after systemic administration of 3-NP. The lesions produced by intrastriatal injection or systemic administration of 3-NP were blocked by prior decortication. However, the NMDA antagonist MK-801 did not block the effects of intrastriatal 3-NP, consistent with a non-NMDA excitotoxic mechanism. In contrast to subacute systemic administration of 3-NP, chronic (1 month) administration produced lesions confined to the striatum in which there was relative sparing of NADPH-diaphorase interneurons, consistent with an NMDA excitotoxic process. Chronic administration showed growth-related proliferative changes in dendrites of spiny neurons similar to changes in Huntington's disease (HD). These results are consistent with in vitro studies showing that mild metabolic compromise can selectively activate NMDA receptors while more severe compromise activates both NMDA and non-NMDA receptors. Chronic administration of 3-NP over 1 month produces selective striatal lesions that replicate many of the characteristic histologic and neurochemical features of HD.


Molecular mechanisms of pulmonary response progression in crystalline silica exposed rats.
An understanding of the mechanisms underlying diseases is critical for their prevention. Excessive exposure to crystalline silica is a risk factor for silicosis, a potentially fatal pulmonary disease. Male Fischer 344 rats were exposed by inhalation to crystalline silica (15 mg/m3, six hours/day, five days) and pulmonary response was determined at 44 weeks following termination of silica exposure. Additionally, global gene expression profiling in lungs and BAL cells and bioinformatic analysis of the gene expression data were done to understand the molecular mechanisms underlying the progression of pulmonary response to silica. A significant increase in lactate dehydrogenase activity and albumin content in BAL fluid (BALF) suggested silica-induced pulmonary toxicity in the rats. A significant increase in the number of alveolar macrophages and infiltrating neutrophils in the lungs and elevation in monocyte chemoattractant protein-1 (MCP-1) in BALF suggested the induction of pulmonary inflammation in the silica exposed rats. Histological changes in the lungs included granuloma formation, type II pneumocyte hyperplasia, thickening of alveolar septa and positive response to Masson's trichrome stain. Microarray analysis of global gene expression detected 94 and 225 significantly differentially expressed genes in the lungs and BAL cells, respectively. Bioinformatic analysis of the gene expression data identified significant enrichment of several disease and biological function categories and canonical pathways related to pulmonary toxicity, especially inflammation. Taken together, these data suggested the involvement of chronic inflammation as a mechanism underlying the progression of pulmonary response to exposure of rats to crystalline silica at 44 weeks following termination of exposure.


Exposure to Cryptococcus neoformans var. gattii--a seroepidemiological study.
An enzyme-linked immunosorbent assay was developed to study prevalence of immunoglobulin G (IgG) antibody to non-capsular Cryptococcus neoformans var. gattii antigen in a population of healthy Papua New Guinea (PNG) controls and patients. Patients with acute C. neoformans var. gattii meningitis had elevated levels of IgG which declined significantly following treatment (P = 0.034). Levels in the sera of convalescent patients were significantly higher than those in PNG controls (P < 0.001), which in turn were significantly higher than in a UK control group (P < 0.001). Clear differences were observed amongst the PNG controls. Adults had significantly higher levels than children (P = 0.002) and men had significantly higher levels than women (P = 0.047). No difference was observed between levels in patient-related and unrelated controls. IgG responses in PNG controls mirror the prevalence of disease in this population. It is postulated that exposure to C. neoformans var. gattii is less common in children and women due to some as yet unidentified behavioural difference and that exposure occurs away from the home environment.


Statistical aspects of quantitative real-time PCR experiment design.
Experiments using quantitative real-time PCR to test hypotheses are limited by technical and biological variability; we seek to minimise sources of confounding variability through optimum use of biological and technical replicates. The quality of an experiment design is commonly assessed by calculating its prospective power. Such calculations rely on knowledge of the expected variances of the measurements of each group of samples and the magnitude of the treatment effect; the estimation of which is often uninformed and unreliable. Here we introduce a method that exploits a small pilot study to estimate the biological and technical variances in order to improve the design of a subsequent large experiment. We measure the variance contributions at several 'levels' of the experiment design and provide a means of using this information to predict both the total variance and the prospective power of the assay. A validation of the method is provided through a variance analysis of representative genes in several bovine tissue-types. We also discuss the effect of normalisation to a reference gene in terms of the measured variance components of the gene of interest. Finally, we describe a software implementation of these methods, powerNest, that gives the user the opportunity to input data from a pilot study and interactively modify the design of the assay. The software automatically calculates expected variances, statistical power, and optimal design of the larger experiment. powerNest enables the researcher to minimise the total confounding variance and maximise prospective power for a specified maximum cost for the large study.


The neural substrate of positive bias in spontaneous emotional processing.
Even in the presence of negative information, healthy human beings display an optimistic tendency when thinking of past success and future chances, giving a positive bias to everyday's cognition. The tendency to actively select positive thoughts suggests the existence of a mechanism to exclude negative content, raising the issue of its dependence on mechanisms like those of effortful control. Using perfusion imaging, we examined how brain activations differed according to whether participants were left to prefer positive thoughts spontaneously, or followed an explicit instruction to the same effect, finding a widespread dissociation of brain perfusion patterns. Under spontaneous processing of emotional material, recruitment of areas associated with effortful attention, such as the dorsolateral prefrontal cortex, was reduced relative to instructed avoidance of negative material (F(1,58) = 26.24, p = 0.047, corrected). Under spontaneous avoidance perfusion increments were observed in several areas that were deactivated by the task, including the perigenual medial prefrontal cortex. Furthermore, individual differences in executive capacity were not associated with positive bias. These findings suggest that spontaneous positive cognitive emotion regulation in health may result from processes that, while actively suppressing emotionally salient information, differ from those associated with effortful and directed control.


A Complete Overhaul of the Electron Energy-Loss Spectroscopy and X-Ray Absorption Spectroscopy Database: eelsdb.eu.
The electron energy-loss spectroscopy (EELS) and X-ray absorption spectroscopy (XAS) database has been completely rewritten, with an improved design, user interface, and a number of new tools. The database is accessible at https://eelsdb.eu/ and can now be used without registration. The submission process has been streamlined to encourage spectrum submissions and the new design gives greater emphasis on contributors' original work by highlighting their papers. With numerous new filters and a powerful search function, it is now simple to explore the database of several hundred EELS and XAS spectra. Interactive plots allow spectra to be overlaid, facilitating online comparison. An application-programming interface has been created, allowing external tools and software to easily access the information held within the database. In addition to the database itself, users can post and manage job adverts and read the latest news and events regarding the EELS and XAS communities. In accordance with the ongoing drive toward open access data increasingly demanded by funding bodies, the database will facilitate open access data sharing of EELS and XAS spectra.


