Glutamate excitoxicity is the key molecular mechanism which is influenced by body temperature during the acute phase of brain stroke.
Glutamate excitotoxicity, metabolic rate and inflammatory response have been associated to the deleterious effects of temperature during the acute phase of stroke. So far, the association of temperature with these mechanisms has been studied individually. However, the simultaneous study of the influence of temperature on these mechanisms is necessary to clarify their contributions to temperature-mediated ischemic damage. We used non-invasive Magnetic Resonance Spectroscopy to simultaneously measure temperature, glutamate excitotoxicity and metabolic rate in the brain in animal models of ischemia. The immune response to ischemia was measured through molecular serum markers in peripheral blood. We submitted groups of animals to different experimental conditions (hypothermia at 33°C, normothermia at 37°C and hyperthermia at 39°C), and combined these conditions with pharmacological modulation of glutamate levels in the brain through systemic injections of glutamate and oxaloacetate. We show that pharmacological modulation of glutamate levels can neutralize the deleterious effects of hyperthermia and the beneficial effects of hypothermia, however the analysis of the inflammatory response and metabolic rate, demonstrated that their effects on ischemic damage are less critical than glutamate excitotoxity. We conclude that glutamate excitotoxicity is the key molecular mechanism which is influenced by body temperature during the acute phase of brain stroke.


Intra-operative assessment of human pulmonary alveoli in vivo using Sidestream Dark Field imaging: a feasibility study.
In vivo videomicroscopy has been used for years to visualize subpleural alveoli in animal studies. This has led to a better understanding of alveolar physiology. We tested the hypothesis whether a novel handheld videomicroscope could be used for intraoperative detection of alveoli in surgical patients during mechanical ventilation. Using Sidestream Dark Field imaging, we observed 6 patients (3 adults and 3 children) who underwent elective cardiac surgery. In each patient, the tip of the microscope was placed on the visceral pleural surface of the left upper pulmonary lobe after weaning from cardiopulmonary bypass. The acquired images were converted into digital signals and captured on a computer. Although cardiac motion artifacts were present, visceral pleural microvascular blood flow could be observed in adults and infants. In infants, sub-pleural cavities (alveoli) were observed. These alveoli were remarkably similar in dimension and structure to those identified previously as true alveoli in animal studies. Quantification of these alveoli demonstrated that mean alveolar diameter, perimeter and area increased with age among the investigated infants (all parameters p<0.001). High-quality images of visceral pleural microvessels as well as subpleural cavities, reflecting superficial alveoli, could be obtained in infants. These findings create the opportunity to begin human intervention studies, which should investigate alveolar dynamics during mechanical ventilation in cardio-thoracic surgery in more detail.


Use of critical access hospital emergency rooms by patients with mental health symptoms.
National data demonstrate that mental health (MH) visits to the emergency room (ER) comprise a small, but not inconsequential, proportion of all visits; however, we lack a rural picture of this issue. This study investigates the use of critical access hospital (CAH) ERs by patients with MH problems to understand the role these facilities play in rural MH needs and the challenges they face. Primary data were collected through the combination of a telephone survey and ER visit logs. Our sampling frame was the universe of CAHs at the time the survey was fielded. About 43% of CAHs surveyed operate in communities with no MH services, while 9.4% of all logged visits were by patients identified as having some type of MH problem. The most common problems identified were affective disorders, substance abuse, anxiety, and psychotic disorders. Only 32% of CAHs have access to on-site detoxification and 2% have inpatient psychiatric services, meaning that patients in need of these services typically must leave their communities to gain treatment. The lack of community resources may impact CAHs' ability to assist patients with MH problems. Among those with a primary MH condition, 21% left the ER with no or unknown treatment, as did 51% of patients whose MH condition was secondary to their emergent problem. Patients in need of detoxification or inpatient psychiatric services often must travel over an hour to obtain these services, potentially creating significant issues for themselves and their families.


Hyperacusis in Children: A Preliminary Study on the Effects of Hypersensitivity to Sound on Speech and Language.
There is a growing awareness that children may experience hyperacusis, a condition that is often associated with behavioral and developmental disorders. This preliminary study was aimed to investigate the effects of hyperacusis alone on various components of speech and language in children without developmental disorders. This study was conducted on 109 children aged between 4 and 7 years attending kindergarten and primary school. Hyperacusis was assessed through behavioral observation of children and questionnaires for parents. Different components of speech and language were assessed through specific tests. Hyperacusis was diagnosed in fifteen children (13.8%); ten (66.7%) were attending primary school and five (33.3%) kindergarten. A significant difference between children with and without hyperacusis was found for tests evaluating the average number of words in a sentence and phonemic fluency; older children appeared to have more difficulties. Several differences in education profiles were found: parents of children with hyperacusis spent less time with their children compared to parents of children without hyperacusis. Our preliminary results suggest some difficulties in lexical access and the use of shorter sentences by children with hypersensitivity to sound; however, the small size of our sample and the largely unknown interactions between hyperacusis and developmental disorders suggest caution when interpreting these results. Further studies on larger samples are necessary to gain additional knowledge on the effects of hyperacusis on speech and language in children without developmental disorders.


[Assessment of microbiological quality of food served in dining rooms of private enterprises].
The aim of this study was to evaluate foods microbiological quality in food-service establishments. A total of 620 food samples were obtained from four establishments and analyzed for aerobic mesophilic (AM), yeasts, moulds, Staphylococcus aureus and Escherichia coli counts and the presence of Salmonella spp. Drinking water, equipment, surfaces, environment and food handlers were also sampled. E. coli was found in raw vegetables (76.2%), cooked vegetables (15.2%), beef and pork (15.9%), poultry (16.7%), fish (11.8%), desserts (27.3%), equipments (57.9%), surfaces and environment (53.6%) and in 21.9% of food handlers. Survey results were compared with the recommended maximum microbial levels. Our results demonstrate the need for the adoption of more effective hygienic measures in this kind of establishment in order to avoid any risk to consumers.


