Zic2 mutation causes holoprosencephaly via disruption of NODAL signalling.
The ZIC2 transcription factor is one of the genes most commonly mutated in Holoprosencephaly (HPE) probands. Studies in cultured cell lines and mice have shown a loss of ZIC2 function is the pathogenic mechanism but the molecular details of this ZIC2 requirement remain elusive. HPE arises when signals that direct morphological and fate changes in the developing brain and facial primordia are not sent or received. One critical signal is sent from the prechordal plate (PrCP) which develops beneath the ventral forebrain. An intact NODAL signal transduction pathway and functional ZIC2 are both required for PrCP establishment. We now show that ZIC2 acts downstream of the NODAL signal during PrCP development. ZIC2 physically interacts with SMAD2 and SMAD3, the receptor activated proteins that control transcription in a NODAL dependent manner. Together SMAD3 and ZIC2 regulate FOXA2 transcription in cultured cells and Zic2 also controls the foxA2 expression during Xenopus development. Variant forms of the ZIC2 protein, associated with HPE in man or mouse, are deficient in their ability to influence SMAD-dependent transcription. These findings reveal a new mechanism of NODAL signal transduction in the mammalian node and provide the first molecular explanation of how ZIC2 loss-of-function precipitates HPE.


[Analysis of water fluoride concentration in Curitiba, Brazil: comparison of techniques].
To evaluate the adequacy of fluoride levels in the public water system in Curitiba, state of Paraná, Brazil, as determined by two techniques (colorimetric and electrometric). Data from independent measurements of fluoride in the public water system in Curitiba routinely performed by the city government were obtained for the period between January 2000 and July 2008. Mean levels of fluoride concentration were calculated for each of these years. After that, fluoride concentrations measured in 1 470 samples by the state water utility (SANEPAR) using the electrometric technique in 2006 and 2007 were compared with the corresponding levels measured by the city using the colorimetric method. The rate of samples meeting the standard for the city (0.8 ppmF), and below and above the standard, was calculated for both methods. Fluoride levels were compared between sanitary districts, months for the period between December 2007 and July 2008, and water treatment facilities. The overall mean fluoride level between 2000 and 2008 was 0.7 ppmF based on the independent measurements. The comparison between techniques showed a higher mean fluoride level with the electrometric technique (0.743 ppmF ± 0.133) vs. the colorimetric technique (0.637 ppmF ± 0.164). The rate of samples meeting the ideal standard of 0.8 ppmF was 15.05% for the colorimetric and 63.97% for the electrometric technique; 62.03% and 22.85% of the samples were below that standard and 21.10% and 13.18% were above that standard, respectively. Fluoride levels were statistically significant (P < 0.001) for the comparison between sanitary districts and months. The choice of technique significantly influences the resulting levels of fluoride. Independent monitoring of fluoride levels should employ the same technique used by the water utility. Further studies should aim at defining which technique is the most adequate to determine fluoride concentration in public water systems.


Chronic condition and risk behaviours in Portuguese adolescents.
Living with a chronic condition (CC) in adolescence has been historically considered protective for risk behaviours. However, research from the last decade suggest that when compared with healthy peers, adolescents living with a chronic condition can engage in risky behaviours in a similar if not higher rate than their counterparts living with out a CC. This study aims to characterize and evaluate the impact of 1) living with a chronic condition (CC), and 2) how the perception of living with a CC affects school participation, and its association with risk/protective behaviours (drunkenness, physical fight, sadness and self-harm). For this purpose 4 groups were identified: adolescents with mostly healthy behaviours, adolescents with mostly risk behaviours, adolescents with mostly risk-internalizing behaviours and adolescents with mostly risk-externalizing behaviours. A large sample was included in this study, composed by 3494 Portuguese adolescents with an average age of 15 years, who participated in the Portuguese Survey of Health Behaviour in School-aged Children/WHO (HBSC). Main results show that adolescents living with a CC have more risk-internalizing behaviours when compared to adolescents without CC, who present more healthy behaviors. Furthermore, adolescents that report that having a CC affects school participation show more risky behaviours than those not affected by a CC who present more healthy behaviours. Boys with a CC show more healthy behaviours, and those who feel that the CC affects school participation present more risky behaviours. On the other hand, girls with a CC have more risk-internalizing behaviours and less healthy behaviours It is important to point out that adolescents living with a CC represent a vulnerable group, and may engage in experimental/risky behaviours as likely as their non CC peers. Thus, potential benefits can arise from reinforcing interventions within protective contexts (family/peers/school setting). Health/education professionals, more than considering risk behaviours as dangerous in themselves, should offer adolescents with a CC an opportunity to reflect on their own decisions. Educational programs would benefit from looking at risk behaviors more from an experimentation perspective, focusing on constructive ways to help adolescents with CC to proceed into adulthood in a more appropriate developmental way.


Randomized study of vaginal misoprostol (PGE(1)) and dinoprostone gel (PGE(2)) for induction of labor at term.
To investigate the efficacy and safety of misoprostol in the induction of labor at term by comparing this agent with the commonly used dinoprostone gel. A randomized clinical trial of vaginal misoprostol, 50 microg 6-hourly, and dinoprostone gel, 1-2 mg 6-hourly, in 435 women undergoing induction of labor at term. The women, 210 in the misoprostol group and 225 in the dinoprostone group, were compared to determine whether there was a significant difference in achieving vaginal delivery within 24 h, the incidence of hyperstimulation syndrome, Cesarean section rate and adverse neonatal outcome. They were also offered the option of preinduction sonographic cervical assessment. Misoprostol, compared to dinoprostone gel, was associated with a significantly shorter median induction-to-delivery interval (14.6 h vs. 19.0 h; P = 0.0014), a higher incidence of vaginal delivery within 24 h of induction (65.7% vs. 54.2%; P = 0.019) and a reduced need for oxytocin augmentation during labor (20.5% vs. 29.8%; P = 0.034). The groups did not differ significantly in the rates of Cesarean section (18.1% vs. 19.1%; P = 0.88) and hyperstimulation syndrome (2.4% vs. 0.9%; P = 0.27). None of the cases of hyperstimulation required treatment with tocolysis. All nine cases of excessive uterine contractility occurred after the first dose of the drug. There were no significant differences in maternal and neonatal morbidity between the two groups. There was a significant association between preinduction cervical length and the induction-to-delivery interval in both those receiving misoprostol and those treated with dinoprostone. The use of misoprostol is associated with a shorter duration of labor and a higher rate of vaginal delivery within 24 h from induction without an increase in maternal and neonatal morbidity. Transvaginal sonographic measurement of cervical length is useful in the prediction of the likelihood of vaginal delivery within 24 h of induction and of the induction-to-delivery interval and may be useful in the stratification of patients participating in randomized studies that examine the effectiveness of inducing agents.


The component polypeptide chains of bovine insulin nucleate or inhibit aggregation of the parent protein in a conformation-dependent manner.
Amyloid fibrils are typically rigid, unbranched structures with diameters of approximately 10 nm and lengths up to several micrometres, and are associated with more than 20 diseases including Alzheimer's disease and type II diabetes. Insulin is a small, predominantly alpha-helical protein consisting of 51 residues in two disulfide-linked polypeptide chains that readily assembles into amyloid fibrils under conditions of low pH and elevated temperature. We demonstrate here that both the A-chain and the B-chain of insulin are capable of forming amyloid fibrils in isolation under similar conditions, with fibrillar morphologies that differ from those composed of intact insulin. Both the A-chain and B-chain fibrils were found to be able to cross-seed the fibrillization of the parent protein, although these reactions were substantially less efficient than self-seeding with fibrils composed of full-length insulin. In both cases, the cross-seeded fibrils were morphologically distinct from the seeding material, but shared common characteristics with typical insulin fibrils, including a very similar helical repeat. The broader distribution of heights of the cross-seeded fibrils compared to typical insulin fibrils, however, indicates that their underlying protofilament hierarchy may be subtly different. In addition, and remarkably in view of this seeding behavior, the soluble forms of the A-chain and B-chain peptides were found to be capable of inhibiting insulin fibril formation. Studies using mass spectrometry suggest that this behavior might be attributable to complex formation between insulin and the A-chain and B-chain peptides. The finding that the same chemical form of a polypeptide chain in different physical states can either stimulate or inhibit the conversion of a protein into amyloid fibrils sheds new light on the mechanisms underlying fibril formation, fibril strain propagation and amyloid disease initiation and progression.


High-dose inhaled steroids in asthmatics: moderate efficacy gain and suppression of the hypothalamic-pituitary-adrenal (HPA) axis. Research Council of the Norwegian Thoracic Society.
We wanted to evaluate the improvement in efficacy when increasing the daily dose of inhaled steroids and to compare the efficacy, safety, and tolerance of 1.6 mg beclomethasone dipropionate (BDP) with that of 2.0 mg fluticasone propionate (FP). The study was a randomized, double-blind, 3 month, multicentre study. One hundred and thirty four asthmatics currently using inhaled steroids (0.4-1.6 mg BDP or budesonide (BUD)) were stratified according to pretrial daily steroid use. Within each stratum they were randomized to either 1.6 mg BDP or 2.0 mg FP. A significant increase in the primary efficacy variables, i.e. mean morning and evening peak expiratory flow (PEF) (approximately 20 l.min-1) during the treatment period, was found for both treatments. No significant differences between the drugs were revealed for these primary or any other secondary efficacy variables (use of beta 2-agonists, symptom scores, and PEF, forced vital capacity (FVC), forced expiratory volume in one second (FEV1) recorded at the clinical visits). However, significant differences between treatments occurred regarding decrease of serum cortisol and adrenocorticotropic hormone. We conclude that, although both treatments gave statistically significant increases in efficacy parameters when compared with baseline, the increases were so small that they can be regarded as being clinically unimportant. Daily doses of BDP, 1.6 mg, and FP, 2.0 mg, had comparable effects on lung function. A suppression of the hypothalamic pituitary adrenal (HPA) axis was only found with a daily dose of 2 mg FP.


