Influence of morphometric factors on quantitation of paracellular permeability of intestinal epithelia in vitro.
The relative contribution of the small and large intestine to paracellular absorption is a subject of some controversy. Direct comparison of paracellular permeability in different epithelia is complicated by variations in junctional density and/or the absorptive surface area. This study used a combination of morphometric analyses and in vitro absorption studies to define permeability characteristics in relation to the amount of paracellular pathway present in rat ileum, colon and the model epithelium, Caco-2. Mucosal to serosal amplification was higher in ileum (3.9) than colon (1.9) or Caco-2 (1). Tight junctional density (lp) of ileal crypts was approximately 3 fold greater (91 m/cm2) than that measured in ileal villi, colonic surface and crypt cells or Caco-2 monolayers (34-37 m/cm2). However, when the relative contributions of the crypts and villi was taken into account there was no significant difference in the mean lp per mucosal area for the three epithelia studied. Using these data to correct for morphometric differences the permeabilities of a range of small hydrophilic molecules (atenolol, D-PheAsp and PEG oligomers MW 282-634) was measured. Permeability of rat ileum and colon were virtually identical for all compounds studied. In contrast, Caco-2 monolayers showed a significantly lower permeability than intestinal tissues with the difference increasing markedly with molecular size. These studies suggest the importance of accounting for morphological variation when comparing the permeability characteristics of different epithelial systems.


Altered arousal response in infants exposed to cigarette smoke.
A failure of the arousal mechanism is a key feature in the apnoea theory for sudden infant death syndrome (SIDS). In infants studied at an age when the incidence of SIDS is highest, we evaluated whether in utero smoke exposed infants have altered arousal response to standardised auditory stimuli, and/or sleep pattern, as recorded on overnight complex sleep polysomnography. A standardised sequence of audiology stimuli was applied binaurally to 20 in utero smoke and non-smoke exposed infants aged 8-12 weeks during a rapid eye movement (REM) and NREM epoch, in a controlled (temperature, position, pacifier use, noise) sleep environment. Infants were monitored for 10-12 hours using complex sleep polysomnography. Five infants exposed to in utero tobacco smoke did not have behavioural arousal response, whereas all non-smoke exposed infants aroused during NREM (p = 0.016). There was, however, no difference in REM sleep, and the groups did not differ in routine overnight complex sleep polysomnography parameters. At the age when the incidence of SIDS is at its peak, infants of smoking mothers are less rousable than those of non-smoking mothers in NREM sleep; this may partly explain why such infants are more at risk of SIDS.


Toward an improved structural model of the frog-skin antimicrobial peptide esculentin-1b(1-18).
Antimicrobial peptides (AMPs) are found in various classes of organisms as part of the innate immune system. Despite high sequence variability, they share common features such as net positive charge and an amphipathic fold when interacting with biologic membranes. Esculentin-1b is a 46-mer frog-skin peptide, which shows an outstanding antimicrobial activity. Experimental studies revealed that the N-terminal fragment encompassing the first 18 residues, Esc(1-18), is responsible for the antimicrobial activity of the whole peptide, with a negligible toxicity toward eukaryotic cells, thus representing an excellent candidate for future pharmaceutical applications. Similarly to most of the known AMPs, Esc(1-18) is expected to act by destroying/permeating the bacterial plasma-membrane but, to date, its 3D structure and the detailed mode of action remains unexplored. Before an in-depth investigation on peptide/membranes interactions could be undertaken, it is necessary to characterize peptide's folding propensity in solution, to understand what is intrinsically due to the peptide sequence, and what is actually driven by the membrane interaction. Circular dichroism and nuclear magnetic resonance spectroscopy were used to determine the structure adopted by the peptide, moving from water to increasing amounts of trifluoroethanol. The results showed that Esc(1-18) has a clear tendency to fold in a helical conformation as hydrophobicity of the environment increases, revealing an intriguing amphipathic structure. The helical folding is adopted only by the N-terminal portion of the peptide, while the rest is unstructured. The presence of a hydrophobic cluster of residues in the C-terminal portion suggests its possible membrane-anchoring role.


The dependence of the growth rate and meat content of young boars on semen parameters and conception rate.
Boars have a decisive impact on the progress in pig production, however, there is no recent information about the optimal growth parameters during the rearing period for modern breed later used in artificial insemination (AI) stations. Therefore, the objective of the research was to conduct semen parameter and conception rate analyses on the basis of growth rate and meat content assessments made during the rearing of AI boars of different genotypes. The study was carried out between 2010 and 2014 and included 184 boars in five breed combinations: 46 Polish Large White, 50 Polish Landrace, 27 Pietrain, 36 Duroc×Pietrain and 25 Hampshire×Pietrain. Boars were qualified by daily gains and meat content assessment (between 170 and 210 days of life). A total number of 38 272 ejaculates were examined (semen volume (ml), spermatozoa concentration (×106 ml-1), total number of spermatozoa (×109) and number of insemination doses from one ejaculate (n)). The fertility was determined by the conception rate (%). Semen volume, spermatozoa concentration and conception rate (P<0.01), followed by the total number of spermatozoa and insemination doses (P<0.05) were characterized by the highest variability in relation to breed of boars. The effect of daily gains was reported for spermatozoa concentration, number of insemination doses, conception rate (all P<0.01) and total number of spermatozoa (P<0.05). The peak of growth for spermatozoa concentration, total number of spermatozoa, insemination doses and conception rate was achieved for 800 to 850 g gains. Meat content affected semen volume, number of insemination doses and conception rate (P<0.05). Rearing boars while maintaining daily gains at the 800 to 850 g level and 62.5% to 65% meat content helps AI stations to increase the efficiency and economic profitability, and the number of insemination doses to increase by up to 300 doses/boar within a year. The analyses of growth parameters may help increase the efficiency and economic viability of AI stations.


The role of the 2H4 molecule in the generation of suppressor function in Con A-activated T cells.
The molecular basis for the suppression generated in a concanavalin A (Con A)-activated T cell culture remains unknown. In this study, we have attempted to determine whether the 2H4 and 4B4 molecules on Con A-activated T cells play some role in the generation of suppression by such cells. We have shown that Con A-activated suppressor cells belong to the 2H4+ subset of T cells but not the 4B4+ (2H4-) subset. Con A-activated T cells exerted their optimal suppressor function on day 2 in culture, a time at which the expression of 2H4 on such cells was maximal and 4B4 was minimal. Furthermore, the stimulation of T cells with the higher concentration of Con A generated the stronger suppressor function. At the same time, both 2H4 expression and density were increased and 4B4 expression and density were decreased on such Con A-activated T cells. More importantly, the treatment of Con A-activated T cells with anti-2H4 antibody but not with anti-4B4, anti-TQ1, or anti-T4 antibodies can block the suppressor function of such cells. Taken together, the above results strongly suggest that the 2H4 molecule itself may be involved in the generation of suppressor function in Con A-activated T cells. The 2H4 antigen on such cells was shown to be comprised of 220,000 and 200,000 m.w. glycoproteins. Thus this study indicates that the 220,000 and 200,000 m.w. structure of the 2H4 molecule may itself play a crucial role in the generation of suppressor signals of Con A-activated cells.


Specialty supplement use and biologic measures of oxidative stress and DNA damage.
Oxidative stress and resulting cellular damage have been suggested to play a role in the etiology of several chronic diseases, including cancer and cardiovascular disease. Identifying factors associated with reduced oxidative stress and resulting damage may guide future disease-prevention strategies. In the VITamins And Lifestyle (VITAL) biomarker study of 209 persons living in the Seattle area, we examined the association between current use of several specialty supplements and oxidative stress, DNA damage, and DNA repair capacity. Use of glucosamine, chondroitin, fish oil, methylsulfonylmethane (MSM), coenzyme Q10 (CoQ10), ginseng, ginkgo, and saw palmetto was ascertained by a supplement inventory/interview, whereas the use of fiber supplements was ascertained by questionnaire. Supplements used by more than 30 persons (glucosamine and chondroitin) were evaluated as the trend across number of pills/week (non-use, <14 pills/week, 14+ pills/week), whereas less commonly used supplements were evaluated as use/non-use. Oxidative stress was measured by urinary 8-isoprostane and PGF2α concentrations using enzyme immunoassays (EIA), whereas lymphocyte DNA damage and DNA repair capacity were measured using the Comet assay. Multivariate-adjusted linear regression was used to model the associations between supplement use and oxidative stress/DNA damage. Use of glucosamine (Ptrend: 0.01), chondroitin (Ptrend: 0.003), and fiber supplements (P: 0.01) was associated with reduced PGF2α concentrations, whereas CoQ10 supplementation was associated with reduced baseline DNA damage (P: 0.003). Use of certain specialty supplements may be associated with reduced oxidative stress and DNA damage. Further research is needed to evaluate the association between specialty supplement use and markers of oxidative stress and DNA damage.


Optical imaging with phase-coded aperture.
Experimental results are shown for an integrated computational imaging system with a phase-coded aperture. A spatial light modulator works as a phase screen that diffracts light from a point object into a uniformly redundant array (URA). Excellent imaging results are achieved after correlation processing. The system has the same depth of field as a diffraction-limited lens. Potential applications are discussed.


How effective is school-based deworming for the community-wide control of soil-transmitted helminths?
The London Declaration on neglected tropical diseases was based in part on a new World Health Organization roadmap to "sustain, expand and extend drug access programmes to ensure the necessary supply of drugs and other interventions to help control by 2020". Large drug donations from the pharmaceutical industry form the backbone to this aim, especially for soil-transmitted helminths (STHs) raising the question of how best to use these resources. Deworming for STHs is often targeted at school children because they are at greatest risk of morbidity and because it is remarkably cost-effective. However, the impact of school-based deworming on transmission in the wider community remains unclear. We first estimate the proportion of parasites targeted by school-based deworming using demography, school enrolment, and data from a small number of example settings where age-specific intensity of infection (either worms or eggs) has been measured for all ages. We also use transmission models to investigate the potential impact of this coverage on transmission for different mixing scenarios. In the example settings <30% of the population are 5 to <15 years old. Combining this demography with the infection age-intensity profile we estimate that in one setting school children output as little as 15% of hookworm eggs, whereas in another setting they harbour up to 50% of Ascaris lumbricoides worms (the highest proportion of parasites for our examples). In addition, it is estimated that from 40-70% of these children are enrolled at school. These estimates suggest that, whilst school-based programmes have many important benefits, the proportion of infective stages targeted by school-based deworming may be limited, particularly where hookworm predominates. We discuss the consequences for transmission for a range of scenarios, including when infective stages deposited by children are more likely to contribute to transmission than those from adults.