Reducing central line-associated bloodstream infections in the blood and marrow transplantation population: a review of the literature.
Healthcare-associated infections are a significant cause of patient morbidity and mortality. Most healthcare-associated bloodstream infections are related to the presence of central venous catheters and are called central line-associated bloodstream infections (CLABSIs). A review of the literature showed a decrease in CLABSIs in intensive care units (ICUs) since the implementation of guidelines from the Centers for Disease Control and Prevention. This review demonstrated the effectiveness of daily chlorhexidine gluconate (CHG) bathing toward reducing CLABSIs as well as infections from methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococci in this setting. A need exists for implementation of a similar intervention for patients undergoing blood and marrow transplantation, and CHG bathing would be a logical choice because of its proven efficacy in the ICU, its safety, and its cost affordability.


Nigella sativa: reduces the risk of various maladies.
Coinage of terms like nutraceuticals, functional, and pharma foods has diverted the attention of human beings to where they are seeking more natural cures. Though pharmaceutical drugs have been beneficial for human health and have cured various diseases but they also impart some side effects. Numerous plants have been tested for their therapeutic potential; Nigella sativa, commonly known as black cumin, is one of them. It possesses a nutritional dense profile as its fixed oil (lipid fraction), is rich in unsaturated fatty acids while essential oil contains thymoquinone and carvacrol as antioxidants. N. sativa seeds also contain proteins, alkaloids (nigellicines and nigelledine), and saponins (alpha-hederin) in substantial amounts. Recent pharmacological investigations suggested its potential role, especially for the amelioration of oxidative stress through free radical scavenging activity, the induction of apoptosis to cure various cancer lines, the reduction of blood glucose, and the prevention of complications from diabetes. It regulates hematological and serological aspects and can be effective in dyslipidemia and respiratory disorders. Moreover, its immunopotentiating and immunomodulating role brings balance in the immune system. Evidence is available supporting the utilization of Nigella sativa and its bioactive components in a daily diet for health improvement. This review is intended to focus on the composition of Nigella sativa and to elaborate its possible therapeutic roles as a functional food to prevent an array of maladies.


Childhood blindness in a rural population of southern India: prevalence and etiology.
To determine the prevalence and etiology of childhood blindness in a rural population in southern India through a population based study. A cross sectional, house-to-house survey to screen for childhood blindness included 14,423 children < 16 years. Blindness was defined as best corrected visual acuity < 3/60 in the better eye. The first stage of screening for blindness was done by interns under supervision of ophthalmology residents. Senior residents examined the referred cases from the first stage. Those detected to be blind were brought to a tertiary care hospital for detailed examination. Fifty-four children were referred after first stage of screening of 13,241 children. Of these 14 were bilaterally blind giving a prevalence of 1.06/1000 (95% confidence interval (CI), 0.50 to 1.61); 6 (42.9%) had lens and related complications, 4 (28.6%) had globe anomalies (2 Microphthalmos and 2 Anophthalmos), 2 (14.3%) had retinal dystrophy and 1 (7.1%) each of glaucoma and optic atrophy. Among the parents of blind children, 71.4% (p = 0.002) had consanguineous marriage (83.3% in cataract blind children). More than half of the blindness detected was potentially avoidable. Genetic counseling, early identification and access to tertiary care would reduce the burden of childhood blindness in the local community.


Chronic nephropathies of cocaine and heroin abuse: a critical review.
Renal disease in cocaine and heroin users is associated with the nephrotic syndrome, acute glomerulonephritis, amyloidosis, interstitial nephritis, and rhabdomyolysis. The pathophysiologic basis of cocaine-related renal injury involves renal hemodynamic changes, glomerular matrix synthesis and degradation, and oxidative stress and induction of renal atherogenesis. Heroin is the most commonly abused opiate in the United States. Previous studies identified a spectrum of renal diseases in heroin users. The predominant renal lesion in black heroin users is focal segmental glomerulosclerosis and in white heroin users is membranoproliferative glomerulonephritis. Although the prevalence of heroin use in the United States has increased, the incidence of "heroin nephropathy" has declined. Because reports of heroin nephropathy predated the surveillance of hepatitis C virus and HIV, the varied findings might be related to the spectrum of viral illnesses that are encountered in injection drug users. Socioeconomic conditions, cultural and behavioral practices, or differences in genetic susceptibilities may be more associated with the development of nephropathy in heroin users than the drug's pharmacologic properties. Administration of cocaine in animal models results in nonspecific glomerular, interstitial, and tubular cell lesions, but there is no animal model of heroin-associated renal disease. The heterogeneity of responses that are associated with heroin is not consistent with a single or simple notion of nephropathogenesis. There are no well-designed, prospective, epidemiologic studies to assess the incidence and the prevalence of renal disease in populations of opiate users and to establish the validity of a syndrome such as heroin nephropathy. It is concluded although there is a paucity of evidence to support a heroin-associated nephropathy, the evidence from in vitro cellular and animal studies to support the existence of cocaine-induced renal changes is more convincing.


Marked endotheliotropism of highly pathogenic avian influenza virus H5N1 following intestinal inoculation in cats.
Highly pathogenic avian influenza virus (HPAIV) H5N1 can infect mammals via the intestine; this is unusual since influenza viruses typically infect mammals via the respiratory tract. The dissemination of HPAIV H5N1 following intestinal entry and associated pathogenesis are largely unknown. To assess the route of spread of HPAIV H5N1 to other organs and to determine its associated pathogenesis, we inoculated infected chicken liver homogenate directly into the intestine of cats by use of enteric-coated capsules. Intestinal inoculation of HPAIV H5N1 resulted in fatal systemic disease. The spread of HPAIV H5N1 from the lumen of the intestine to other organs took place via the blood and lymphatic vascular systems but not via neuronal transmission. Remarkably, the systemic spread of the virus via the vascular system was associated with massive infection of endothelial and lymphendothelial cells, resulting in widespread hemorrhages. This is unique for influenza in mammals and resembles the pathogenesis of HPAIV infection in terrestrial poultry. It contrasts with the pathogenesis of systemic disease from the same virus following entry via the respiratory tract, where lesions are characterized mainly by necrosis and inflammation and are associated with the presence of influenza virus antigen in parenchymal, not endothelial cells. The marked endotheliotropism of the virus following intestinal inoculation indicates that the pathogenesis of systemic influenza virus infection in mammals may differ according to the portal of entry.