Cross-amplification and characterization of microsatellite markers in Alcantarea patriae Versieux & Wand.
The Alcantarea patriae is a Bromeliaceae endemic to the inselbergs of the Atlantic Forest. This taxon, described in the year of 2007 by Versieux & Wanderley, presents restricted and fragmented distribution outside conservation units. Studies to evaluate the genetic structure of its populations can contribute to the conservation and management strategies for the species. In this study, 31 microsatellite markers, descriptive to six different Bromeliaceae species, were evaluated by cross-amplification tests in 20 individuals of A. patriae. The individuals were collected in the district of Vila Cruzeiro, in the municipality of Jerônimo Monteiro. Twelve markers were polymorphic and 10 monomorphic, with an amplification success rate of 71%. The displayed polymorphism information content was considered high, indicating that the selected markers are informative. The values found for the fixation index were positive and indicated the occurrence of inbreeding. The mean number of alleles was 4.66 (3-6), the mean expected and observed heterozygosities were 0.6605 and 0.4618, respectively. The detection of polymorphic markers was important for future studies of diversity and genetic structuring of natural populations and for germplasm bank creation aiming to contribute to in situ and ex situ conservations of A. patriae.


A systematic review of socio-economic assessments in support of coastal zone management (1992-2011).
Cooperation between the social and natural sciences has become essential in order to encompass all the dimensions of coastal zone management. Socio-economic approaches are increasingly recommended to complement integrated assessment in support of these initiatives. A systematic review of the academic literature was carried out in order to analyze the main types of socio-economic assessments used to inform the coastal zone management process as well as their effectiveness. A corpus of 1682 articles published between 1992 and 2011 was identified by means of the representative coverage approach, from which 170 were selected by applying inclusion/exclusion criteria and then classified using a content analysis methodology. The percentage of articles that mention the use of socio-economic assessment in support of coastal zone management initiatives is increasing but remains relatively low. The review examines the links between the issues addressed by integrated assessments and the chosen analytical frameworks as well as the various economic assessment methods which are used in the successive steps of the coastal zone management process. The results show that i) analytical frameworks such as 'risk and vulnerability', 'DPSIR', 'valuation', 'ecosystem services' and 'preferences' are likely to lead to effective integration of social sciences in coastal zone management research while 'integration', 'sustainability' and 'participation' remain difficult to operationalize, ii) risk assessments are insufficiently implemented in developing countries, and iii) indicator systems in support of multi-criteria analyses could be used during more stages of the coastal zone management process. Finally, it is suggested that improved collaboration between science and management would require that scientists currently involved in coastal zone management processes further educate themselves in integrated assessment approaches and participatory methodologies.


Systemic and mucosal humoral immune responses induced by the JY-adjuvanted nasal spray H7N9 vaccine in mice.
Since the first case of human avian influenza A (H7N9) virus infection in 2013, five H7N9 epidemics have occurred in China, all of which caused severe diseases, including pneumonia and acute respiratory distress syndrome, and the fatality rates of these epidemics were as high as 30-40%. To control the prevalence of H7N9 influenza, an effective vaccine is urgently needed. In the present study, we used chitosan and recombinant human interleukin-2 as an adjuvant (JY) to promote the systemic and mucosal immune responses induced by the H7N9 vaccine. Mice were immunized intranasally with the inactivated split influenza A (H7N9) virus (A/Shanghai/02/2013) vaccine with or without JY. The hemagglutination inhibition (HI) titers of mice immunized with the JY-adjuvanted vaccine were significantly higher than those of mice immunized with the vaccine without adjuvant (21.11 ± 9.58 vs. 5.04 ± 3, P < 0.05). The JY-adjuvanted H7N9 nasal spray vaccine induced higher HI titers (8 ± 0.82 vs. 6.7 ± 0.67, P = 0.0035) than those did the poly (I:C)-adjuvanted H7N9 vaccine or the LTB-adjuvanted H7N9 vaccine (8 ± 0.82 vs. 6.9 ± 0.88, P = 0.0186). The optimal immunization regimen for the nasal spray H7N9 vaccine was determined to be a 21-day interval between the primary immunization and booster, with a dose of 4.5 μg hemagglutinin per mouse. The immunogenicities of the nasal spray H7N9 vaccine and intramuscular vaccine (containing only the inactivated split virus) were compared in mice. Two doses of the nasal spray H7N9 vaccine induced higher titers of HI (6.7 ± 0.67 vs. 5.3 ± 1.16, P = 0.004) and anti-HA IgG in sera (19.26 ± 0.67 vs. 13.97 ± 0.82, P < 0.0001) and of anti-HA sIgA (7.13 ± 2.54 vs. 0, P = 0.0000) in bronchoalveolar lavage fluid (BALF) than one dose of intramuscular H7N9 vaccine 3 weeks after the last immunization. However, when we immunized the mice with two doses of both vaccines separately, the nasal spray H7N9 vaccine induced higher titers of anti-HA IgG (19.26 ± 0.67 vs. 17.56 ± 0.57, P < 0.0001) and anti-HA sIgA (7.13 ± 2.54 vs. 4.02 ± 0.33, P = 0.0026) than did the intramuscular H7N9 vaccine, and there was no difference in HI titer between the two groups (P = 0.3745). This finding indicates that the JY-adjuvanted nasal spray H7N9 vaccine induced not only the systemic immune response but also a local mucosal response, which may improve the efficacy of H7N9 influenza prevention through respiratory tract transmission.


Posterior epistaxis: endonasal exposure and occlusion of the branches of the sphenopalatine artery.
Intractable posterior epistaxis (PE) is a frequent emergency for which different treatment modalities are available. While nasal packing causes extreme discomfort and angiography with consecutive selective embolization is not available everywhere, recent studies emphasize the value of sphenopalatine artery (SPA) occlusion by different techniques and indicate success rates of 13-33%. In our institution, previously endoscopic management of PE consisted either of isolated coagulation of an identified bleeding source (group A) or cutting and coagulation of arterial branches running through the sphenopalatine foramen (SPF) (group B). According to our neuroradiological and rhinological experience we developed a modification of SPA transsection and coagulation following identification of the division in conchal and septal branches of the SPA (group C). During a 26-month period the success rates of these three techniques in 95 patients were compared prospectively. The three modalities revealed a re-bleeding rate of 3 out of 21 (21%) in group A, 1 in 6 (16.7%) in group B and 3 in 69 (4.3%) in group C. At the level of the SPF, 36 out of 69 patients had one conchal branch, whereas 30 (43.5%) had two and 3 (4.4%) had three. If SPA transsection and coagulation for intractable PE is adopted the anatomic varieties of the SPA with its division in conchal and septal branches have to be taken into account. According to our experience the septal branch of the SPA plays a major role in PE. Its occlusion significantly improves the success rate of PE treatment.