Classification and Genomic Diversity of Enterically Transmitted Hepatitis Viruses.
Hepatitis A virus (HAV) and hepatitis E virus (HEV) are significant human pathogens and are responsible for a substantial proportion of cases of severe acute hepatitis worldwide. Genetically, both viruses are heterogeneous and are classified into several genotypes that differ in their geographical distribution and risk group association. There is, however, little evidence that variants of HAV or HEV differ antigenically or in their propensity to cause severe disease. Genetically more divergent but primarily hepatotropic variants of both HAV and HEV have been found in several mammalian species, those of HAV being classified into eight species within the genus Hepatovirus in the virus family Picornaviridae. HEV is classified as a member of the species Orthohepevirus A in the virus family Hepeviridae, a species that additionally contains viruses infecting pigs, rabbits, and a variety of other mammalian species. Other species (Orthohepevirus B-D) infect a wide range of other mammalian species including rodents and bats.


Do all anti-citrullinated protein/peptide antibody tests measure the same? Evaluation of discrepancy between anti-citrullinated protein/peptide antibody tests in patients with and without rheumatoid arthritis.
Different methods exist to demonstrate anti-citrullinated protein/peptide antibodies (ACPA). To evaluate discrepancy between four ACPA tests. Population 1 consisted of patients with a new diagnostic problem, including 86 patients with rheumatoid arthritis (RA) and 450 patients without RA. Population 2 consisted of 155 patients with RA who had long-standing disease. Population 3 consisted of 188 patients with psoriatic arthritis and in population 4 there were 192 patients with systemic lupus erythematosus. Populations 1 and 2 were tested with the anti-human fibrinogen antibody (AhfibA) test, anti-CCP2 from Eurodiagnostica (CCP2-euro), anti-CCP2 from Pharmacia (CCP2-phar) and anti-CCP3 test by Inova (CCP3). Samples were annotated as discrepant if positive in one and negative in at least one other test. Each discrepant sample was re-analysed in a different run. Populations 3 and 4 were analysed in the CCP2-euro and AhFibA test. In population 1, ACPA positivity was found in 17 of 450 (3.8%) patients without RA; 14 (82%) of these 17 samples were discrepant. In contrast, 61 of 86 (70.9%) patients with RA were ACPA positive of whom 18 of 61 (29.5%) were discrepant (70.9% vs. 29.5%, p<0.001). The discrepancies between tests could be partly attributed to borderline results, inter-assay discrepancy and inter-test variability. They were more prevalent in patients with systemic lupus erythematosus who were ACPA positive than in those with psoriatic arthritis who were ACPA positive. Discrepancy between different ACPA tests was observed attributable to the occurrence of borderline results, inter-assay variability and mainly to inter-test variability. The lowest inter-test discrepancy is observed between tests that use the same substrate.


Comparison of Proteomic Measurements Across Platforms in the Atherosclerosis Risk in Communities (ARIC) Study.
The plasma proteome can be quantified using different types of highly multiplexed technologies, including aptamer-based and proximity-extension immunoassay methods. There has been limited characterization of how these protein measurements correlate across platforms and with absolute measures from targeted immunoassays. We assessed the comparability of (a) highly multiplexed aptamer-based (SomaScan v4; Somalogic) and proximity-extension immunoassay (OLINK Proseek® v5003; Olink) methods in 427 Atherosclerosis Risk in Communities (ARIC) Study participants (Visit 5, 2011-2013), and (b) 18 of the SomaScan protein measurements against targeted immunoassays in 110 participants (55 cardiovascular disease cases, 55 controls). We calculated Spearman correlations (r) between the different measurements and compared associations with case-control status. There were 417 protein comparisons (366 unique proteins) between the SomaScan and Olink platforms. The average correlation was r = 0.46 (range: -0.21 to 0.97; 79 [19%] with r ≥ 0.8). For the comparison of SomaScan and targeted immunoassays, 6 of 18 assays (growth differentiation factor 15 [GDF15], interleukin-1 receptor-like 1 [ST2], interstitial collagenase [MMP1], adiponectin, leptin, and resistin) had good correlations (r ≥ 0.8), 2 had modest correlations (0.5 ≤ r < 0.8; osteopontin and interleukin-6 [IL6]), and 10 were poorly correlated (r < 0.5; metalloproteinase inhibitor 1 [TIMP1], stromelysin-1 [MMP3], matrilysin [MMP7], C-C motif chemokine 2 [MCP1], interleukin-10 [IL10], vascular cell adhesion protein 1 [VCAM1], intercellular adhesion molecule 1 [ICAM1], interleukin-18 [IL18], tumor necrosis factor [TNFα], and visfatin) overall. Correlations for SomaScan and targeted immunoassays were similar according to case status. There is variation in the quantitative measurements for many proteins across aptamer-based and proximity-extension immunoassays (approximately 1/2 showing good or modest correlation and approximately 1/2 poor correlation) and also for correlations of these highly multiplexed technologies with targeted immunoassays. Design and interpretation of protein quantification studies should be informed by the variation across measurement techniques for each protein.


4D CT angiography more closely defines intracranial thrombus burden than single-phase CT angiography.
In patients with acute stroke, the location and extent of intravascular thrombi correlate with clinical and imaging outcomes and have been used to predict the success of intravenous thrombolysis. We hypothesized that 4D-CTA reconstructed from whole-brain CTP more closely outlines intracranial thrombi than conventional single-phase CTA. Sixty-seven patients with anterior circulation occlusion were retrospectively analyzed. For 4D-CTA, temporal maximum intensity projections were calculated that combine all 30 spiral scans of the CTP examination through temporal fusion. Thrombus extent was assessed by a semi-quantitative clot burden score (0-10; in which 0 = complete unilateral anterior circulation occlusion and 10 = patent vasculature). In patients with sufficient collateral flow, the length of the filling defect and corresponding hyperdense middle cerebral artery sign on NCCT were measured. Clot burden on temporal maximum intensity projection (median clot burden score, 7.0; interquartile range, 5.1-8.0) was significantly lower than on single-phase CT angiography (median, 6.0; interquartile range, 4.5-7.0; P < .0001). The length of the hyperdense middle cerebral artery sign (14.30 ± 5.93 mm) showed excellent correlation with the filling defect in the middle cerebral artery on temporal maximum intensity projection (13.40 ± 6.40 mm); this filling defect was larger on single-phase CT angiography (18.08 ± 6.54 mm; P = .043). As the result of an increased sensitivity for collateral flow, 4D-CTA temporal maximum intensity projection more closely outlines intracranial thrombi than conventional single-phase CT angiography. Our findings can be helpful when planning acute neurointervention. Further research is necessary to validate our data and assess the use of 4D-CTA in predicting response to different recanalization strategies.


Amino- and ionic liquid-functionalised nanocrystalline ZnO via silane anchoring - an antimicrobial synergy.
The synthesis of highly antimicrobial nanocrystalline zinc oxide and its covalent modifications are presented. In order to achieve further improvement of antimicrobial activity, the surface of ZnO was effectively modified with selected silanes comprising amino- and ionic liquid-functionalities. We demonstrate for the first time ionic liquid surface immobilization on ZnO and the application of this hybrid material for antimicrobial purposes. Different microscopic and spectroscopic techniques as well as size, surface and elemental composition analyses were employed to prove these modifications. Characterization revealed that surface and antimicrobial properties strongly depend on the modification employed. Most of the amino- and ionic liquid-functionalised nanocrystalline ZnO exhibited improved antimicrobial activity compared to commercially available silane-containing antimicrobial agents attached to nanocrystalline ZnO. Bacterial growth reduction was assessed by following the optical density of bacterial growth with different concentrations of the novel antimicrobial nanomaterials. Complete bacterial inactivation was achieved for specific amino- and ionic liquid-modifications at 0.125 g L-1, revealing the synergistic effect of ZnO and its modifications, exhibiting up to 2-fold improvement compared to unmodified ZnO.


Tongue nodules in canine leishmaniosis--a case report.
Canine leishmaniosis (CanL) caused by Leishmania infantum is an endemic zoonosis in southern European countries. Infected dogs can present rare or atypical forms of the disease and diagnosis can be challenging. The present report describes a case of tongue nodules in a 3-year-old neutered female Labrador Retriever dog with leishmaniosis. A fine needle aspiration of the lingual nodules revealed amastigote forms of Leishmania inside macrophages. Differential diagnosis ruled out neoplasia, calcinosis circumscripta, solar glossitis, vasculitis, amyloidosis, eosinophilic granulomas, chemical and electrical burns, uremic glossitis and autoimmune diseases. Combined therapy with antimoniate meglumine and allopurinol for 30 days resulted in the normalization of hematological and biochemical parameters. Two months after diagnosis and the beginning of treatment, a mild inflammatory infiltrate was observed by histopathology, but an anti-Leishmania immunofluorescence antibody test (IFAT) was negative as well as a PCR on both tongue lesions and a bone marrow aspirate. Seven months after diagnosis, the dog's general condition appeared good, there were no tongue lesions and a new IFAT was negative. Fifteen months after diagnosis this clinically favourable outcome continued. The dog could have suffered a relapsing episode of CanL, but a new systemic or local infection cannot be excluded. Regular clinical re-evaluation should be maintained, as a future relapse can potentially occur. In conclusion, CanL should be considered in the differential diagnosis of nodular glossitis in dogs.


Molecular cytogenetic analyses of Epinephelus bruneus and Epinephelus moara (Perciformes, Epinephelidae).
Genus Epinephelus (Perciformes, Epinephelidae), commonly known as groupers, are usually difficult in species identification for the lack and/or change of morphological specialization. In this study, molecular cytogenetic analyses were firstly performed to identify the closely related species Epinephelus bruneus and E. moara in this genus. The species-specific differences of both fish species showed in karyotype, chromosomal distribution of nucleolar organizer regions (NORs) and localization of 18S rDNA. The heterochromatin (interstitial C-bands) and distribution pattern of telomere (TTAGGG) n in E. bruneus revealed the chromosomal rearrangements and different karyotypic evolutionary characteristics compared to those in E. moara. The cytogenetic data suggested that the lineages of E. bruneus and E. moara were recently derived within the genus Epinephelus, and E. moara exhibited more plesiomorphic features than E. bruneus. All results confirmed that E. moara, which has long been considered a synonym of E. bruneus, is a distinct species in the family Epinephelidae. In addition, molecular cytogenetic analyses are useful in species differentiation and phylogenetic reconstruction in groupers.