Role of the Escherichia coli recombination hotspot, chi, in RecABCD-dependent homologous pairing.
Genetic recombination occurring in wild type Escherichia coli is stimulated at DNA sequences known as chi sites, 5'-GCTGGTGG-3'. In vitro, homologous pairing between duplex DNA substrates dependent upon the RecA, RecBCD, and SSB proteins is stimulated by the presence of a chi sequence in the donor linear double-stranded DNA. We show that this stimulation is due to two factors: 1) the enhanced production of chi-specific single-stranded DNA fragments and 2) their preferential use in the RecA protein-promoted pairing step. Furthermore, under conditions of limiting Mg2+ concentration, joint molecule formation does not occur, even though DNA unwinding and chi-specific single-stranded DNA fragment production are observed. Also, under these conditions, chi-specific fragments derived from both the upstream and downstream regions of the DNA strand containing chi and from cleavage of the non-chi-containing DNA strand are detected. Finally, the behavior of mutant RecBCD enzymes (RecBC*D and RecBCD not equal to) in this in vitro reaction is shown to parallel their in vivo phenotypes with respect to chi stimulation of recombination. Thus we suggest that, in addition to its ability to regulate the degradative activities of RecBCD enzyme, chi itself may be a preferred site for initiation of homologous pairing in this concerted process.


Complete Genome Sequence of Burkholderia sp. Strain RPE64, Bacterial Symbiont of the Bean Bug Riptortus pedestris.
We isolated Burkholderia symbiont strain RPE64 from the bean bug Riptortus pedestris. Analysis of the complete 6.96-Mb genome, which consists of three chromosomes and two plasmids, will facilitate further understanding of insect-microbe symbiosis and the development of pest-control technologies.


Codon usage bias covaries with expression breadth and the rate of synonymous evolution in humans, but this is not evidence for selection.
In numerous species, from bacteria to Drosophila, evidence suggests that selection acts even on synonymous codon usage: codon bias is greater in more abundantly expressed genes, the rate of synonymous evolution is lower in genes with greater codon bias, and there is consistency between genes in the same species in which codons are preferred. In contrast, in mammals, while nonequal use of alternative codons is observed, the bias is attributed to the background variance in nucleotide concentrations, reflected in the similar nucleotide composition of flanking noncoding and exonic third sites. However, a systematic examination of the covariants of codon usage controlling for background nucleotide content has yet to be performed. Here we present a new method to measure codon bias that corrects for background nucleotide content and apply this to 2396 human genes. Nearly all (99%) exhibit a higher amount of codon bias than expected by chance. The patterns associated with selectively driven codon bias are weakly recovered: Broadly expressed genes have a higher level of bias than do tissue-specific genes, the bias is higher for genes with lower rates of synonymous substitutions, and certain codons are repeatedly preferred. However, while these patterns are suggestive, the first two patterns appear to be methodological artifacts. The last pattern reflects in part biases in usage of nucleotide pairs. We conclude that we find no evidence for selection on codon usage in humans.


Principlism, medical individualism, and health promotion in resource-poor countries: can autonomy-based bioethics promote social justice and population health?
Through its adoption of the biomedical model of disease which promotes medical individualism and its reliance on the individual-based anthropology, mainstream bioethics has predominantly focused on respect for autonomy in the clinical setting and respect for person in the research site, emphasizing self-determination and freedom of choice. However, the emphasis on the individual has often led to moral vacuum, exaggeration of human agency, and a thin (liberal?) conception of justice. Applied to resource-poor countries and communities within developed countries, autonomy-based bioethics fails to address the root causes of diseases and public health crises with which individuals or communities are confronted. A sociological explanation of disease causation is needed to broaden principles of biomedical ethics and provides a renewed understanding of disease, freedom, medical practice, patient-physician relationship, risk and benefit of research and treatment, research priorities, and health policy.


MMP-13 is constitutively produced in human chondrocytes and co-endocytosed with ADAMTS-5 and TIMP-3 by the endocytic receptor LRP1.
Matrix metalloproteinase 13 (MMP-13) degrades collagenous extracellular matrix and its aberrant activity associates with diseases such as arthritis, cancer, atherosclerosis and fibrosis. The wide range of MMP-13 proteolytic capacity suggests that it is a powerful, potentially destructive proteinase and thus it has been believed that MMP-13 is not produced in most adult human tissues in the steady state. Present study has revealed that human chondrocytes isolated from healthy adults constitutively express and secrete MMP-13, but that it is rapidly endocytosed and degraded by chondrocytes. Both pro- and activated MMP-13 bind to clusters II and III of low-density lipoprotein (LDL) receptor-related protein 1 (LRP1). Domain deletion studies indicated that the hemopexin domain is responsible for this interaction. Binding competition between MMP-13 and ADAMTS-4, -5 or TIMP-3, which also bind to cluster II, further shown that the MMP-13 binding site within cluster II is different from those of ADAMTS-4, -5 or TIMP-3. MMP-13 is therefore co-endocytosed with ADAMTS-5 and TIMP-3 by human chondrocytes. These findings indicate that MMP-13 may play a role on physiological turnover of cartilage extracellular matrix and that LRP1 is a key modulator of extracellular levels of MMP-13 and its internalization is independent of the levels of ADAMTS-4, -5 and TIMP-3.


An illustrated key to the cuckoo wasps (Hymenoptera, Chrysididae) of the Nordic and Baltic countries, with description of a new species.
The Chrysididae are a group of cleptoparasitic and parasitoid aculeate wasps with a large number of rare and endangered species. The taxonomy of this group has long been confusing due to the similarity of species and extensive intraspecific variation. We present for the first time a comprehensive dichotomous key for all 74 species found in the Nordic and Baltic countries. In addition to diagnostic characters, information on the distribution and biology of each species is also presented. A new species, Chrysis borealis Paukkunen, Ødegaard & Soon, sp. n. is described on the basis of specimens collected from Fennoscandia. Chrysis gracillima Förster, 1853 is recorded as new to the Nordic and Baltic countries.


Levels of glutamatergic neurometabolites in patients with severe treatment-resistant schizophrenia: a proton magnetic resonance spectroscopy study.
Approximately 30% of patients with schizophrenia do not respond to antipsychotics and are thus considered to have treatment-resistant schizophrenia (TRS). To date, only four studies have examined glutamatergic neurometabolite levels using proton magnetic resonance spectroscopy (1H-MRS) in patients with TRS, collectively suggesting that glutamatergic dysfunction may be implicated in the pathophysiology of TRS. Notably, the TRS patient population in these studies had mild-to-moderate illness severity, which is not entirely reflective of what is observed in clinical practice. In this present work, we compared glutamate + glutamine (Glx) levels in the dorsal anterior cingulate cortex (dACC) and caudate among patients with TRS, patients with non-TRS, and healthy controls (HCs), using 3T 1H-MRS (PRESS, TE = 35 ms). TRS criteria were defined by severe positive symptoms (i.e., ≥5 on 2 Positive and Negative Syndrome Scale (PANSS)-positive symptom items or ≥4 on 3 PANSS-positive symptom items), despite standard antipsychotic treatment. A total of 95 participants were included (29 TRS patients [PANSS = 111.2 ± 20.4], 33 non-TRS patients [PANSS = 49.8 ± 13.7], and 33 HCs). dACC Glx levels were higher in the TRS group vs. HCs (group effect: F[2,75] = 4.74, p = 0.011; TRS vs. HCs: p = 0.012). No group differences were identified in the caudate. There were no associations between Glx levels and clinical severity in either patient group. Our results are suggestive of greater heterogeneity in TRS relative to non-TRS with respect to dACC Glx levels, necessitating further research to determine biological subtypes of TRS.


Malarone treatment failure and in vitro confirmation of resistance of Plasmodium falciparum isolate from Lagos, Nigeria.
We report the first in vitro and genetic confirmation of Malarone (GlaxoSmithKline; atovaquone and proguanil hydrochloride) resistance in Plasmodium falciparum acquired in Africa. On presenting with malaria two weeks after returning from a 4-week visit to Lagos, Nigeria without prophylaxis, a male patient was given a standard 3-day treatment course of Malarone. Twenty-eight days later the parasitaemia recrudesced. Parasites were cultured from the blood and the isolate (NGATV01) was shown to be resistant to atovaquone and the antifolate pyrimethamine. The cytochrome b gene of isolate NGATV01 showed a single mutation, Tyr268Asn which has not been seen previously.


A topical desiccant agent in association with ultrasonic debridement in the initial treatment of chronic periodontitis: a clinical and microbiological study.
Effective sub-gingival debridement is crucial to prevent serious systemic infections in hospitalized patients. Lack of compliance and the impracticality of repeated treatment in a short span of time are identified barriers to the performance of full mouth scaling and root planing (SRP). The aim of this randomized study was to evaluate the clinical and microbiological effects of the adjunctive administration of a locally delivered desiccant liquid with molecular hygroscopic properties (HYBENX® Oral Tissue Decontaminant™; HBX) in association with sub-gingival ultrasonic debridement (UD) in a hospital setting. Sixteen patients presenting moderate to severe chronic periodontitis were followed in a randomized 3 month, split mouth, single-blind, prospective study. At baseline (T1) control and test sides were treated with supra and subgingival UD with or without the association of a locally delivered desiccant liquid (HBX). Treatment was repeated after 6 weeks (T2). Clinical and microbiological parameters were assessed at T1, T2 and at 3 months (T3). The test group sites presented a significantly greater reduction in visible plaque index (VPI), bleeding on probing scores (BOP) and gingival index (GI) at T2 and T3 compared to the control group sites. HBX as monotherapy reached the same bacterial load reduction as UD. Compared to UD, a combined HBX-UD treatment resulted in a statistically significant greater bacterial load reduction immediately after treatment. A significantly lower anaerobic bacterial load was still present at T2. Data obtained show that decreased inflammatory signs and reduction of the bacterial load can be obtained in the short term by topical association of the desiccant agent HBX with UD.