Histochemical aspects of the vascular invasion at the erosion zone of the epiphyseal cartilage in MMP-9-deficient mice.
We have histologically examined vascular invasion and calcification of the hypertrophic zone during endochondral ossification in matrix metalloproteinase (MMP)-9 deficient (MMP-9-/-) mice and in their littermates at 3 days, 3 weeks and 6 weeks after birth. Capillaries and osteoclasts at the chondro-osseous junction showed an intense MMP-9 immunopositivity, suggesting that they recognize chemical properties of cartilaginous matrices, and then release MMP-9 for cartilage degradation. CD31-positive capillaries and tartrate-resistant acid phosphatase-reactive osteoclasts could be found in the close proximity in the region of chondro-osseous junction in MMP-9-/- mice, while in wild-type mice, vascular invasion preceded osteoclastic migration into the epiphyseal cartilage. Although MMP-9-/- mice revealed larger hypertrophic zones, the index of calcified area was significantly smaller in MMP-9-/- mice. Interestingly, the lower layer of the MMP-9-/- hypertrophic zone showed intense MMP-13 staining, which could not be observed in wild-type mice. This indicates that MMP-13 may compensate for MMP-9 deficiency at that specific region, but not to a point at which the deficiency could be completely rescued. In conclusion, it seems that MMP-9 is the optimal enzyme for cartilage degradation during endochondral ossification by controlling vascular invasion and subsequent osteoclastic migration.


Increased risk of ischaemic heart disease in shift workers.
504 papermill workers were followed up for 15 years and the incidence of ischaemic heart disease (IHD) in shift workers was compared with that in day workers. The relative risk (RR) of IHD rose with increasing duration of reported exposure to shift work. A significant risk of IHD was associated with an exposure of 11 - 15 years (RR = 2.2, p less than 0.04) and of 16 to 20 years (RR = 2.8, p less than 0.03. The association was independent of age and smoking history. The RR of IHD fell sharply after 20 years of shift work. This was ascribed to the pronounced positive selection that had taken place in this group.


Synergistic improvements in cell and axonal migration across sciatic nerve lesion gaps using bioresorbable filaments and heregulin-beta1.
The success of entubulation for peripheral nerve regeneration is still limited, especially with long lesion gaps. In this study, we examined if regeneration could be enhanced by constructing implants to both align axonal growth and promote Schwann cell proliferation and migration. Silicone implants were used to bridge a 1.4-cm gap in the rat sciatic nerve. Adult female Sprague-Dawley rats were divided into four groups of tubes containing either 1) Matrigel; 2) Matrigel and heregulin; 3) Matrigel and poly(L-lactic acid) (PLLA) microfilaments; or 4) Matrigel, PLLA microfilaments, and heregulin. Ten weeks postimplantation, the number of axons and Schwann cells were measured at the distal end of implants. Implants with microfilaments displayed better tissue cable formation, increased Schwann cell migration, and regeneration of anti-calcitonin gene-related peptide-positive axons, but not RMDO95-positive axons compared with nonfilament-containing groups. Heregulin treatment caused an increase in Schwann cell number, but it demonstrated no significant improvement in either tissue cable formation or axon number. Extensive regeneration was observed through implants containing Matrigel, microfilaments, and heregulin, which induced significant improvements in the number and longitudinal organization of both Schwann cells and axons. These results indicate that physical guidance of microfilaments and the Schwann cell growth factor, heregulin, act synergistically to improve nerve regeneration across long lesion gaps.


Pneumonectomy is a valuable treatment option after neoadjuvant therapy for stage III non-small-cell lung cancer.
The mortality of pneumonectomy after chemotherapy or chemoradiotherapy for locally advanced non-small-cell lung cancer is reported to be as high as 26%. We retrospectively reviewed the medical records of patients undergoing these procedures in 2 specialized thoracic centers to determine the outcome. Retrospective analyses were performed of all patients who underwent pneumonectomy after neoadjuvant chemotherapy or chemoradiotherapy for locally advanced non-small-cell lung cancer from 1998 to 2007. Presurgical treatment consisted of 3-4 platin-based doublets alone in 20% of patients or combined with radiotherapy (45Gy) to the tumor and mediastinum in 80% of patients. Of 827 patients who underwent neoadjuvant therapy, 176 pneumonectomies were performed, including 138 (78%) extended resections. Post-induction pathologic stages were 0 in 36 patients (21%), I in 33 patients (19%), II in 38 patients (21%), III in 57 patients (32%), and IV in 12 patients (7%). Three patients died of pulmonary embolism, 2 patients of respiratory failure, and 1 patient of cardiac failure, resulting in a 90 postoperative day mortality rate of 3%; 23 major complications occurred in 22 patients (13%). For the overall population, 3-year survival was 43% and 5-year survival was 38%. Pneumonectomy after neoadjuvant therapy for non-small-cell lung cancer can be performed with a perioperative mortality rate of 3%. Thus, the need of a pneumonectomy for complete resection alone should not be a reason to exclude patients from a potentially curative procedure if done in an experienced center. The 5-year survival of 38%, which can be achieved, justifies extended surgery within a multimodality concept for selected patients with locally advanced non-small-cell lung cancer.


Membrane vesiculation function and exocytosis of wild-type and mutant matrix proteins of vesicular stomatitis virus.
Transfection of mammalian CV1 cells with a recombinant M-gene pTM1 plasmid, driven by vaccinia virus-expressed phage T7 polymerase, resulted in the expression of matrix (M) protein, which is progressively released from the exterior surface of the transfected-cell plasma membrane. Exocytosis of M protein begins 2 to 4 h posttransfection and reaches a peak by 10 to 16 h posttransfection; dye uptake studies reveal that > 97% of cells are alive and have intact membranes at 16 h posttransfection. Density gradient centrifugation and labeling with radioactive palmitic acid revealed that the M protein is released from cells in association with lipid vesicles. Expression of M-gene deletion mutants suggests that exocytosis of M protein requires the presence of a membrane-binding site at N-terminal amino acids 1 to 50. Cells transfected with the pTM1 plasmid containing the M gene of the temperature-sensitive mutant tsO23 expressed ample quantities of the mutant M protein at permissive (31 degrees C) and restrictive (39 degrees C) temperatures, but the exocytosis of the mutant M protein occurred only at the permissive temperature. The tsO23 M gene has three site-specific mutations resulting in amino acid substitutions at residues 21, 111, and 227. Expression of wild-type and mutant M genes with mutations or revertants at each of these sites resulted in exocytosis of M protein at the nonpermissive temperature only when wild-type leucine was present at residue 111, but M-protein exocytosis was restricted (to some extent even at the permissive temperature) when mutant phenylalanine was present at residue 111. Past and present data indicate that a specific structural conformation of the M protein is responsible for the formation and budding of vesicles, a property of the M protein which probably also promotes vesicular stomatitis virus assembly and budding of virions from host cells.