Unique role of the complement receptor CR1 in the degradation of C3b associated with immune complexes.
The main finding of this paper is that CR1, the membrane receptor for C3b and C4b, together with C3b/C4b-inactivator (I), degrades C3b bound to immune complexes (C3b*). Two fragments are generated: C3c, which is released from the immune complexes, and C3d*. The C3c fragment released from the cell intermediate EAC1423b prepared with 125I-C3 was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and radioautography. It has a 135,000 mol wt and contains disulfide bonded labeled polypeptide chains of 75,000 and 31,000 mol wt, which presumably represent the beta and a fragment of the alpha-chain of C3b*. Silver staining of the SDS-PAGE gels revealed other C3-derived bands with 39-42,000 mol wt. Human erythrocytes + I also cleave C3b* into C3c and C3d*. The activity of the erythrocytes is CR1 mediated because it can be totally inhibited by monoclonal antibodies to CR1. In contrast with these results, I together with the serum protein beta 1H (H) transform EAC1423b into hemolytically inactive EAC1423bi and cleave the alpha' chain of C3b* into fragments of 70,000 and 40,000 mol wt. Small amounts of C3c are also released at relatively high concentrations of H. On a molar basis, the efficiency of CR1 in the generation of C3c and C3d is 10(4)-10(5) greater than H. An additional observation was that C3c could be released by treating EAC1423bi with CR1 + I and that this reaction was also inhibited by monoclonal antibodies to CR1. Therefore, it is likely that CR1 has binding affinity for iC3b and that the degradation of C3b* proceeds as follows: C3b (formula, see text) C3c + C3d*. Taken together, our findings argue that the processing of C3b* in vivo occurs in solid phase, that is, on the surface of cells bearing CR1.


The energy expenditure of snowshoeing in packed vs. unpacked snow at low-level walking speeds.
Snowshoeing is currently ranked as one of the top 20 participatory sports in the United States, and the number of participants almost tripled, from 440,000 to 1.2 million in 1998. Despite this large increase in participation, no scientific evidence exists to quantify any physiologic response to the activity. Therefore, the purpose of this investigation was to assess the energy expenditure of snowshoeing at selected low-level speeds and evaluate its acceptability as a form of aerobic conditioning exercise. Ten habitually active subjects (7 men, 3 women, mean age = 24 +/- 3.9 years, mass = 76.6 +/- 14.5 kg, height = 173.7 +/- 9.6 cm) were recruited. Steady state heart rate data were determined from 2 treadmill tests at 4 and 6 mph. Steady state heart rates at 4 mph and 6 mph from treadmill speeds were then reproduced outdoors under 2 snow conditions, packed, and unpacked snow, while caloric expenditure and speed were determined. Expired gases were collected in Douglas bags for both snowshoe and treadmill trials and then analyzed and corrected indoors for the fractional concentrations of carbon dioxide and oxygen. Data analyses indicate that caloric expenditure during snowshoeing may be considerably higher than previously reported. Snowshoeing on packed snow at 2.95 mph elicited a similar heart rate and energy expenditure response as walking on a treadmill at 4 mph or snowshoeing in unpacked snow at 2.04 mph (Vo(2) = 18.18 +/- 0.8 ml x kg(-1) x min(-1)). Snowshoeing on packed snow at 3.97 mph elicited the same heart rate and energy expenditure response as walking on a treadmill at 6 mph or snowshoeing on unpacked snow at 2.87 mph (Vo(2) = 36.72 +/- 0.8 ml x kg(-1) x min(-1)). Furthermore, increasing walking speed on snow by just 1 mph at slow speeds (2 and 3 mph) resulted in approximately twice the energy expenditure. Our data indicate that current estimates of energy expenditure while snowshoeing underestimate by greater than 50%. Apparently the energy expenditure during snowshoeing is much higher than previously considered and varies considerably because of snow terrain. Furthermore, energy expenditure levels similar to walking can be achieved on snowshoes at much slower speeds. This study represents an original investigation into energy expenditure during snowshoeing.


Reversible doxorubicin-induced congestive heart failure.
Doxorubicin hydrochloride is a chemotherapeutic agent highly effective against a wide range of neoplasms. A prime limiting factor to the administration of this drug is cardiotoxicity, which frequently develops when the cumulative dose exceeds 500 mg/sq m. Late cardiomyopathy, which may develop up to a year after therapy has been discontinued, was thought to be rapidly progressive and unresponsive to standard cardiac therapy. An adult who received 475 mg/sq m of doxorubicin hydrochloride experienced a cardiotoxic reaction one year after the completion of therapy. The patient responded to standard cardiac therapy. Resolution of left ventricular dysfunction was verified by echocardiography and radionuclide angiocardiography.


Acanthocephalan parasites of the flounder species Paralichthys isosceles, Paralichthys patagonicus and Xystreurys rasile from Brazil.
Flounders are commercially and economically important fish. A total of 120 specimens of flounders (60 Paralichthys isosceles, 30 Paralichthys patagonicus and 30 Xystreurys rasile) were collected off the coast of the state of Rio de Janeiro, Brazil. The fish were measured, necropsied and filleted, and then had their organs investigated for acanthocephalans. Taxonomic identification of the parasites was based on morphological, morphometric and genetic characters. Paralichthys isosceles and P. patagonicus were parasitized by juveniles of Serrasentis sagittifer, Bolbosoma turbinella, Corynosoma australe and C. cetaceum; Xystreurys rasile was parasitized by C. australe. Genetic characterization confirmed the identification of specimens of Bolbosoma turbinella and Corynosoma australe, as demonstrated by phylogenetic analyses using both ITS and cox1 molecular targets. Parasite indices of prevalence, intensity, mean intensity, abundance, mean abundance, and range of infection, as well as infection site, were evaluated for each parasite species. This is the first report of S. sagittifer parasitizing P. isosceles and P. patagonicus, and B. turbinella parasitizing P. patagonicus.


Integrin-linked kinase tunes cell-cell and cell-matrix adhesions to regulate the switch between apoptosis and EMT downstream of TGFβ1.
Epithelial-mesenchymal transition (EMT) is a morphogenetic process that endows epithelial cells with migratory and invasive potential. Mechanical and chemical signals from the tumor microenvironment can activate the EMT program, thereby permitting cancer cells to invade the surrounding stroma and disseminate to distant organs. Transforming growth factor β1 (TGFβ1) is a potent inducer of EMT that can also induce apoptosis depending on the microenvironmental context. In particular, stiff microenvironments promote EMT while softer ones promote apoptosis. Here, we investigated the molecular signaling downstream of matrix stiffness that regulates the phenotypic switch in response to TGFβ1 and uncovered a critical role for integrin-linked kinase (ILK). Specifically, depleting ILK from mammary epithelial cells precludes their ability to sense the stiffness of their microenvironment. In response to treatment with TGFβ1, ILK-depleted cells undergo apoptosis on both soft and stiff substrata. We found that knockdown of ILK decreases focal adhesions and increases cell-cell adhesions, thus shifting the balance from cell-matrix to cell-cell adhesion. High cell-matrix adhesion promotes EMT whereas high cell-cell adhesion promotes apoptosis downstream of TGFβ1. These results highlight an important role for ILK in controlling cell phenotype by regulating adhesive connections to the local microenvironment.


Current management of primary central nervous system lymphoma.
Primary central nervous system lymphoma (PCNSL) is a rare but treatable disease. The hallmark therapy is based on the use of high-dose intravenous methotrexate (HDMTX), with clinical trial data suggesting that a more aggressive and targeted chemoimmunotherapeutic approach, to include the use of targeted therapy with rituximab (RTX) and the addition of other chemotherapeutic agents, may improve prognosis and reduce or defer the need for radiation therapy. Understanding of the molecular basis of tumor growth and proliferation may allow for the use of new targeted agents.


Serological evidence of HTLV-I and HTLV-II coinfections in HIV-1 positive patients in Belém, state of Pará, Brazil.
The occurrence of HTLV-I/II and HIV-1 coinfections have been shown to be frequent, probably in consequence of their similar modes of transmission. This paper presents the prevalence of coinfection of HTLV among HIV-1 infected and AIDS patients in Belém, State of Pará, Brazil. A group of 149 patients attending the AIDS Reference Unit of the State Department of Health was tested for the presence of antibodies to HTLV-I/II using an enzyme immunoassay and the positive reactions were confirmed with a Western blot that discriminates between HTLV-I and HTLV-II infections. Four patients (2.7%) were positive to HTLV-I, seven (4.7%) to HTLV-II and one (0.7%) showed an indeterminate pattern of reaction. The present results show for the first time in Belém not only the occurrence of HTLV-II/HIV-1 coinfections but also a higher prevalence of HTLV-II in relation to HTLV-I. Furthermore, it also enlarges the geographical limits of the endemic area for HTLV-II in the Amazon region of Brazil.


Tamm-Horsfall protein excretion during chronic alterations in urinary concentration and protein intake in the rat.
Tamm-Horsfall protein (THP), a normal constituent of mammalian urine, has been determined in rat urine under various conditions in an attempt to elucidate the physiological role of this glycoprotein. Experiments were designed to assess whether THP production is related to the process of urine concentration or to the transport activity of the thick ascending limb of the loop of Henle (TAL), the nephron segment where it is produced. For this purpose, THP excretion was measured, by radioimmunoassay, in adult male rats under 4 different conditions induced by the following chronic treatments: (1) furosemide (12 mg/day in osmotic minipumps); (2) increased water intake; (3) antidiuretic hormone (ADH) infusion (50 ng DDAVP/day in osmotic minipumps) in rats of the Brattleboro strain with hereditary hypothalamic diabetes insipidus; (4) high-protein (32% casein) versus low-protein diet (10% casein). Each experiment included 6 experimental and 6 control rats. After treatment for 1-3 weeks, 24-h urines were collected for determination of urine flow rate, osmolality, and creatinine and THP concentrations. No significant changes in THP excretion were observed in experiments (1) and (2) despite 5- to 7-fold-differences in urine flow rate. Antidiuretic hormone treatment in (3) slightly lowered THP excretion (287 +/- 53 vs. 367 +/- 41 micrograms/day per 100 g body weight; p less than 0.005), whereas high-protein diet, in experiment (4), led to a 50% increase in THP excretion (446 +/- 57 vs. 304 +/- 79 micrograms/day per 100 g body weight; p less than 0.001). Expressing THP excretion relative to that of creatine did not change these findings. These results show (1) that chronically established changes in the level of diuresis, chronic furosemide-induced blockade of the Na,K,Cl-cotransporter or the absence of ADH in Brattleboro rats have little or no impact on the level of THP production, and (2) that THP production is independent of the intensity of transport in the TAL, since two conditions which both are known to increase the transport rate of solutes in the TAL (ADH infusion and high-protein diet), resulted in opposite changes in THP excretion. It is concluded that the rate of THP synthesis is neither linked to the process of urine concentration nor to the ion transport activity of the TAL.


