Role of microsomal and cytosolic glutathione S-transferases in the conjugation of hexachloro-1:3-butadiene and its possible relevance to toxicity.
The mechanism of hexachloro-1:3-butadiene (HCBD)-induced glutathione depletion in male and female rats has been investigated in rat liver and kidney preparations in vitro in order to characterize the enzymes involved and to study the relationship between this effect and the nephrotoxic action of this compound. HCBD caused a marked reduction in glutathione concentration when incubated with male or female hepatic microsomal or cytosolic fractions fortified with glutathione. In contrast with that reported for other halo-olefin's, the depletion of glutathione in the microsomal fraction is not related to the formation of metabolites via cytochrome P-450. The microsomal rate of depletion appeared to be due to a direct reaction catalyzed by a microsomal glutathione S-transferase. A glutathione adduct of HCBD was isolated by thin-layer chromatography and mass spectral analysis strongly indicates the structure to be as S-(1,1,2,3,4-pentachloro-1,3-butadienyl)glutathione, confirming a direct substitution reaction without prior oxidation. This conjugate was formed at a faster rate by the hepatic microsomal fraction than by the cytosolic fraction suggesting a major role for the microsomal glutathione S-transferases in the disposition of this compound. A second more polar glutathione-dependent adduct which may be a double conjugate was formed with cytosol. Glutathione adducts were also formed by male and female kidney cytosol and microsomal fractions but at a slower rate than in liver fractions. It is suggested that the glutathione conjugate of HCBD may be converted to the cysteine derivative, the structure of which is similar to that of S-dichloro-vinyl-L-cysteine and therefore may be nephrotoxic by a similar mechanism.


Effects of the DSP-toxic dinoflagellate Dinophysis acuta on clearance and respiration rate of the blue mussel, Mytilus edulis.
Diarrheic Shellfish Poisoning toxins (DST) are a severe health risk to shellfish consumers and can be a major problem for the shellfish industry. Bivalve molluscs can accumulate DST via ingestion of toxic dinoflagellates like Dinophysis spp., which are the most prominent producers of DST. The effects of DST-containing dinoflagellate Dinophysis acuta on bivalve clearance and respiration rate were investigated in the blue mussel (Mytilus edulis) exposed to different algal densities in a controlled laboratory study. Results showed that M. edulis exposed to D. acuta displayed a reduced clearance rate compared to M. edulis exposed to equivalent bio-volumes of the non-toxic cryptophyte Rhodomonas salina. Furthermore, M. edulis ceased to feed on D. acuta after 1 to 4 h, depending on D. acuta densities. The quickest response was observed at the highest densities of D. acuta. The estimated total amount of DST accumulated in the M. edulis exceeded the regulatory limit for human consumption and furthermore, intoxication of the M. edulis seemed to occur faster at high cell toxicity rather than at high cell density. However, respiration rates were, similar, irrespective of whether M. edulis were fed single diets of R. salina, D. acuta or a mixed diet of both algal species. In conclusion, the DST-containing D. acuta had a severe negative effect on the clearance of M. edulis, which can affect the conditions of the M. edulis negatively. Hence, DST may cause low quality M. edulis, due to reduced feeding when exposed to DST-containing D. acuta.


HIBCH deficiency in a patient with phenotypic characteristics of mitochondrial disorders.
HIBCH (3-hydroxyisobutyryl-CoA hydrolase) deficiency (MIM #250620) is a rare autosomal recessive inborn error of metabolism, leading to a block in the catabolic pathway of the amino acid valine and presumably to accumulation of toxic valine metabolites in mitochondria. Only three families with HIBCH deficiency and biallelic HIBCH mutations have been described. We report on a further patient, first child of healthy consanguineous parents, with severe developmental delay, seizures, hyperintensities of the basal ganglia on magnetic resonance imaging (MRI), progressive brain atrophy, optic nerve atrophy, repeatedly elevated blood lactate, and respiratory chain complexes I, I + III and cytochrome c oxidase deficiencies with borderline depletion of mitochondrial DNA in muscle tissue. Laboratory findings in blood and skeletal muscle were inconsistent and did not allow a definite diagnosis, but supported the hypothesis of mitochondrial dysfunction. Homozygosity mapping and whole-exome sequencing revealed a homozygous one-base pair insertion in HIBCH. Deficiency of enzyme activity was confirmed in cultured fibroblasts. Although relatively unspecific, the clinical features were similar to those of the previously reported cases. Given the clinical variability and large number of differential diagnoses, the prevalence of HIBCH deficiency is probably underestimated. Next-generation sequencing approaches are an effective tool for identifying the underlying genetic basis in patients suspected of mitochondrial disorders.


Girls on the Run: a quasi-experimental evaluation of a developmentally focused youth sport program.
Evidence supporting the effectiveness of a developmental-focused youth sport (DYS) program designed exclusively for elementary school aged girls is mounting. The purpose of this study was to evaluate the impact of on the Girls on the Run program on psychological and physical assets among 3rd- to 5th-grade girls. A longitudinal quasi-experimental study was conducted to evaluate intervention effects among 877 participants categorized into 1 of 3 groups (never, newly, and previously exposed). A 64-item self-report survey measured developmental assets at 3 time-points. Nested random effects ANOVA models were used to compare demographic factors and psychological and physical assets between exposure groups and to compare longitudinal differences in these assets. After adjustment for multiple comparisons, previous program participants had significantly higher physical activity commitment (P = .006) and physical activity levels (P = .047) at preintervention than never exposed. From pre- to postintervention body image improved in newly exposed participants (P = .03). Physical activity increased from preintervention to follow-up among never and newly exposed participants (all P < .05). Although we were unable to fully confirm the study hypotheses, the results of the current study provide new evidence to support future long-term studies examining the effectiveness of an innovative DYS program for 3rd- to 5th-grade girls.


The effects of changing dairy intake on trans and saturated fatty acid levels- results from a randomized controlled study.
Dairy food is an important natural source of saturated and trans fatty acids in the human diet. This study evaluates the effect of dietary advice to change dairy food intake on plasma fatty acid levels known to be present in milk in healthy volunteers. Twenty one samples of whole fat dairy milk were analyzed for fatty acids levels. Changes in levels of plasma phospholipid levels were evaluated in 180 healthy volunteers randomized to increase, not change or reduce dairy intake for one month. Fatty acids were measured by gas chromatography-mass spectrometry and levels are normalized to d-4 alanine. The long chain fatty acids palmitic (13.4%), stearic (16.7%) and myristic (18.9%) acid were most common saturated fats in milk. Four trans fatty acids constituted 3.7% of the total milk fat content. Increased dairy food intake by 3.0 (± 1.2) serves/ day for 1 month was associated with small increases in plasma levels of myristic (+0.05, 95% confidence level-0.08 to 0.13, p = 0.07), pentadecanoic (+0.014, 95% confidence level -0.016 to 0.048, p = 0.02) and margaric acid (+0.02, -0.03 to 0.05, p = 0.03). There was no significant change in plasma levels of 4 saturated, 4 trans and 10 unsaturated fatty acids. Decreasing dairy food intake by 2.5 (± 1.2) serves per day was not associated with change in levels of any plasma fatty acid levels. Dietary advice to change dairy food has a minor effect on plasma fatty acid levels. ACTRN12612000574842.


Investigating socio-economic inequity in access to and expenditures on routine immunization services in Anambra state.
Addressing existing inequities in the utilization of priority health services such as routine immunization is a current public health priority. Increasing access to routine immunization from the current low levels amongst all socio-economic status groups in Nigeria is challenging. However, little is known on the level of SES inequity in utilization of routine immunization services and such information which will inform the development of strategies for ensuring equitable provision of routine immunization services in the country. The study was a cross sectional household survey, which was undertaken in two randomly selected communities in Anambra State, southeast Nigeria. A pre-tested interviewer administered questionnaire was used to collect data on levels of access to RI by children under-2 years from randomly selected households. In each household, data was collected from the primary care givers or their representative (in their absence). The relationship between access to routine immunization and socio-economic status of households and other key variables was explored in data analysis. Households from high socio-economic status (well-off) groups utilized routine immunization services more than those that belong to low socio-economic status (poor) groups (X2 = 9.97, p < 0.002). It was found that higher percentage of low socio-economic status households compared to the high socio-economic status households received routine immunization services at public health facilities. Households that belong to low socio-economic status groups had to travel longer distance to get to health facilities consequently incurring some transportation cost. The mean expenditures on service charge for routine immunization services (mostly informal payments) and transportation were US$1.84 and US$1.27 respectively. Logistic regression showed that access to routine immunization was positively related to socio-economic status and negatively related to distant of a household to a health facility. Ability to pay affects access to services, even when such services are free at point of consumption with lower socio-economic status groups having less access to services and also having other constraints such as transportation. Hence, innovative provision methods that will bring routine immunization services closer to the people and eliminate all formal and informal user fees for routine immunization will help to increase and improve equitable coverage with routine immunization services.


Intervertebral disc tissue engineering: A brief review.
Intervertebral disc (IVD) degeneration (IDD) is associated with low back pain and significantly affects the patient's quality of life. Degeneration of the IVD alters disk height and the mechanics of the spine, leading to chronic segmental spinal instability. The pathophysiology of IVD disease is still not well understood. Current therapies for IDD include conservative and invasive approaches, but none of those treatments are able to restore the disc structure and function. Recently, tissue engineering techniques emerged as a possible approach to treat IDD, by replacing a damaged IVD with scaffolds and appropriate cells. Advances in manufacturing techniques, material processing and development, surface functionalization, drug delivery systems and cell incorporation furthered the development of tissue engineering therapies. In this review, biomaterial scaffolds and cell-based therapies for IVD regeneration are briefly discussed.