The Differential Effect of Toxoplasma Gondii Infection on the Stability of BCL2-Family Members Involves Multiple Activities.
The regulation of mitochondrial permeability, a key event in the initiation of apoptosis is governed by the opposing actions of the pro- and anti-apoptotic members of the BCL2-family of proteins. The BCL2-family can be classified further based on the number of BCL-homology (BH) domains they encode. Pathogen mediated modulation of BCL2-family members play a significant role in their ability to affect the apoptotic pathways in the infected host cell. The protozoan parasite Toxoplasma gondii establishes a profound blockade of apoptosis noted by a requirement for host NFκB activity and correlating with the selective degradation of pro-apoptotic BCL2-family members. In this study, we explore the potential activities associated with the inherent stability of the anti-apoptotic BCL2 as well as the selective degradation of the pro-apoptotic proteins BAX, BAD, and BID. We find that multiple activities govern the relative stability of BCL2-family members suggesting a complex and balanced network of stability-enhancing and-destabilizing activities are perturbed by parasite infection. The data leave open the possibility for both parasite induced host activities as well as the direct consequence of parasite effectors in governing the relative levels of BCL2-proteins in the course of infection.


Frequency of CHEK2*1100delC in New York breast cancer cases and controls.
The 1100delC CHEK2 allele has been associated with a 1.4-4.7 fold increased risk for breast cancer in women carrying this mutation. While the frequency of 1100delC was 1.1-1.4% in healthy Finnish controls, the frequency of this allele in a North American control population and in North American breast cancer kindreds remains unclear. We genotyped 1665 healthy New York volunteers and 300 cases of breast cancer for the CHEK2*1100delC. The overall frequency of the 1100delC was 3/300 (1.0%) among all cases with either a family history of breast cancer (n = 192) or a personal history of breast cancer (n = 108, of which 46 were bilateral, 46 unilateral, and 16 were male breast cancer cases), compared to a frequency of 5/1665 (0.3%) in healthy controls (p = 0.1). There was no difference in allele frequency among Ashkenazi and non-Ashkenazi controls. The relatively low breast cancer penetrance of this allele, along with the low population frequency, will limit the clinical applicability of germline testing for CHEK2*1100delC in North American kindreds.


Psychometric Properties of Child (0-5 Years) Outcome Measures as used in Randomized Controlled Trials of Parent Programs: A Systematic Review.
This systematic review is one of the three which sought to identify measures commonly implemented in parenting program research, and to assess the level of psychometric evidence available for their use with this age group. This review focuses specifically on measures of child social-emotional and behavioral outcomes. Two separate searches of the same databases were conducted; firstly to identify eligible instruments, and secondly to identify studies reporting on the psychometric properties of the identified measures. Five commercial platforms hosting 19 electronic databases were searched from their inception to conducted search dates. Twenty-four measures were identified from Search 1: a systematic search of randomized controlled trial evaluations of parenting programs. For Search 2, inclusion/exclusion criteria were applied to 21,329 articles that described the development and/or validation of the 24 measures identified in Search 1. Thirty articles met the inclusion criteria. resulting in 11 parent report questionnaires and three developmental assessment measures for review. Data were extracted and synthesized to describe the methodological quality of each article using the COSMIN checklist alongside the overall quality rating of the psychometric property reported for each measure. Measure reliability was categorized into four domains (internal consistency, test-re-test, inter-rater, and intra-rater). Measure validity was categorized into four domains (content, structural, convergent/divergent, and discriminant). Results indicated that supporting evidence for included measures is weak. Further work is required to improve the evidence base for those measures designed to assess children's social-emotional and behavioral development in this age group. PROSPERO Registration number: CRD42016039600.


Bacillus subtilis natto: a non-toxic source of poly-γ-glutamic acid that could be used as a cryoprotectant for probiotic bacteria.
It is common practice to freeze dry probiotic bacteria to improve their shelf life. However, the freeze drying process itself can be detrimental to their viability. The viability of probiotics could be maintained if they are administered within a microbially produced biodegradable polymer - poly-γ-glutamic acid (γ-PGA) - matrix. Although the antifreeze activity of γ-PGA is well known, it has not been used for maintaining the viability of probiotic bacteria during freeze drying. The aim of this study was to test the effect of γ-PGA (produced by B. subtilis natto ATCC 15245) on the viability of probiotic bacteria during freeze drying and to test the toxigenic potential of B. subtilis natto. 10% γ-PGA was found to protect Lactobacillus paracasei significantly better than 10% sucrose, whereas it showed comparable cryoprotectant activity to sucrose when it was used to protect Bifidobacterium breve and Bifidobacterium longum. Although γ-PGA is known to be non-toxic, it is crucial to ascertain the toxigenic potential of its source, B. subtilis natto. Presence of six genes that are known to encode for toxins were investigated: three component hemolysin (hbl D/A), three component non-haemolytic enterotoxin (nheB), B. cereus enterotoxin T (bceT), enterotoxin FM (entFM), sphingomyelinase (sph) and phosphatidylcholine-specific phospholipase (piplc). From our investigations, none of these six genes were present in B. subtilis natto. Moreover, haemolytic and lecithinase activities were found to be absent. Our work contributes a biodegradable polymer from a non-toxic source for the cryoprotection of probiotic bacteria, thus improving their survival during the manufacturing process.


Who wears the pants in a mute swan pair? Deciphering the effects of male and female age and identity on breeding success.
Traditionally, many breeding traits (e.g. the timing and size of clutches) were considered to be female-only traits in that males played little-to-no role in their expression. Although the contribution of males to such breeding traits, as well as other aspects of reproduction, is increasingly recognized, few studies have demonstrated the effects of male age and life history on breeding traits and, importantly, whether these effects are underlined by additive-genetic variation. Here, we take advantage of a long-term data set on mute swans (Cygnus olor) to demonstrate that the ages of both the male and female parents play significant roles in the timing and size of clutches, although recruitment success did not show similar effects. Individual males varied significantly in their influence on the timing of egg laying. We decomposed this variation using an 'animal model'; competing models that were the source of this variation as additive-genetic or permanent-environmental variation was not statistically distinguishable. Our results add to the growing evidence that reproductive performance should be considered as a product of the identity and condition of both parents.