Differences between immigrant and non-immigrant groups in the use of primary medical care; a systematic review.
Studies on differences between immigrant and non-immigrant groups in health care utilization vary with respect to the extent and direction of differences in use. Therefore, our study aimed to provide a systematic overview of the existing research on differences in primary care utilization between immigrant groups and the majority population. For this review PubMed, PsycInfo, Cinahl, Sociofile, Web of Science and Current Contents were consulted. Study selection and quality assessment was performed using a predefined protocol by 2 reviewers independently of each other. Only original, quantitative, peer-reviewed papers were taken into account. To account for this hierarchical structure, logistic multilevel analyses were performed to examine the extent to which differences are found across countries and immigrant groups. Differences in primary care use were related to study characteristics, strength of the primary care system and methodological quality. A total of 37 studies from 7 countries met all inclusion criteria. Remarkably, studies performed within the US more often reported a significant lower use among immigrant groups as compared to the majority population than the other countries. As studies scored higher on methodological quality, the likelihood of reporting significant differences increased. Adjustment for health status and use of culture-/language-adjusted procedures during the data collection were negatively related to reporting significant differences in the studies. Our review underlined the need for careful design in studies of differences in health care use between immigrant groups and the majority population. The results from studies concerning differences between immigrant and the majority population in primary health care use performed within the US might be interpreted as a reflection of a weaker primary care system in the US compared to Europe and Canada.


Multiplex amplicon sequencing for microbe identification in community-based culture collections.
Microbiome analysis using metagenomic sequencing has revealed a vast microbial diversity associated with plants. Identifying the molecular functions associated with microbiome-plant interaction is a significant challenge concerning the development of microbiome-derived technologies applied to agriculture. An alternative to accelerate the discovery of the microbiome benefits to plants is to construct microbial culture collections concomitant with accessing microbial community structure and abundance. However, traditional methods of isolation, cultivation, and identification of microbes are time-consuming and expensive. Here we describe a method for identification of microbes in culture collections constructed by picking colonies from primary platings that may contain single or multiple microorganisms, which we named community-based culture collections (CBC). A multiplexing 16S rRNA gene amplicon sequencing based on two-step PCR amplifications with tagged primers for plates, rows, and columns allowed the identification of the microbial composition regardless if the well contains single or multiple microorganisms. The multiplexing system enables pooling amplicons into a single tube. The sequencing performed on the PacBio platform led to recovery near-full-length 16S rRNA gene sequences allowing accurate identification of microorganism composition in each plate well. Cross-referencing with plant microbiome structure and abundance allowed the estimation of diversity and abundance representation of microorganism in the CBC.


Optical and SPION-enhanced MR imaging shows that trans-stilbene inhibitors of NF-κB concomitantly lower Alzheimer's disease plaque formation and microglial activation in AβPP/PS-1 transgenic mouse brain.
Alzheimer's disease (AD) is associated with a microglia-dependent neuroinflammatory response against plaques containing the fibrous protein amyloid-β (Aβ). Activation of microglia, which closely associate with Aβ plaques, engenders the release of pro-inflammatory cytokines and the internalization of Aβ fibrils. Since the pro-inflammatory transcription factor NF-κB is one of the major regulators of Aβ-induced inflammation, we treated transgenic amyloid-β protein protein/presenilin-1 (AβPP/PS1) mice for one year with a low dose (0.01% by weight in the diet) of either of two trans-stilbene NF-κB inhibitors, resveratrol or a synthetic analog LD55. The 3D distribution of Aβ plaques was measured ex vivo in intact brains at 60 μm resolution by quantitative magnetic resonance imaging (MRI) using blood-brain barrier-permeable, anti-AβPP-conjugated superparamagentic iron oxide nanoparticles (SPIONs). The MRI measurements were confirmed by optical microscopy of thioflavin-stained brain tissue sections and indicated that supplementation with either of the two trans-stilbenes lowered Aβ plaque density in the cortex, caudoputamen, and hippocampus by 1.4 to 2-fold. The optical measurements also included the hippocampus and indicated that resveratrol and LD55 reduced average Aβ plaque density by 2.3-fold and 3.1-fold, respectively. Ex vivo measurements of the regional distribution of microglial activation by Iba-1 immunofluorescence of brain tissue sections showed that resveratrol and LD55 reduced average microglial activation by 4.2- fold and 3.5-fold, respectively. Since LD55 lacked hydroxyl groups but both resveratrol and LD55 concomitantly reduced both Aβ plaque burden and neuroinflammation to a similar extent, it appears that the antioxidant potential of resveratrol is not an important factor in plaque reduction.


Long-term outcomes of induction chemotherapy and neoadjuvant stereotactic body radiotherapy for borderline resectable and locally advanced pancreatic adenocarcinoma.
Limited data are available to guide neoadjuvant treatment of borderline resectable (BRPC) and locally advanced (LAPC) pancreatic cancer. We updated our institutional outcomes with a neoadjuvant chemotherapy and stereotactic body radiotherapy (SBRT) approach. An IRB-approved analysis was performed of all BRPC and LAPC patients treated with our departmental treatment protocol. After staging, medically fit patients underwent chemotherapy for 2-3 months, with regimen at the discretion of the treating medical oncologist. Patients then received SBRT delivered in five consecutive daily fractions with median total radiation doses of 30 Gy to tumor and 40 Gy dose painted to tumor-vessel interfaces. This was followed by restaging imaging for possible resection. Overall survival (OS), event free survival (EFS), and locoregional control (LRC) rates were estimated and compared by Kaplan-Meier and log-rank methods. We identified 159 patients, 110 BRPC and 49 LAPC, with 14.0 months median overall follow-up. The resection and margin negative (R0) rate for BRPC patients who completed neoadjuvant therapy was 51% and 96%, respectively. Estimated median OS was 19.2 months for BRPC patients and 15.0 months for LAPC patients (p = 0.402). Median OS was 34.2 months for surgically resected patients versus 14.0 months for unresected patients (p < 0.001). Five of 21 (24%) LAPC patients receiving FOLFIRINOX chemotherapy underwent R0 resection. In LAPC, FOLFIRINOX recipients underwent R0 resection more often than other chemotherapy recipients (5 of 21 vs. 0 of 28, p = 0.011). There was a trend for improved survival in those resected LAPC patients (p = 0.09). For those not undergoing resection, one year LRC was 78%. Any grade ≥ 3 potentially radiation-related toxicity rate was 7%. These data underscore the feasibility, safety, and effectiveness of neoadjuvant SBRT and chemotherapy for BRPC and LAPC.


Helical computed tomographic portography in ten normal dogs and ten dogs with a portosystemic shunt.
Contrast enhanced helical computed tomography (CT) of the liver and portal system is routinely performed in human patients. The purpose of this project is to develop a practical protocol for helical CT portography in the dog. Ten clinically normal dogs were initially evaluated to develop a protocol. Using this protocol, ten dogs with confirmed portosystemic shunts (PSS) were then evaluated. Each patient was anesthetized, and a test dose of sodium iothalamate (400 mg I/ml) at 0.55 ml/kg was injected. Serial images were acquired at the level of T12-13 or T13-L1. The time to maximum enhancement of the portal vein was determined. This time period was used as the period between the second injection (2.2 ml/kg) and the start of the helical examination of the cranial abdomen. Delay times for normal dogs ranged from 34.5 s-66.0 s (median: 43.5 s) or 1.41 s/kg-4.12 s/kg (median: 2.09 s/kg). For patients with a PSS, the delay times were 16.5-70.5 s (median: 34.5 s) or 1.47-19.17 s/kg (median: 3.39 s/kg). The aorta, caudal vena cava, portal vein, shunt vessels, and their respective branches were well visualized on the CT images. Clinical case results were surgically confirmed. The surgeons reported that the information gained from the CT portography resulted in a subjective decrease in surgical time and degree of dissection necessary compared with similar surgeries performed without angiographic information. We believe that helical CT portography in the dog will be a useful adjunct in the diagnosis of PSS. The use of helical CT portography may allow clinicians to give clients a more accurate prognosis prior to surgery and will allow patients with lesions that are not surgically correctable to avoid a costly and invasive procedure.


Involvement of inducible nitric oxide synthase in radiation-induced vascular endothelial damage.
The use of radiation therapy has been linked to an increased risk of cardiovascular disease. To understand the mechanisms underlying radiation-induced vascular dysfunction, we employed two models. First, we examined the effect of X-ray irradiation on vasodilation in rabbit carotid arteries. Carotid arterial rings were irradiated with 8 or 16 Gy using in vivo and ex vivo methods. We measured the effect of acetylcholine-induced relaxation after phenylephrine-induced contraction on the rings. In irradiated carotid arteries, vasodilation was significantly attenuated by both irradiation methods. The relaxation response was completely blocked by 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, a potent inhibitor of soluble guanylate cyclase. Residual relaxation persisted after treatment with L-N(ω)-nitroarginine (L-NA), a non-specific inhibitor of nitric oxide synthase (NOS), but disappeared following the addition of aminoguanidine (AG), a selective inhibitor of inducible NOS (iNOS). The relaxation response was also affected by tetraethylammonium, an inhibitor of endothelium-derived hyperpolarizing factor activity. In the second model, we investigated the biochemical events of nitrosative stress in human umbilical-vein endothelial cells (HUVECs). We measured iNOS and nitrotyrosine expression in HUVECs exposed to a dose of 4 Gy. The expression of iNOS and nitrotyrosine was greater in irradiated HUVECs than in untreated controls. Pretreatment with AG, L-N(6)-(1-iminoethyl) lysine hydrochloride (a selective inhibitor of iNOS), and L-NA attenuated nitrosative stress. While a selective target of radiation-induced vascular endothelial damage was not definitely determined, these results suggest that NO generated from iNOS could contribute to vasorelaxation. These studies highlight a potential role of iNOS inhibitors in ameliorating radiation-induced vascular endothelial damage.