Implications for introgression: has selection for fast growth altered the size threshold for precocious male maturation in domesticated Atlantic salmon?
Mature male parr (MMP) represent an important alternative life-history strategy in Atlantic salmon populations. Previous studies indicate that the maturation size threshold for male parr varies among wild populations and is influenced by individual growth, environmental conditions, and genetics. More than ten generations of breeding have resulted in domesticated salmon displaying many genetic differences to wild salmon, including greatly increased growth rates. This may have resulted in domesticated fish with the potential to outgrow the size threshold for early maturation, or evolution of the size threshold of the trait itself. To investigate this, we performed a common-garden experiment under farming conditions using 4680 salmon from 39 families representing four wild, two wild-domesticated hybrid, and two domesticated strains. Domesticated salmon outgrew wild salmon 2-5-fold, and hybrids displayed intermediate growth. Overall, the numbers of MMP varied greatly among families and strains: averaging 4-12% in domesticated, 18-25% in hybrid, and 43-74% in the wild populations. However, when the influence of growth was accounted for, by dividing fish into lower and upper size modes, no difference in the incidence of MMP was detected among domesticated and wild strains in either size mode. In the lower size mode, hybrids displayed significantly lower incidences of mature males than their wild parental strains. No consistent differences in the body size of MMP, connected to domestication, was detected. Our data demonstrate: 1- no evidence for the evolution of the size threshold for MMP in domesticated salmon, 2- the vastly lower incidence of MMP in domesticated strains under aquaculture conditions is primarily due to their genetically increased growth rate causing them to outgrow the size threshold for early maturation, 3- the incidence of MMP is likely to overlap among domesticated and wild salmon in the natural habitat where they typically display overlapping growth, although hybrid offspring may display lower incidences of mature male parr. These results have implications for wild salmon populations that are exposed to introgression from domesticated escapees.


[Floral biology and pollinators of Trichocereus pasacana (Cactaceae) in Parque Nacional Los Cardones, Argentina].
Many columnar cacti are bat pollinated. It has been suggested that this kind of pollination would be more important in tropical than in temperate regions where flowers are open only one night. Thrichocereus pasacana produces big and resistant white flowers. We analyzed flower characteristics, floral cycle, stigmatic receptivity, nectar production, pollen presence and floral visitors in a T. pasacana population at National Park Los Cardones (Salta, Argentina) in November 1997. Flower features were constant between individuals of the population. Flowers start opening at evening and anthesis time is from 18 to 40 hs. The estigma was receptive throughout the floral cycle. Anther dehiscence occurs with flower opening. Nectar production was highest between 18 to 24 hs. Although T. pasacana are open during the night, floral visitors are diurnal. The most frequent was Xylocopa sp. In the study area, nectarivorous bats were not detected. The morphological features of T. pasacana flowers were similar but bigger compared to other columnar cacti. Anthesis time was also longer while nectar production was lower. T. pasacana pollination at National Park Los Cardones is done by bees.


Initial Patterns of Prescription Opioid Supply and Risk of Mortality Among Insured Adults in the United States.
To examine the association between initial patterns of prescription opioid supply (POS) and risk of all-cause mortality among an insured opioid-naïve patient population in the United States (US). This retrospective observational cohort study used de-identified, administrative health care claims data from a large national insurer (Optum Clinformatics Data Mart) from 2010 to 2015. Participants included insured, cancer-free adults prescribed opioid analgesics. Prescription opioids received during the first 6 months of therapy were used to categorize initial patterns of POS as daily or nondaily. Cox regression was used to estimate the association of initial patterns of POS with all-cause mortality within one year of follow-up, adjusting for baseline covariates to control for confounding. A total of 4,054,417 patients were included, of which 2.75% had incident daily POS; 54.8% were female; median age was 50 years; mean Charlson comorbidity index (CCI) was 0.21 (standard deviation = 0.77); and mean daily morphine milligram equivalent was 34.61 (95% confidence intervals: 34.59, 34.63). There were 2068 more deaths per 100,000 person-years among patients who were prescribed opioids daily than nondaily. After adjusting for baseline covariates, the hazard of all-cause mortality among patients with incident daily POS was nearly twice that among those prescribed nondaily (hazard ratio [HR] = 1.94; 95% confidence intervals: 1.84, 2.04). Among insured adult patients with noncancer pain, incident chronic POS was associated with a significantly increased risk of all-cause mortality over at most 1 year of follow-up. Because these results may be susceptible to bias, more research is needed to establish causality.


Epidemiology of moderate-to-severe penetrating versus closed traumatic brain injury in the Iraq and Afghanistan wars.
US combat operations in Iraq and Afghanistan have resulted in a greater proportion of service members with head and neck wounds caused by explosions compared with that of previous wars. Although penetrating traumatic brain injury (TBI) is frequently associated with these wounds, the epidemiology of penetrating TBI from these conflicts has not been well described. The Joint Theater Trauma Registry was queried for January 2003 through December 2010 to identify all patients with moderate-to-severe brain injury with a maximum Abbreviated Injury Scale (AIS) score of the head of 3 or greater and a diagnosis of penetrating or closed TBI in accordance with the Department of Defense Traumatic Brain Injury Surveillance definition. The epidemiology of these injuries was examined, including demographics, TBI severity, overall injury severity, and surgical interventions provided. A total of 1,255 TBI patients (774 penetrating, 481 closed) meeting criteria were identified. Penetrating brain injuries were more severe, more likely to be battle related, and less likely to be isolated injuries than a group of moderate-to-severe closed TBIs within the same range of anatomic injury severity. During the 5-year period of the Iraq war with the largest numbers of TBIs (2004-2008), the numbers of penetrating TBIs exceeded closed TBIs by a ratio of 2:1. During the 3-year period of the Afghanistan war with the greatest numbers of TBIs (2008-2010), the ratio of penetrating to closed TBIs was substantially lower, approximately 1.3:1. This study represents the first comprehensive report on the epidemiology of moderate-to-severe penetrating and closed TBIs resulting from the wars in Iraq and Afghanistan using Joint Theater Trauma Registry data. With the maturing theater of conflicts, penetrating TBIs were substantially less predominant compared with closed TBIs. While this finding may reflect changes in the use of protective measures and tactics or improvements in diagnosis of closed TBIs, additional research is needed to identify the reason for this shift and the subsequent effect on outcome after combat-related TBIs. Epidemiologic study, level III.


A gene necessary for reproductive suppression in termites.
A major transition in evolution is the origin of a division between reproduction and work among individuals. Nowhere is this divide more striking than in social insects, where workers rarely produce offspring even though they are often capable of reproduction should the queen or king die. The molecular mechanisms that control worker reproduction remain largely unknown. We used a combination of behavioral assays and RNA interference (RNAi) to identify a gene required for the reproductive division of labor between the queen and the workers.


Comment on "Tumor growth need not be driven by rare cancer stem cells".
Kelly et al. (Brevia, 20 July 2007, p. 337) questioned xenotransplant experiments supporting the cancer stem cell (CSC) hypothesis because they found a high frequency of leukemia-initiating cells (L-IC) in some transgenic mouse models. However, the CSC hypothesis depends on prospective purification of cells with tumor-initiating capacity, irrespective of frequency. Moreover, we found similar L-IC frequencies in genetically comparable leukemias using syngeneic or xenogeneic models.


Effectiveness of base in prism for presbyopes with convergence insufficiency.
Although the treatment of symptomatic convergence insufficiency (CI) with base-in (BI) prism in adults has a strong theoretical foundation, there have been very few studies addressing its efficacy. The purpose of this study was to investigate whether the application of BI prism, using a novel progressive addition lens design which incorporates BI prism in the near portion only, could help alleviate the symptoms in presbyopes with CI. A total of 29 symptomatic CI subjects aged 45 to 68 years were examined. All subjects took the CI Symptom Survey V-15 (CISS) and scored >or=16 points. Each subject was assigned two pairs of progressive addition glasses made by the same manufacturer in a randomized sequence, one with BI prism and one with the same lens prescription but no prism (placebo). Subjects wore each pair of glasses for 3 weeks and completed the CISS at the end of the 3rd week. Symptom level measured with CISS was the major outcome measure. The mean (standard deviation) CISS score was 30.21 (9.30) at baseline and decreased to 13.38 (9.44) with the BI-prism glasses, vs. 23.62 (10.76) with the placebo glasses. There were significant differences between the baseline survey score and the score with the BI-prism glasses (p < 0.0001) and between the score with placebo glasses and the one with BI-prism glasses (p = 0.001). The progressive addition glasses with BI-prism were found to be effective in alleviating symptoms of presbyopes with symptomatic CI.


Time to Review Authorisation and Funding for New Cancer Medicines in Europe? Inferences from the Case of Olaratumab.
The potential benefits of early patient access to new medicines in areas of high unmet medical need are recognised, but uncertainties concerning effectiveness, safety and added value when new medicines are authorised, and subsequently funded based on initial preliminary data only, have important implications. In 2016 olaratumab received accelerated conditional approval from both the European Medicines Agency and the US Food and Drug Administration for the treatment of soft-tissue sarcoma, based on the claims of a substantial reduction in the risk of death with an 11.8-month improvement in median overall survival in a phase II trial in combination with doxorubicin vs. doxorubicin alone. The failure to confirm these benefits in the post-authorisation pivotal trial has highlighted key concerns regarding early access and conditional approvals for new medicines. Concerns include potentially considerable clinical and economic costs, so that patients may have received suboptimal treatment and any money spent has foregone the opportunity to improve access to effective treatments. As a result, it seems reasonable to reconsider current marketing authorisation models and approaches. Potential pathways forward include closer collaboration between regulators, pharmaceutical companies and payers to enhance the generation of rapid and comparative confirmatory trials in a safe and fair manner, with minimal patient exposure as required to achieve robust evidence. Additionally, it may be time to review early access systems, and to explore new avenues regarding who should pay or part pay for new treatments whilst information is being collected as part of any obligations for conditional marketing authorisation. Greater co-operation between countries regarding the collection of data in routine clinical care, and further research on post-marketing data analysis and interpretation, may also contribute to improved appraisal and continued access to new innovative cancer treatments.