Determination of Aflatoxin M1 in Raw Milk from Different Provinces of Ecuador.
Aflatoxin M1 (AFM1) is a mycotoxin from Aspergillus flavus and A. parasiticus, classified as carcinogenic and hepatotoxic. The objective of the present investigation was to determine its presence in raw milk from north-central Ecuador, constituted by the provinces of Pichincha, Manabí, and Santo Domingo de los Tsáchilas. These areas represent approximately 30% of Ecuadorian milk production. By the end of the investigation, a total of 209 raw milk samples were collected, obtained both during the dry (June and August) and rainy seasons (April and November) of 2019. AFM1 concentrations were measured with lateral flow immunochromatographic assays, and 100% of the samples were positive for this mycotoxin, presenting a mean value of 0.0774 μg/kg with a range of 0.023 to 0.751 μg/kg. These AFM1 levels exceeded the European Union regulatory limit of 0.05 μg/kg in 59.3% (124/209) of samples, while only 1.9% (4/209) exceeded the Ecuadorian legal limit of 0.5 μg/kg. By using non-parametric tests, significant differences were determined (p ≤ 0.05) between the provinces for months of study, climatic season (being higher in the dry season), and climatic region (greater in the coast region). On the other hand, there were no significant differences (p ≥ 0.05) between the types of producers or between production systems. Therefore, AFM1 contamination in raw milk does not present a serious public health problem in Ecuador, but a monitoring and surveillance program for this mycotoxin in milk should be developed to prevent consumer health problems.


Hormonal changes during salinity-induced leaf senescence in tomato (Solanum lycopersicum L.).
Leaf senescence is one of the most limiting factors to plant productivity under salinity. Both the accumulation of specific toxic ions (e.g. Na+) and changes in leaf hormone relations are involved in the regulation of this process. Tomato plants (Solanum lycopersicum L. cv Moneymaker) were cultivated for 3 weeks under high salinity (100 mM NaCl) and leaf senescence-related parameters were studied during leaf development in relation to Na+ and K+ contents and changes in abscisic acid (ABA), cytokinins, the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC), and the auxin indole-3-acetic acid (IAA). Na+ accumulated to a similar extent in both leaves 4 and 5 (numbering from the base of the plant) and more quickly during the third week, while concurrently K+ contents sharply decreased. However, photosystem II efficiency, measured as the F(v)/F(m) ratio, decreased from the second week of salinization in leaf 4 but only at the end of the third week in the younger leaf 5. In the prematurely senescent leaf 4, ABA content increased linearly while IAA strongly decreased with salinization time. Although zeatin (Z) levels were scarcely affected by salinity, zeatin-riboside (ZR) and the total cytokinin content (Z+ZR) progressively decreased by 50% from the imposition of the stress. ACC was the only hormonal compound that increased in leaf tissue coincident with the onset of oxidative damage and the decline in chlorophyll fluorescence, and prior to massive Na+ accumulation. Indeed, (Z+ZR) and ACC contents and their ratio (Z+ZR/ACC) were the hormonal parameters best correlated with the onset and progression of leaf senescence. The influence of different hormonal changes on salt-induced leaf senescence is discussed.


Uncommon localizations of hydatid cyst. Review of the literature.
Hydatid disease is an endemic anthropozoonosis with usual localization in liver and lungs. Rarely it localizes in uncommon sites as spleen, skeleton, kidney, brain, cardiac muscle, peritoneum, sub cutis. Complications of uncommon localizations are the same that for usual ones. Review of the literature on rare and atypical localization of hydatid cysts in soft tissues. Key-words used on Pub-Med [(echinococ OR hydatid) AND (soft tissue OR subcutaneous OR cutaneous)] without time limit. There were found 282 articles; 242 were excluded because of muscular or bone localizations. 40 were coherent. Different variables are taken into account: age, sex, geographic area, anatomic localization of the cyst, dimension, symptoms, signs, mobility, blood exams and specific serological tests, imaging techniques for diagnosis, existing of septa in the structure, treatment, anaesthesia, spillage, neo-adjuvant and adjuvant treatment, follow-up period, recurrent lesions. It would be useful create an homogeneous and standardized collection of data of these rare and potentially life-threatening conditions in order to create guide-line of diagnostic and therapeutic process and create (or adopt) unique classification of the lesions.


Sympatric incipient speciation of spiny mice Acomys at "Evolution Canyon," Israel.
Does the paucity of empirical evidence of sympatric speciation in nature reflect reality, despite theoretical support? Or is it due to inappropriate searches in nature with overly restrictive assumptions and an incorrect null hypothesis? Spiny mice, Acomys, described here at Evolution Canyon (EC) incipiently and sympatrically speciate owing to microclimatic interslope divergence. The opposite slopes at EC vary dramatically, physically and biotically, representing the dry and hot south-facing slope savannoid-African continent ["African" slope (AS)], abutting with the north-facing slope forested south-European continent ["European" slope (ES)]. African-originated spiny mice, of the Acomys cahirinus complex, colonized Israel 30,000 y ago based on fossils. Genotypically, we showed significantly higher genetic diversity of mtDNA and amplified fragment length polymorphism of Acomys on the AS compared with the ES. This is also true regionally across Israel. In complete mtDNA, 25% of the haplotypes at EC were slope-biased. Phenotypically, the opposite slope's populations also showed adaptive morphology, physiology, and behavior divergence paralleling regional populations across Israel. Preliminary tests indicate slope-specific mate choices. Colonization of Acomys at the EC first occurred on the AS and then moved to the ES. Strong slope-specific natural selection (both positive and negative) overrules low interslope gene flow. Both habitat slope selection and mate choices suggest ongoing incipient sympatric speciation. We conclude that Acomys at the EC is ecologically and genetically adaptively, incipiently, sympatrically speciating on the ES owing to adaptive microclimatic natural selection.