Wolbachia lipoprotein stimulates innate and adaptive immunity through Toll-like receptors 2 and 6 to induce disease manifestations of filariasis.
Wolbachia endosymbiotic bacteria have been implicated in the inflammatory pathogenesis of filariasis. Inflammation induced by Brugia malayi female worm extract (BMFE) is dependent on Toll-like receptors 2 and 6 (TLR2/6) with only a partial requirement for TLR1. Removal of Wolbachia, lipids, or proteins eliminates all inflammatory activity. Wolbachia bacteria contain the lipoprotein biosynthesis genes Ltg and LspA but not Lnt, suggesting Wolbachia proteins cannot be triacylated, accounting for recognition by TLR2/6. Lipoprotein databases revealed 3-11 potential lipoproteins from Wolbachia. Peptidoglycan-associated lipoprotein (PAL) and Type IV secretion system-VirB6 were consistently predicted, and B. malayi Wolbachia PAL (wBmPAL) was selected for functional characterization. Diacylated 20-mer peptides of wBmPAL (Diacyl Wolbachia lipopeptide (Diacyl WoLP)) showed a near identical TLR2/6 and TLR2/1 usage compared with BMFE and bound directly to TLR2. Diacyl WoLP induced systemic tumor necrosis factor-alpha and neutrophil-mediated keratitis in mice. Diacyl WoLP activated monocytes induce up-regulation of gp38 on human lymphatic endothelial cells and induced dendritic cell maturation and activation. Dendritic cells primed with BMFE generated a non-polarized Th1/Th2 CD4+ T cell profile, whereas priming with Wolbachia depleted extracts (following tetracycline treatment; BMFEtet) polarized to a Th2 profile that could be reversed by reconstitution with Diacyl WoLP. BMFE generated IgG1 and IgG2c antibody responses, whereas BMFEtet or inoculation of TLR2 or MyD88-/- mice produced defective IgG2c responses. Thus, in addition to innate inflammatory activation, Wolbachia lipoproteins drive interferon-gamma-dependent CD4+ T cell polarization and antibody switching.


The low molecular weight heparin Enoxaparin inhibits the consumption of factor VII and prothrombin activation in vivo associated with elective knee replacement surgery.
Patients over 40 years of age who undergo elective orthopaedic surgery have a relatively high risk for developing post-surgical deep vein thrombosis (DVT). Prophylactic use of heparin or low molecular weight heparins can reduce the incidence of post-operative DVT by up to 80%. It is not known whether prophylaxis is achieved by inhibition of prothrombin activation or catalysis of thrombin inhibition in vivo. We determined the changes in concentrations of factor VII zymogen and thrombin-antithrombin III (the latter as an index of prothrombin activation) in the plasmas of 129 patients randomized to receive two daily subcutaneous injections of placebo or 30 mg of Enoxaparin after elective knee surgery. Enoxaparin reduced the frequency of post-surgical DVT by 70%. The concentration of factor VII zymogen had decreased by approximately 50% within 24 h after the knee surgery, followed by a gradual increase to near presurgical values. Additionally, post-Enoxaparin plasmas had statistically significant higher concentrations of factor VII zymogen than post-placebo plasmas. Post-Enoxaparin plasmas had significantly lower concentrations of endogenous thrombin-antithrombin III than comparable post-placebo plasmas. Finally, post-Enoxaparin plasmas inactivated exogenous factor Xa and thrombin more effectively than comparable post-placebo plasmas. As Enoxaparin moderated the generation of endogenous thrombin-antithrombin III after elective knee surgery, inhibition of prothrombin activation in vivo by Enoxaparin may be important for its prophylactic antithrombotic effect.


Gesturing and naming: the use of functional knowledge in object identification.
Studies using functional imaging show reliable activation of premotor cortex when observers view manipulable objects. This result has led to the view that knowledge of object function, particularly the actions associated with the typical use of objects, may play a causal role in object identification. To obtain relevant evidence regarding this causal role, we asked subjects to learn gesture-color associations and then attempt to identify objects presented in colors denoting functional gestures that were congruent or incongruent with the objects' use. A strong congruency effect was observed when subjects gestured the use of an object, but not when they named an object. We conclude that our procedure constitutes a sensitive measure of the recruitment and causal role of functional knowledge and that this recruitment is not present during object naming. Preliminary evidence, however, indicates that gestures evoked by the volumetric shape of an object do contribute to object naming.


Indonesian mitochondrial DNA and its opposition to a Pleistocene era origin of proto-Polynesians in island southeast Asia.
The origin of modern Polynesians, the route of their expansion into the Pacific Ocean, and the timing of their movements all remain contentious topics in modern anthropology. Numerous studies have used molecular data to elucidate settlement patterns in the Indo-Pacific region, but the same evidence is often interpreted in opposing ways by different researchers. Above all, mitochondrial DNA (mtDNA) diversity has been used to discriminate between competing migration models and has narrowed the probable source of proto-Polynesian peoples to southern China and Taiwan or eastern Indonesia. Richards et al. (1998) used a dating method employing the p statistic to argue for an origin of Polynesian peoples in eastern Indonesia during the Pleistocene (> 10,000 years ago). Here, the time to the most recent common ancestor (TMRCA) is recalculated for a new series of Indonesian mtDNA sequences with Polynesian affinities. These data, which incorporate additional sequences published after 1998, produce dates that cannot rule out the possibility of a common ancestor for these sequences during the Holocene (< 10,000 years ago). This implies that previous estimates of TMRCA dates for Indonesian sequences lacked the statistical robustness necessary for replicability. The extant mtDNA evidence can no longer be viewed as favoring a Polynesian origin in eastern Indonesia, but instead remains consistent with an origin of proto-Polynesian peoples in southern China and Taiwan.


Mumps epidemiology in Cuba between 2004 and 2015.
Vaccination with the measles, mumps and rubella vaccine decreased the mumps incidence in Cuba, but in 2006 and 2007 an outbreak with more than 1000 laboratory confirmed cases occurred, mainly among high school and university students. The objective of the study was to investigate mumps epidemiology in Cuba between 2004 and 2015 and provide an in-depth laboratory characterization of selected samples from mumps patients. Samples from 116 cases (throat swabs, urines, paired acute and convalescent serum samples) were tested for mumps-specific IgM antibodies by ELISA, in a hemagglutination inhibition assay (HIA) or by RT-PCR. IgM antibodies were found in 80.2% of cases. 48.3% of first sera were positive, 30 of which were collected within two days after symptom onset. Testing of all 116 paired sera by HIA showed seroconversion in 55.2% individuals and an at least fourfold increase in antibodies in 44.8% of cases. In 18 out of the 111 vaccinated people (16.2%) no IgM antibodies were detected, neither in the acute nor the convalescent sera, but 14 of them showed seroconversion by HIA and 4 had an at least fourfold increase of hemagglutinin antibody titers. In the RT-PCR, 23 acute phase sera, 4 throat swabs and 5 urines were positive. Detection of mumps-specific IgM antibodies by ELISA and additional diagnostic methods may be required in settings with high vaccination coverage rates.


Determinants and patterns of service utilization and recourse to professionals for mental health reasons.
This study has a dual purpose: 1) identify determinants of healthcare service utilization for mental health reasons (MHR) in a Canadian (Montreal) catchment area; 2) determine the patterns of recourse to healthcare professionals in terms of frequency of visits and type of professionals consulted, and as it relates to the most prevalent mental disorders (MD) and psychological distress. Data was collected from a random sample of 1,823 individuals interviewed after a two-year follow-up period. A regression analysis was performed to identify variables associated with service utilization and complementary analyses were carried out to better understand participants' patterns of healthcare service utilization in relation to the most prevalent MD. Among 243 individuals diagnosed with a MD in the 12 months preceding an interview, 113 (46.5%) reported having used healthcare services for MHR. Determinants of service utilization were emotional and legal problems, number of MD, higher personal income, lower quality of life, inability of individuals to influence events occurring in their neighborhood, female gender and, marginally, lack of alcohol dependence in the past 12 months. Emotional problems were the most significant determinant of healthcare service utilization. Frequent visits with healthcare professionals were more likely associated with major depression and number of MD with or without dependence to alcohol or drugs. People suffering from major depression, psychological distress and social phobia were more likely to consult different professionals, while individuals with panic disorders relied on their family physician only. Concerning social phobia, panic disorders and psychological distress, more frequent visits with professionals did not translate into involvement of a higher number of professionals or vice-versa. This study demonstrates the impact of emotional problems, neighborhood characteristics and legal problems in healthcare service utilization for MHR. Interventions based on inter-professional collaboration could be prioritized to increase the ability of healthcare services to take care especially of individuals suffering from social phobia, panic disorders and psychological distress. Others actions that could be prioritized are training of family physicians in the treatment of MD, use of psychiatric consultants, internet outreach, and reimbursement of psychological consultations for individuals with low income.


The insulin-like growth factor II receptor gene is mutated in genetically unstable cancers of the endometrium, stomach, and colorectum.
Disruption of the DNA mismatch repair system, characterized by microsatellite instability (MI), plays an important role in the course of human carcinogenesis. Repetitive sequences constitute targets for mutation in MI+ cells, and frequent mutations have indeed been reported in such regions within the transforming growth factor beta receptor II (RII) gene in genetically unstable colorectal and gastric cancers. However, other genes that are targets for mutations in MI+ cells during the course of carcinogenesis have proven elusive. Because the insulin-like growth factor II receptor (IGFIIR) gene contains several repetitive sequences within its coding region, we examined mutations of this gene in MI+ cancers occurring at various primary sites. We found frameshift mutations in the poly(G)8 tract of IGFIIR in eight tumors, all of which were MI+: 4 of 26 (15%) MI+ endometrial cancers, 3 of 12 (25%) MI+ gastric cancers, and 1 of 18 (6%) MI+ colorectal cancers. In contrast, no mutation was found in 51 pancreatic cancers, 7 of which (14%) were MI+. These results implicate abnormal IGFIIR-mediated growth control in carcinogenesis involving the endometrium, stomach, and colorectum but not the pancreas.