Determination and correction of position detection nonlinearity in single particle tracking and three-dimensional scanning probe microscopy.
A general method is presented for determining and correcting nonlinear position detector responses in single particle tracking as used in three-dimensional scanning probe microscopy based on optical tweezers. The method uses locally calculated mean square displacements of a Brownian particle to detect spatial changes in the sensitivity of the detector. The method is applied to an optical tweezers setup, where the position fluctuations of a microsphere within the optical trap are measured by an interferometric detection scheme with nanometer precision and microsecond temporal resolution. Detector sensitivity profiles were measured at arbitrary positions in solution with a resolution of approximately 6 nm and 20 nm in the lateral and axial directions, respectively. Local detector sensitivities are used to reconstruct the real positions of the particle from the measured position signals.


Our journey, our story: a study protocol for the evaluation of a co-design framework to improve services for Aboriginal youth mental health and well-being.
Mainstream Australian mental health services are failing Aboriginal young people. Despite investing resources, improvements in well-being have not materialised. Culturally and age appropriate ways of working are needed to improve service access and responsiveness. This Aboriginal-led study brings Aboriginal Elders, young people and youth mental health service staff together to build relationships to co-design service models and evaluation tools. Currently, three Western Australian youth mental health services in the Perth metropolitan area and two regional services are working with local Elders and young people to improve their capacity for culturally and age appropriate services. Further Western Australian sites will be engaged as part of research translation. Relationships ground the study, which utilises Indigenous methodologies and participatory action research. This involves Elders, young people and service staff as co-researchers and the application of a decolonising, strengths-based framework to create the conditions for engagement. It foregrounds experiential learning and Aboriginal ways of working to establish relationships and deepen non-Aboriginal co-researchers' knowledge and understanding of local, place-based cultural practices. Once relationships are developed, co-design workshops occur at each site directed by local Elders and young people. Co-designed evaluation tools will assess any changes to community perceptions of youth mental health services and the enablers and barriers to service engagement. The study has approval from the Kimberley Aboriginal Health Planning Forum Kimberley Research Subcommittee, the Western Australian Aboriginal Health Ethics Committee, and the Curtin University Human Research Ethics Committee. Transferability of the outcomes across the youth mental health sector will be directed by the co-researchers and is supported through Aboriginal and non-Aboriginal organisations including youth mental health services, peak mental health bodies and consumer groups. Community reports and events, peer-reviewed journal articles, conference presentations and social and mainstream media will aid dissemination.


Rethinking Giftedness and Gifted Education: A Proposed Direction Forward Based on Psychological Science.
For nearly a century, scholars have sought to understand, measure, and explain giftedness. Succeeding theories and empirical investigations have often built on earlier work, complementing or sometimes clashing over conceptions of talent or contesting the mechanisms of talent development. Some have even suggested that giftedness itself is a misnomer, mistaken for the results of endless practice or social advantage. In surveying the landscape of current knowledge about giftedness and gifted education, this monograph will advance a set of interrelated arguments: The abilities of individuals do matter, particularly their abilities in specific talent domains; different talent domains have different developmental trajectories that vary as to when they start, peak, and end; and opportunities provided by society are crucial at every point in the talent-development process. We argue that society must strive to promote these opportunities but that individuals with talent also have some responsibility for their own growth and development. Furthermore, the research knowledge base indicates that psychosocial variables are determining influences in the successful development of talent. Finally, outstanding achievement or eminence ought to be the chief goal of gifted education. We assert that aspiring to fulfill one's talents and abilities in the form of transcendent creative contributions will lead to high levels of personal satisfaction and self-actualization as well as produce yet unimaginable scientific, aesthetic, and practical benefits to society. To frame our discussion, we propose a definition of giftedness that we intend to be comprehensive. Giftedness is the manifestation of performance that is clearly at the upper end of the distribution in a talent domain even relative to other high-functioning individuals in that domain. Further, giftedness can be viewed as developmental in that in the beginning stages, potential is the key variable; in later stages, achievement is the measure of giftedness; and in fully developed talents, eminence is the basis on which this label is granted. Psychosocial variables play an essential role in the manifestation of giftedness at every developmental stage. Both cognitive and psychosocial variables are malleable and need to be deliberately cultivated. Our goal here is to provide a definition that is useful across all domains of endeavor and acknowledges several perspectives about giftedness on which there is a fairly broad scientific consensus. Giftedness (a) reflects the values of society; (b) is typically manifested in actual outcomes, especially in adulthood; (c) is specific to domains of endeavor; (d) is the result of the coalescing of biological, pedagogical, psychological, and psychosocial factors; and (e) is relative not just to the ordinary (e.g., a child with exceptional art ability compared to peers) but to the extraordinary (e.g., an artist who revolutionizes a field of art). In this monograph, our goal is to review and summarize what we have learned about giftedness from the literature in psychological science and suggest some directions for the field of gifted education. We begin with a discussion of how giftedness is defined (see above). In the second section, we review the reasons why giftedness is often excluded from major conversations on educational policy, and then offer rebuttals to these arguments. In spite of concerns for the future of innovation in the United States, the education research and policy communities have been generally resistant to addressing academic giftedness in research, policy, and practice. The resistance is derived from the assumption that academically gifted children will be successful no matter what educational environment they are placed in, and because their families are believed to be more highly educated and hold above-average access to human capital wealth. These arguments run counter to psychological science indicating the need for all students to be challenged in their schoolwork and that effort and appropriate educational programing, training and support are required to develop a student's talents and abilities. In fact, high-ability students in the United States are not faring well on international comparisons. The scores of advanced students in the United States with at least one college-educated parent were lower than the scores of students in 16 other developed countries regardless of parental education level. In the third section, we summarize areas of consensus and controversy in gifted education, using the extant psychological literature to evaluate these positions. Psychological science points to several variables associated with outstanding achievement. The most important of these include general and domain-specific ability, creativity, motivation and mindset, task commitment, passion, interest, opportunity, and chance. Consensus has not been achieved in the field however in four main areas: What are the most important factors that contribute to the acuities or propensities that can serve as signs of potential talent? What are potential barriers to acquiring the "gifted" label? What are the expected outcomes of gifted education? And how should gifted students be educated? In the fourth section, we provide an overview of the major models of giftedness from the giftedness literature. Four models have served as the foundation for programs used in schools in the United States and in other countries. Most of the research associated with these models focuses on the precollegiate and early university years. Other talent-development models described are designed to explain the evolution of talent over time, going beyond the school years into adult eminence (but these have been applied only by out-of-school programs as the basis for educating gifted students). In the fifth section we present methodological challenges to conducting research on gifted populations, including definitions of giftedness and talent that are not standardized, test ceilings that are too low to measure progress or growth, comparison groups that are hard to find for extraordinary individuals, and insufficient training in the use of statistical methods that can address some of these challenges. In the sixth section, we propose a comprehensive model of trajectories of gifted performance from novice to eminence using examples from several domains. This model takes into account when a domain can first be expressed meaningfully-whether in childhood, adolescence, or adulthood. It also takes into account what we currently know about the acuities or propensities that can serve as signs of potential talent. Budding talents are usually recognized, developed, and supported by parents, teachers, and mentors. Those individuals may or may not offer guidance for the talented individual in the psychological strengths and social skills needed to move from one stage of development to the next. We developed the model with the following principles in mind: Abilities matter, domains of talent have varying developmental trajectories, opportunities need to be provided to young people and taken by them as well, psychosocial variables are determining factors in the successful development of talent, and eminence is the aspired outcome of gifted education. In the seventh section, we outline a research agenda for the field. This agenda, presented in the form of research questions, focuses on two central variables associated with the development of talent-opportunity and motivation-and is organized according to the degree to which access to talent development is high or low and whether an individual is highly motivated or not. Finally, in the eighth section, we summarize implications for the field in undertaking our proposed perspectives. These include a shift toward identification of talent within domains, the creation of identification processes based on the developmental trajectories of talent domains, the provision of opportunities along with monitoring for response and commitment on the part of participants, provision of coaching in psychosocial skills, and organization of programs around the tools needed to reach the highest possible levels of creative performance or productivity.