Use of therapies other than disease-modifying agents, including complementary and alternative medicine, by patients with multiple sclerosis: a survey study.
Many patients with multiple sclerosis use complementary and alternative medicine (CAM) to supplement their traditional treatment. To identify both the prevalence and frequency of use of therapies other than disease-modifying agents (DMAs), including CAM, among patients with multiple sclerosis. The authors administered a 13-question survey regarding patients' current use of non-DMA therapies-including dietary supplements, exercise, and "true" CAM (eg, acupuncture, chiropractic, massage)-and mainstream treatments, including physical therapy and osteopathic manipulative treatment. Patients rated their level of disability on a scale of 1 to 10 (with 10 being most severe). A hospital outpatient clinic in Philadelphia, Pennsylvania. Inclusion criteria were physician-confirmed multiple sclerosis (either relapsing-remitting or progressive), regardless of sex, duration of disease, age at onset, disability level, or type of disease. Patients were excluded if they were younger than 18 years. Patient-reported use of non-DMA therapies and perception of disability. A total of 111 patients with multiple sclerosis completed the survey properly. All respondents used non-DMA therapies. Twenty-three patients (20.7%) used these therapies without concomitantly taking a DMA. A plurality (34.8%) of those patients reported a disability score of 7 or 8. Sixty-two of the 88 participants (70.5 %) who used DMAs reported disability scores of 5 or less. Sixty-five patients (58.6.%) reported exercising on a weekly basis. Among those patients, 47 (72.3%) reported a disability score of 5 or less. Sixty-four patients (57.7%) used such CAM therapies as acupuncture and massage, or such other non-DMA treatments as osteopathic manipulative treatment and psychotherapy. Among those patients, 37 (64.9%) reported a disability score of 5 or less. Many patients with multiple sclerosis are seeking more than traditional medical treatment. Physicians and other health care professionals must be aware of the extensive use of alternative modalities among these patients, and these professionals must provide guidance and monitoring in use of these therapies to improve outcomes.


Attenuation of glycation-induced multiple protein modifications by Indian antidiabetic plant extracts.
Protein glycation is the major contributing factor in the development of diabetic complications. The antiglycation potential of medicinal plants provides a promising opportunity as complementary interventions for complications. To investigate the antiglycation potential of 19 medicinal plants extracts using albumin by estimating different indicators: (1) glycation (early and late), (2) albumin oxidation, and (3) amyloid aggregation. The effect of aqueous plant extracts (1% w/v) on protein glycation was assessed by incubating albumin (10 mg/mL) with fructose (250 mM) for 4 days. Degree of protein glycation in the absence and presence of plant extracts was assessed by estimating fructosamine, advanced glycation end products (AGEs), carbonyls, free thiol group and β-amyloid aggregation. Petroselinum crispum, Boerhavia diffusa, Terminalia chebula, Swertia chirayita and Glycyrrhiza glabra showed significant antiglycating activity. P. crispum and A. barbadensis inhibited the carbonyl stress and protected the thiol group from oxidative damage. There was significant correlation between protein thiols and amyloid inhibition (R = -.69, p < .001). P. crispum, B. diffusa and T. chebula had the most potent antiglycation activity. These plant exerted noticeable antiglycation activity at different glycation modifications of albumin. These findings are important for identifying plants with potential to combat diabetic complications.


Prevalence of comorbidities and evaluation of their screening in spondyloarthritis: results of the international cross-sectional ASAS-COMOSPA study.
Increased risk of some comorbidities has been reported in spondyloarthritis (SpA). Recommendations for detection/management of some of these comorbidities have been proposed, and it is known that a gap exists between these and their implementation in practice. To evaluate (1) the prevalence of comorbidities and risk factors in different countries worldwide, (2) the gap between available recommendations and daily practice for management of these comorbidities and (3) the prevalence of previously unknown risk factors detected as a result of the present initiative. Cross-sectional international study with 22 participating countries (from four continents), including 3984 patients with SpA according to the rheumatologist. The prevalence of comorbidities (cardiovascular, infection, cancer, osteoporosis and gastrointestinal) and risk factors; percentage of patients optimally monitored for comorbidities according to available recommendations and percentage of patients for whom a risk factor was detected due to this study. The most frequent comorbidities were osteoporosis (13%) and gastroduodenal ulcer (11%). The most frequent risk factors were hypertension (34%), smoking (29%) and hypercholesterolaemia (27%). Substantial intercountry variability was observed for screening of comorbidities (eg, for LDL cholesterol measurement: from 8% (Taiwan) to 98% (Germany)). Systematic evaluation (eg, blood pressure (BP), cholesterol) during this study unveiled previously unknown risk factors (eg, elevated BP (14%)), emphasising the suboptimal monitoring of comorbidities. A high prevalence of comorbidities in SpA has been shown. Rigorous application of systematic evaluation of comorbidities may permit earlier detection, which may ultimately result in an improved outcome of patients with SpA.


Molecular typing methods for outbreak detection and surveillance of invasive disease caused by Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae, a review.
Invasive disease caused by the encapsulated bacteria Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae remains an important cause of morbidity and mortality worldwide, despite the introduction of successful conjugate polysaccharide vaccines that target disease-associated strains. In addition, resistance, or more accurately reduced susceptibility, to therapeutic antibiotics is spreading in populations of these organisms. There is therefore a continuing requirement for the surveillance of vaccine and non-vaccine antigens and antibiotic susceptibilities among isolates from invasive disease, which is only partially met by conventional methods. This need can be met with molecular and especially nucleotide sequence-based typing methods, which are fully developed in the case of N. meningitidis and which could be more widely deployed in clinical laboratories for S. pneumoniae and H. influenzae.


Biochemical and spectroscopic characterization of Escherichia coli aconitases (AcnA and AcnB).
Escherichia coli contains two major aconitases (Acns), AcnA and AcnB. They are distantly related monomeric Fe-S proteins that contain different arrangements of four structural domains. On the basis of the differential expression of the acnA and acnB genes, AcnA has been designated as an aerobic-stationary-phase enzyme that is specifically induced by iron and oxidative stress, whereas AcnB functions as the major citric-acid-cycle enzyme during exponential growth. The biochemical and kinetic properties of the purified enzymes have now shown that AcnA is more stable than AcnB, has a higher affinity for citrate, and operates optimally over a wider pH range, consistent with its role as a maintenance or survival enzyme during nutritional or oxidative stress. In contrast, the better performance at high substrate concentrations and greater instability of AcnB indicate that AcnB is specifically adapted to function as the main catabolic enzyme and, by inactivation, to rapidly modulate energy metabolism in response to oxidative or pH stress, either directly or indirectly by regulating post-transcriptional gene expression. EPR and magnetic-CD spectroscopy showed that the iron-sulphur clusters of the bacterial Acns (and their binding sites) strongly resemble those of the mammalian enzymes. The EPR and MCD spectra of the oxidized inactive form of AcnB confirmed the presence of a [3Fe-4S](1+) (S=1/2) cluster. Comparisons showed that the EPR spectrum of AcnB more closely resembled that of mammalian mitochondrial Acn (m-Acn), whereas the spectrum of AcnA more closely resembled that of the cytoplasmic enzyme (c-Acn). The MCD spectra revealed spectroscopic signatures similar to that of m-Acn. Reconstitution of the active [4Fe-4S](2+) forms followed by one-electron reduction gave rise to EPR spectra that are almost identical with those reported for the mammalian enzymes.


Structure and assembly of cargo Rubisco in two native α-carboxysomes.
Carboxysomes are a family of bacterial microcompartments in cyanobacteria and chemoautotrophs. They encapsulate Ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) and carbonic anhydrase catalyzing carbon fixation inside a proteinaceous shell. How Rubisco complexes pack within the carboxysomes is unknown. Using cryo-electron tomography, we determine the distinct 3D organization of Rubisco inside two distant α-carboxysomes from a marine α-cyanobacterium Cyanobium sp. PCC 7001 where Rubiscos are organized in three concentric layers, and from a chemoautotrophic bacterium Halothiobacillus neapolitanus where they form intertwining spirals. We further resolve the structures of native Rubisco as well as its higher-order assembly at near-atomic resolutions by subtomogram averaging. The structures surprisingly reveal that the authentic intrinsically disordered linker protein CsoS2 interacts with Rubiscos in native carboxysomes but functions distinctively in the two α-carboxysomes. In contrast to the uniform Rubisco-CsoS2 association in the Cyanobium α-carboxysome, CsoS2 binds only to the Rubiscos close to the shell in the Halo α-carboxysome. Our findings provide critical knowledge of the assembly principles of α-carboxysomes, which may aid in the rational design and repurposing of carboxysome structures for new functions.


Does breast size affect how women participate in physical activity?
Research has shown that women with large breasts experience increased breast motion, which can act as a barrier to participating in physical activity. Despite this evidence, limited research has investigated the effect of breast size on physical activity participation. Cross-sectional study. 355 Australian women (aged 18-75years) who volunteered as participants were divided into four breast size groups (small, medium, large and hypertrophic). All participants completed the Active Australia Survey to assess their participation in physical activity in the previous week, and were asked to indicate whether they agreed or disagreed that their breast size affected their participation in physical activity. Participant responses to time spent in each type of activity were compared across the four breast size groups using an ANCOVA design, controlling for age. Participants with hypertrophic breasts participated in significantly less total physical activity per week, particularly less vigorous-intensity physical activity, compared to their counterparts with smaller breasts. Participants with large breasts also participated in less vigorous-intensity physical activity compared to participants with small and with medium breasts, and a high percentage of women with large and with hypertrophic breasts perceived their breast size to affect the amount and intensity of physical activity they performed. Breast size should be acknowledged as a potential barrier to women participating in physical activity. Strategies to assist women with large and hypertrophic breasts participate in all types and intensities of physical activity are needed so women can enjoy the health benefits associated with an active lifestyle.