The neuroanatomical organization of projection neurons associated with different olfactory bulb pathways in the sea lamprey, Petromyzon marinus.
Although there is abundant evidence for segregated processing in the olfactory system across vertebrate taxa, the spatial relationship between the second order projection neurons (PNs) of olfactory subsystems connecting sensory input to higher brain structures is less clear. In the sea lamprey, there is tight coupling between olfaction and locomotion via PNs extending to the posterior tuberculum from the medial region of the olfactory bulb. This medial region receives peripheral input predominantly from the accessory olfactory organ. However, the axons from olfactory sensory neurons residing in the main olfactory epithelium extend to non-medial regions of the olfactory bulb, and the non-medial bulbar PNs extend their axons to the lateral pallium. It is not known if the receptive fields of the PNs in the two output pathways overlap; nor has the morphology of these PNs been investigated. In this study, retrograde labelling was utilized to investigate the PNs belonging to medial and non-medial projections. The dendrites and somata of the medial PNs were confined to medial glomerular neuropil, and dendrites of non-medial PNs did not enter this territory. The cell bodies and dendrites of the non-medial PNs were predominantly located below the glomeruli (frequently deeper in the olfactory bulb). While PNs in both locations contained single or multiple primary dendrites, the somal size was greater for medial than for non-medial PNs. When considered with the evidence-to-date, this study shows different neuroanatomical organization for medial olfactory bulb PNs extending to locomotor control centers and non-medial PNs extending to the lateral pallium in this vertebrate.


The antioxidant protein peroxiredoxin 4 is epigenetically down regulated in acute promyelocytic leukemia.
The antioxidant peroxiredoxin (PRDX) protein family comprises 6 members, which are implicated in a variety of cellular responses, including growth factor signal transduction. PRDX4 resides in the endoplasmic reticulum (ER), where it locally controls oxidative stress by reducing H(2)O(2) levels. We recently provided evidence for a regulatory function of PRDX4 in signal transduction from a myeloid growth factor receptor, the granulocyte colony-stimulating factor receptor (G-CSFR). Upon activation, the ligand-induced G-CSFR undergoes endocytosis and routes via the early endosomes where it physically interacts with ER-resident PRDX4. PRDX4 negatively regulates G-CSFR mediated signaling. Here, we investigated whether PRDX4 is affected in acute myeloid leukemia (AML); genomic alterations and expression levels of PRDX4 were investigated. We show that genomic abnormalities involving PRDX4 are rare in AML. However, we find a strong reduction in PRDX4 expression levels in acute promyelocytic leukemia (APL) compared to normal promyelocytes and different molecular subtypes of AML. Subsequently, the possible role of DNA methylation and histone modifications in silencing of PRDX4 in APLs was investigated. We show that the reduced expression is not due to methylation of the CpG island in the promoter region of PRDX4 but correlates with increased trimethylation of histone 3 lysine residue 27 (H3K27me3) and lysine residue 4 (H3K4me3) at the transcriptional start site (TSS) of PRDX4, indicative of a bivalent histone code involved in transcriptional silencing. These findings suggest that the control of G-CSF responses by the antioxidant protein PRDX4 may be perturbed in APL.


Sky radiance and spectral gradient are orienting cues for the sandhopper Talitrus saltator (Crustacea, Amphipoda).
The sandhopper Talitrus saltator relies on both the sun and the moon compasses to return to the belt of damp sand on the beach in which it lives buried during the day. In addition to the sun, the gradient of radiance and the spectral distribution across the sky could provide directional information that T. saltator can potentially use to orient itself during the day even when the sun is not visible (e.g. cloudy sky). The scope of this work was (1) to determine the intensity levels of sky radiance that the sandhoppers use in their zonal recovery and (2) to investigate whether this species relies on the celestial spectral gradient in its zonal recovery. Sandhoppers were tested in the laboratory under artificial radiance or spectral gradients. Our results show that under an artificial sky simulating the natural radiance gradient on a cloudless day, sandhoppers orientated toward the correct seaward direction of their home beach; however, individuals lost their ability to use the intensity gradient as an orientation cue when the radiance was attenuated by at least 40%. Sandhoppers were also able to head in the correct seaward direction of their home beach at any time of the day by using the spectral gradient as their only source of visual orientation reference.


Case definitions for chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME): a systematic review.
To identify case definitions for chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME), and explore how the validity of case definitions can be evaluated in the absence of a reference standard. Systematic review. International. A literature search, updated as of November 2013, led to the identification of 20 case definitions and inclusion of 38 validation studies. Validation studies were assessed for risk of bias and categorised according to three validation models: (1) independent application of several case definitions on the same population, (2) sequential application of different case definitions on patients diagnosed with CFS/ME with one set of diagnostic criteria or (3) comparison of prevalence estimates from different case definitions applied on different populations. A total of 38 studies contributed data of sufficient quality and consistency for evaluation of validity, with CDC-1994/Fukuda as the most frequently applied case definition. No study rigorously assessed the reproducibility or feasibility of case definitions. Validation studies were small with methodological weaknesses and inconsistent results. No empirical data indicated that any case definition specifically identified patients with a neuroimmunological condition. Classification of patients according to severity and symptom patterns, aiming to predict prognosis or effectiveness of therapy, seems useful. Development of further case definitions of CFS/ME should be given a low priority. Consistency in research can be achieved by applying diagnostic criteria that have been subjected to systematic evaluation.


Effects of Age Discrimination on Self-perceptions of Aging and Cancer Risk Behaviors.
Almost one-third of older adults report experiencing age discrimination. We hypothesized sequential links between older adults' everyday experiences of age discrimination and future health behaviors related to cancer risk through self-perceptions of aging (SPA). Participants were community-dwelling respondents (age: 51-96 years) from the 2008, 2012, and 2014 waves of the Health and Retirement Study (N = 4,467). Generalized path models estimated the immediate and enduring effects of age discrimination in 2008 on proximal SPA in 2012 and distal health behaviors in 2014. Age discrimination was associated with lower positive SPA and higher negative SPA in 2012. The effect of age discrimination on physical activity, smoking, and drinking in 2014 was mediated by positive and negative SPA in 2012. Through subsequent SPA, those who experienced age discrimination in 2008 were less likely to engage in regular moderate physical activity, more likely to smoke, and less likely to drink more than 3 times per week in 2014. Analysis of change in positive and negative SPA showed the effect of age discrimination on physical activity to be mediated by change in positive, but not negative, SPA. The enduring effects of age discrimination were found through a reduction in positive SPA. Elevating positive SPA could be as important as reducing negative SPA for future health behaviors related to cancer risk.


The effects of socioeconomic status, race, and parenting on language development in early childhood.
The authors examined the associations between socioeconomic status (SES), race, maternal sensitivity, and maternal negative-intrusive behaviors and language development in a sample selected to reduce the typical confound between race and SES (n = 146). Mother-child interactions were observed at 12 and 24 months (coded by randomly assigned African American and European American coders); language abilities were assessed at 18, 24, 30, and 36 months. For receptive language, race was associated with ability level, and maternal sensitivity and negative-intrusive parenting were related to rate of growth. For expressive communication, race, SES, and maternal sensitivity were associated with rate of growth; race moderated the association between negative-intrusive parenting and rate of growth such that the relation was weaker for African American than for European American children. The results highlight the importance of sensitive parenting and suggest that the association between negative-intrusive parenting and language development may depend upon family context. Future work is needed concerning the race differences found, including examining associations with other demographic factors and variations in language input experienced by children, using culturally and racially validated indices of language development.


Gambling warning messages: The impact of winning and losing on message reception across a gambling session.
Gambling warning messages have been shown to lead to prevention and modification of risk-taking behaviors. Laboratory studies have shown messages can increase a player's knowledge about gambling specific risks, modify their gambling-related cognitive distortions, and even change play. In the present laboratory study, participants were randomly assigned to a winning or losing slot machine gambling experience where they either viewed periodic warning messages or not. It was hypothesized that those in the message conditions would place smaller bets, spend more time considering bets, and spend less time gambling than those in the control conditions. We also hypothesized participants would play differently across the contexts of winning or losing. The results showed those who received warning messages while winning made the fewest number of spins and did not speed up their bet rate over the course of play as much as those in other conditions. Players who received warning messages while losing decreased the size of their bets over the course of play compared to those who received messages while winning. Despite receiving warning messages, losing players did not decrease their number of spins or rate of betting. Winning or losing during slot machine play appears to have significant consequences on the impact of a warning message. Whereas a message to change gambling behavior may encourage a winning gambler to stop play, the same message for a losing player may lead to a small minimization in harm by helping them to decrease bet size, though not their rate of betting. (PsycINFO Database Record


Renal sodium-glucose cotransporter inhibition in the management of type 2 diabetes mellitus.
Hyperglycemia is the primary factor responsible for the microvascular, and to a lesser extent macrovascular, complications of diabetes. Despite this well-established relationship, approximately half of all type 2 diabetic patients in the US have a hemoglobin A1c (HbA1c) ≥7.0%. This is associated in part with the side effects, i.e., weight gain and hypoglycemia, of currently available antidiabetic agents and in part with the failure to utilize medications that reverse the basic pathophysiological defects present in patients with type 2 diabetes. The kidney has been shown to play a central role in the development of hyperglycemia by excessive production of glucose throughout the sleeping hours and enhanced reabsorption of filtered glucose by the renal tubules secondary to an increase in the threshold at which glucose spills into the urine. Recently, a new class of antidiabetic agents, the sodium-glucose cotransporter 2 (SGLT2) inhibitors, has been developed and approved for the treatment of patients with type 2 diabetes. In this review, we examine their mechanism of action, efficacy, safety, and place in the therapeutic armamentarium. Since the SGLT2 inhibitors have a unique mode of action that differs from all other oral and injectable antidiabetic agents, they can be used at all stages of the disease and in combination with all other antidiabetic medications.


Expression levels of MHC class I molecules are inversely correlated with promiscuity of peptide binding.
Highly polymorphic major histocompatibility complex (MHC) molecules are at the heart of adaptive immune responses, playing crucial roles in many kinds of disease and in vaccination. We report that breadth of peptide presentation and level of cell surface expression of class I molecules are inversely correlated in both chickens and humans. This relationship correlates with protective responses against infectious pathogens including Marek's disease virus leading to lethal tumours in chickens and human immunodeficiency virus infection progressing to AIDS in humans. We propose that differences in peptide binding repertoire define two groups of MHC class I molecules strategically evolved as generalists and specialists for different modes of pathogen resistance. We suggest that differences in cell surface expression level ensure the development of optimal peripheral T cell responses. The inverse relationship of peptide repertoire and expression is evidently a fundamental property of MHC molecules, with ramifications extending beyond immunology and medicine to evolutionary biology and conservation.