Lack of run-down of smooth muscle P2X receptor currents recorded with the amphotericin permeabilized patch technique, physiological and pharmacological characterization of the properties of mesenteric artery P2X receptor ion channels.
Immunoreactivity for P2X(1), P2X(4) and P2X(5) receptor subtypes was detected in the smooth muscle cell layer of second and third order rat mesenteric arteries immunoreactivity, for P2X(2), P2X(3), P2X(6) and P2X(7) receptors was below the level of detection in the smooth muscle layer. P2X receptor-mediated currents were recorded in patch clamp studies on acutely dissociated mesenteric artery smooth muscle cells. Purinergic agonists evoked transient inward currents that decayed rapidly in the continued presence of agonist (tau approximately 200 ms). Standard whole cell responses to repeated applications of agonist at 5 min intervals ran down. Run-down was unaffected by changes in extracellular calcium concentration, intracellular calcium buffering or the inclusion of ATP and GTP in the pipette solution. Run-down was overcome and reproducible responses to purinergic agonists were recorded using the amphotericin permeabilized patch recording configuration. The rank order of potency at the P2X receptor was ATP=2 methylthio ATP>alpha, beta-methylene ATP>CTP=l-beta,gamma-methylene ATP. Only ATP and 2meSATP were full agonists. The P2 receptor antagonists suramin and PPADS inhibited P2X receptor-mediated currents with IC(50)s of 4 microM and 70 nM respectively. These results provide further characterization of artery P2X receptors and demonstrate that the properties are dominated by a P2X(1)-like receptor phenotype. No evidence could be found for a phenotype corresponding to homomeric P2X(4) or P2X(5) receptors or to heteromeric P2X(1/5) receptors and the functional role of these receptors in arteries remains unclear.


Cross-reactions of sera from dogs infected with Angiostrongylus vasorum in commercially available Dirofilaria immitis test kits.
Dirofilaria immitis and Angiostrongylus vasorum are both important potentially fatal canine nematodes with overlapping endemic areas, especially in Europe. The preadult and adult stages of both species are living in the Arteria pulmonalis and the right heart, and diagnostically detectable circulating parasite antigens have been demonstrated for both species. For the detection of D. immitis infections, a variety of commercial tests have been developed, however, they have not been evaluated for cross-reactions against circulating antigens of A. vasorum. In this study, potential cross-reactions of sera from 16 dogs, which were experimentally infected with A. vasorum and which had circulating antigens as confirmed by a species-specific ELISA, were evaluated for the detection of A. vasorum antigen in six commercially available D. immitis test kits. In three fast tests (Witness® Dirofilaria, SensPERT® Canine Heartworm, SNAP® 4Dx® Plus), all sera were negative. One fast membrane ELISA (SNAP® HTWM RT Test) was positive with four sera (25%), and one serum delivered a non-valid result twice. In the PetChek® HTWM PF Test, depending on the interpretation protocol, 5 or 8 dogs (31.2 - 50%) were positive. With the DiroCHEK®-ELISA, a single A. vasorum-infected dog (6.2%) tested positive. Due to potential cross-reactions with A. vasorum in commercially available test kits for the detection of D. immitis antigen, the simultaneous use of highly specific diagnostic methods for the differentiation of these two canine heart worms is recommended.


SIRT1 pathway dysregulation in the smoke-exposed airway epithelium and lung tumor tissue.
Cigarette smoke produces a molecular field of injury in epithelial cells lining the respiratory tract. However, the specific signaling pathways that are altered in the airway of smokers and the signaling processes responsible for the transition from smoking-induced airway damage to lung cancer remain unknown. In this study, we use a genomic approach to study the signaling processes associated with tobacco smoke exposure and lung cancer. First, we developed and validated pathway-specific gene expression signatures in bronchial airway epithelium that reflect activation of signaling pathways relevant to tobacco exposure, including ATM, BCL2, GPX1, NOS2, IKBKB, and SIRT1. Using these profiles and four independent gene expression datasets, we found that SIRT1 activity is significantly upregulated in cytologically normal bronchial airway epithelial cells from active smokers compared with nonsmokers. In contrast, this activity is strikingly downregulated in non-small cell lung cancer. This pattern of signaling modulation was unique to SIRT1, and downregulation of SIRT1 activity is confined to tumors from smokers. Decreased activity of SIRT1 was validated using genomic analyses of mouse models of lung cancer and biochemical testing of SIRT1 activity in patient lung tumors. Together, our findings indicate a role of SIRT1 in response to smoke and a potential role in repressing lung cancer. Furthermore, our findings suggest that the airway gene expression signatures derived in this study can provide novel insights into signaling pathways altered in the "field of injury" induced by tobacco smoke and thus may impact strategies for prevention of tobacco-related lung cancer.


Controlling high harmonics generation by spatial shaping of high-energy femtosecond beam.
We demonstrate controlled high-order harmonic generation in gas using high-energy femtosecond pulses (50 fs-50 mJ on target) by performing spatial shaping of the terrawatt fundamental laser beam. We have developed a two optical paths mirror that can withstand high power and shape the pump beam into a quasi-flat-top profile (super Gaussian) near focus. We observe clear signatures of the spatial shaping on the harmonic beam in terms of profile, divergence, level of signal, and spectrum. The harmonic generation in neon with a quasi-flat-top beam results in a broadband extreme UV beam with extremely low divergence (~340 μrad).


The role of procalcitonin outside of the Intensive Care Unit (ICU): a multidisciplinary approach.
Biochemical markers are commonly used in medicine to guide diagnostic investigation or therapy duration and/or monitor treatment efficacy. Due to the emergence and spread of antimicrobial resistance, markers able to prompt a more rational use of antimicrobial therapy are regarded with the greatest attention. Procalcitonin (PCT) certainly stands out among others, yet its role must be better established especially outside of the critical care area. Data about PCT utilization in non-critical patients, optimal negativity cut-offs as well as a protocol for measurement timing are all lacking. To address these issues, a focus group was set up to propose and endorse shared statements regarding the most beneficial use of PCT in real life as infection marker for non-critical patients, based on the authors' experience and a review of recent literature. A group of nine experts in the fields of Infectious Diseases, Internal Medicine, Microbiology, Clinical Chemistry, Surgery and Medical Economics participated in the discussion of nine pre-specified statements. The potential role for PCT in differentiating infectious and non-infectious clinical syndromes and guiding antimicrobial therapy discontinuation was acknowledged. Moreover, a shared measurement protocol and desirable cut-offs for the non-critical area were proposed. Finally, observations were made about a reasonable selection of the patient population to be tested.


D1 and D2 dopamine receptor-regulated gene expression of striatonigral and striatopallidal neurons.
The striatum, which is the major component of the basal ganglia in the brain, is regulated in part by dopaminergic input from the substantia nigra. Severe movement disorders result from the loss of striatal dopamine in patients with Parkinson's disease. Rats with lesions of the nigrostriatal dopamine pathway caused by 6-hydroxydopamine (6-OHDA) serve as a model for Parkinson's disease and show alterations in gene expression in the two major output systems of the striatum to the globus pallidus and substantia nigra. Striatopallidal neurons show a 6-OHDA-induced elevation in their specific expression of messenger RNAs (mRNAs) encoding the D2 dopamine receptor and enkephalin, which is reversed by subsequent continuous treatment with the D2 agonist quinpirole. Conversely, striatonigral neurons show a 6-OHDA-induced reduction in their specific expression of mRNAs encoding the D1 dopamine receptor and substance P, which is reversed by subsequent daily injections of the D1 agonist SKF-38393. This treatment also increases dynorphin mRNA in striatonigral neurons. Thus, the differential effects of dopamine on striatonigral and striatopallidal neurons are mediated by their specific expression of D1 and D2 dopamine receptor subtypes, respectively.


Secretory immunoglobulin A antibodies against the sigma1 outer capsid protein of reovirus type 1 Lang prevent infection of mouse Peyer's patches.
Reovirus type 1 Lang (T1L) adheres to M cells in the follicle-associated epithelium of mouse intestine and exploits the transport activity of M cells to enter and infect the Peyer's patch mucosa. Adult mice that have previously cleared a reovirus T1L infection have virus-specific immunoglobulin G (IgG) in serum and IgA in secretions and are protected against reinfection. Our aim in this study was to determine whether secretory IgA is sufficient for protection of Peyer's patches against oral reovirus challenge and, if so, against which reovirus antigen(s) the IgA may be directed. Monoclonal antibodies (MAbs) of the IgA isotype, directed against the sigma1 protein of reovirus T1L, the viral adhesin, were produced and tested along with other, existing IgA and IgG MAbs against reovirus T1L outer capsid proteins. Anti-sigma1 IgA and IgG MAbs neutralized reovirus T1L in L cell plaque reduction assays and inhibited T1L adherence to L cells and Caco-2(BBe) intestinal epithelial cells in vitro, but MAbs against other proteins did not. Passive oral administration of anti-sigma1 IgA and IgG MAbs prevented Peyer's patch infection in adult mice, but other MAbs did not. When anti-sigma1 IgA and IgG MAbs were produced in mice from hybridoma backpack tumors, however, the IgA prevented Peyer's patch infection, but the IgG did not. The results provide evidence that neutralizing IgA antibodies specific for the sigma1 protein are protective in vitro and in vivo and that the presence of these antibodies in intestinal secretions is sufficient for protection against entry of reovirus T1L into Peyer's patches.


The Kv2.1 C terminus can autonomously transfer Kv2.1-like phosphorylation-dependent localization, voltage-dependent gating, and muscarinic modulation to diverse Kv channels.
Modulation of K+ channels is widely used to dynamically regulate neuronal membrane excitability. The voltage-gated K+ channel Kv2.1 is an abundant delayed rectifier K+ (IK) channel expressed at high levels in many types of mammalian central neurons where it regulates diverse aspects of membrane excitability. Neuronal Kv2.1 is constitutively phosphorylated, localized in high-density somatodendritic clusters, and has a relatively depolarized voltage dependence of activation. Here, we show that the clustering and voltage-dependent gating of endogenous Kv2.1 in cultured rat hippocampal neurons are modulated by cholinergic stimulation, a common form of neuromodulation. The properties of neuronal Kv2.1 are recapitulated in recombinant Kv2.1 expressed in human embryonic kidney 293 (HEK293) cells, but not COS-1 cells, because of cell background-specific differences in Kv2.1 phosphorylation. As in neurons, Kv2.1 in HEK293 cells is dynamically regulated by cholinergic stimulation, which leads to Ca2+/calcineurin-dependent dephosphorylation of Kv2.1, dispersion of channel clusters, and hyperpolarizing shifts in the voltage-dependent gating properties of the channel. Immunocytochemical, biochemical, and biophysical analyses of chimeric Kv channels show that the Kv2.1 cytoplasmic C-terminal domain can act as an autonomous domain sufficient to transfer Kv2.1-like clustering, voltage-dependent activation, and cholinergic modulation to diverse Kv channels. These findings provide novel mechanistic insights into cholinergic modulation of ion channels and regulation of the localization and voltage-dependent gating properties of the abundant neuronal Kv2.1 channel by cholinergic and other neuromodulatory stimuli.