GenGIS: A geospatial information system for genomic data.
The increasing availability of genetic sequence data associated with explicit geographic and ecological information is offering new opportunities to study the processes that shape biodiversity. The generation and testing of hypotheses using these data sets requires effective tools for mathematical and visual analysis that can integrate digital maps, ecological data, and large genetic, genomic, or metagenomic data sets. GenGIS is a free and open-source software package that supports the integration of digital map data with genetic sequences and environmental information from multiple sample sites. Essential bioinformatic and statistical tools are integrated into the software, allowing the user a wide range of analysis options for their sequence data. Data visualizations are combined with the cartographic display to yield a clear view of the relationship between geography and genomic diversity, with a particular focus on the hierarchical clustering of sites based on their similarity or phylogenetic proximity. Here we outline the features of GenGIS and demonstrate its application to georeferenced microbial metagenomic, HIV-1, and human mitochondrial DNA data sets.


Inferring infection hazard in wildlife populations by linking data across individual and population scales.
Our ability to infer unobservable disease-dynamic processes such as force of infection (infection hazard for susceptible hosts) has transformed our understanding of disease transmission mechanisms and capacity to predict disease dynamics. Conventional methods for inferring FOI estimate a time-averaged value and are based on population-level processes. Because many pathogens exhibit epidemic cycling and FOI is the result of processes acting across the scales of individuals and populations, a flexible framework that extends to epidemic dynamics and links within-host processes to FOI is needed. Specifically, within-host antibody kinetics in wildlife hosts can be short-lived and produce patterns that are repeatable across individuals, suggesting individual-level antibody concentrations could be used to infer time since infection and hence FOI. Using simulations and case studies (influenza A in lesser snow geese and Yersinia pestis in coyotes), we argue that with careful experimental and surveillance design, the population-level FOI signal can be recovered from individual-level antibody kinetics, despite substantial individual-level variation. In addition to improving inference, the cross-scale quantitative antibody approach we describe can reveal insights into drivers of individual-based variation in disease response, and the role of poorly understood processes such as secondary infections, in population-level dynamics of disease.


Halogenated pollutants in terrestrial and aquatic bird eggs: converging patterns of pollutant profiles, and impacts and risks from high levels.
We investigated the presence, levels, relationships, and risks of HCHs, DDTs, chlordanes, mirex, PCBs, and brominated flame retardants (BFRs) in terrestrial and aquatic bird eggs from an area in South Africa where DDT is used for malaria control. We found one of the highest ΣDDT levels reported this century; 13,000 ng/g wm (wet mass) in Grey Heron eggs which exceeds critical levels for reproductive success (3000 ng/g wm) calculated for Brown Pelicans, with a no-effect level estimated at 500 ng/g wm. Even higher ΣDDT levels at 16,000 ng/g wm were found in House Sparrow eggs (possibly the highest ever recorded for sparrows), with a maximum of 24,400 ng/g wm. Significant eggshell thinning in Cattle Egrets (33% between thickest and thinnest) was associated with increased levels of p,p'-DDT and p,p'-DDE. There were indications of unknown use of DDT and lindane. Relative to DDT, PCBs and BFRs levels were quite low. Ordinated data showed that different terrestrial pollutant profiles converged to a homogenised aquatic profile. Converging profiles, high levels of DDT in heron and sparrow eggs, and thinning eggs shells, indicate risk and impacts at release, in the aquatic environment, and in between. If characteristic life-strategies of birds in warm areas (e.g. longer-lived and fewer eggs per clutch) increases the risk compared with similar birds living in colder regions when both experience the same environmental pollutant levels, then malaria control using DDT probably has more significant impacts on biota than previously realised. Therefore, risk assessment and modelling without hard data may miss crucial impacts and risks, as the chemical use patterns and ecologies in Africa and elsewhere may differ from the conditions and assumptions of existing risk assessment and modelling parameters. Consideration of other findings associated with DDT from the same area (intersex in fish and urogental birth defects in baby boys), together with the findings of this study (high levels of DDT in bird eggs, eggshell thinning in the Cattle Egrets, and the apparent absence of breeding piscivore birds in the sprayed area) are strongly suggestive of negative impacts from DDT spraying for Malaria control. Our data presents strong arguments for an expedited process of replacing DDT with sustainable methods.