Climatic influences on wood anatomy and tree-ring features of Great Basin conifers at a new mountain observatory.
A network of mountain observing stations has been installed in the Great Basin of North America. NevCAN (Nevada Climate-ecohydrological Assessment Network), which spans a latitudinal range of 2.5° and two elevation ranges of about 2000 m each, enabled us to investigate tree growth in relation to climate. • We analyzed wood anatomy and tree-ring characteristics of four conifer species in response to different levels of water availability by comparing a low- and a high-elevation population. Chronologies of earlywood and latewood widths, as well as cellular parameters, were developed from the year 2000 to 2012. • At the southern (drier and warmer) sites, Pinus monophylla had smaller cell lumen, tracheid diameter, and cell wall thickness. Pinus monophylla and P. flexilis showed bigger cellular elements at the higher elevations, whereas the opposite pattern was found in Picea engelmannii and Pinus longaeva. When all species and sites were pooled together, stem diameter was positively related with earlywood anatomical parameters. • We have provided a glimpse of the applications that NevCAN, as a new scientific tool, could allow in the general field of botany. In particular, we were able to investigate how differences in water stress related to elevation lead to changes in xylem anatomy.


Interposition Dermal Matrix Xenografts: A Successful Alternative to Traditional Treatment of Massive Rotator Cuff Tears.
Management of massive rotator cuff tears in shoulders without glenohumeral arthritis remains problematic for surgeons. Repairs of massive rotator cuff tears have failure rates of 20% to 94% at 1 to 2 years postoperatively as demonstrated with arthrography, ultrasound, and magnetic resonance imaging. Additionally, inconsistent outcomes have been reported with debridement alone of massive rotator cuff tears, and limitations have been seen with other current methods of operative intervention, including arthroplasty and tendon transfers. The use of interposition porcine acellular dermal matrix xenograft in patients with massive rotator cuff tears will result in improved subjective outcomes, postoperative pain, function, range of motion, and strength. Case series; Level of evidence, 4. Sixty patients (61 shoulders) were prospectively observed for a mean of 50.3 months (range, 24-63 months) after repair of massive rotator cuff tears with porcine acellular dermal matrix xenograft as an interposition graft. Subjective outcome data were obtained with visual analog scale for pain score (0-10, 0 = no pain) and Modified American Shoulder and Elbow Surgeons (MASES) score. Active range of motion in flexion, external rotation, and internal rotation were recorded. Strength in the supraspinatus and infraspinatus muscles was assessed manually on a 10-point scale and by handheld dynamometer. Ultrasound was used to assess the integrity of the repair during latest follow-up. Mean visual analog scale pain score decreased from 4.0 preoperatively to 1.0 postoperatively ( P < .001). Mean active forward flexion improved from 140.7° to 160.4° ( P < .001), external rotation at 0° of abduction from 55.6° to 70.1° ( P = .001), and internal rotation at 90° of abduction from 52.0° to 76.2° ( P < .001). Supraspinatus manual strength increased from 7.7 to 8.8 ( P < .001) and infraspinatus manual strength from 7.7 to 9.3 ( P < .001). Mean dynamometric strength in forward flexion was 77.7 N in nonoperative shoulders (shoulder that did not undergo surgery) and 67.8 N ( P < .001) in operative shoulders (shoulder that underwent rotator cuff repair with interposition porcine dermal matrix xenograft). Mean dynamometric strength in external rotation was 54.5 N in nonoperative shoulders and 50.1 N in operative shoulders ( P = .04). Average postoperative MASES score was 87.8. Musculoskeletal ultrasound showed that 91.8% (56 of 61) of repairs were fully intact; 3.3% (2 of 61), partially intact; and 4.9% (3 of 61), not intact. Patients who underwent repair of massive rotator cuff tears with interposition porcine acellular dermal matrix graft have good subjective function as assessed by the MASES score. Patients have significant improvement in pain, range of motion, and manual muscle strength. Postoperative ultrasound demonstrated that the repair was completely intact in 91.8% of patients, a vast improvement compared with results previously reported for primary repairs of massive rotator cuff tears.


Employment and disability pension after central nervous system infections in adults.
In this nationwide population-based cohort study using national Danish registries, in the period 1980-2008, our aim was to study employment and receipt of disability pension after central nervous system infections. All patients diagnosed between 20 and 55 years of age with meningococcal (n = 451), pneumococcal (n = 553), or viral (n = 1,433) meningitis or with herpes simplex encephalitis (n = 115), who were alive 1 year after diagnosis, were identified. Comparison cohorts were drawn from the general population, and their members were individually matched on age and sex to patients. Five years after diagnosis, the differences in probability of being employed as a former patient with pneumococcal meningitis or herpes simplex encephalitis versus being a member of the comparison cohorts were -19.9% (95% confidence interval (CI): -24.7, -15.1) and -21.1% (95% CI: -33.0, -9.3), respectively, and the corresponding differences in probability of receiving disability pension were 20.2% (95% CI: 13.7, 26.7) and 16.2% (95% CI: 6.2, 26.3). The differences in probability of being employed or receiving disability pension in former meningococcal or viral meningitis patients versus members of the comparison cohorts were small. In conclusion, pneumococcal meningitis and herpes simplex encephalitis were associated with substantially decreased employment and increased need for disability pension. These associations did not seem to apply to meningococcal meningitis or viral meningitis.


CXCL12 and its receptors regulate granulosa cell apoptosis in PCOS rats and human KGN tumor cells.
Polycystic ovary syndrome (PCOS) is a common endocrine disorder accompanied by chronic low-grade inflammation; its etiology is still undefined. This study investigated the expression of CXCL12, CXCR4, and CXCR7 in PCOS rats and their role in regulation of apoptosis. To accomplish this, we established an in vivo PCOS rat model and studied KGN cells (human ovarian granulosa cell line) in vitro. In PCOS rats, the ovarian expression of CXCL12, CXCR4, and CXCR7 was reduced, and the apoptosis rate of granulosa cells was increased, accompanied by decreased expression of BCL2 and increased expression of BAX and cleaved CASPASE3 (CASP3). We further showed that recombinant human CXCL12 treatment upregulated BCL2, downregulated BAX, and cleaved CASP3 in KGN cells to inhibit their apoptosis in a concentration-dependent manner; moreover, the effect of CXCL12 was weakened by CXCR4 antagonist AMD3100 and anti-CXCR7 neutralizing antibody. In conclusion, PCOS rats showed decreased CXCL12, CXCR4, and CXCR7 expression and increased apoptosis rate of ovarian granulosa cells. Further, in human KGN cells, CXCL12 regulated the expression of BAX, BCL2, and cleaved CASP3 to inhibit apoptosis through CXCR4- and CXCR7-mediated signal transmission. These findings may provide a theoretical and practical basis for illuminating the role of proinflammatory cytokines in the pathogenesis of PCOS.