The temporoparietal fascial flap is an alternative to free flaps for orbitomaxillary reconstruction.
The temporoparietal fascial flap is a thin, pliable, well-vascularized, locoregional flap that can be a reasonable alternative to traditional free flap reconstruction, but its utility for reconstruction of orbitomaxillary defects is often overlooked. The authors investigated the rationale for and benefits of the use of this flap over free tissue transfer in a well-defined subset of head and neck cancer cases. The records of all patients who underwent temporoparietal fascial reconstruction for orbitomaxillectomy between 1993 and 2008 were reviewed. Demographic data, preoperative plans, operative details, and outcomes were assessed to (1) determine the overall outcomes for the temporoparietal fascial flap and (2) analyze preoperative and intraoperative factors that led to the choice of this flap instead of the originally planned free flap. Nineteen patients (mean age, 56 years) were included in this study. Ten (53 percent) were initially considered for free flap reconstruction. Among them, the principal reasons for electing the temporoparietal fascial flap were (1) smaller-than-anticipated extent of resection, (2) need for coverage of implant or bone or thin prosthesis support, and (3) patient comorbidities. Nine resections (47 percent) involved the orbit alone, two (11 percent) involved the maxilla alone, and eight (42 percent) involved a combination of the two. Five patients (26 percent) received neoadjuvant radiation. Four (21 percent) experienced complications (flap failure, ectropion, enophthalmos with intraoral mesh extrusion, and partial skin graft loss). Mean follow-up time was 36 +/- 27 months. The temporoparietal fascial flap is a viable alternative to free flaps for orbitomaxillary defects when orbital resection is limited, when obturator reconstruction is desired, or when comorbidities preclude microvascular surgery.


Neuropsychological functioning of U.S. Gulf War veterans 10 years after the war.
Many U.S. Gulf War-era veterans complained of poor cognition following the war. This study assessed neuropsychological functioning in veterans 10 years after the war through objective tests. 2189 Gulf War-era veterans (1061 deployed, 1128 non-deployed) were examined at 1 of 16 U.S. Veterans Affairs medical centers. Outcomes included neuropsychological domains derived from factor analysis and individual test scores. Deployed veterans performed significantly worse than non-deployed veterans on 2 of 8 factors (motor speed & sustained attention, analysis not corrected for multiple comparisons) and on 4 of 27 individual test variables (Trails A & B, California Verbal Learning Test-List B, and Continuous Performance Test sensitivity, with only Trails B surviving Bonferroni correction). Within deployed veterans, Khamisiyah exposure was negatively correlated with motor speed after controlling for emotional distress. Depressive symptoms and self-reported exposure to toxicants were independently and significantly associated with worse sustained attention. Other factors were also associated with self-reported exposures. The findings were not a result of differential effort across groups. Gulf War deployment is associated with subtle declines of motor speed and sustained attention, despite overall intact neuropsychological functioning. Evidence suggests that toxicant exposures influence both these functions, and depressive symptoms also influence attention.


Immunocytochemical localization of the mGluR1 alpha metabotropic glutamate receptor in the dorsal cochlear nucleus.
We demonstrate that the metabotropic glutamate receptor mGluR1 alpha is enriched in two interneuron cell populations in the dorsal division of the cochlear nucleus. Electron microscopic analysis confirms that mGluR1 alpha immunoreactivity is concentrated in the dendritic spines of cartwheel cells and in dendrites of the recently described unipolar brush cells. The cartwheel cells, which have many similarities to the Purkinje cells of the cerebellum, participate in a local neuronal circuit that modulates the output of the dorsal cochlear nucleus. Immunostained unipolar brush cells were observed in granule cell regions of the cochlear nucleus and the vestibulocerebellum. The presence of analogous cell types with similar patterns of immunolabeling in the cerebellum and in the dorsal cochlear nucleus suggests that a shared but as yet unknown mode of processing may occur in both structures.


Deeply Supervised Discriminative Learning for Adversarial Defense.
Deep neural networks can easily be fooled by an adversary with minuscule perturbations added to an input image. The existing defense techniques suffer greatly under white-box attack settings, where an adversary has full knowledge of the network and can iterate several times to find strong perturbations. We observe that the main reason for the existence of such vulnerabilities is the close proximity of different class samples in the learned feature space of deep models. This allows the model decisions to be completely changed by adding an imperceptible perturbation to the inputs. To counter this, we propose to class-wise disentangle the intermediate feature representations of deep networks, specifically forcing the features for each class to lie inside a convex polytope that is maximally separated from the polytopes of other classes. In this manner, the network is forced to learn distinct and distant decision regions for each class. We observe that this simple constraint on the features greatly enhances the robustness of learned models, even against the strongest white-box attacks, without degrading the classification performance on clean images. We report extensive evaluations in both black-box and white-box attack scenarios and show significant gains in comparison to state-of-the-art defenses.


Enhanced biogas recovery by applying post-digestion in large-scale centralized biogas plants.
The main objective of this study was to investigate the degradation efficiency of centralized biogas plants and provide guidance for the design of more efficient digester and post-digestion systems. These centralized biogas plants in Denmark digest manure together with organic waste from the food industry to generate biogas, which is used for electricity and thermal energy. A total of 20 such plants are currently active in Denmark, most of which were included in the investigation. From the plants, samples were obtained from various steps of the process. Samples were analysed and the residual biogas potential determined by batch post-digestion at various temperature levels. Results were correlated with plant characteristics and production statistics in order to judge the efficiency of various digestion concepts. A simplified model based on a two-step biogas production process was developed and experimental data were used to determine kinetic constants. Experimental results and analysis combined with model simulations showed that the residual biogas potential in the main digestion step effluent is originating mainly from undegraded particulate matter in the biomass. For thermophilic plants 93% of the residual biogas potential was originating from particulate matter and 88% for the mesophilic biogas plants. This indicates that the residual biogas potential is mainly due to insufficient retention time in the main digestion step for hydrolysis of particulate material and that the hydrolysis step is the methane yield limiting factor, while conversion of soluble material such as VFA is the rate limiting factor critical for achieving a stable process.


Phytoestrogen signaling and symbiotic gene activation are disrupted by endocrine-disrupting chemicals.
Some organochlorine pesticides and other synthetic chemicals mimic hormones in representatives of each vertebrate class, including mammals, reptiles, amphibians, birds, and fish. These compounds are called endocrine-disrupting chemicals (EDCs). Similarly, hormonelike signaling has also been observed when vertebrates are exposed to plant chemicals called phytoestrogens. Previous research has shown the mechanism of action for EDCs and phytoestrogens is as unintended ligands for the estrogen receptor (ER). Although pesticides have been synthesized to deter insects and weeds, plants produce phytoestrogens to deter herbivores, as attractant cues for insects, and as recruitment signals for symbiotic soil bacteria. Our data present the first evidence that some of the same organochlorine pesticides and EDCs known to disrupt endocrine signaling through ERs in exposed wildlife and humans also disrupt the phytoestrogen signaling that leguminous plants use to recruit Sinorhizobium meliloti soil bacteria for symbiotic nitrogen fixation. Here we report that a variety of EDCs and pesticides commonly found in agricultural soils interfere with the symbiotic signaling necessary for nitrogen fixation, suggesting that the principles underlying endocrine disruption may have more widespread biological and ecological importance than had once been thought.


Sample size considerations of prediction-validation methods in high-dimensional data for survival outcomes.
A variety of prediction methods are used to relate high-dimensional genome data with a clinical outcome using a prediction model. Once a prediction model is developed from a data set, it should be validated using a resampling method or an independent data set. Although the existing prediction methods have been intensively evaluated by many investigators, there has not been a comprehensive study investigating the performance of the validation methods, especially with a survival clinical outcome. Understanding the properties of the various validation methods can allow researchers to perform more powerful validations while controlling for type I error. In addition, sample size calculation strategy based on these validation methods is lacking. We conduct extensive simulations to examine the statistical properties of these validation strategies. In both simulations and a real data example, we have found that 10-fold cross-validation with permutation gave the best power while controlling type I error close to the nominal level. Based on this, we have also developed a sample size calculation method that will be used to design a validation study with a user-chosen combination of prediction. Microarray and genome-wide association studies data are used as illustrations. The power calculation method in this presentation can be used for the design of any biomedical studies involving high-dimensional data and survival outcomes.


The use of fluorescein-conjugated lectins for visualizing atypical mycobacteria.
We investigated the feasibility of using fluorescein-conjugated lectins for visualizing and differentiating two species of atypical mycobacteria. Pure cultures of Mycobacterium fortuitum and Mycobacterium chelonei were established, as was an experimental model of infectious keratitis involving these two organisms. Samples from the pure cultures and corneal scrapings were placed on glass slides, fixed, and incubated with one of a panel of 22 fluorescein-conjugated lectins. The slides were examined using an epifluorescence microscope. Fluorescein-conjugated concanavalin A brightly stained both species of atypical mycobacteria, in both the pure culture and experimental keratitis samples. Several additional fluorescein-conjugated lectins (wheat germ agglutinin, succinylated wheat germ agglutinin, Phaseolus vulgaris erythroagglutinin, and Psophocarpus tetragonolobus agglutinin) brightly stained M chelonei, but only moderately stained M fortuitum. These staining patterns are consistent with the known carbohydrate compositions of the cell walls of atypical mycobacteria and suggest that fluorescein-conjugated lectins may be useful for the visualization of these organisms in corneal infections.


Direct testing of blood culture for detection of the serotype 5 and 8 capsular polysaccharides of Staphylococcus aureus.
Monoclonal antibodies (MAbs) reactive with serotype 5 and 8 capsular polysaccharides of Staphylococcus aureus have been used to test, by enzyme-linked immunosorbent assay (ELISA), blood culture fluids for the presence of S. aureus. A total of 748 blood cultures from 665 patients yielding 706 bacterial isolates belonging to more than 26 bacterial species were studied. All blood cultures containing bacterial strains belonging to species other than S. aureus were negative in ELISA. All 23 blood cultures containing serotype 5 S. aureus were positive in ELISA with the corresponding MAb. Out of 20 blood cultures containing serotype 8 S. aureus, 19 were positive with the corresponding MAb. All 5 blood cultures containing nontypeable S. aureus were negative in ELISA with both MAbs. This method provides reliable identification of serotype 5 or serotype 8 S. aureus by direct testing of blood culture fluids with ELISA.