Effects of glenoid inclination and acromion index on humeral head translation and glenoid articular cartilage strain.
Previous clinical studies have reported associations between glenoid inclination (GI), the acromion index (AI), and the critical shoulder angle (CSA) on the one hand and the occurrence of glenohumeral osteoarthritis and supraspinatus tendon tears on the other hand. The objective of this work was to analyze the correlations and relative importance of these different anatomic parameters. Using a musculoskeletal shoulder model developed from magnetic resonance imaging scans of 1 healthy volunteer, we varied independently GI from 0° to 15° and AI from 0.5 to 0.8. The corresponding CSA varied from 20.9° to 44.1°. We then evaluated humeral head translation and critical strain volume in the glenoid articular cartilage at 60° of abduction in the scapular plane. These values were correlated with GI, AI, and CSA. Humeral head translation was positively correlated with GI (R = 0.828, P < .0001), AI (R = 0.539, P < .0001), and CSA (R = 0.964, P < .0001). Glenoid articular cartilage strain was also positively correlated with GI (R = 0.489, P = .0004) but negatively with AI (R = -0.860, P < .0001) and CSA (R = -0.285, P < .0473). The biomechanical shoulder model is consistent with clinical observations. The prediction strength of CSA is confirmed for humeral head translation and thus presumably for rotator cuff tendon tears, whereas the AI seems more appropriate to evaluate the risk of glenohumeral osteoarthritis caused by excessive articular cartilage strain. As a next step, we should corroborate these theoretical findings with clinical data.


Kinetics of primary SIV infection in lymph nodes.
The very early stages of SIV infection in lymph nodes of rhesus monkeys were characterized by in situ hybridization. Massive viral replication was detected in macrophages and lymphocytes during the first week of infection. In a second phase, SIV RNA concentrated in the developing germinal centers, and colocalized with the follicular dendritic cells. The down-regulation of the viral load in lymph nodes varied depending on the animal, indicating early differences in the susceptibility to SIV infection.


Transfer of information from personal health records: a survey of veterans using My HealtheVet.
Personal health records provide patients with ownership of their health information and allow them to share information with multiple healthcare providers. However, the usefulness of these records relies on patients understanding and using their records appropriately. My HealtheVet is a Web-based patient portal containing a personal health record administered by the Veterans Health Administration. The goal of this study was to explore veterans' interest and use of My HealtheVet to transfer and share information as well as to identify opportunities to increase veteran use of the My HealtheVet functions. Two waves of data were collected in 2010 through an American Customer Satisfaction Index Web-based survey. A random sample of veterans using My HealtheVet was invited to participate in the survey conducted on the My HealtheVet portal through a Web-based pop-up browser window. Wave One results (n=25,898) found that 41% of veterans reported printing information, 21% reported saving information electronically, and only 4% ever sent information from My HealtheVet to another person. In Wave Two (n=18,471), 30% reported self-entering medication information, with 18% sharing this information with their Veterans Affairs (VA) provider and 9.6% sharing with their non-VA provider. Although veterans are transferring important medical information from their personal health records, increased education and awareness are needed to increase use. Personal health records have the potential to improve continuity of care. However, more research is needed on both the barriers to adoption as well as the actual impact on patient health outcomes and well-being.


In vivo biological response to extracorporeal shockwave therapy in human tendinopathy.
Extracorporeal shock wave therapy (ESWT) is a non-invasive treatment for chronic tendinopathies, however little is known about the in-vivo biological mechanisms of ESWT. Using microdialysis, we examined the real-time biological response of healthy and pathological tendons to ESWT. A single session of ESWT was administered to the mid-portion of the Achilles tendon in thirteen healthy individuals (aged 25.7 ± 7.0 years) and patellar or Achilles tendon of six patients with tendinopathies (aged 39.0 ± 14.9 years). Dialysate samples from the surrounding peri-tendon were collected before and immediately after ESWT. Interleukins (IL)-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-17A, vascular endothelial growth factor and interferon-γ were quantified using a cytometric bead array while gelatinase activity (MMP-2 and -9) was examined using zymography. There were no statistical differences between the biological tissue response to ESWT in healthy and pathological tendons. IL-1β, IL-2, IL-6 and IL-8 were the cytokines predominantly detected in the tendon dialysate. IL-1β and IL-2 did not change significantly with ESWT. IL-6 and IL-8 concentrations were elevated immediately after ESWT and remained significantly elevated for four hours post-ESWT (p < 0.001). Pro-forms of MMP-2 and -9 also increased after ESWT (p < 0.003), whereas there were no significant changes in active MMP forms. In addition, the biological response to ESWT treatment could be differentiated between possible responders and non-responders based on a minimum 5-fold increase in any inflammatory marker or MMP from pre- to post-ESWT. Our findings provide novel evidence of the biological mechanisms underpinning ESWT in humans in vivo. They suggest that the mechanical stimulus provided by ESWT might aid tendon remodelling in tendinopathy by promoting the inflammatory and catabolic processes that are associated with removing damaged matrix constituents. The non-response of some individuals may help to explain why ESWT does not improve symptoms in all patients and provides a potential focus for future research.


Bodily differences?: Gender, race, and class in Hans Sloane's Jamaican medical practice, 1687-1688.
Despite the multitude of seventeenth- and early eighteenth-century British medical publications regarding empire and health, Hans Sloane's A Voyage To the Islands [of] Madera, Barbados, Nieves, S. Christophers and Jamaica (1707) was the first to incorporate significant numbers of female and African patients among its printed case histories. Comprising some sixty-four pages of the introduction, this unique set of records affords scholars the rare opportunity to examine how patients (of both sexes and races, various ages, and all social levels) residing in a "torrid zone" were diagnosed and treated by an English physician during the 1680s. Sloane had expected to encounter illnesses vastly different from those found in England when he arrived in Jamaica, but after practicing medicine in Jamaica for over a year, he concluded that there existed very little difference in the manifestation of illnesses in different climates. Although some ailments were sex-specific and culture-specific, for the most part Sloane transgressed categories of gender and race by diagnosing and treating all his patients according to the same medical ideology. And although it did not directly challenge accepted medical views, Sloane's Voyage revealed incongruities in dealing with such categories within the context of early imperial medicine.


Reverse Iontophoresis: Noninvasive Assessment of Topical Drug Bioavailability.
Assessing drug disposition in the skin after the application of a topical formulation is difficult. It is hypothesized that reverse iontophoresis (RI), which can extract charged/polar molecules for monitoring purposes, may provide a noninvasive approach for the assessment of local drug bioavailability. The passive and RI extraction of salicylic acid (SA) and nicotine (NIC) from porcine skin in vitro was assessed after a simple solution of the former and a transdermal patch of the latter had been applied for 24 and 8 h, respectively. Immediately after this "passive skin loading", the amount of drug in the stratum corneum (SC) and "viable" tissue (VT) was measured either (a) after tape-stripping and subsequent solvent extraction of both skin layers or (b) following RI extraction over 4 h. Parallel experiments were then performed in vivo in healthy volunteers; in this case, the VT was not sampled and the skin loading period for NIC was only 4 h. RI extraction of both drugs was significantly higher (in vitro and in vivo) than that achieved passively, and the cumulative RI extraction profiles as a function of time were mathematically analyzed using a straightforward compartmental model. Best-fit estimates of drug amounts in the SC and VT (ASC,0 and AVT,0, respectively) at the end of "loading" and two first-order rate constants describing transfer between the model compartments were then determined. The in vitro predictions of ASC,0 and AVT,0 were in excellent agreement with the experimental results, as was the value of the former in vivo. The rate constants derived from the in vitro and in vivo results were also similar. In summary, the results provide proof-of-concept that the RI method has the potential to noninvasively assess relevant metrics of drug bioavailability in the skin.


A systematic review of the effect of community treatment orders on service use.
The evidence regarding community treatment order effectiveness has been conflicting. This systematic review aims to bring up to date the review performed by Churchill and colleagues in 2005 by assessing and interpreting evidence of CTO effectiveness defined by admission rates, number of inpatient days, community service use, and medication adherence published since 2006. Databases were searched to obtain relevant studies published from January 2006 to March 2013. 18 studies including one randomised controlled trial were included. There remains lack of evidence from randomised and non-randomised studies that CTOs are associated with or affected by admission rates, number of inpatient days or community service use. The most recent and largest RCT is included in this review and found no significant impact on admission rate (RR = 1.0, 95% CI 0.75-1.33) or number of days in hospital (IR = 0.90, 95% CI 0.65-1.26). Results from the two largest longitudinal datasets included in this review do not concur. Studies using the New York dataset found that CTOs were associated with reduced admission rates and inpatient days, while studies using the Victoria dataset generally found that they were associated with increased admission rates and inpatient days. There is now robust evidence in the literature that CTOs have no significant effects on hospitalisation and other service use outcomes. Non-randomised studies continue to report conflicting results. Distinguishing between CTO recall and revocation and different patterns of community contact is needed in future research to ensure differentiation between CTO process and outcome.


The role of responsibility and fear of guilt in hypothesis-testing.
Recent theories argue that both perceived responsibility and fear of guilt increase obsessive-like behaviours. We propose that hypothesis-testing might account for this effect. Both perceived responsibility and fear of guilt would influence subjects' hypothesis-testing, by inducing a prudential style. This style implies focusing on and confirming the worst hypothesis, and reiterating the testing process. In our experiment, we manipulated the responsibility and fear of guilt of 236 normal volunteers who executed a deductive task. The results show that perceived responsibility is the main factor that influenced individuals' hypothesis-testing. Fear of guilt has however a significant additive effect. Guilt-fearing participants preferred to carry on with the diagnostic process, even when faced with initial favourable evidence, whereas participants in the responsibility condition only did so when confronted with an unfavourable evidence. Implications for the understanding of obsessive-compulsive disorder (OCD) are discussed.


Vowels in infant-directed speech: More breathy and more variable, but not clearer.
Infant-directed speech (IDS) is known to differ from adult-directed speech (ADS) in a number of ways, and it has often been argued that some of these IDS properties facilitate infants' acquisition of language. An influential study in support of this view is Kuhl et al. (1997), which found that vowels in IDS are produced with expanded first and second formants (F1/F2) on average, indicating that the vowels are acoustically further apart in IDS than in ADS. These results have been interpreted to mean that the way vowels are produced in IDS makes infants' task of learning vowel categories easier. The present paper revisits this interpretation by means of a thorough analysis of IDS vowels using a large-scale corpus of Japanese natural utterances. We will show that the expansion of F1/F2 values does occur in spontaneous IDS even when the vowels' prosodic position, lexical pitch accent, and lexical bias are accounted for. When IDS vowels are compared to carefully read speech (CS) by the same mothers, however, larger variability among IDS vowel tokens means that the acoustic distances among vowels are farther apart only in CS, but not in IDS when compared to ADS. Finally, we will show that IDS vowels are significantly more breathy than ADS or CS vowels. Taken together, our results demonstrate that even though expansion of formant values occurs in spontaneous IDS, this expansion cannot be interpreted as an indication that the acoustic distances among vowels are farther apart, as is the case in CS. Instead, we found that IDS vowels are characterized by breathy voice, which has been associated with the communication of emotional affect.