Microbial diversity in a hydrocarbon- and chlorinated-solvent-contaminated aquifer undergoing intrinsic bioremediation.
A culture-independent molecular phylogenetic approach was used to survey constituents of microbial communities associated with an aquifer contaminated with hydrocarbons (mainly jet fuel) and chlorinated solvents undergoing intrinsic bioremediation. Samples were obtained from three redox zones: methanogenic, methanogenic-sulfate reducing, and iron or sulfate reducing. Small-subunit rRNA genes were amplified directly from aquifer material DNA by PCR with universally conserved or Bacteria- or Archaea-specific primers and were cloned. A total of 812 clones were screened by restriction fragment length polymorphisms (RFLP), approximately 50% of which were unique. All RFLP types that occurred more than once in the libraries, as well as many of the unique types, were sequenced. A total of 104 (94 bacterial and 10 archaeal) sequence types were determined. Of the 94 bacterial sequence types, 10 have no phylogenetic association with known taxonomic divisions and are phylogenetically grouped in six novel division level groups (candidate divisions WS1 to WS6); 21 belong to four recently described candidate divisions with no cultivated representatives (OP5, OP8, OP10, and OP11); and 63 are phylogenetically associated with 10 well-recognized divisions. The physiology of two particularly abundant sequence types obtained from the methanogenic zone could be inferred from their phylogenetic association with groups of microorganisms with a consistent phenotype. One of these sequence types is associated with the genus Syntrophus; Syntrophus spp. produce energy from the anaerobic oxidation of organic acids, with the production of acetate and hydrogen. The organism represented by the other sequence type is closely related to Methanosaeta spp., which are known to be capable of energy generation only through aceticlastic methanogenesis. We hypothesize, therefore, that the terminal step of hydrocarbon degradation in the methanogenic zone of the aquifer is aceticlastic methanogenesis and that the microorganisms represented by these two sequence types occur in syntrophic association.


Differential regulation of nuclear and mitochondrial Bcl-2 in T cell apoptosis.
Activated T cells require anti-apoptotic cytokines for their survival. The anti-apoptotic effects of these factors are mediated by their influence on the balance of expression and localisation of pro- and anti-apoptotic members of the Bcl-2 family. Among the anti-apoptotic Bcl-2 family members, the expression level of Bcl-2 itself and its interaction with the pro-apoptotic protein Bim are now regarded as crucial for the regulation of survival in activated T cells. We studied the changes in Bcl-2 levels and its subcellular distribution in relation to mitochondrial depolarisation and caspase activation in survival factor deprived T cells. Intriguingly, the total Bcl-2 level appeared to remain stable, even after caspase 3 activation indicated entry into the execution phase of apoptosis. However, cell fractionation experiments showed that while the dominant nuclear pool of Bcl-2 remained stable during apoptosis, the level of the smaller mitochondrial pool was rapidly downregulated. Signals induced by anti-apoptotic cytokines continuously replenish the mitochondrial pool, but nuclear Bcl-2 is independent of such signals. Mitochondrial Bcl-2 is lost rapidly by a caspase independent mechanism in the absence of survival factors, in contrast only a small proportion of the nuclear pool of Bcl-2 is lost during the execution phase and this loss is a caspase dependent process. We conclude that these two intracellular pools of Bcl-2 are regulated through different mechanisms and only the cytokine-mediated regulation of the mitochondrial pool is relevant to the control of the initiation of apoptosis.


Common themes in microbial pathogenicity revisited.
Bacterial pathogens employ a number of genetic strategies to cause infection and, occasionally, disease in their hosts. Many of these virulence factors and their regulatory elements can be divided into a smaller number of groups based on the conservation of similar mechanisms. These common themes are found throughout bacterial virulence factors. For example, there are only a few general types of toxins, despite a large number of host targets. Similarly, there are only a few conserved ways to build the bacterial pilus and nonpilus adhesins used by pathogens to adhere to host substrates. Bacterial entry into host cells (invasion) is a complex mechanism. However, several common invasion themes exist in diverse microorganisms. Similarly, once inside a host cell, pathogens have a limited number of ways to ensure their survival, whether remaining within a host vacuole or by escaping into the cytoplasm. Avoidance of the host immune defenses is key to the success of a pathogen. Several common themes again are employed, including antigenic variation, camouflage by binding host molecules, and enzymatic degradation of host immune components. Most virulence factors are found on the bacterial surface or secreted into their immediate environment, yet virulence factors operate through a relatively small number of microbial secretion systems. The expression of bacterial pathogenicity is dependent upon complex regulatory circuits. However, pathogens use only a small number of biochemical families to express distinct functional factors at the appropriate time that causes infection. Finally, virulence factors maintained on mobile genetic elements and pathogenicity islands ensure that new strains of pathogens evolve constantly. Comprehension of these common themes in microbial pathogenicity is critical to the understanding and study of bacterial virulence mechanisms and to the development of new "anti-virulence" agents, which are so desperately needed to replace antibiotics.


Sarcoplasmic Reticulum Structure and Functional Properties that Promote Long-Lasting Calcium Sparks.
Calcium (Ca) sparks are the fundamental sarcoplasmic reticulum (SR) Ca release events in cardiac myocytes, and they have a typical duration of 20-40 ms. However, when a fraction of ryanodine receptors (RyRs) are blocked by tetracaine or ruthenium red, Ca sparks lasting hundreds of milliseconds have been observed experimentally. The fundamental mechanism underlying these extremely prolonged Ca sparks is not understood. In this study, we use a physiologically detailed mathematical model of subcellular Ca cycling to examine how Ca spark duration is influenced by the number of functional RyRs in a junctional cluster (which is reduced by tetracaine or ruthenium red) and other SR Ca handling properties. One RyR cluster contains a few to several hundred RyRs, and we use a four-state Markov RyR gating model. Each RyR opens stochastically and is regulated by cytosolic and luminal Ca. We varied the number of functional RyRs in the single cluster, diffusion within the SR network, diffusion between network and junctional SR, cytosolic Ca diffusion, SERCA uptake activity, and RyR open probability. For long-lasting Ca release events, opening events within the cluster must occur continuously because the typical open time of the RyR is only a few milliseconds. We found the following: 1) if the number of RyRs is too small, it is difficult to maintain consecutive openings and stochastic attrition terminates the release; 2) if the number of RyRs is too large, the depletion of Ca from the junctional SR terminates the release; and 3) very long release events require relatively small-sized RyR clusters (reducing flux as seen experimentally with tetracaine) and sufficiently rapid intra-SR Ca diffusion, such that local junctional intra-SR [Ca] can be maintained by intra-SR diffusion and overall SR Ca reuptake.