Mineral metabolism and bone mass at peripheral and axial skeleton in diabetes mellitus.
Bone mineral content (BMC), mineral homeostasis, and diabetes control were evaluated in 31 Caucasian insulin-dependent diabetic patients (disease duration 18.3 +/- 7.7 yr, mean +/- SD) with normal kidney function. To evaluate bone mass, we performed radiogrammetry and single- and dual-photon absorptiometry. In women, a significantly lower mean BMC was found in the distal radius, at a mixed trabecular-cortical (P less than .01) and a cortical (P less than .05) site, as well as in the lumbar spine (P less than .02). In diabetic men, mean BMC was significantly reduced at the trabecularcortical (P less than .01) and cortical (P less than .05) sites of the radius but not in the lumbar spine. When expressed as densities (i.e., BMC/width or lumbar BMC/area), only the BMC/width at the radius cortical area was significantly reduced in women (P less than .05). The results of the radiogrammetry showed a larger endosteal diameter in the diabetic women, resulting in a significantly lower cortical thickness (P less than .05). Diabetic men did not show abnormalities on radiogrammetry. Diabetic patients had diminished serum calcium and phosphorus concentrations (P less than .001), whereas serum parathyroid, 25-hydroxyvitamin D3, and concentrations of both total and free 1,25-dihydroxyvitamin D3 were normal. No correlation between parameters of diabetes control (HbA1, insulin dose, and triglycerides) or calcium-regulating hormones and BMC were found. These data confirm that, despite large overlap of individual values, mean bone mass at the peripheral skeleton is significantly decreased in diabetic patients. Moreover, we report that the BMC of the lumbar spine is significantly reduced in female diabetic patients.


The Association of Vitamin D Status in Lower Extremity Muscle Strains and Core Muscle Injuries at the National Football League Combine.
To evaluate the association between serum vitamin D level and the prevalence of lower extremity muscle strains and core muscle injuries in elite level athletes at the National Football League (NFL) combine. During the 2015 NFL combine, all athletes with available serum vitamin D levels were included for study. Baseline data were collected, including age, race, body mass index, position, injury history specific to lower extremity muscle strain or core muscle injury, and Functional Movement Screen scores. Serum 25-hydroxyvitamin D was collected and defined as normal (≥32 ng/mL), insufficient (20-31 ng/mL), and deficient (<20 ng/mL). Univariate regression analysis was used to examine the association of vitamin D level and injury history. Subsequent multivariate regression analysis was used to examine this relation with adjustment for collected baseline data variables. The study population included 214 athletes, including 78% African American athletes and 51% skilled position players. Inadequate vitamin D was present in 59%, including 10% with deficient levels. Lower extremity muscle strain or core muscle injury was present in 50% of athletes, which was associated with lower vitamin D levels (P = .03). Athletes with a positive injury history also showed significantly lower vitamin D levels as compared with uninjured athletes (P = .03). African American/black race (P < .001) and injury history (P < .001) was associated with lower vitamin D. Vitamin D groups showed no differences in age (P = .9), body mass index (P = .9), or Functional Movement Screen testing (P = .2). Univariate analysis of inadequate vitamin D levels showed a 1.86 higher odds of lower extremity strain or core muscle injury (P = .03), and 3.61 higher odds of hamstring injury (P < .001). Multivariate analysis did not reach an independent association of low vitamin D with injury history (P = .07). Inadequate vitamin D levels are a widespread finding in athletes at the NFL combine. Players with a history of lower extremity muscle strain and core muscle injury had a higher prevalence of inadequate vitamin D. Level IV, retrospective study-case series.


Effects of dopamine and epinephrine infusions on renal hemodynamics in severe malaria and severe sepsis.
To describe and compare the effects of dopamine and epinephrine in various doses on renal hemodynamics and oxygen transport in patients with severe malaria and severe sepsis. Prospective, controlled, crossover trial. The intensive care unit of an infectious diseases hospital in Viet Nam. Fourteen patients with severe falciparum malaria and five with severe sepsis. In an open, crossover design, we observed the effects on renal and systemic hemodynamics and oxygen transport of separate stepped infusions of epinephrine and dopamine. We measured renal blood flow (RBF) and cardiac output by the thermodilution method using fluoroscopically guided catheters. Creatinine clearance at each time point was calculated from the renal plasma flow and the renal arteriovenous difference in plasma creatinine. Dopamine at a "renal" dose (2.5 microg/kg/min) was associated with a mean (95% confidence interval) fractional increase in the absolute renal blood flow index (RBFI) of 37% (13% to 61%) and in RBF as a fraction of cardiac output (RBF/CO) of 35% (10% to 59%; p = .007 and p = .014, respectively). The consequent 39% (14% to 64%) increase in renal oxygen supply (p = .002) was accompanied by a 32% (20% to 44%) decrease in the renal oxygen extraction ratio (p = .0003), leading to no net change in renal oxygen consumption. At higher doses (10 microg/kg/min), both RBF and RBF/CO were not significantly different from baseline values and decreased further as the dose was reduced again. There was no obvious explanation for this hysteresis. There was no change in renal oxygen consumption throughout the study. Because lactic acidosis developed, epinephrine was only given to eight of the 19 patients, and the full stepped epinephrine infusion was given to four patients. Epinephrine infusion was associated, both in absolute terms and when compared with dopamine, with a significant increase in renal vascular resistance (p = .0008 and .0005, respectively), a decrease in RBF/CO (p = .002 and .03), and a compensatory increase in the renal oxygen extraction ratio (p = .005 and .0001). RBFI and renal oxygen consumption remained constant throughout the epinephrine infusion profile. Neither epinephrine nor dopamine significantly affected creatinine clearance or urine output. Twelve patients (63%) were in established renal failure (plasma creatinine, >3 mg/dL) at the time of the study, although the presence or absence of renal failure did not significantly influence the effects of the study drugs. However, overall, the presence of renal failure was associated with a lower mean renal oxygen consumption, a lower mean renal oxygen consumption as a fraction of systemic oxygen consumption, and a higher mean renal vascular resistance. Although dopamine increased and epinephrine decreased fractional renal blood flow, there was no evidence that either drug produced either a beneficial or a deleterious effect on renal oxygen metabolism or function at any of the doses investigated.


Accurate post-fabrication trimming of ultra-compact resonators on silicon.
One of the challenges of the high refractive index contrast of silicon photonics platform is the high sensitivity of the resonance wavelength of resonators to dimensional variations caused by fabrication process variations. In this work, we have experimentally demonstrated an accurate post-fabrication trimming technique for optical devices that is robust to process variations. Using this technique, we have reduced the random variation of the resonance wavelength of 4 µm diameter resonators by a factor of 6 to below 50 pm. The level of accuracy achieved in this work is adequate for most of the RF-photonic, interconnect, and optical signal processing applications. We also discuss the throughput of this technique and its viability for wafer-scale post-fabrication trimming of silicon photonic chips.


The polymorphism of YWHAE, a gene encoding 14-3-3epsilon, and brain morphology in schizophrenia: a voxel-based morphometric study.
YWHAE is a possible susceptibility gene for schizophrenia that encodes 14-3-3epsilon, a Disrupted-in-Schizophrenia 1 (DISC1)-interacting molecule, but the effect of variation in its genotype on brain morphology remains largely unknown. In this voxel-based morphometric magnetic resonance imaging study, we conducted whole-brain analyses regarding the effects of YWHAE single-nucleotide polymorphisms (SNPs) (rs28365859, rs11655548, and rs9393) and DISC1 SNP (rs821616) on gray matter volume in a Japanese sample of 72 schizophrenia patients and 86 healthy controls. On the basis of a previous animal study, we also examined the effect of rs28365859 genotype specifically on hippocampal volume. Whole-brain analyses showed no significant genotype effect of these SNPs on gray matter volume in all subjects, but we found significant genotype-by-diagnosis interaction for rs28365859 in the left insula and right putamen. The protective C allele carriers of rs28365859 had a significantly larger left insula than the G homozygotes only for schizophrenia patients, while the controls with G allele homozygosity had a significantly larger right putamen than the C allele carriers. The C allele carriers had a larger right hippocampus than the G allele homozygotes in schizophrenia patients, but not in healthy controls. No significant interaction was found between rs28365859 and DISC1 SNP on gray matter volume. These different effects of the YWHAE (rs28365859) genotype on brain morphology in schizophrenia and healthy controls suggest that variation in its genotype might be, at least partly, related to the abnormal neurodevelopment, including in the limbic regions, reported in schizophrenia. Our results also suggest its specific role among YWHAE SNPs in the pathophysiology of schizophrenia.


The effect of drugs on the cardiovascular response to exercise.
Anti-hypertensives, diuretics, digitalis, beta-blockers, nitrates, anti-arrhythmics, and psychiatric drugs are reviewed for their effects on the cardiovascular response to exercise, in particular work capacity, heart rate, blood pressure, anginal threshold, and exertional arrhythmias. The limitations of these measurements to assess changes in the various aspects of cardiovascular performance in the presence of drugs becomes evident, emphasizing the need for an understanding of the relationship between drug effects on specific target organs and the altered integrated exercise response. The variable response of individuals to drug therapy, depending on the pathophysiological effects of their disease, demonstrates the value of exercise testing in selecting optimal drug regimens.


beta-TrCP binding and processing of NF-kappaB2/p100 involve its phosphorylation at serines 866 and 870.
Processing of the NF-kappaB2 precursor protein p100 is a major step in noncanonical NF-kappaB signaling. This signaling step requires the NF-kappaB inducing kinase (NIK) and its downstream kinase, IkappaB kinase alpha (IKKalpha). We show here that p100 undergoes phosphorylation at serines 866, 870, and possibly 872, in cells stimulated with noncanonical NF-kappaB stimuli or transfected with NIK and IKKalpha. Phosphorylation of this serine cluster creates a binding site for beta-TrCP, the receptor subunit of the beta-TrCP(SCF) ubiquitin ligase. Mutation of either serine 866 or serine 870 abolishes the beta-TrCP recruitment and ubiquitination of p100. The functional significance of p100 phosphorylation is further supported by the finding that this molecular event occurs in a NIK- and IKKalpha-dependent manner. Additionally, induction of p100 phosphorylation can be blocked by a protein synthesis inhibitor, suggesting the requirement of de novo protein synthesis. These data suggest that p100 processing involves its phosphorylation at specific terminal serines, which form a binding site for beta-TrCP thereby regulating p100 ubiquitination.