Cytokine networks in multiple sclerosis: lost in translation.
This review will discuss aspects of cytokine networks in neuroinflammatory diseases and attempt to provide some explanation for our failures and successes in translating preclinical data to benefit patients with multiple sclerosis (MS). We will discuss innate cytokines such as tumor necrosis factor alpha and interferon (IFN) beta and will then go on to cover recent findings on the role of interleukin-23 and the so-called T(H)17 cells and how they are implicated in the pathogenesis of neuroinflammation. Even though IFN-beta has been used for the treatment of MS for many years, it is only recently that the mechanistic underpinnings of the IFN-beta-mediated immune modulation was discovered in preclinical models. The timeline is at odds with the idea that preclinical data should shape the design of therapeutic strategies in the clinic. Conversely, the discovery of the so-called T(H)17 cells and their association with neuroinflammation has broken the dogma that IFN-gamma-producing T(H)1 cells have the exclusive capacity to invade and destroy the central nervous system tissue. So why then did a clinical trial targeting the T(H)17-promoting cytokine interleukin-23 fail? Preclinical studies using the animal models for MS have yielded promising results, but unfortunately the translation into the clinic is often disappointing. The reason for this may be the complex nature of the pathogenesis of autoimmune neuroinflammation, but more often an oversimplified interpretation of preclinical observations appears to hinder our progress.


Negative Hydration Expansion in ZrW_{2}O_{8}: Microscopic Mechanism, Spaghetti Dynamics, and Negative Thermal Expansion.
We use a combination of x-ray diffraction, total scattering, and quantum mechanical calculations to determine the mechanism responsible for hydration-driven contraction in ZrW_{2}O_{8}. The inclusion of H_{2}O molecules within the ZrW_{2}O_{8} network drives the concerted formation of new W─O bonds to give one-dimensional (─W─O─)_{n} strings. The topology of the ZrW_{2}O_{8} network is such that there is no unique choice for the string trajectories: the same local changes in coordination can propagate with a large number of different periodicities. Consequently, ZrW_{2}O_{8}·H_{2}O is heavily disordered, with each configuration of strings forming a dense aperiodic "spaghetti." This new connectivity contracts the unit cell via large shifts in the Zr and W atom positions. Fluctuations of the undistorted parent structure towards this spaghetti phase emerge as the key negative thermal expansion (NTE) phonon modes in ZrW_{2}O_{8} itself. The large relative density of NTE phonon modes in ZrW_{2}O_{8} actually reflects the degeneracy of volume-contracting spaghetti excitations, itself a function of the particular topology of this remarkable material.


Both TLR2 and TRIF contribute to interferon-β production during Listeria infection.
Synthesis of interferon-β (IFN-β) is an innate response to cytoplasmic infection with bacterial pathogens. Our recent studies showed that Listeria monocytogenes limits immune detection and IFN-β synthesis via deacetylation of its peptidoglycan, which renders the bacterium resistant to lysozyme degradation. Here, we examined signaling requirements for the massive IFN-β production resulting from the infection of murine macrophages with a mutant strain of L. monocytogenes, ΔpgdA, which is unable to modify its peptidoglycan. We report the identification of unconventional signaling pathways to the IFN-β gene, requiring TLR2 and bacterial internalization. Induction of IFN-β was independent of the Mal/TIRAP adaptor protein but required TRIF and the transcription factors IRF3 and IRF7. These pathways were stimulated to a lesser degree by wild-type L. monocytogenes. They operated in both resident and inflammatory macrophages derived from the peritoneal cavity, but not in bone marrow-derived macrophages. The novelty of our findings thus lies in the first description of TLR2 and TRIF as two critical components leading to the induction of the IFN-β gene and in uncovering that individual macrophage populations adopt different strategies to link pathogen recognition signals to IFN-β gene expression.


Active transport of the survival motor neuron protein and the role of exon-7 in cytoplasmic localization.
Spinal muscular atrophy (SMA) is a neurodegenerative disease caused by deletion and/or mutation of the survival motor neuron protein Gene (SMN1) that results in the expression of a truncated protein lacking the C terminal exon-7. Whereas SMN has been shown to be an important component of diverse ribonucleoprotein (RNP) complexes, its function in neurons is unknown. We hypothesize that the active transport of SMN may be important for neurite outgrowth and that disruption of exon-7 could impair its normal intracellular trafficking. SMN was localized in granules that were associated with cytoskeletal filament systems and distributed throughout neurites and growth cones. Live cell imaging of enhanced green fluorescent protein (EGFP)-SMN granules revealed rapid, bidirectional and cytoskeletal-dependent movements. Exon-7 was necessary for localization of SMN into the cytoplasm but was not sufficient for granule formation and transport. A cytoplasmic targeting signal within exon-7 was identified that could completely redistribute the nuclear protein D-box binding factor 1 into the cytoplasm. Neurons transfected with SMN lacking exon-7 had significantly shorter neurites, a defect that could be rescued by redirecting the exon-7 deletion mutant into neurites by a targeting sequence from growth-associated protein-43. These findings provide the first demonstration of cytoskeletal-based active transport of SMN in neuronal processes and the function of exon-7 in cytoplasmic localization. Such observations provide motivation to investigate possible transport defects or inefficiency of SMN associated RNPs in motor neuron axons in SMA.


Parenting as a "package deal": relationships, fertility, and nonresident father involvement among unmarried parents.
Fatherhood has traditionally been viewed as part of a "package deal" in which a father's relationship with his child is contingent on his relationship with the mother. We evaluate the accuracy of this hypothesis in light of the high rates of multiple-partner fertility among unmarried parents using the Fragile Families and Child Wellbeing Study, a recent longitudinal survey of nonmarital births in large cities. We examine whether unmarried mothers' and fathers' subsequent relationship and parenting transitions are associated with declines in fathers' contact with their nonresident biological children. We find that father involvement drops sharply after relationships between unmarried parents end. Mothers 'transitions into new romantic partnerships and new parenting roles are associated with larger declines in involvement than fathers' transitions. Declines in fathers' involvement following a mother's relationship or parenting transition are largest when children are young. We discuss the implications of our results for the well-being of nonmarital children and the quality of nonmarital relationships faced with high levels of relationship instability and multiple-partner fertility.


The analgesic efficacy of intrathecal D-Ala2-D-Leu5-enkephalin in cancer patients with chronic pain.
D-Ala-D-Leu-enkephalin (DADL) is a pentapeptide which, compared to morphine, preferentially binds to the delta receptor. We compared the analgesic and side effects of intrathecal (i.t.) DADL and i.t. morphine sulfate (MS) in 10 tolerant cancer patients with chronic pain at or below the T12 level who were receiving inadequate relief or unacceptable side effects from systemic opiates. These patients were given i.t. DADL and i.t. MS in a randomized, double-blind, cross-over study on separate days at least 1 day apart. I.t. DADL produced analgesia in all patients tested. Total pain relief was greater with DADL than MS in 6 patients, equal in 1 patient and less with DADL in 3. Side effects, most commonly drowsiness, were similar with both MS and DADL and suggest supraspinal effects by both drugs. At the doses given i.t. DADL produced effective pain relief in patients tolerant to systemic opiates although no significant difference in analgesic efficacy between MS and DADL was observed. Studies of the relative analgesic potency of i.t. DADL in man are necessary to fully assess its value in those patients tolerant to systemic or i.t. opiates.


Biomarkers of oxidative and nitrosative damage in Alzheimer's disease and mild cognitive impairment.
Alzheimer's disease (AD) is the most common type of dementia in the elderly. Products of oxidative and nitrosative stress (OS and NS, respectively) accumulate with aging, which is the main risk factor for AD. This provides the basis for the involvement of OS and NS in AD pathogenesis. OS and NS occur in biological systems due to the dysregulation of the redox balance, caused by a deficiency of antioxidants and/or the overproduction of free radicals. Free radical attack against lipids, proteins, sugars and nucleic acids leads to the formation of bioproducts whose detection in fluids and tissues represents the currently available method for assessing oxidative/nitrosative damage. Post-mortem and in-vivo studies have demonstrated an accumulation of products of free radical damage in the central nervous system and in the peripheral tissues of subjects with AD or mild cognitive impairment (MCI). In addition to their individual role, biomarkers for OS and NS in AD are associated with altered bioenergetics and amyloid-beta (Abeta) metabolism. In this review we discuss the main results obtained in the field of biomarkers of oxidative/nitrosative stress in AD and MCI in humans, in addition to their potential role as a tool for diagnosis, prognosis and treatment efficacy in AD.


The natural history of acute cough in children aged 0 to 4 years in primary care: a systematic review.
Professional and parental uncertainty regarding the natural history of cough and respiratory tract infection (R77) in pre-school children may in part be responsible for the high consultation, reconsultation, and antibiotic prescribing rates in this age group. The aim of the study was to review the evidence about the natural history of acute cough in children aged between 0 and 4 years presenting to primary care in terms of illness duration and complications. The study was a systematic review, with qualitative and quantitative data synthesis, of control and placebo arms of systematic reviews, randomised controlled trials (RCTs), and cohort studies set in primary care. Searches were done of MEDLINE (between 1966 and June 1998), EMBASE (between 1988 and September 1998), and the Cochrane Library databases, using the MeSH terms 'respiratory tract infection, 'cough, and 'bronchitis, and the textwords 'cough' 'bronchitis, and 'chest infection, limited to children aged between 0 and 4years, and English language articles. Eight RCTs and two cohort studies met the review criteria. At one week, 75% of children may have improved but 50% may be still coughing and/or have a nasal discharge. At two weeks up to 24% of children may be no better. Within two weeks of presentation, 12% of children may experience one or more complication, such as rash, painful ears, diarrhoea, vomiting, or progression to bronchitis/pneumonia. This review offers parents and clinicians more prognostic information about acute cough in pre-school children. Illness duration may be longer and complications higher than many parents and clinicians expect. This may help to set more realistic expectations of the illness and help parents to decide when and if to reconsult. This information may be useful to those designing patient information and self-help resources.