Factors influencing treatment selection and survival in advanced lung cancer.
Despite numerous breakthrough therapies, inoperable lung cancer still places a heavy burden on patients who might not be candidates for chemotherapy. To identify potential candidates for the newly emerging immunotherapy-based treatment paradigms, we explored the clinical and biologic factors affecting treatment decisions. We retrospectively reviewed the records of patients diagnosed at our university-affiliated cancer centre between 1 January 2011 and 31 December 2013. Patient demographics, systemic treatment, and survival were examined. During the 3-year study period, 683 patients fitting the inclusion criteria were identified. First-line therapy was administered in 49.5% of patients; only 22.4% received further lines of therapy. The main reasons for withholding therapy were poor performance status [ps (43.2%)], rapidly deteriorating ps (31.9%), patient refusal of therapy (20.9%), and associated comorbidities (4%). Older age, the presence of brain metastasis at diagnosis, and non-small-cell histology were also associated with therapeutic restraint. Oncology referrals were infrequent in patients who did not receive therapy (32.2%). Older patients and those with a poor ps experienced superior survival when treatment was administered (hazard ratio: 0.25; 95% confidence interval: 0.16 to 0.38; and hazard ratio: 0.44; 95% confidence interval: 0.23 to 0.87 respectively; p < 0.001). Advanced lung cancer still poses a therapeutic challenge, with a high proportion of patients being deemed unfit for therapy. This issue cannot be resolved until appropriate measures are taken to ensure the inclusion of older patients and those with a relatively poor ps in large clinical trials. Immunotherapy might be interesting in this setting, given that it appears to be more tolerable. Another consequential undertaking would be the deployment of strategies to reduce wait times during the diagnostic process for patients with a high index of suspicion for lung cancer.


Collagen I gel promotes homogenous osteogenic differentiation of adipose tissue-derived mesenchymal stem cells in serum-derived albumin scaffold.
Repair of bone defects is a difficult clinical problem for reconstructive surgeons. Bone tissue engineering using an appropriate scaffold with cells is a new therapy for the repair of bone defects. The aim of this study was to evaluate the in vitro osteogenesis of canine adipose tissue-derived mesenchymal stem cells (Ad-MSCs) cultured in a combination of collagen I gel and a porous serum-derived albumin scaffold. A serum-derived albumin scaffold was prepared with canine serum by cross-linking and freeze-drying procedures. Ad-MSCs were seeded into serum-derived albumin scaffolds with or without collagen I gel, and were exposed to osteogenic differentiation conditions in vitro. After 28 days of in vitro culture, the distribution and osteogenic differentiation of Ad-MSCs cultured in the scaffold were evaluated by scanning electron microscopy, histology, immunohistochemistry, alkaline phosphatase (ALP) activity assay, and calcium colorimetric assay. Ad-MSCs showed more homogeneous distribution and osteogenic differentiation in the scaffold with collagen I gel than without collagen I gel. ALP activity and extracellular matrix mineralization in the construct with type I collagen were significantly higher than in the construct without type I collagen (p < 0.05). In conclusion, the combination of collagen I gel and the serum-derived albumin scaffold enhanced osteogenic differentiation and homogenous distribution of Ad-MSCs.


Chromosome microdissection: a brief overview.
Chromosome microdissection arose as a means of facilitating long range physical mapping of chromosome regions involved in either a genetic or malignant disorder. However, with the rapid development of improved techniques for mapping and sequencing the human genome, microdissection is considered by many investigators to be a cumbersome and time consuming procedure. Nonetheless, based on the impressive number of informative diagnostic DNA markers that are now available as a result of this technology, microdissection still must be considered one of the most rapid and direct methods available for generating new DNA markers from any chromosome region, irrespective of its sequence composition. In addition, it remains an important means to dissect DNA markers from any organism, eukaryotic and prokaryotic, and has resulted in generating disease associated DNA sequences from both human and animal genomes. Recently, microdissection of single cells has emerged as a viable alternative for isolating pure populations of specific cell types, especially tumor cells, which can then be studied without background contamination from any other cellular constituents. This overview will provide a glimpse into the present applications of the microdissection technology, as well as the importance this technology will have for future exploration into the human genome.


Sarcoidosis.
Sarcoidosis is a multisystemic disorder of unknown cause characterized by the formation of immune granulomas in involved organs. It is an ubiquitous disease with incidence (varying according to age, sex, race and geographic origin) estimated at around 16.5/100,000 in men and 19/100,000 in women. The lung and the lymphatic system are predominantly affected but virtually every organ may be involved. Other severe manifestations result from cardiac, neurological, ocular, kidney or laryngeal localizations. In most cases, sarcoidosis is revealed by persistent dry cough, eye or skin manifestations, peripheral lymph nodes, fatigue, weight loss, fever or night sweats, and erythema nodosum. Abnormal metabolism of vitamin D3 within granulomatous lesions and hypercalcemia are possible. Chest radiography is abnormal in about 90% of cases and shows lymphadenopathy and/or pulmonary infiltrates (without or with fibrosis), defining sarcoidosis stages from I to IV. The etiology remains unknown but the prevailing hypothesis is that various unidentified, likely poorly degradable antigens of either infectious or environmental origin could trigger an exaggerated immune reaction in genetically susceptible hosts. Diagnosis relies on compatible clinical and radiographic manifestations, evidence of non-caseating granulomas obtained by biopsy through tracheobronchial endoscopy or at other sites, and exclusion of all other granulomatous diseases. The evolution and severity of sarcoidosis are highly variable. Mortality is estimated at between 0.5-5%. In most benign cases (spontaneous resolution within 24-36 months), no treatment is required but a regular follow-up until recovery is necessary. In more serious cases, a medical treatment has to be prescribed either initially or at some point during follow-up according to clinical manifestations and their evolution. Systemic corticosteroids are the mainstay of treatment of sarcoidosis. The minimal duration of treatment is 12 months. Some patients experience repeated relapses and may require long-term low-dose corticosteroid therapy during years. Other treatments (immunosuppressive drugs and aminoquinolins) may be useful in case of unsatisfactory response to corticosteroids, poor tolerance and as sparing agents when high doses of corticosteroids are needed for a long time. In some strictly selected cases refractory to standard therapy, specific antiTNF-alpha agents may offer precious improvement. Some patients benefit from topical corticosteroids.


Intraspecific body size frequency distributions of insects.
Although interspecific body size frequency distributions are well documented for many taxa, including the insects, intraspecific body size frequency distributions (IaBSFDs) are more poorly known, and their variation among mass-based and linear estimates of size has not been widely explored. Here we provide IaBSFDs for 16 species of insects based on both mass and linear estimates and large sample sizes (n ≥ 100). In addition, we review the published IaBSFDs for insects, though doing so is complicated by their under-emphasis in the literature. The form of IaBSFDs can differ substantially between mass-based and linear measures. Nonetheless, in non-social insects they tend to be normally distributed (18 of 27 species) or in fewer instances positively skewed. Negatively skewed distributions are infrequently reported and log transformation readily removes the positive skew. Sexual size dimorphism does not generally cause bimodality in IaBSFDs. The available information on IaBSFDs in the social insects suggests that these distributions are usually positively skewed or bimodal (24 of 30 species). However, only c. 15% of ant genera are polymorphic, suggesting that normal distributions are probably more common, but less frequently investigated. Although only 57 species, representing seven of the 29 orders of insects, have been considered here, it appears that whilst IaBSFDs are usually normal, other distribution shapes can be found in several species, though most notably among the social insects. By contrast, the interspecific body size frequency distribution is typically right-skewed in insects and in most other taxa.