Phenotypic proteomic profiling identifies a landscape of targets for circadian clock-modulating compounds.
Determining the exact targets and mechanisms of action of drug molecules that modulate circadian rhythms is critical to develop novel compounds to treat clock-related disorders. Here, we have used phenotypic proteomic profiling (PPP) to systematically determine molecular targets of four circadian period-lengthening compounds in human cells. We demonstrate that the compounds cause similar changes in phosphorylation and activity of several proteins and kinases involved in vital pathways, including MAPK, NGF, B-cell receptor, AMP-activated protein kinases (AMPKs), and mTOR signaling. Kinome profiling further indicated inhibition of CKId, ERK1/2, CDK2/7, TNIK, and MST4 kinases as a common mechanism of action for these clock-modulating compounds. Pharmacological or genetic inhibition of several convergent kinases lengthened circadian period, establishing them as novel circadian targets. Finally, thermal stability profiling revealed binding of the compounds to clock regulatory kinases, signaling molecules, and ubiquitination proteins. Thus, phenotypic proteomic profiling defines novel clock effectors that could directly inform precise therapeutic targeting of the circadian system in humans.


Effect of tightening torque on the marginal adaptation of cement-retained implant-supported fixed dental prostheses.
The final position of the abutment changes with the amount of tightening torque. This could eventually lead to loss of passivity and marginal misfit of prostheses. The aim of this study was to evaluate the effect of three different tightening torques on the marginal adaptation of 3-unit cement-retained implant-supported fixed dental prostheses (FDPs). Two implants (Straumann) were inserted in an acrylic block so that one of the implants was placed vertically and the other at a 15° vertical angle. A straight abutment and a 15° angulated abutment were connected to the vertically and obliquely installed implants, respectively, so that the two abutments were parallel. Then, 10 cement-retained FDPs were waxed and cast. Abutments were tightened with 10, 20, and 35 Ncm torques, respectively. Following each tightening torque, FDPs were luted on respective abutments with temporary cement. The marginal adaptation of the retainers was evaluated using stereomicroscope. FDPs were then removed from the abutments and were sectioned at the connector sites. The retainers were luted again on their respective abutments. Luting procedures and marginal adaptation measurement were repeated. Data were analyzed by ANOVA and least significant difference tests (α = 0.05). After cutting the FDP connectors, the independent samples t-test was used to compare misfit values (α = 0.05). Following 10, 20, and 35 Ncm tightening torques, the marginal discrepancy of the retainers of FDPs significantly increased (P < 0.05). There was no significant difference between the marginal discrepancies of these two retainers (P > 0.05). The marginal gap values of angulated abutment retainers (ANRs) were significantly higher than those of the straight abutment after cutting the connectors (P = 0.026). Within the limitations of this study, the marginal misfit of cement-retained FDPs increased continuously when the tightening torque increased. After cutting the connectors, the marginal misfit of the ANRs was higher than those of the straight abutment retainers.


The prediction of risk of recurrence and time to recurrence of hepatocellular carcinoma after orthotopic liver transplantation: a pilot study.
Orthotopic liver transplantation (OLTx) in the presence of hepatocellular carcinoma (HCC) has been complicated by high recurrence rates. The ability to determine the risk and timing of HCC recurrence on an individual basis would greatly aid in the candidate selection process resulting in a more efficient use of donated organs and allow the individualization and better evaluation of adjuvant chemotherapy. The 214 patients who underwent OLTx in the presence of HCC were analyzed. From the 178 patients who survived more than 150 days, 71 (40%) have suffered HCC recurrence. Based on five risk factors, that is, gender, tumor number, lobar tumor distribution, tumor size, grade of vascular invasion, artificial neural network models predicting the likelihood of HCC recurrence within 1, 2, and 3 consecutive years after transplantation were developed. Based on model predictions, those combinations of risk factors that should/should not lead to recurrence were generated, allowing stratification of patients into the following three groups: 1) patients who should not suffer HCC recurrence and who should not need adjuvant therapy, 2) patients who will suffer recurrence and for whom postoperative chemotherapy significantly prolonged survival (but did not prevent recurrence), and 3) patients who may or may not suffer HCC recurrence and whose recurrence may be prevented by adjuvant chemotherapy. The outcome of OLTx for patients with HCC can be prognosticated based on a number of clinical variables. If verified through multicenter trials, these models could be made available to transplantation programs performing OLTx in the presence of HCC.


dbPSHP: a database of recent positive selection across human populations.
The dbPSHP database (http://jjwanglab.org/dbpshp) aims to help researchers to efficiently identify, validate and visualize putative positively selected loci in human evolution and further discover the mechanism governing these natural selections. Recent evolution of human populations at the genomic level reflects the adaptations to the living environments, including climate change and availability and stability of nutrients. Many genetic regions under positive selection have been identified, which assist us to understand how natural selection has shaped population differences. Here, we manually collect recent positive selections in different human populations, consisting of 15,472 loci from 132 publications. We further compiled a database that used 15 statistical terms of different evolutionary attributes for single nucleotide variant sites from the HapMap 3 and 1000 Genomes Project to identify putative regions under positive selection. These attributes include variant allele/genotype properties, variant heterozygosity, within population diversity, long-range haplotypes, pairwise population differentiation and evolutionary conservation. We also provide interactive pages for visualization and annotation of different selective signals. The database is freely available to the public and will be frequently updated.


Standardization of pathologic evaluation and reporting of postneoadjuvant specimens in clinical trials of breast cancer: recommendations from an international working group.
Neoadjuvant systemic therapy is being used increasingly in the treatment of early-stage breast cancer. Response, in the form of pathological complete response, is a validated and evaluable surrogate end point of survival after neoadjuvant therapy. Thus, pathological complete response has become a primary end point for clinical trials. However, there is a current lack of uniformity in the definition of pathological complete response. A review of standard operating procedures used by 28 major neoadjuvant breast cancer trials and/or 25 sites involved in such trials identified marked variability in specimen handling and histologic reporting. An international working group was convened to develop practical recommendations for the pathologic assessment of residual disease in neoadjuvant clinical trials of breast cancer and information expected from pathology reports. Systematic sampling of areas identified by informed mapping of the specimen and close correlation with radiological findings is preferable to overly exhaustive sampling, and permits taking tissue samples for translational research. Controversial areas are discussed, including measurement of lesion size, reporting of lymphovascular space invasion and the presence of isolated tumor cells in lymph nodes after neoadjuvant therapy, and retesting of markers after treatment. If there has been a pathological complete response, this must be clearly stated, and the presence/absence of residual ductal carcinoma in situ must be described. When there is residual invasive carcinoma, a comment must be made as to the presence/absence of chemotherapy effect in the breast and lymph nodes. The Residual Cancer Burden is the preferred method for quantifying residual disease in neoadjuvant clinical trials in breast cancer; other methods can be included per trial protocols and regional preference. Posttreatment tumor staging using the Tumor-Node-Metastasis system should be included. These recommendations for standardized pathological evaluation and reporting of neoadjuvant breast cancer specimens should improve prognostication for individual patients and allow comparison of treatment outcomes within and across clinical trials.


Abortion laws and medical developments: a medico-legal anomaly in Queensland.
In October 2010 the District Court sitting in Cairns, Queensland, found Tegan Leach not guilty of attempting to procure her own abortion and Sergie Brennan not guilty of supplying Leach with the drugs Mifepristone and Misoprostol to procure an abortion. Brennan obtained the drugs from his sister in the Ukraine through the regular postal system. R v Brennan and Leach was the first case in Queensland's history where a woman was charged with procuring her own abortion. The drugs are accepted by the medical profession worldwide for medical abortions. A prosecution witness gave evidence that Mifepristone is not harmful or injurious to the health of a woman and it is listed as an essential medicine by the World Health Organisation and approved for use by the Australian Therapeutic Goods Administration. The jury found the defendants not guilty because they were not satisfied beyond reasonable doubt that the combination of the drugs Mifepristone and Misoprostol was a "noxious" substance under the Criminal Code (Old). This article concludes that there is no regulatory miracle which will stop the traffic of Mifepristone and Misoprostol into Australia and therefore an intelligent regulatory response is required which would make it unnecessary for women to seek Mifepristone and Misoprostol from overseas networks and the internet. Among other things, this would include the repeal of confusing, inappropriate and ineffective abortion laws.


Chondrocyte allografts for repair of full-thickness defects in the condylar articular cartilage of rabbits.
This study investigated the feasibility of repairing defects of the condylar articular cartilage by chondrocyte allotransplantation. A full-thickness defect (2 mm diameter) was made in the condylar articular cartilage of 6-month-old rabbits with a No. 701 dental fissure bur that penetrated both the articular cartilage and the subchondral bone, and entered the marrow cavity. Sixty-four animals were divided into 5 groups. In the cell transplantation group (20 rabbits), the defect was filled with a collagen membrane embedded with chondrocytes of neonate rabbits cultured in vitro for 1 week. In control group 1 (18 rabbits), the defect was left untreated. In control group 2 (18 rabbits), the defect was filled with a collagen membrane without chondrocytes. In sham-operation control group (6 rabbits), the condyle surface was exposed but left unchanged. Two rabbits were added as a normal control group. Mandibular movement was not restricted postoperatively. The macroscopic and microscopic features of the condyles were observed at 1, 2, 4, 8, 12, and 20 weeks following surgery. The defects in the condylar articular cartilage were repaired with cartilage tissue after cell transplantation. A defect in the condylar articular cartilage can be repaired with articular cartilage-like tissue by allotransplantation of chondrocytes.