Characterization of hepatitis C virus resistance to grazoprevir reveals complex patterns of mutations following on-treatment breakthrough that are not observed at relapse.
A detailed analysis of hepatitis C virus (HCV) resistance-associated substitutions (RASs) is required to understand why people fail direct-acting antiviral therapies. This study was conducted to assess RASs in an analysis of 2 trials evaluating the second-generation NS3/4A protease inhibitor grazoprevir (GZR) in combination with peginterferon/ribavirin. From a total of 113 participants with HCV genotype 1 infection, RASs were evaluated in 25 patients who relapsed and 6 patients with on-treatment virologic breakthrough using consensus Sanger and clonal sequence analysis of NS3/NS4a genes, with in vitro testing of replicon mutants. Next-generation sequencing (NGS) was used in a subset of participants to assess minority variants and the evolution of the whole viral genome. Baseline RASs did not predict treatment failure. Relapse was most commonly associated with RASs at D168, although additional RASs (Y56, R155 and A156) were also detected, particularly in participants with on-treatment breakthrough. Treatment-emergent RASs usually reverted to wild-type (WT), suggesting these mutations were associated with a negative fitness cost (confirmed using in vitro assays). NGS was the most sensitive assay for the detection of minor variants. Significant viral sequence divergence (up to 5.9% codons) was observed across whole genomes in association with the acquisition and reversion of RASs. Relapse with GZR and peginterferon/ribavirin is commonly associated with single RASs in NS3 that generally revert to WT, while breakthrough follows more complex patterns of viral resistance. NGS suggests that large diverse pools of viral quasispecies that emerge with RASs facilitate rapid viral evolution.


Lower body negative pressure: a method to differentiate vasodilators in the intact rat.
1. In rats lower body negative pressure induced venous pooling in a reproducible manner. The response to LBNP in the pentobarbitone-anaesthetized rat appeared not to be influenced by the autonomic nervous system. 2. The hypotensive effect of arterial vasodilators was diminished at 5 mmHg LBNP, compared with effects at 3 mmHg, whereas the hypotensive effect of venous vasodilators was not changed at 5 mmHg LBNP, compared with that at 3 mmHg LBNP. 3. In the pithed rat dihydroergotamine caused venoconstriction. In the phenobarbitone-anaesthetized rat the hypotensive effect of venous vasodilators was decreased significantly by pretreatment with this drug, whereas it did not influence the hypotensive effect of arterial vasodilators. 4. Thus venodilatation by LBNP and venoconstriction by dihydroergotamine provide a method for differentiating arterial and venous sites of action of vasodilators in intact rats.


Excitatory sympathetic reflex in NaCl-sensitive spontaneously hypertensive rats.
We have previously demonstrated blunted reflex responses of lumbar sympathetic nerve activity during volume expansion in NaCl-sensitive spontaneously hypertensive rats maintained on basal (1% NaCl) diets compared with NaCl-resistant spontaneously hypertensive rats, Wistar-Kyoto rats, and Sprague-Dawley rats. The current study tested the hypothesis that chronic ingestion of a high (8%) NaCl diet further blunts cardiopulmonary reflex function in the NaCl-sensitive spontaneously hypertensive rat. After 3 weeks of a 1% or 8% NaCl diet, male rats of all four strains were instrumented with femoral arterial and venous cannulas and lumbar nerve recording electrodes at 10 weeks of age. Two days later, conscious rats were infused with whole blood to expand blood volume. NaCl-sensitive spontaneously hypertensive rats maintained on a 1% NaCl diet had blunted responses of nerve activity to acute volume expansion compared with control strains. NaCl-sensitive spontaneously hypertensive rats maintained on an 8% NaCl diet had increases in nerve activity responses to volume expansion. In a second experiment, the volume expansion protocol was repeated in anesthetized NaCl-sensitive spontaneously hypertensive rats that had been subjected to sinoaortic denervation after 3 weeks of a 1% or 8% NaCl diet. After sinoaortic denervation, an increase in nerve activity was again observed during volume expansion in animals fed the 8% NaCl diet. In animals fed the 1% NaCl diet, changes in nerve activity were variable. The excitatory response was significantly reduced after bilateral vagotomy. These studies suggest that blood pressure regulation in NaCl-sensitive spontaneously hypertensive rats is a complex interaction of excitatory and inhibitory sympathetic reflex systems that is altered by high dietary NaCl exposure.


Dental caries in diabetes mellitus: role of salivary flow rate and minerals.
This study was designed to evaluate the possible protective role of salivary factors like salivary flow rate and adequate level of calcium, phosphate, and fluoride in diabetes mellitus type 2 patients with dental caries. A total of 398 diabetes mellitus type 2 patients with dental caries and 395 age- and sex-matched non-diabetic subjects with dental caries were included as controls, all of whom gave informed consent. All subjects were divided into four groups according to their age. Decayed, missed, and filled teeth (DMFT) were scored to indicate the severity of dental caries. Saliva was collected, flow rate was noted, and calcium, phosphate, and fluoride were analyzed. The blood glucose, HbA1c, and DMFT indices were found to be significantly high in diabetic patients as compared to controls. The salivary flow rate, calcium, phosphate, and fluoride were found to be significantly low whereas no significant difference was found in salivary magnesium in patients as compared to controls. Optimum salivary flow rate is responsible for establishing protective environment against dental caries. Adequate level of salivary calcium, phosphate, and fluoride is also involved in significant deposition of these minerals in plaque, which greatly reduces the development of caries in the adjacent enamel of teeth.


Are T cells helpful for COVID-19: the relationship between response and risk.
The disease spectrum of coronavirus disease 2019 (COVID-19) ranges from no symptoms to multisystem failure and death. Characterization of virus-specific immune responses to severe acute respiratory coronavirus 2 (SARS-CoV-2) is key to understanding disease pathogenesis, but few studies have evaluated T cell immunity. In this issue of the JCI, Sattler and Angermair et al. sampled blood from subjects with COVID-19 and analyzed the activation and function of virus antigen-specific CD4+ T cells. T cells that failed to respond to peptides from the membrane, spike, or nucleocapsid proteins were more common in subjects who died. In those whose T cells had the capacity to respond, older patients with comorbidity had larger numbers of activated T cells compared with patients who had fewer risk factors, but these cells showed impaired IFN-γ production. This cross-sectional study relates activated T cell responses to patient risk factors and outcome. However, T cell response trajectory over the disease course remains an open question.


Screening of a large cohort of leber congenital amaurosis and retinitis pigmentosa patients identifies novel LCA5 mutations and new genotype-phenotype correlations.
This study was undertaken to investigate the prevalence of sequence variants in LCA5 in patients with Leber congenital amaurosis (LCA), early-onset retinal dystrophy (EORD), and autosomal recessive retinitis pigmentosa (arRP); to delineate the ocular phenotypes; and to provide an overview of all published LCA5 variants in an online database. Patients underwent standard ophthalmic evaluations after providing informed consent. In selected patients, optical coherence tomography (OCT) and fundus autofluorescence imaging were possible. DNA samples from 797 unrelated patients with LCA and 211 with the various types of retinitis pigmentosa (RP) were screened by Sanger sequence analysis of all LCA5 exons and intron/exon junctions. Some LCA patients were prescreened by APEX technology or selected based on homozygosity mapping. In silico analyses were performed to assess the pathogenicity of the variants. Segregation analysis was performed where possible. Published and novel LCA5 variants were collected, amended for their correct nomenclature, and listed in a Leiden Open Variation Database (LOVD). Sequence analysis identified 18 new probands with 19 different LCA5 variants. Seventeen of the 19 LCA5 variants were novel. Except for two missense variants and one splice site variant, all variants were protein-truncating mutations. Most patients expressed a severe phenotype, typical of LCA. However, some LCA subjects had better vision and intact inner segment/outer segment (IS/OS) junctions on OCT imaging. In two families with LCA5 variants, the phenotype was more compatible with EORD with affected individuals displaying preserved islands of retinal pigment epithelium. One of the families with a milder phenotype harbored a homozygous splice site mutation; a second family was found to have a combination of a stop mutation and a missense mutation. This is the largest LCA5 study to date. We sequenced 1,008 patients (797 with LCA, 211 with arRP) and identified 18 probands with LCA5 mutations. Mutations in LCA5 are a rare cause of childhood retinal dystrophy accounting for ∼2% of disease in this cohort, and the majority of LCA5 mutations are likely null. The LCA5 protein truncating mutations are predominantly associated with LCA. However, in two families with the milder EORD, the LCA5 gene analysis revealed a homozygous splice site mutation in one and a stop mutation in combination with a missense mutation in a second family, suggesting that this milder phenotype is due to residual function of lebercilin and expanding the currently known phenotypic spectrum to include the milder early onset RP. Some patients have remaining foveal cone structures (intact IS/OS junctions on OCT imaging) and remaining visual acuities, which may bode well for upcoming treatment trials.


The Acute Potentiating Effects of Heavy Sled Pulls on Sprint Performance.
This study examined the acute potentiating effects of heavy sprint-style sled pulls on sprint performance. Twenty-two experienced resistance-trained rugby athletes performed 2 heavy sprint-style sled pull training protocols on separate occasions using a randomized, crossover, and counterbalanced design. The protocols consisted of 2-baseline 15 m sprints followed by 15 m sprints at 4, 8, and 12 minutes after completing 15 and 7.5 m heavy sled pulls with loads of 75 and 150% body mass (respectively). A significantly faster (p ≤ 0.05) 15 m sprint time was observed at 12 minutes for the 75% body mass load. Small nonsignificant improvements (effect size [ES] = 0.22-0.33) in 5, 10, and 15 m sprint times were observed at 8 and 12 minutes after the 75% body mass sled pull. No significant changes were observed for any sprint time after the 150% body mass sled pull. Significant differences in the percentage of change in sprint times between the 2 sled pull conditions were observed at 4 (ES = 0.44-0.52), 8 (ES = 0.59), and 12 minutes (ES = 0.64). It would seem that the 75% body mass sled pull can be an effective preload stimulus for improving subsequent sprint performance provided that adequate recovery (8-12 minutes) is allowed. Practitioners should be advised that prescription of training load based on decrement in sprint velocity may be the best approach to determine loading for athletes.