DAFS: simultaneous aligning and folding of RNA sequences via dual decomposition.
It is well known that the accuracy of RNA secondary structure prediction from a single sequence is limited, and thus a comparative approach that predicts a common secondary structure from aligned sequences is a better choice if homologous sequences with reliable alignments are available. However, correct secondary structure information is needed to produce reliable alignments of RNA sequences. To tackle this dilemma, we require a fast and accurate aligner that takes structural information into consideration to yield reliable structural alignments, which are suitable for common secondary structure prediction. We develop DAFS, a novel algorithm that simultaneously aligns and folds RNA sequences based on maximizing expected accuracy of a predicted common secondary structure and its alignment. DAFS decomposes the pairwise structural alignment problem into two independent secondary structure prediction problems and one pairwise (non-structural) alignment problem by the dual decomposition technique, and maintains the consistency of a pairwise structural alignment by imposing penalties on inconsistent base pairs and alignment columns that are iteratively updated. Furthermore, we extend DAFS to consider pseudoknots in RNA structural alignments by integrating IPknot for predicting a pseudoknotted structure. The experiments on publicly available datasets showed that DAFS can produce reliable structural alignments from unaligned sequences in terms of accuracy of common secondary structure prediction.


Evaluation of a Clinical Protocol to Assess and Diagnose Neuropathic Pain During Acute Hospital Admission: Results From Traumatic Spinal Cord Injury.
A clinical protocol was developed for clinicians to routinely assess and initiate treatment for patients with neuropathic pain (NP) in an acute care setting. The objectives of this study were to: (1) determine the incidence and onset of NP in patients with traumatic spinal cord injury during acute care and (2) describe how the implementation of a clinical protocol impacts the assessment and diagnosis of NP. The study was a cohort analysis with a pre-post-test utilizing a historical control. Data were retrospectively collected from a patient registry and charts. Participants were randomly selected in cohort 1 (control) and cohort 2 (NP clinical protocol). The incidence of NP was 56% without significant difference between the cohorts (P=0.3). Onset of NP was 8 days (SD=14) across the study and >85% of the participants with NP were diagnosed within 2 weeks. Participants with incomplete injuries had a significant earlier onset than participants with complete injuries (6.2±12.8, 10.9±15.8 d; P=0.003). The mean number of days from hospital admission to initial assessment decreased with use of the NP clinical protocol (3.7±5.7 d; P=0.02). This study demonstrates a high incidence and early onset of NP in traumatic spinal cord injury during acute hospital care, with an earlier emergence in participants with incomplete injury. The NP clinical protocol ensured continuous assessment and documentation of NP while decreasing the time to an initial screen, but did not impact diagnosis.


Rhodol-based thallium sensors for cellular imaging of potassium channel activity.
Thallium (Tl+) flux assays enable imaging of potassium (K+) channel activity in cells and tissues by exploiting the permeability of K+ channels to Tl+ coupled with a fluorescent Tl+ sensitive dye. Common Tl+ sensing dyes utilize fluorescein as the fluorophore though fluorescein exhibits certain undesirable properties in these assays including short excitation wavelengths and pH sensitivity. To overcome these drawbacks, the replacement of fluorescein with rhodols was investigated. A library of 13 rhodol-based Tl+ sensors was synthesized and their properties and performance in Tl+ flux assays evaluated. The dimethyl rhodol Tl+ sensor emerged as the best of the series and performed comparably to fluorescein-based sensors while demonstrating greater pH tolerance in the physiological range and excitation and emission spectra 30 nm red-shifted from fluorescein.


Association of low serum magnesium with diabetes and hypertension: Findings from Qatar Biobank study.
We aimed to examine the association between serum magnesium and diabetes and hypertension among Qatari adults. In the cross-sectional study, we used data from 9693 Qatari participants aged 20 years and above attending the Qatar Biobank (QBB) Study. Blood samples were analyzed in a central lab. Habitual food consumption was assessed by a food frequency questionnaire. Reduced rank regression was used to construct magnesium related dietary pattern (MRDP) using serum magnesium as a response variable. Diabetes was defined by blood glucose, HbA1c or known diabetes. Prediabetes was defined as HbA1c between 5.7% and 6.4%. Subclinical magnesium deficiency was defined as serum magnesium <0.85 mmol/L. The prevalence of diabetes, prediabetes and subclinical magnesium deficiency was 18.9%, 11.5% and 59.5%, respectively. Across the quartiles of serum magnesium from high to low, the prevalence ratios (PR 95%CI) for diabetes were 1.00, 1.35, 1.88, and 2.70 (95%CI 2.38-3.05), respectively (p for trend <0.001). The presence of hypertension significantly increased the probability of diabetes along a wide range of low serum magnesium. A low intake of MRDP was also positively associated with diabetes and high HbA1c. Subclinical magnesium deficiency is common in Qatar and associates with diabetes, prediabetes and hypertension in Qatari adults.


Development of a measure of mental health stigma including police behaviors.
Various stigmatizing notions are associated with mental illness, resulting in negative personal (e.g. employment discrimination) and societal (e.g. public treatment of the mentally ill as 'dangerous' and/or 'criminal') outcomes. This study develops and validates a new multi-scale assessment tool to assess several dimensions of mental illness stigma, including perceived dangerousness, self-care, social distance, treatment amenability and predicted police behavior. A total of 641 undergraduate students from various American universities completed the new stigma measure along with two other existing measures. The results indicate that the new stigma measure has an acceptable three-factor solution consisting of self-care, dangerousness and police behavior. The self-care and dangerousness factors were found to have concurrent validity with the corresponding scales of the existing measures. Future research involving different populations, as well as the policy implications of the new police behavior factor, are discussed.