A new diagnostic work-up for defining anemia etiologies: a cohort study in patients ≥ 50 years in general practices.
To study etiologies of anemia using an extensive laboratory analysis in general practices. An extensive laboratory analysis was performed in blood of newly diagnosed anemia patients aged ≥50 years from the general population in the city of Dordrecht area, the Netherlands. Eight laboratory-orientated etiologies of anemia were defined. Patients were assigned one or more of these etiologies on the basis of their test results. Blood of 4152 patients (median age 75 years; 49% male) was analyzed. The anemia etiology was unclear in 20%; a single etiology was established in 59%; and multiple etiologies in 22% of the patients. The most common etiologies were anemia of chronic disease (ACD) (54.5%), iron deficiency anemia (IDA) (19.1%) and renal anemia (13.8%). The most common single etiologies were IDA (82%) and ACD (68%), while the multiple etiologies most commonly included folic acid deficiency (94%) and suspected bone marrow disease (88%). Older age was associated with a lower incidence of IDA and a higher incidence of renal anemia. Mild anemia was more often associated with ACD and uncertain anemia, while severe anemia was mainly seen in patients with IDA. Extensive laboratory analysis in anemic patients from the general population helped clarify the etiology of anemia and revealed many various combinations of etiologies in a significant proportion of patients. Age, sex and the severity of anemia are predictive of the underlying etiology.


Developing a new measure of dissociation: The Dissociative Experiences Measure, Oxford (DEMO).
The field of dissociation is receiving increasing attention, despite a lack of coherent conceptualisation of the construct. Advances in the field would be aided by a measure that reflects service user experiences of dissociative experiences and can be easily scored. The current study describes the development of a new measure of dissociation (Dissociative Experiences Measure, Oxford; DEMO) that aims to fulfil these criteria. The study follows an exploratory, data-driven, measure development design. Resource searching and feedback from clinicians (n = 3) and service users (n = 6) were used to develop an extensive item pool. An online sample (n = 691) provided data for a factor analysis of the item pool. Factor analysis produced a measure with five subscales: 'unreality', 'numb and disconnected', 'memory blanks', 'zoned out', and 'vivid internal world'. Further analysis indicated that the new measure has high internal consistency, and high convergent, divergent, and discriminant validity. The DEMO shows promise as an up-to-date clinical and research tool for the assessment of dissociative experiences. These results are preliminary, and further validation of the measure with a clinical sample is required.


Universal test and treat and the HIV epidemic in rural South Africa: a phase 4, open-label, community cluster randomised trial.
Universal antiretroviral therapy (ART), as per the 2015 WHO recommendations, might reduce population HIV incidence. We investigated the effect of universal test and treat on HIV acquisition at population level in a high prevalence rural region of South Africa. We did a phase 4, open-label, cluster randomised trial of 22 communities in rural KwaZulu-Natal, South Africa. We included individuals residing in the communities who were aged 16 years or older. The clusters were composed of aggregated local areas (neighbourhoods) that had been identified in a previous study in the Hlabisa subdistrict. The study statisticians randomly assigned clusters (1:1) with MapInfo Pro (version 11.0) to either the control or intervention communities, stratified on the basis of antenatal HIV prevalence. We offered residents repeated rapid HIV testing during home-based visits every 6 months for about 4 years in four clusters, 3 years in six clusters, and 2 years in 12 clusters (58 cluster-years) and referred HIV-positive participants to trial clinics for ART (fixed-dose combination of tenofovir, emtricitabine, and efavirenz) regardless of CD4 cell count (intervention) or according to national guidelines (initially ≤350 cells per μL and <500 cells per μL from January, 2015; control). Participants and investigators were not masked to treatment allocation. We used dried blood spots once every 6 months provided by participants who were HIV negative at baseline to estimate the primary outcome of HIV incidence with cluster-adjusted Poisson generalised estimated equations in the intention-to-treat population after 58 cluster-years of follow-up. This study is registered with ClinicalTrials.gov, number NCT01509508, and the South African National Clinical Trials Register, number DOH-27-0512-3974. Between March 9, 2012, and June 30, 2016, we contacted 26 518 (93%) of 28 419 eligible individuals. Of 17 808 (67%) individuals with a first negative dried blood spot test, 14 223 (80%) had subsequent dried blood spot tests, of whom 503 seroconverted after follow-up of 22 891 person-years. Estimated HIV incidence was 2·11 per 100 person-years (95% CI 1·84-2·39) in the intervention group and 2·27 per 100 person-years (2·00-2·54) in the control group (adjusted hazard ratio 1·01, 95% CI 0·87-1·17; p=0·89). We documented one case of suicidal attempt in a woman following HIV seroconversion. 128 patients on ART had 189 life-threatening or grade 4 clinical events: 69 (4%) of 1652 in the control group and 59 (4%) of 1367 in the intervention group (p=0·83). The absence of a lowering of HIV incidence in universal test and treat clusters most likely resulted from poor linkage to care. Policy change to HIV universal test and treat without innovation to improve health access is unlikely to reduce HIV incidence. ANRS, GiZ, and 3ie.


Current and innovative approaches in the treatment of non-muscle invasive bladder cancer: the role of transurethral resection of bladder tumor and organoids.
Background Bladder cancer is the 7th most common cancer in men. About 75% of all bladder cancer are non-muscle invasive (NMIBC). The golden standard for definite diagnosis and first-line treatment of NMIBC is transurethral resection of bladder tumour (TURB). Historically, the monopolar current was used first, today bipolar current is preferred by most urologists. Following TURB, depending on the tumour grade, additional intravesical chemo- or/and immunotherapy is indicated, in order to prevent recurrence and need for surgical resection. Development of new technologies, molecular and cell biology, enabled scientists to develop organoids - systems of human cells that are cultivated in the laboratory and have characteristics of the tissue from which they were harvested. In the field of urologic cancers, the organoids are used mainly for studying the course of different diseases, however, in the field of bladder cancer the data are scarce. Conclusions Different currents - monopolar and bipolar, have different effect on urothelium, that is important for oncological results and pathohistological interpretation. Specimens of bladder cancer can be used for preparation of organoids that are further used for studying carcinogenesis. Bladder organoids are step towards personalised medicine, especially for testing effectiveness of chemo-/immunotherapeutics.


Using an Ontology to Facilitate More Accurate Coding of Social Prescriptions Addressing Social Determinants of Health: Feasibility Study.
National Health Service (NHS) England supports social prescribing in order to address social determinants of health, which account for approximately 80% of all health outcomes. Nevertheless, data on ongoing social prescribing activities are lacking. Although NHS England has attempted to overcome this problem by recommending 3 standardized primary care codes, these codes do not capture the social prescribing activity to a level of granularity that would allow for fair attribution of outcomes to social prescribing. In this study, we explored whether an alternative approach to coding social prescribing activity, specifically through a social prescribing ontology, can be used to capture the social prescriptions used in primary care in greater detail. The social prescribing ontology, implemented according to the Web Ontology Language, was designed to cover several key concepts encompassing social determinants of health. Readv2 and Clinical Terms Version 3 codes were identified using the NHS Terms Browser. The Royal College of General Practitioners Research Surveillance Centre, a sentinel network of over 1000 primary care practices across England covering a population of more than 4,000,000 registered patients, was used for data analyses for a defined period (ie, January 2011 to December 2019). In all, 668 codes capturing social prescriptions addressing different social determinants of health were identified for the social prescribing ontology. For the study period, social prescribing ontology codes were used 5,504,037 times by primary care practices of the Royal College of General Practitioners Research Surveillance Centre as compared to 29,606 instances of use of social prescribing codes, including NHS England's recommended codes. A social prescribing ontology provides a powerful alternative to the codes currently recommended by NHS England to capture detailed social prescribing activity in England. The more detailed information thus obtained will allow for explorations about whether outputs or outcomes of care delivery can be attributed to social prescriptions, which is essential for demonstrating the overall value that social prescribing can deliver to the NHS and health care systems.


Prospective memory impairment in former users of methamphetamine.
Considerable research indicates that methamphetamine use is associated with neurocognitive impairment, but no empirical study to date has assessed whether these difficulties extend to memory for future intentions (prospective memory). The present study assessed prospective performance on a laboratory measure of prospective memory that closely represents the types of prospective memory tasks that actually occur in everyday life and provides an opportunity to investigate the different sorts of prospective memory failures that occur ("Virtual Week"). Twenty adults with confirmed history of methamphetamine use and dependence, currently engaged in rehabilitation and confirmed to be abstinent for an average period of 6 months, and 20 methamphetamine-naive participants were tested on Virtual Week. Various other aspects of cognitive function were also assessed, including retrospective memory and executive functioning. Methamphetamine users were significantly impaired on Virtual Week, and these deficits did not vary as a function of specific prospective memory task demands. Of all the cognitive measures, cognitive inhibition shared greatest variance with group effects on the prospective memory measure. Prospective memory performance is sensitive to prior methamphetamine use even well into abstinence. Methamphetamine users experience generalized difficulties with prospective memory, suggesting that these deficits are likely to have important implications for day-to-day functioning.


Human platelets inhibit liver fibrosis in severe combined immunodeficiency mice.
To investigate the role of human platelets in liver fibrosis. Severe combined immunodeficiency (SCID) mice were administered CCl4 and either phosphate-buffered saline (PBS group) or human platelet transfusions (hPLT group). Concentrations of hepatocyte growth factor (HGF), matrix metallopeptidases (MMP)-9, and transforming growth factor-β (TGF-β) in the liver tissue were compared between the PBS and the hPLT groups by enzyme-linked immunosorbent assay (ELISA) and Western blotting. The effects of a human platelet transfusion on liver fibrosis included the fibrotic area, hydroxyproline content, and α-smooth muscle actin (α-SMA) expression, which were evaluated by picrosirius red staining, ELISA, and immunohistochemical staining using an anti-mouse α-SMA antibody, respectively. Phosphorylations of mesenchymal-epithelial transition factor (Met) and SMAD3, downstream signals of HGF and TGF-β, were compared between the two groups by Western blotting and were quantified using densitometry. Hepatocyte apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling. Furthermore, the accumulation of human platelets in the liver 2 h after platelet transfusion was compared between normal and fibrotic livers by immunohistochemical staining using an anti-human CD41 antibody. The fibrotic area and hydroxyproline content in the liver were both significantly lower in the hPLT group when compared to the PBS group (fibrotic area, 1.7% ± 0.6% vs 2.5% ± 0.6%, P = 0.03; hydroxyproline content, 121 ± 26 ng/g liver vs 156 ± 47 ng/g liver, P = 0.04). There was less α-smooth muscle actin staining in the hPLT group than in the PBS group (0.5% ± 0.1% vs 0.8% ± 0.3%, P = 0.02). Hepatic expression levels of mouse HGF and MMP-9 were significantly higher in the hPLT group than in the PBS group (HGF, 109 ± 13 ng/g liver vs 88 ± 22 ng/g liver, P = 0.03; MMP-9, 113% ± 7%/GAPDH vs 92% ± 11%/GAPDH, P = 0.04). In contrast, the concentration of mouse TGF-β in the liver tissue was significantly lower in the hPLT group than in the PBS group (22 ± 5 ng/g liver vs 39 ± 6 ng/g liver, P = 0.02). Phosphorylation of Met was more prevalent in the hPLT group than in the PBS group (37% ± 4%/GAPDH vs 20% ± 8%/GAPDH, P = 0.03). Phosphorylation of SMAD3 was weaker in the hPLT group than in the PBS group (60% ± 12%/GAPDH vs 84% ± 12%/GAPDH, P = 0.1), although this difference was not significant. Furthermore, a lower rate of hepatocyte apoptosis was observed in the hPLT group than in the PBS group (5.9% ± 1.7% vs 2.9% ± 2.1%, P = 0.02). Significant human platelet accumulation was observed in the fibrotic liver tissues, whereas few platelets accumulated in the normal liver. Human platelets inhibit liver fibrosis in SCID mice. Increased concentration of HGF in the liver suppresses hepatic stellate cell activation, induces MMPs, and inhibits hepatocyte apoptosis.