Evaluation of a 5-day education programme in type 1 diabetes: achieving individual targets with a patient-centred approach.
To evaluate if a single inpatient education training programme can achieve individualized therapeutic targets. Patients with Type 1 diabetes participating in a flexible intensive therapy programme were consecutively included in a prospective monocentric study. They all participated in the same education programme which had a patient-centred approach. Before the intervention, patients were divided into three groups according to their main therapeutic target: Group 1, to decrease HbA1c concentration in patients with baseline HbA1c ≥ 58 mmol/mol (7.5%); Group 2, to improve quality of life and satisfaction with treatment in patients with baseline HbA1c < 58 mmol/mol (7.5%); and Group 3, to decrease the frequency of hypoglycaemic episodes in patients with severe or frequent hypoglycaemic episodes. Therapeutic targets were evaluated at 12 months. Quality of life and treatment satisfaction were evaluated with validated questionnaires completed at baseline and 6 months. In Group 1 (n = 74), the mean ± sd HbA1c concentration decreased from 75 ± 15 mmol/mol (9.0 ±1.4%) to 68 ±15 mmol/mol (8.4 ± 1.4%; P < 0.001), with 53% of patients experiencing a decrease in HbA1c concentration of at least 6 mmol/mol (0.5%), without weight gain or more frequent hypoglycaemia. In Group 2 (n = 12), patient satisfaction with treatment improved significantly (P < 0.0001). In Group 3 (n = 35), minor hypoglycaemia significantly decreased from a mean ± sd of 6.6 ± 4.7 to 3.2 ± 3.0 hypoglycaemic episodes/week (P < 0.001) and the incidence of severe hypoglycaemia dropped significantly from a mean ± sd of 2.31 ± 3.07 to 0.86 ± 2.46 episodes/patient/year (P < 0.001). Many patients with different needs, who attended the same flexible intensive therapy education programme, which had a patient-centred approach, were able to achieve their individual therapeutic targets.


Dimensions of Reproductive Attitudes and Knowledge Related to Unintended Childbearing Among U.S. Adolescents and Young Adults.
Measures of attitudes and knowledge predict reproductive behavior, such as unintended fertility among adolescents and young adults. However, there is little consensus as to the underlying dimensions these measures represent, how to compare findings across surveys using different measures, or how to interpret the concepts captured by existing measures. To guide future research on reproductive behavior, we propose an organizing framework for existing measures. We suggest that two overarching multidimensional concepts-reproductive attitudes and reproductive knowledge-can be applied to understand existing research using various measures. We adapt psychometric analytic techniques to analyze two data sets: the National Longitudinal Survey of Adolescent to Adult Health (Add Health) and the Relationship Dynamics and Social Life study (RDSL). Although the specific survey measures and sample composition of the two data sets are different, the dimensionality of the concepts and the content of the items used to measure their latent factors are remarkably consistent across the two data sets, and the factors are predictive of subsequent contraceptive behavior. However, some survey items do not seem strongly related to any dimension of either construct, and some dimensions of the two concepts appear to be poorly measured with existing survey questions. Nonetheless, we argue that the concepts of reproductive attitudes and reproductive knowledge are useful for categorizing and analyzing social psychological measures related to unintended fertility. The results can be used to guide secondary data analyses to predict reproductive behavior, compare results across data sets, and structure future data collection efforts.


Metastatic parathyroid carcinoma in the MEN2A syndrome.
We report a patient with a metastatic parathyroid carcinoma and medullary carcinoma of the thyroid. This patient represents a variation of the multiple endocrine neoplasia syndrome (MEN) type 2A. There was no evidence of a phaeochromocytoma. The case illustrates the difficulties that may be encountered in localising the source of PTH secretion; the patient underwent four unsuccessful exploratory operations of the neck and mediastinum before further investigations revealed a single metastatic deposit of parathyroid carcinoma involving the first thoracic vertebra. PCR amplification and sequencing of the RET oncogene from the metastatic parathyroid carcinoma and genomic DNA revealed a heterozygous mutation (Cys634Tyr) in exon 11, as has previously been described to occur in MEN 2A. In addition, loss of tumour heterozygosity was demonstrated at loci from chromosomes 1, 2, 3p, 13q and 16p. This represents the first report of a parathyroid carcinoma in a MEN2A patient, in which the multiple allelic deletions are consistent with the generalised losses observed in aggressive tumours.


In vivo characterization and dynamic receptor occupancy imaging of TPA023B, an alpha 2/alpha 3/alpha 5 subtype selective gamma-aminobutyric acid-a partial agonist.
A novel, high-affinity (.7-2.0 nmol) compound that selectively activates the alpha2, alpha 3, and alpha 5 (but not alpha1) gamma-aminobutyric acid-A (GABA(A)) receptor subtypes, TPA023B (2',6-difluoro-5'-[3-(1-hydroxy-1-methylethyl) imidazo[1,2-b][1,2,4]triazin-7-yl][1,1'-biphenyl]-2-carbonitrile) was pharmacologically characterized and studied by means of positron emission tomography (PET) to determine dynamic occupancies of the benzodiazepine binding site of human brain GABA(A) receptors after a single oral dose. Four healthy male volunteers were studied in a double-blind, randomized placebo-controlled study of which three were given a single dose of 1.5 mg TPA023B and the fourth received placebo. The time course of GABA(A) receptor occupancy was determined with multiple dynamic [(11)C]flumazenil PET studies at pre-dose baseline and 5 and 24 hours after dose. Arterial sampling and full kinetic modeling with a two-compartment model was used to calculate parametric maps of receptor availability (distribution volume V(T)) and of occupancy. The GABA(A) receptor occupancy as determined from [(11)C]flumazenil V(T) values in all brain regions was reduced homogeneously, on average by 52.5 +/- 1.2% after 5 hours and 46.4 +/- 6.0% after 24 hours. No serious adverse events were encountered in humans. Single oral doses of 1.5 mg of TPA023B correspond to average receptor occupancies in neocortical regions of 52% and 46% after 5 and 24 hours, respectively. Provided suitable ligands and quantification methods are available for the appropriate target, quantitative PET offers a unique tool for dynamic in vivo measurement of relevant on-site receptor occupancy.


PAX immunoreactivity identifies alveolar rhabdomyosarcoma.
PAX5 is a member of the paired box transcription factors involved in development and its expression has been well characterized among hematopoietic malignancies of B-cell lineage. Its expression has also been reported in a subset of neuroendocrine carcinomas, urothelial tumors, Merkel cell carcinoma, glioblastoma, and neuroblastoma cell lines. As such, we sought to assess it as a diagnostic marker in the evaluation of pediatric small round blue cell tumors. Tumors selected for evaluation included embryonal rhabdomyosarcoma (55 cases), alveolar rhabdomyosarcoma (ARMS) (51 cases), neuroblastoma (22 cases), Wilms tumor (18 cases), Ewing Family of Tumors (11 cases), lymphoblastic lymphoma (8 cases), hepatoblastoma (6 cases), and granulocytic sarcoma (3 cases) as either cores in a tissue microarray or whole mount sections. All cases were immunostained using an antibody directed toward PAX5 and immunoreactivity was scored semiquantitatively according to percentage of nuclear staining. As expected, all B-cell lymphoblastic lymphomas were strongly immunoreactive against PAX5. Additionally, all Wilms tumors showed staining of variable intensity, most intensely in the epithelial component. Of the rhabdomyosarcoma cases, 34 of 51 (67%) ARMS were immunoreactive whereas none of the 55 embryonal rhabdomyosarcoma cases stained. No other tumor type on the array was immunoreactive toward PAX5. Genetic information was available on 7 ARMS, 5 of which had characteristic translocations involving PAX genes, either t(2:13) or t(1;13). Of the translocation-positive cases, all showed nuclear reactivity toward PAX5, and both the translocation-negative cases did not. Possible explanations of PAX5 staining include aberrant expression of the PAX5 transcription factor, PAX5 expression in normal tissue at the time the tumors most closely recapitulates in development or crossreactivity with another member of the PAX family. PAX3 and PAX7 fusion genes characterize the majority of ARMS making crossreactivity with these proteins an attractive theory, and suggest that PAX5 immunoreactivity may be specific for translocation-positive ARMS. Further study in a larger series of rhabdomyosarcomas is warranted to assess the sensitivity and specificity of PAX5 immunoreactivity for the ARMS variant.


Clinical diagnosis of acute bacterial rhinosinusitis, typical of experts.
Clinical diagnosis of acute bacterial sinusitis (ABS) is a concern when a patient presents with nasal discharge of recent onset together with facial pain or pressure. Given this presentation, the doctor would benefit from having access to software that specifies, first, what diagnostic indicators experts typically use in that diagnosis and then, upon entry of those facts, what experts' typical probability of ABS is in such a case. We specified a set of 23 hypothetical presentations of this type by patients 20-75 years of age, involving a comprehensive set of clinical-diagnostic indicators. Members of an international expert panel independently set the probability of ABS in each of these cases. A logistic function of the diagnostic indicators was fitted to the medians of the probabilities. The fitting led to an expression of the experts' median probability of ABS as a joint function of the duration of the patient's facial pain/pressure, and indicators of the location(s) of this; indicators of exacerbation of the pain/pressure on bending forward, nasal obstruction, maxillary and/or frontal tenderness, pus from middle meatus, purulent postnasal drip, and fever; and indicators of recent upper respiratory tract infection, nasal polyposis and status post sinus surgery. This probability function is accessible at http://www.evimed.ch/ABS. That probability function, made readily accessible, provides for expertly probability setting in clinical diagnosis of ABS, relevant for decisions about further diagnostics or treatment without further tests.


Challenges Confronting the Practice of Nursing in Singapore.
Singapore, a young nation like many developed countries, faced a shortage of nurses. Attempts to resolve the workforce shortage through the employment of foreign nurses started in the mid-1980s. Over the years, workforce recruitment from traditional sources nearby, namely Malaysia and Philippines, has expanded to include nurses from countries such as People Republic of China, India, and Myanmar. Attempts have also been made to train, recruit, and retain local nurses such as improving working conditions and remunerations, raising the profile of nursing, improving career recognition and progression, and encouraging nonpracticing nurses back to the workforce. However, the institutions' and the government's attempts to ameliorate the nursing shortage were met with limited success. Even with the recruitment of foreign nurses, the shortage of workforce persists. The shortage is compounded by the three major health-care challenges confronting Singapore: (1) rapid growth in population; (2) rapid aging of the population; and (3) increasing burden of chronic diseases. As the population continues to grow and as more of the population ages, as life expectancy increases and the burden of chronic diseases increases, not only will the demand for nurses continue, but the intensity and the nursing care they require will also increase. This article describes the challenges confronting the practice of nursing in Singapore and their implications. Although these challenges are daunting, they offer nursing the unprecedented opportunities to shape health-care delivery systems and increase nursing influences everywhere across settings and along the delivery continuum.