Glucokinase regulatory protein: complexity at the crossroads of triglyceride and glucose metabolism.
Glucokinase regulator (GCKR) encodes glucokinase regulatory protein (GKRP), a hepatocyte-specific inhibitor of the glucose-metabolizing enzyme glucokinase (GCK). Genome-wide association studies have identified a common coding variant within GCKR associated with multiple metabolic traits. This review focuses on recent insights into the critical role of GKRP in hepatic glucose metabolism that have stemmed from the study of human genetics. This knowledge has improved our understanding of glucose and lipid physiology and informed the development of targeted molecular therapeutics for diabetes. Rare GCKR variants have effects on GKRP expression, localization, and activity. These variants are collectively associated with hypertriglyceridaemia but are not causal. Crystal structures of GKRP and the GCK-GKRP complex have been solved, providing greater insight into the molecular interactions between these proteins. Finally, small molecules have been identified that directly bind GKRP and reduce blood glucose levels in rodent models of diabetes. GCKR variants across the allelic spectrum have effects on glucose and lipid homeostasis. Functional analysis has highlighted numerous molecular mechanisms for GKRP dysfunction. Hepatocyte-specific GCK activation via small molecule GKRP inhibition may be a new avenue for type 2 diabetes treatment, particularly considering evidence indicating GKRP loss-of-function alone does not cause hypertriglyceridaemia.


Modulation by cyclic AMP of beta adrenergic receptor-stimulated prostacyclin synthesis in rabbit ventricular myocytes.
The purpose of the present study was to determine the possible interaction of cyclic AMP (cAMP) and the synthesis of prostacyclin [measured as immunoreactive 6-keto-prostaglandin (PG)F1 alpha] elicited by the beta adrenergic receptor agonist isoproterenol (ISOP), in freshly dissociated rabbit ventricular myocytes. ISOP (10(-13) to 10(-11) M) increased 6-keto-PGF1 alpha synthesis without altering the level of cAMP. Increasing the concentration of ISOP from 10(-10) to 10(-7) M enhanced accumulation of cAMP, which was associated with a decline in 6-keto-PGF1 alpha synthesis. Forskolin (10(-6) M), an activator of adenylyl cyclase, and 3-isobutyl-1-methylxanthine (10(-5) M), an inhibitor of cAMP phosphodiesterase, increased cAMP accumulation and inhibited ISOP-induced 6-keto-PGF1 alpha synthesis. 8-(4-chlorophenylthio) (cpt)-cAMP (10(-7) M) also inhibited ISOP-induced 6-keto-PGF1 alpha production. On the other hand, miconazole (10(-4) M), an inhibitor of adenylyl cyclase, reduced cAMP accumulation and enhanced ISOP-induced 6-keto-PGF1 alpha synthesis in myocytes. Miconazole also attenuated ISOP-, forskolin- and cpt-cAMP-induced increases in protein kinase A activity. The protein kinase A inhibitor H-89 {N-[2-(p-bromocinnamylamino)ethyl] -5-isoquinolinesulfonamide} attenuated the ISOP (10(-7) M)-induced increase in the activity of this enzyme and minimized the decline in 6-keto-PGF1 alpha synthesis produced by 10(-7) M ISOP and the inhibitory effect of cpt-cAMP and forskolin on 6-keto-PGF1 alpha production. 3-Isobutyl-1-methylxanthine, forskolin and cpt-cAMP did not alter the conversion of exogenous arachidonic acid to 6-keto-PGF1 alpha. These data indicate that beta adrenergic receptor activation promotes prostacyclin synthesis in rabbit ventricular myocytes and that cAMP acts as an inhibitory modulator. This action is mediated via activation of protein kinase A, probably by decreasing the activity of the lipase, involved in beta adrenergic receptor-induced arachidonic acid release.


Telomerase mRNA-transfected dendritic cells stimulate antigen-specific CD8+ and CD4+ T cell responses in patients with metastatic prostate cancer.
Telomerase reverse transcriptase (hTERT) represents an attractive target for cancer immunotherapy because hTERT is reactivated in most human tumors. A clinical trial was initiated in which hTERT mRNA-transfected dendritic cells (DC) were administered to 20 patients with metastatic prostate cancer. Nine of these subjects received DC transfected with mRNA encoding a chimeric lysosome-associated membrane protein-1 (LAMP) hTERT protein, allowing for concomitant induction of hTERT-specific CD8+ and CD4+ T cell responses. Treatment was well tolerated. Intense infiltrates of hTERT-specific T cells were noted at intradermal injection sites after repeated vaccination. In 19 of 20 subjects, expansion of hTERT-specific CD8+ T cells was measured in the peripheral blood of study subjects, with 0.9-1.8% of CD8+ T cells exhibiting Ag specificity. Patients immunized with the chimeric LAMP hTERT vaccine developed significantly higher frequencies of hTERT-specific CD4+ T cells than subjects receiving DC transfected with the unmodified hTERT template. Moreover, CTL-mediated killing of hTERT targets was enhanced in the LAMP hTERT group, suggesting that an improved CD4+ response could augment a CTL response. Vaccination was further associated with a reduction of prostate-specific Ag velocity and molecular clearance of circulating micrometastases. Our findings provide a rationale for further development of hTERT-transfected DC vaccines in the treatment of prostate and other cancers.


Amino acid interactions that facilitate enzyme catalysis.
Interactions in enzymes between catalytic and neighboring amino acids and how these interactions facilitate catalysis are examined. In examples from both natural and designed enzymes, it is shown that increases in catalytic rates may be achieved through elongation of the buffer range of the catalytic residues; such perturbations in the protonation equilibria are, in turn, achieved through enhanced coupling of the protonation equilibria of the active ionizable residues with those of other ionizable residues. The strongest coupling between protonation states for a pair of residues that deprotonate to form an anion (or a pair that accept a proton to form a cation) is achieved when the difference in the intrinsic pKas of the two residues is approximately within 1 pH unit. Thus, catalytic aspartates and glutamates are often coupled to nearby acidic residues. For an anion-forming residue coupled to a cation-forming residue, the elongated buffer range is achieved when the intrinsic pKa of the anion-forming residue is higher than the intrinsic pKa of the (conjugate acid of the) cation-forming residue. Therefore, the high pKa, anion-forming residues tyrosine and cysteine make good coupling partners for catalytic lysine residues. For the anion-cation pairs, the optimum difference in intrinsic pKas is a function of the energy of interaction between the residues. For the energy of interaction ε expressed in units of (ln 10)RT, the optimum difference in intrinsic pKas is within ∼1 pH unit of ε.