Palaeopathology: Current challenges and medical impact.
Palaeopathology is the science which studies ancient human diseases. Throughout its relatively young history it underwent tremendous technological and methodological improvements (from pure morphology and histology to CT scanning) that have constantly reshaped its scientific rationale. Among other achievements, the study of mummies and fossilized hominids has allowed to effectively extract ancient DNA, prove the existence of atherosclerosis in ancient times, demonstrate the presence of disease vectors, better clarify the etiology of infectious diseases otherwise only postulated on the basis of ancient accounts as well as to show the presence of spine pathology in our hominid ancestors. The research levels in this discipline are three: basic research, individual cases, population. The first and the third levels contribute most to the discipline, while the second is the one more appealing to the general public on account of its description of important cases reports. In addition, a recently introduced sub-specialty of palaeopathology, pathography is aiming to use an interdisciplinary approach to find traces of diseases in ancient literary sources and artistic representations. In spite of its discoveries, palaeopathology is not always viewed positively by clinicians because certain old-fashioned techniques are still due to technical restrictions. The authors provide a set of suggestions on how to strengthen the scientific recognition of this subject and explain at length how it could contribute to the progress of medical research. Clin. Anat. 29:816-822, 2016. © 2016 Wiley Periodicals, Inc.


During latency, herpes simplex virus type 1 DNA is associated with nucleosomes in a chromatin structure.
Latent herpes simplex virus type 1 DNA has a nucleosomal structure similar to that of cellular chromatin, as determined by micrococcal nuclease digestion. All of the major regions, including the transcriptionally active region of the genome, were found to be associated with nucleosomes. Such a chromatin structure is likely to be an important element in the control of herpes simplex virus type 1 gene expression during latency.


Protocol for a prospective, longitudinal cohort of people with COVID-19 and their household members to study factors associated with disease severity: the Predi-COVID study.
A few major clinical factors such as sex, obesity or comorbidities have already been associated with COVID-19 severity, but there is a need to identify new epidemiological, clinical, digital and biological characteristics associated with severity and perform deep phenotyping of patients according to severity. The objectives of the Predi-COVID study are (1) to identify new determinants of COVID-19 severity and (2) to conduct deep phenotyping of patients by stratifying them according to risk of complications, as well as risk factors for infection among household members of Predi-COVID participants (the Predi-COVID-H ancillary study). Predi-COVID is a prospective, hybrid cohort study composed of laboratory-confirmed COVID-19 cases in Luxembourg who will be followed up remotely for 1 year to monitor their health status and symptoms. Predi-COVID-H is an ancillary cohort study on household members of index cases included in Predi-COVID to monitor symptoms and household clusters in this high-risk population. A subcohort of up to 200 Predi-COVID and 300 Predi-COVID-H participants with biological samples will be included. Severity of infection will be evaluated by occurrence and duration of hospitalisation, admission and duration of stay in intensive care units or equivalent structures, provision of and duration of supplemental oxygen and ventilation therapy, transfer to another hospital, as well as the impact of infection on daily activities following hospital discharge. The study has been approved by the National Research Ethics Committee of Luxembourg (study number 202003/07) in April 2020. An informed consent is signed by study participants. Scientific articles will be submitted to international peer-reviewed journals, along with press releases for lay audience for major results. NCT04380987.


Cytogenetic effects of grape extracts (Vitis vinifera) and polyphenols on mitomycin C-induced sister chromatid exchanges (SCEs) in human blood lymphocytes.
In the present study, the effects of extracts and polyphenol-rich fractions as well as monomer polyphenols identified in them, from both red and white grapes, on mitomycin C (MMC) induced sister chromatid exchanges (SCEs) in human peripheral blood lymphocytes were investigated. The grape extracts and two of the three polyphenol-rich fractions promoted MMC-induced SCEs at concentrations from 75 to 300 microg/mL. However, none of the extracts or fractions alone induced SCEs. Thus, these results suggest caution especially with regard to the use of grape extracts as dietary supplements. On the other hand, the fact that these extracts were not genotoxic alone may indicate a selective activity against genetically damaged cells. This is the first study regarding the clastogenic effects of grape extracts in human cells. Moreover, from the tested polyphenols, caffeic acid, gallic acid, and rutin hydrate enhanced MMC-induced clastogenicity, whereas ferulic acid, protocatechuic acid, (+)-catechin, (-)-epicatechin, and trans-resveratrol had no effect at concentrations between 5 and 100 microM. The differences in the chemical structures of the tested polyphenols may account for their differential effects on MMC clastogenicity.


The influence of diet and nutrients on platelet function.
Cardiovascular disease (CVD) is the leading cause of death worldwide. Platelet activation and aggregation play an integral role in hemostasis and thrombosis. Diets and nutrients play a potential role in modifying CVD progression, particularly in platelet function, and have the potential of altering platelet function tests. Diets such as Mediterranean diet, high in omega-3 polyunsaturated fatty acids (PUFA), and vegetarian diets have inverse relationships with CVD. Dark chocolate, foods with low glycemic index, garlic, ginger, omega-3 PUFA, onion, purple grape juice, tomato, and wine all reduce platelet aggregation. Dark chocolate and omega-3 PUFA also reduce P-selectin expression. In addition, dark chocolate reduces PAC-1 binding and platelet microparticle formation. Berries inhibit platelet function (PFA-100). Energy drinks have been shown to increase platelet aggregation and caffeine increases platelet microparticle formation. Therefore, repeat testing of platelet function may be required, not only after exclusion of known antiplatelet medications but also potentially after exclusion of dietary substances/nutrients that could have plausibly affected initial test data.


Development of liposome gel based formulations for intravaginal delivery of the recombinant HIV-1 envelope protein CN54gp140.
Mucosally-administered vaccine strategies are widely investigated as a promising means of preventing HIV infection. This study describes the development of liposomal gel formulations, and novel lyophilised variants, comprising HIV-1 envelope glycoprotein, CN54gp140, encapsulated within neutral, positively charged or negatively charged liposomes. The CN54gp140 liposomes were evaluated for mean vesicle diameter, polydispersity, morphology, zeta potential and antigen encapsulation efficiency before being incorporated into hydroxyethyl cellulose (HEC) aqueous gel and subsequently lyophilised to produce a rod-shaped solid dosage form for practical vaginal application. The lyophilised liposome-HEC rods were evaluated for moisture content and redispersibility in simulated vaginal fluid. Since these rods are designed to revert to gel form following intravaginal application, mucoadhesive, mechanical (compressibility and hardness) and rheological properties of the reformed gels were evaluated. The liposomes exhibited good encapsulation efficiency and the gels demonstrated suitable mucoadhesive strength. The freeze-dried liposome-HEC formulations represent a novel formulation strategy that could offer potential as stable and practical dosage form.


Opsin-like immunoreactivity in the circadian pacemaker neurons and photoreceptors of the eye of the opisthobranch mollusc Bulla gouldiana.
Circadian pacemaker cells in the eyes of the opisthobranch mollusc Bulla gouldiana generate a near 24-h rhythm in the frequency of optic nerve impulses. Previous electrophysiological studies suggest that these basal retinal neurons are intrinsically photosensitive and transduce light signals that shift the phase of their pacemaker mechanism. To test whether the pacemaker neurons contain opsin-like proteins, several polyclonal antibodies that recognize opsins of vertebrate photoreceptors have been tested on histological sections of the eye and on the neurons in primary cell culture. The antibodies label both the pacemaker cells and the large distal photoreceptors that surround the lens. Immunoblot analyses of the proteins of the eye have identified a single band at 62+/-4 kDa. These opsin antibodies may label the photopigment used in the entrainment of the circadian pacemaker.


Disulfide bonds within the C2 domain of RAGE play key roles in its dimerization and biogenesis.
The receptor for advanced glycation end products (RAGE) on the cell surface transmits inflammatory signals. A member of the immunoglobulin superfamily, RAGE possesses the V, C1, and C2 ectodomains that collectively constitute the receptor's extracellular structure. However, the molecular mechanism of RAGE biogenesis remains unclear, impeding efforts to control RAGE signaling through cellular regulation. METHODOLOGY AND RESULT: We used co-immunoprecipitation and crossing-linking to study RAGE oligomerization and found that RAGE forms dimer-based oligomers. Via non-reducing SDS-polyacrylamide gel electrophoresis and mutagenesis, we found that cysteines 259 and 301 within the C2 domain form intermolecular disulfide bonds. Using a modified tripartite split GFP complementation strategy and confocal microscopy, we also found that RAGE dimerization occurs in the endoplasmic reticulum (ER), and that RAGE mutant molecules without the double disulfide bridges are unstable, and are subjected to the ER-associated degradation. Disulfide bond-mediated RAGE dimerization in the ER is the critical step of RAGE biogenesis. Without formation of intermolecular disulfide bonds in the C2 region, RAGE fails to reach cell surface. This is the first report of RAGE intermolecular disulfide bond.