Post-Golgi traffic in plants.
Secretory and endocytic traffic through the post-Golgi endomembrane system regulates the abundance of plasma-membrane proteins such as receptors, transporters and ion channels, modulating the ability of a cell to communicate with its neighbours and to adapt to a changing environment. The major post-Golgi compartments are numerous and appear to be similar to their counterparts in animals. However, endosomes are rather ill defined morphologically but seem to be involved in specific trafficking pathways. Many plasma-membrane proteins cycle constitutively via endosomal compartments. The trans-Golgi network (TGN) appears to be an early endosome where secretory and endocytic traffic meet. Endocytosed proteins that are to be degraded are targeted to the vacuole via the multivesiculate prevacuolar compartment (PVC) whereas cycling proteins pass through recycling endosomes. The trafficking machinery involves the same classes of proteins as in other eukaryotes. However, there are modifications that match the specifics of post-Golgi traffic in plants. Although plants lack epithelia, some plasma-membrane proteins are located on specific faces of the cell which reflects polarized traffic and influences the physiological performance of the tissue. Plants also differentiate highly polarized tip-growing cells in which post-Golgi traffic is adapted to very high rates of targeted exocytosis, endocytosis and recycling.


Sex Differences in Countermovement Jump Phase Characteristics.
The countermovement jump (CMJ) is commonly used to explore sex differences in neuromuscular function, but previous studies have only reported gross CMJ measures or have partly examined CMJ phase characteristics. The purpose of this study was to explore differences in CMJ phase characteristics between male and female athletes by comparing the force-, power-, velocity-, and displacement-time curves throughout the entire CMJ, in addition to gross measures. Fourteen men and fourteen women performed three CMJs on a force platform from which a range of kinetic and kinematic variables were calculated via forward dynamics. Jump height (JH), reactive strength index modified, relative peak concentric power, and eccentric and concentric displacement, velocity, and relative impulse were all greater for men (g = 0.58⁻1.79). Relative force-time curves were similar between sexes, but relative power-, velocity-, and displacement-time curves were greater for men at 90%⁻95% (immediately before and after peak power), 47%⁻54% (start of eccentric phase) and 85%⁻100% (latter half of concentric phase), and 65%⁻87% (bottom of countermovement and initial concentric phase) of normalized jump time, respectively. The CMJ distinguished between sexes, with men demonstrating greater JH through applying a larger concentric impulse and, thus, achieving greater velocity throughout most of the concentric phase, including take-off.


Age-related response to an acute innate immune challenge in mice: proteomics reveals a telomere maintenance-related cost.
Ageing is characterized by the impairment of the acute innate immune response and the upregulation of low-grade inflammation, i.e. inflammaging. At the cellular level, telomeres are considered as a marker of biological ageing as their length is progressively eroded in the absence of repair mechanisms. However, the link between telomeres and inflammaging remains underexplored. We aimed to identify proteins that are differentially expressed between age classes in response to an acute inflammatory challenge. We challenged young (two months) and old (12 months) C57BL/6 mice using bacterial lipopolysaccharide (LPS) and measured telomere length and proteomic profiles in splenocytes. In total, 233 out of the 1966 proteins we quantified differed among experimental groups. A hierarchical clustering analysis revealed that nine of those 233 proteins were differently expressed among the experimental groups. Young mice responded to LPS by increasing the expression of proteins involved in the innate immune response, and interestingly, in telomere length maintenance. However, this regulation was impaired at older ages. These results are in agreement with the assumption that the strength of selection declines with age, potentially explaining the maintenance of costly, dysregulated, immune responses at old age. We suggest that the immune response is competing with the telomere maintenance process, highlighting how telomeres reflect the ageing trade-off even in a species where telomere length is not related to lifespan.


Functional characterization of aquaporin-4 specific T cells: towards a model for neuromyelitis optica.
Antibodies to the water channel protein aquaporin-4 (AQP4), which is expressed in astrocytic endfeet at the blood brain barrier, have been identified in the serum of Neuromyelitis optica (NMO) patients and are believed to induce damage to astrocytes. However, AQP4 specific T helper cell responses that are required for the generation of anti-AQP4 antibodies and most likely also for the formation of intraparenchymal CNS lesions have not been characterized. Using overlapping 15-meric peptides of AQP4, we identified the immunogenic T cell epitopes of AQP4 that are restricted to murine major histocompatibility complex (MHC) I-A(b). The N-terminal region of AQP4 was highly immunogenic. More precisely, the intracellular epitope AQP4(22-36) was detected as major immunogenic determinant. AQP4(82-108) (located in the second transmembrane domain), AQP4(139-153) (located in the second extracellular loop), AQP4(211-225) (located in the fifth transmembrane domain), AQP4(235-249) (located in the sixth transmembrane domain), as well as AQP4(289-306) in the intracellular C-terminal region were also immunogenic epitopes. AQP4(22-36) and AQP4(289-303) specific T cells were present in the natural T cell repertoire of wild type C57BL/6 mice and T cell lines were raised. However, active immunization with these AQP4 peptides did not induce signs of spinal cord disease. Rather, sensitization with AQP4 peptides resulted in production of IFN-γ, but also IL-5 and IL-10 by antigen-specific T cells. Consistent with this cytokine profile, the AQP4 specific antibody response upon immunization with full length AQP4 included IgG1 and IgG2, which are associated with a mixed Th2/Th1 T cell response. AQP4 is able to induce an autoreactive T cell response. The identification of I-A(b) restricted AQP4 specific T cell epitopes will allow us to investigate how AQP4 specific autoimmune reactions are regulated and to establish faithful mouse models of NMO that include both cellular and humoral responses against AQP4.


Disentangling the nature of the nicotine stimulus.
Learning involving interoceptive stimuli likely plays an important role in many diseases and psychopathologies. Within this area, there has been extensive research investigating the interoceptive stimulus effects of abused drugs. In this pursuit, behavioral pharmacologists have taken advantage of what is known about learning processes and adapted the techniques to investigate the behavioral and receptor mechanisms of drug stimuli. Of particular interest is the nicotine stimulus and the use of the two-lever operant drug discrimination task and the Pavlovian drug discriminated goal-tracking task. There is strong concordance between the two methods when using "standard" testing protocols that minimize learning on test days. For example, ABT-418, nornicotine, and varenicline all fully evoked nicotine-appropriate responding. Notably, research from our laboratory with the discriminated goal-tracking task has used an alternative testing protocol. This protocol assesses stimulus substitution based on how well extinction learning using a non-nicotine ligand transfers back to the nicotine stimulus. These findings challenge conclusions based on more "standard" testing procedures (e.g., ABT-418 is not nicotine-like). As a starting point, we propose Thurstone scaling as a quantitative method for more precisely comparing transfer of extinction across doses, experiments, and investigators. We close with a discussion of future research directions and potential implications of the research for understanding interoceptive stimuli.


Social, behavioral, and biological factors, and sex differences in mortality.
Few studies have examined whether sex differences in mortality are associated with different distributions of risk factors or result from the unique relationships between risk factors and mortality for men and women. We extend previous research by systematically testing a variety offactors, including health behaviors, social ties, socioeconomic status, and biological indicators of health. We employ the National Health and Nutritional Examination Survey III Linked Mortality File and use Cox proportional hazards models to examine sex diferences in adult mortality in the United States. Our findings document that social and behavioral characteristics are key factors related to the sex gap in mortality. Once we controlfor women's lower levels of marriage, poverty, and exercise, the sex gap in mortality widens; and once we control for women 's greater propensity to visit with friends and relatives, attend religious services, and abstain from smoking, the sex gap in mortality narrows. Biological factors-including indicators of inflammation and cardiovascular risk-also inform sex differences in mortality. Nevertheless, persistent sex differences in mortality remain: compared with women, men have 30% to 83% higher risks of death over the follow-up period, depending on the covariates included in the model. Although the prevalence of risk factors difers by sex, the impact of those risk factors on mortality is similar for men and women.


The evolution of host protection by vertically transmitted parasites.
Hosts are often infected by a variety of different parasites, leading to competition for hosts and coevolution between parasite species. There is increasing evidence that some vertically transmitted parasitic symbionts may protect their hosts from further infection and that this protection may be an important reason for their persistence in nature. Here, we examine theoretically when protection is likely to evolve and its selective effects on other parasites. Our key result is that protection is most likely to evolve in response to horizontally transmitted parasites that cause a significant reduction in host fecundity. The preponderance of sterilizing horizontally transmitted parasites found in arthropods may therefore explain the evolution of protection seen by their symbionts. We also find that protection is more likely to evolve in response to highly transmissible parasites that cause intermediate, rather than high, virulence (increased death rate when infected). Furthermore, intermediate levels of protection select for faster, more virulent horizontally transmitted parasites, suggesting that protective symbionts may lead to the evolution of more virulent parasites in nature. When we allow for coevolution between the symbiont and the parasite, more protection is likely to evolve in the vertically transmitted symbionts of longer lived hosts. Therefore, if protection is found to be common in nature, it has the potential to be a major selective force on host-parasite interactions.