Human infection with an unknown species of Dracunculus in Vietnam.
Guinea worm (GW) disease, caused by Dracunculus medinensis, is an almost eradicated waterborne zoonotic disease. The World Health Organization (WHO) currently lists GW as endemic in only five African countries. In July 2020, the Vietnamese public health surveillance system detected a hanging worm in a 23-year-old male patient, who did not report any travel to Africa or any country previously endemic for GW. The patient was hospitalized with symptoms of fatigue, anorexia, muscle aches, and abscesses, with worms hanging out of the skin in the lower limbs. The worms were retrieved from the lesions and microscopically examined in Vietnam, identifying structures compatible with Dracunculus spp. and L1-type larvae. A section of this parasite was sent to the Centers for Disease Control and Prevention (CDC) in Atlanta, United States, for confirmatory diagnosis of GW. The adult worm had cuticle structures compatible with Dracunculus parasites, although the length of L1 larvae was about 339 μm, substantially shorter than D. medinensis. DNA sequence analysis of the 18S small subunit rRNA gene confirmed that this parasite was not GW, and determined that the sample belonged to a Dracunculus sp. not previously reported in GenBank that clustered with the animal-infective Dracunculus insignis and Dracunculus lutrae, located in a different clade than D. medinensis. This study highlights the importance of effective public health surveillance systems and the collaborative work of local public health authorities from Vietnam with the WHO and CDC in efforts to achieve the eradication of GW.


Possible implication of satellite cells in regenerative motoneuritogenesis: HGF upregulates neural chemorepellent Sema3A during myogenic differentiation.
Regenerative coordination and remodeling of the intramuscular motoneuron network and neuromuscular connections are critical for restoring skeletal muscle function and physiological properties. The regulatory mechanisms of such coordination remain unclear, although both attractive and repulsive axon guidance molecules may be involved in the signaling pathway. Here we show that expression of a neural secreted chemorepellent semaphorin 3A (Sema3A) is remarkably upregulated in satellite cells of resident myogenic stem cells that are positioned beneath the basal lamina of mature muscle fibers, when treated with hepatocyte growth factor (HGF), established as an essential cue in muscle fiber growth and regeneration. When satellite cells were treated with HGF in primary cultures of cells or muscle fibers, Sema3A message and protein were upregulated as revealed by reverse transcription-polymerase chain reaction and immunochemical studies. Other growth factors had no inductive effect except for a slight effect of epidermal growth factor treatment. Sema3A upregulation was HGF dose dependent with a maximum (about 7- to 8-fold units relative to the control) at 10-25 ng/ml and occurred exclusively at the early-differentiation stage, as characterized by the level of myogenin expression and proliferation (bromodeoxyuridine incorporation) of the cells. Neutralizing antibody to the HGF-specific receptor, c-met, did not abolish the HGF response, indicating that c-met may not mediate the Sema3A expression signaling. Finally, in vivo Sema3A was upregulated in the differentiation phase of satellite cells isolated from muscle regenerating following crush injury. Overall, the data highlight a heretofore unexplored and active role for satellite cells as a key source of Sema3A expression triggered by HGF, hence suggesting that regenerative activity toward motor innervation may importantly reside in satellite cells and could be a crucial contributor during postnatal myogenesis.


Local GABA concentration is related to network-level resting functional connectivity.
Anatomically plausible networks of functionally inter-connected regions have been reliably demonstrated at rest, although the neurochemical basis of these 'resting state networks' is not well understood. In this study, we combined magnetic resonance spectroscopy (MRS) and resting state fMRI and demonstrated an inverse relationship between levels of the inhibitory neurotransmitter GABA within the primary motor cortex (M1) and the strength of functional connectivity across the resting motor network. This relationship was both neurochemically and anatomically specific. We then went on to show that anodal transcranial direct current stimulation (tDCS), an intervention previously shown to decrease GABA levels within M1, increased resting motor network connectivity. We therefore suggest that network-level functional connectivity within the motor system is related to the degree of inhibition in M1, a major node within the motor network, a finding in line with converging evidence from both simulation and empirical studies. DOI: http://dx.doi.org/10.7554/eLife.01465.001.


Preparation, characterization and pharmacokinetic evaluation of rosuvastatin calcium incorporated cyclodextrin-polyanhydride nanoparticles.
Objective: The aim of the study was to formulate, cyclodextrin (CD)-polyanhydride (PA) nanoparticles (CPNs) with rosuvastatin calcium (RCa) in order to enhance the poor oral bioavailability. Methods: CPNs containing RCa/CD complexes were prepared by a modified solvent displacement method and morphological analyses, particle size (PS), polydispersity index (PDI), zeta potential (ZP), encapsulation efficiency (EE), DSC, FT-IR, XRD, 1H-NMR analyses were performed. In vitro release properties, release kinetics, cytotoxicity, in vitro permeability and pharmacokinetic studies were also studied. The stability of the formulations were evaluated during the storage period of 3 months. Results: The physicochemical studies showed that the RCa/CD complexes were well incorporated into CPNs resulted in nanosized particles (215.22 and 189.13 nm) with homogenous size distribution (PDI: 0.203 and 0.182) with relatively high incorporation capacity (76.11 and 68.18%) for the CPN1 and CPN2 formulations respectively. Sustained release of RCa from CPNs were achieved. The cytotoxicity values showed that the safety of the formulations. According to permeability studies, pure RCa had lowest permeability data (3.08 × 10-7 cm⋅s-1 Papp value) while CPNs gained higher permeability data (1.36 × 10-5 and 1.12 × 10-5 cm⋅s-1 Papp values) for the CPN1 and CPN2 formulations respectively. CPN2 formulation was selected for pharmacokinetic studies and analyses results demonstrated that approximately 8-fold relative oral bioavailability enhancement compared to the pure RCa was achieved. Conclusion: Considering the analyses results of the study, CPNs can be regarded as suitable, safe, functional oral delivery systems for RCa with enhanced oral bioavailability.