Bacterial Leaf Spot of Zinnia Caused by Xanthomonas campestris pv. zinniae, a New Disease in Korea.
In September 2011, bacterial leaf spot was observed on zinnia plants (Zinnia elegans L.) grown in a garden in Suwon, Korea. Leaf symptoms included angular lesions that were yellow or brown-to-reddish brown in the center. Bacterial isolates (BC3293 to BC3299) were recovered on trypticase soy agar from lesions surface-sterilized in 70% ethyl alcohol for 1 min. Pathogenicity of the isolates was confirmed by spray inoculation with a bacterial suspension (106 CFU/ml) prepared in sterile distilled water and applied to zinnia plants at the four- to five-leaf growth stage (two plants per isolate). Sterile distilled water was used as the negative control. The inoculated plants were incubated in a greenhouse at 26 to 30°C and 95% relative humidity. Characteristic leaf spot symptoms developed on inoculated zinnia plants 5 days after inoculation. No symptoms were observed on the negative control plants. The bacterium reisolated from the inoculated leaves was confirmed through gyrB gene sequence analysis (3). All isolates were gram-negative, aerobic rods, each with a single flagellum. Isolates were positive for catalase and negative for oxidase. The biochemical and physiological tests for differentiation of Xanthomonas were performed using methods described by Shaad et al. (2). The isolates were positive for mucoid growth on yeast extract-dextrose-calcium carbonate agar, growth at 35°C, hydrolysis of starch and esculin, protein digestion, acid production from arabitol, and utilization of glycerol and melibiose. Colonies were negative for ice nucleation, and alkaline in litmus milk. The gyrB gene (870 bp) and the 16S-23S rRNA internal transcribed spacer (ITS) regions (884 bp) were sequenced to aid in identification of the original field isolates using published PCR primer sets Xgyr1BF/Xgyr1BR (3) and A1/B1 (1), respectively. Sequence of the gyrB gene (GenBank Accession Nos. JQ665732 to JQ665738) from the zinnia field isolates shared 100% sequence identity with the reference strain of Xanthomonas campestris pv. zinniae (GenBank Accession No. EU285210), and the ITS sequences (GenBank Accession Nos. JQ665725 to JQ665731) had 99.9% sequence identity with X. campestris pv. zinnia XCZ-1 (GenBank Accession No. EF514223). On the basis of the pathogenicity assays, biochemical and physiological tests, and sequence analyses, the isolates were identified as X. campestris pv. zinniae. To our knowledge, this is the first report of bacterial leaf spot of zinnia caused by X. campestris pv. zinniae in Korea. The disease is expected to result in economic and aesthetic losses to plants in Korean landscapes. Thus, seed treatment with bactericides will be required to control the bacterial leaf spot of zinnia before planting. References: (1) T. Barry et al. The PCR Methods Appl. 1:51, 1991. (2) N. W. Schaad et al. Page 189 in: Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. N. W. Schaad et al., eds. The American Phytopathological Society, St. Paul, MN, 2001. (3) J. M. Young et al. Syst. Appl. Microbiol. 31:366, 2008.


A mechanism for exocytotic arrest by the Complexin C-terminus.
ComplexinII (CpxII) inhibits non-synchronized vesicle fusion, but the underlying mechanisms have remained unclear. Here, we provide evidence that the far C-terminal domain (CTD) of CpxII interferes with SNARE assembly, thereby arresting tonic exocytosis. Acute infusion of a CTD-derived peptide into mouse chromaffin cells enhances synchronous release by diminishing premature vesicle fusion like full-length CpxII, indicating a direct, inhibitory function of the CTD that sets the magnitude of the primed vesicle pool. We describe a high degree of structural similarity between the CpxII CTD and the SNAP25-SN1 domain (C-terminal half) and show that the CTD peptide lowers the rate of SDS-resistant SNARE complex formation in vitro. Moreover, corresponding CpxII:SNAP25 chimeras do restore complexin's function and even 'superclamp' tonic secretion. Collectively, these results support a so far unrecognized clamping mechanism wherein the CpxII C-terminus hinders spontaneous SNARE complex assembly, enabling the build-up of a release-ready pool of vesicles for synchronized Ca2+-triggered exocytosis.


Reproductive aging patterns in primates reveal that humans are distinct.
Women rarely give birth after ∼45 y of age, and they experience the cessation of reproductive cycles, menopause, at ∼50 y of age after a fertility decline lasting almost two decades. Such reproductive senescence in mid-lifespan is an evolutionary puzzle of enduring interest because it should be inherently disadvantageous. Furthermore, comparative data on reproductive senescence from other primates, or indeed other mammals, remains relatively rare. Here we carried out a unique detailed comparative study of reproductive senescence in seven species of nonhuman primates in natural populations, using long-term, individual-based data, and compared them to a population of humans experiencing natural fertility and mortality. In four of seven primate species we found that reproductive senescence occurred before death only in a small minority of individuals. In three primate species we found evidence of reproductive senescence that accelerated throughout adulthood; however, its initial rate was much lower than mortality, so that relatively few individuals experienced reproductive senescence before death. In contrast, the human population showed the predicted and well-known pattern in which reproductive senescence occurred before death for many women and its rate accelerated throughout adulthood. These results provide strong support for the hypothesis that reproductive senescence in midlife, although apparent in natural-fertility, natural-mortality populations of humans, is generally absent in other primates living in such populations.


Global and country-specific mainstreaminess measures: Definitions, analysis, and usage for improving personalized music recommendation systems.
Popularity-based approaches are widely adopted in music recommendation systems, both in industry and research. These approaches recommend to the target user what is currently popular among all users of the system. However, as the popularity distribution of music items typically is a long-tail distribution, popularity-based approaches to music recommendation fall short in satisfying listeners that have specialized music preferences far away from the global music mainstream. Addressing this gap, the contribution of this article is three-fold. First, we provide several quantitative measures describing the proximity of a user's music preference to the music mainstream. Assuming that there is a difference between the global music mainstream and a country-specific one, we define the measures at two levels: relating a listener's music preferences to the global music preferences of all users, or relating them to music preferences of the user's country. To quantify such music preferences, we define a music item's popularity in terms of artist playcounts (APC) and artist listener counts (ALC). Moreover, we adopt a distribution-based and a rank-based approach as means to decrease bias towards the head of the long-tail distribution. This eventually results in a framework of 6 measures to quantify music mainstream. Second, we perform in-depth quantitative and qualitative studies of music mainstream in that we (i) analyze differences between countries in terms of their level of mainstreaminess, (ii) uncover both positive and negative outliers (substantially higher and lower country-specific popularity, respectively, compared to the global mainstream), analyzing these with a mixed-methods approach, and (iii) investigate differences between countries in terms of listening preferences related to popular music artists. We conduct our studies and experiments using the standardized LFM-1b dataset, from which we analyze about 800,000,000 listening events shared by about 53,000 users (from 47 countries) of the music streaming platform Last.fm. We show that there are substantial country-specific differences in listeners' music consumption behavior with respect to the most popular artists listened to. Third, we demonstrate the applicability of our study results to improve music recommendation systems. To this end, we conduct rating prediction experiments in which we tailor recommendations to a user's level of preference for the music mainstream using the proposed 6 mainstreaminess measures: defined by a distribution-based or rank-based approach, defined on a global level or on a country level (for the user's country), and for APC or ALC. Our approach roughly equals a hybrid recommendation approach in which a demographic filtering strategy is implemented before collaborative filtering is performed. Results suggest that, in terms of rating prediction accuracy, each of the presented mainstreaminess definitions has its merits.


Photoreceptor mitochondrial tyrosine nitration in experimental uveitis.
In experimental autoimmune uveitis (EAU), phagocytes are thought to be the primary cells in the initiation and maintenance of pathologic tissue damage through the release of cytotoxic agents. Recently, the presence of nitric oxide synthase has been shown in mammalian mitochondria. In this study, the effect of mitochondrial peroxynitrite on the modification of cellular proteins was evaluated in the early phase of uveitis, before the infiltration of leukocytes. Tyrosine nitration in proteins was detected by UV/Vis (visible) absorption and Western blot analysis. The identity of the nitrated protein was obtained by liquid chromatography-tandem mass spectrometry. The release of cytochrome c was assessed in whole retinal extract and in isolated mitochondria. The protein nitration in the inflamed retina was also localized by immunohistochemistry. Before the leukocyte infiltration in the early phase of EAU, the mitochondria-originated peroxynitrite initiated the inflammatory insult by specifically nitrating three mitochondrial proteins. In vitro nitration of the control retina by peroxynitrite donor resulted in nonspecific nitration of all major retinal proteins. After nitration, cytochrome c was displaced from its original binding site in the respiratory chain. Further, the nitration appeared to commence in the early phase of inflammation, on postimmunization day 5, long before the peak of inflammation on day 14. Immunohistochemically, tyrosine-nitrated proteins were localized exclusively in the photoreceptor inner segments, which are known to be densely populated with mitochondria. These data indicate that mitochondrial proteins are the prime targets of inactivation by the mitochondrial peroxynitrite and that photoreceptor mitochondria initiate the subsequent irreversible retinal damage in experimental uveitis.


Activation of fusiform face area by Greebles is related to face similarity but not expertise.
Some of the brain areas in the ventral temporal lobe, such as the fusiform face area (FFA), are critical for face perception in humans, but what determines this specialization is a matter of debate. The face specificity hypothesis claims that faces are processed in a domain-specific way. Alternatively, the expertise hypothesis states that the FFA is specialized in processing objects of expertise. To disentangle these views, some previous experiments used an artificial class of novel objects called Greebles. These experiments combined a learning and fMRI paradigm. Given the high impact of the results in the literature, we replicated and further investigated this paradigm. In our experiment, eight participants were trained for ten 1-hr sessions at identifying Greebles. We scanned participants before and after training and examined responses in FFA and lateral occipital complex. Most importantly and in contrast to previous reports, we found a neural inversion effect for Greebles before training. This result suggests that people process the "novel" Greebles as faces, even before training. This prediction was confirmed in a postexperimental debriefing. In addition, we did not find an increase of the inversion effect for Greebles in the FFA after training. This indicates that the activity in the FFA for Greebles does not depend on the degree of expertise acquired with the objects but on the interpretation of the stimuli as face-related.


Safety assessment of alkyl PEG ethers as used in cosmetics.
The CIR Expert Panel assessed the safety of Alkyl PEG Ethers as used in cosmetics. These ingredients primarily function in cosmetics as surfactants, and some have additional functions as skin-conditioning agents, fragrance ingredients, and emulsion stabilizers. The Panel reviewed available relevant animal and clinical data, as well as information from previous CIR reports; when data were not available for individual ingredients, the Panel extrapolated from the existing data to support safety. The Panel concluded that the Alkyl PEG ethers are safe as used when formulated to be nonirritating, and the same applies to future alkyl PEG ether cosmetic ingredients that vary from those ingredients recited herein only by the number of ethylene glycol repeat units.


Allergic Rhinitis and its Impact on Asthma (ARIA) Phase 4 (2018): Change management in allergic rhinitis and asthma multimorbidity using mobile technology.
Allergic Rhinitis and its Impact on Asthma (ARIA) has evolved from a guideline by using the best approach to integrated care pathways using mobile technology in patients with allergic rhinitis (AR) and asthma multimorbidity. The proposed next phase of ARIA is change management, with the aim of providing an active and healthy life to patients with rhinitis and to those with asthma multimorbidity across the lifecycle irrespective of their sex or socioeconomic status to reduce health and social inequities incurred by the disease. ARIA has followed the 8-step model of Kotter to assess and implement the effect of rhinitis on asthma multimorbidity and to propose multimorbid guidelines. A second change management strategy is proposed by ARIA Phase 4 to increase self-medication and shared decision making in rhinitis and asthma multimorbidity. An innovation of ARIA has been the development and validation of information technology evidence-based tools (Mobile Airways Sentinel Network [MASK]) that can inform patient decisions on the basis of a self-care plan proposed by the health care professional.


Priming of neutrophil oxidative burst in diabetes requires preassembly of the NADPH oxidase.
Hyperglycemia associated with diabetes mellitus results in the priming of neutrophils leading to oxidative stress that is, in part, responsible for diabetic complications. p47phox, a NADPH oxidase cytosolic subunit, is a key protein in the assembly of the NADPH oxidase leading to superoxide generation. Little is known about the priming mechanism of oxidative pathways in neutrophils of people with diabetes. In this study, the kinetics of p47phox activation was investigated by comparing neutrophils from diabetic and healthy subjects, and the mechanism of hyperglycemia-induced changes was studied by using neutrophil-like HL-60 cells as a model. In resting neutrophils from diabetic subjects, p47phox prematurely translocates to the cell membrane and preassembles with p22phox, a NADPH oxidase membrane subunit. This premature p47phox translocation and preassembly with p22phox were also observed in HL-60 cells cultured with high glucose (HG; 25 mM) and with the specific ligand for the receptor for advanced glycation end products (RAGE), S100B. Phosphorylation of ERK1/2, but not p38 MAPK, was the primary signaling pathway, as evidenced by PD98059 suppressing the translocation of p47phox in HL-60 cells incubated with HG and S100B. HL-60 cells cultured in HG and S100B exhibited a 1.8-fold increase in fMLP-induced superoxide generation compared with those cultured in normal glucose (5.5 mM). These data suggest that HG and increased AGE prime neutrophils and increase oxidative stress inducing the translocation of p47phox to the cell membrane and preassembly with p22phox by stimulating a RAGE-ERK1/2 pathway.


Lessons learned on lead poisoning in children: one-hundred years on from Turner's declaration.
There is significant emerging evidence showing life-long negative health, intellectual and socio-behavioural impacts as a result of childhood blood lead concentrations well below the widely used intervention level of 10 µg/dL. This issue raises serious health concerns for children in several Australian smelting and mining towns. Routine educational and home cleanliness advice to wet mop floors rather than to use a brush and pan to reduce lead exposure risks have been shown to have limited efficacy. This paper argues, as advocated 100 years ago by Queensland doctor Alfred Jefferis Turner, that childhood lead poisoning can only be mitigated via primary prevention and reduction of contaminants at source. Given that the effects of lead exposure are irreversible, there is a strong argument for the application of the precautionary principle to dealing with childhood lead exposure. There is a clear need to improve regulatory controls and emissions management to reduce environmental lead exposure risks.


Use of Noninvasive Ventilation with Volume-Assured Pressure Support in Neuralgic Amyotrophy with Bilateral Diaphragmatic Paralysis.
Neuralgic Amyotrophy (NA) is a rare, acute onset inflammatory brachial plexopathy that frequently presents with acute pain followed by shoulder girdle muscle weakness. Phrenic nerve involvement affecting the diaphragms occurs in 7-10% of cases. We present the case of a 52-year-old man with neuralgic amyotrophy with phrenic nerve involvement and bilateral diaphragmatic paralysis with marked respiratory symptoms and sleep hypoventilation, who was treated with non-invasive ventilation with volume assured pressure support mode. By 21 months post disease onset, the patient had experienced marked improvement in orthopnea, sleep quality and functional status. This is the first reported case of the use of this mode of noninvasive ventilation in neuralgic amyotrophy.


Does Short-Term High Dose Probiotic Supplementation Containing Lactobacillus casei Attenuate Exertional-Heat Stress Induced Endotoxaemia and Cytokinaemia?
The study aimed to determine if short-term high dose probiotic supplementation containing Lactobacillus casei (L.casei) attenuates the commonly reported exertional-heat stress (EHS) induced endotoxinaemia and cytokinaemia. Eight endurance trained male volunteers (mean± SD: age 26 ± 6 y, nude body mass 70.2 ± 8.8 kg, height 1.75 ± 0.05 m, VO2max 59 ± 5 ml·kg-1·min-1) completed a blinded randomized cross-over design, whereby oral ingestion of a commercially available probiotic beverage containing L.casei (volume equivalent for ×1011 colony forming units·day-1) (PRO) or placebo (PLA) was consumed for 7 consecutive days before exposure to EHS, which comprised of 2h running exercise at 60% VO2max in hot ambient conditions (34.0 °C and 32% RH). Blood samples were collected at baseline (7 days before EHS), pre-EHS, post-EHS (1 hr, 2 hr, 4 hr, and at 24 hr). Plasma samples were analyzed for gram-negative bacterial endotoxin, cytokine profile (IL-6, IL-1β, TNF-α, IFN-γ, IL-8, and IL-10) and plasma osmolality. Plasma osmolality did not differ between trials. Seven days of L.casei supplementation did not show significant changes in resting circulatory endotoxin concentration or plasma cytokine profile compared with PLA. A main effect of time was observed for IL-6, TNF-α, IL-10 and IL-8; whereby levels increased in response to EHS (p < .05). Relative to pre-EHS concentrations, higher plasma concentrations of endotoxin (p = .05), and a trend for higher plasma TNF-α concentration (p = .09) was observed on PRO compared with PLA throughout recovery. Short-term high dose supplementation of a probiotic beverage containing L.casei before EHS did not attenuate EHS induced endotoxaemia and cytokinaemia; nor is it more positively favorable over a placebo.


Using "mother trainers" for malaria control: the Nigerian experience.
Studies of care seeking in Nigeria show that a tremendous amount of treatment for malaria takes place at home and, in most instances, such treatments are incorrect. This deficiency is attributed to caregivers' poor knowledge of treatment. This study was designed to empower households to treat malaria correctly in partnership with community members. Selected mothers from study communities were trained as "mother trainers" and were expected to train other members of their communities using a treatment protocol. "Mother trainers" were acceptable to most communities and judged to be effective. They were enthusiastic and their participation in the study boosted their ego and status in the community. Drop-out-rate of "mother trainers" was 24.2%. A few limitations to the use of mothers as trainers that were identified are discussed. It is concluded that mothers have good potential to effectively carry out health education activities in the community if appropriately selected, trained, and supervised.


Epilysin (MMP-28) restrains early macrophage recruitment in Pseudomonas aeruginosa pneumonia.
Several members of the matrix metalloproteinase (MMP) family function in various processes of innate immunity, particularly in controlling leukocyte influx. Epilysin (MMP-28) is expressed in numerous tissues and, in adult mice, it has the highest expression in lung, where it is detected in bronchial epithelial cells (Clara cells). Epilysin is also expressed by bone marrow-derived macrophages, but not by alveolar macrophages, suggesting that its expression by macrophages is dependent on localization and differentiation. To assess the role of this MMP, we generated epilysin-null (Mmp28(-/-)) mice. Although epilysin is constitutively expressed in normal tissues, Mmp28(-/-) mice have no overt phenotype. However, using a murine model of Pseudomonas aeruginosa pneumonia, we found that Mmp28(-/-) mice had an early increase in macrophage recruitment into the lungs, as well as enhanced bacterial clearance and reduced pulmonary neutrophilia, which we predicted were due to accelerated macrophage influx. Macrophage depletion in WT and Mmp28(-/-) mice confirmed a role for macrophages in clearing P. aeruginosa and regulating neutrophil recruitment. Furthermore, we observed that macrophages derived from Mmp28(-/-) mice migrated faster than did wild-type cells to bronchoalveolar lavage fluid from P. aeruginosa-treated mice of either genotype. These observations indicate that epilysin functions as an intrinsic negative regulator of macrophage recruitment by retarding the chemotaxis of these cells.


Early phase cancer clinical trials: design, ethics and future directions.
The main role of early phase clinical trials in cancer is to determine the dose to take forward for future clinical study. However, study design is changing in order to account for the change in focus of drug development toward molecularly targeted agents.


ApoE production in human monocytes and its regulation by inflammatory cytokines.
The apoE production by tissue macrophages is crucial for the prevention of atherosclerosis and the aim of this study was to further elucidate how this apolipoprotein is regulated by cytokines present during inflammation. Here we studied apoE production in peripheral blood mononuclear cells (PBMC) and analysis was made with a newly developed apoE ELISpot assay. In PBMC, apoE secretion was restricted to monocytes with classical (CD14(++)CD16(-)) and intermediate (CD14(+)CD16(+)) monocytes being the main producers. As earlier described for macrophages, production was strongly upregulated by TGF-β and downregulated by bacterial lipopolysaccharide (LPS) and the inflammatory cytokines IFN-γ, TNF-α and IL-1β. We could here show that a similar down-regulatory effect was also observed with the type I interferon, IFN-α, while IL-6, often regarded as one of the more prominent inflammatory cytokines, did not affect TGF-β-induced apoE production. The TNF-α inhibitor Enbrel could partly block the down-regulatory effect of IFN-γ, IFN-α and IL-1β, indicating that inhibition of apoE by these cytokines may be dependent on or synergize with TNF-α. Other cytokines tested, IL-2, IL-4, IL-12, IL-13, IL-17A and IL-23, had no inhibitory effect on apoE production. In contrast to the effect on monocytes, apoE production by primary hepatocytes and the hepatoma cell line HepG2 was more or less unaffected by treatment with cytokines or LPS.