Calcium-Phosphate-Osteopontin Particles Reduce Biofilm Formation and pH Drops in in situ Grown Dental Biofilms.
This 2-period crossover study investigated the effect of calcium-phosphate-osteopontin particles on biofilm formation and pH in 48-h biofilms grown in situ. Bovine milk osteopontin is a highly phosphorylated glycoprotein that has been shown to interfere with bacterial adhesion to salivary-coated surfaces. Calcium-phosphate-osteopontin particles have been shown to reduce biofilm formation and pH drops in a 5-species laboratory model of dental biofilm without affecting bacterial viability. Here, smooth surface biofilms from 10 individuals were treated ex vivo 6 times/day for 30 min with either calcium-phosphate-osteopontin particles or sterile saline. After growth, the amount of biofilm formed was determined by confocal microscopy, and pH drops upon exposure to glucose were monitored using confocal-microscopy-based pH ratiometry. A total of 160 biofilms were analysed. No adverse effects of repeated ex vivo treatment with calcium-phosphate-osteopontin particles were observed. Particle treatment resulted in a 32% lower amount of biofilm formed (p < 0.05), but large inter-individual differences could be observed. Biofilm pH was significantly higher upon particle treatment, both shortly after the addition of glucose and after 30 min of incubation with glucose (p < 0.05). Calcium-phosphate-osteopontin particles may represent a new therapeutic approach to caries control and aim at directly targeting virulence factors involved in the caries process. Further studies are required to determine the effect of particle treatment on more acidogenic/aciduric biofilms as well as the remineralizing potential of the particles.


Comparison of thermophilic and mesophilic one-stage, batch, high-solids anaerobic digestion.
The concept of starting up a batch, high-solids anaerobic digester by simply flooding the bed with a pH-buffer solution was tested using a mixture of vegetable waste and wood chips as feedstock at mesophilic (38 degrees C) and thermophilic (55 degrees C) conditions. At both temperatures stable and balanced methanogenesis was rapidly established within four days and was sustained until substrate was exhausted. Methanogenesis was more rapidly initiated in the thermophilic digester than in the mesophilic digester. Acetic, propionic and butyric acids accumulated in the leachate of both digesters during the start-up of digestion of uninoculated batch of waste. Thereafter all acids were degraded; which was quicker in the thermophilic digester. The accumulation and degradation of these acids was slower in the mesophilic digester. These studies showed that inoculum for carrying out thermophilic and mesophilic anaerobic digestion is readily available within the waste and its activity for complete mineralization of organic matter can be enhanced and sustained by providing adequate alkalinity. By employing a process in which anaerobic digestion of subsequent batches of waste was carried out by flooding with leachate drained from the digestion of a previous batch of waste, the volatile organic acid accumulation was maintained low and 95% of the methane yield potential of the waste was produced in 11 days under thermophilic conditions as opposed to 27 days under mesophilic conditions.


Purification of antibodies to O antigen of Salmonella Typhimurium from human serum by affinity chromatography.
Nontyphoidal Salmonellae (NTS) are a common cause of bacteraemia in children and HIV-infected adults in Sub-Saharan Africa. We have previously shown that antibodies play a key role in both bactericidal and cellular mechanisms of immunity to NTS, but found that high concentrations of antibody to Salmonella Typhimurium O antigen (OAg) in the serum of some HIV-infected African adults is associated with impaired killing of NTS. To further investigate the function of antibodies to the OAg of NTS, we developed a method to purify these antibodies from human serum by affinity chromatography. Purified Salmonella Typhimurium OAg was activated with adipic acid dihydrazide (ADH) via two different chemistries before linking to N-hydroxysuccinamide-Sepharose resin: one ADH molecule was introduced per OAg chain on its terminal 3-deoxy-D-manno-octulosonic acid sugar (OAg-ADH), or multiple ADH molecules were attached along the OAg chain after oxidation with sodium periodate (OAgoxADH). Both resulting columns worked well when tested with commercial polyclonal anti-O:4,5 antibodies from rabbit serum. Over 90% of the applied antibodies bound to the resin and 89% of these antibodies were then eluted as detected by ELISA. OAg-ADH was preferred as the method for OAg derivatisation as it does not modify the saccharide chain and can be applied to OAg from different bacteria. Both columns were able to bind OAg-specific antibodies in human serum, but antibody recovery was initially low. Different elution buffers were tested and different amounts of OAg-ADH were linked to the resin to improve the yield. Optimal recovery (51%) was obtained by loading 1mg of activated OAg per ml of resin and eluting with 0.1M glycine, 0.1M NaCl pH2.4. The column matrix could be regenerated following elution with no detectable loss in performance for over ten uses. This method offers the potential to purify antibodies to Salmonella OAg from polyclonal serum following vaccination or natural exposure to Salmonella and so investigate the functionality and diversity of the antibody response to OAg.


Mutagenesis of the catalytic triad of tissue transglutaminase abrogates coeliac disease serum IgA autoantibody binding.
Tissue transglutaminase (tTG) is an autoantigen in coeliac disease and the related disorder, dermatitis herpetiformis. The detection of autoantibodies directed against tTG is a highly specific marker of coeliac disease; however, it is unclear if there is a role for these autoantibodies in the disease process. The aim of this study was to investigate whether the catalytic triad of tTG is targeted by coeliac disease autoantibodies. A full-length wild-type recombinant tTG and a novel site-directed mutagenic variant lacking the catalytic triad were produced in Escherichia coli. Serum samples from 61 biopsy-proven coeliac disease and 10 dermatitis herpetiformis patients were tested for their recognition of both antigens in enzyme-linked immunosorbent assay. Although IgA autoantibodies from sera of patients with coeliac disease and dermatitis herpetiformis bound wild-type tTG well, a dramatic decrease in binding to the mutant tTG was observed with a mean reduction of 79% in coeliac disease and 58% in dermatitis herpetiformis samples. IgG anti-tTG antibodies did not show a similar pattern of reduction, with no overall difference in recognition of the wild-type or mutant tTGs. These results suggest that the IgA anti-tTG response in coeliac disease and dermatitis herpetiformis is focused on the region of tTG responsible for its transamidation and deamidation reactions, whereas the IgG response may target other regions of the enzyme.


Characterizing the Natural History of Visual Function in Choroideremia Using Microperimetry and Multimodal Retinal Imaging.
Centripetal retinal degeneration in choroideremia (CHM) leads to early visual field restriction and late central vision loss. The latter marks an acute decline in quality of life but visual prognostication remains challenging. We investigated visual function in CHM by correlating best-corrected visual acuity (BCVA), microperimetry and multimodal imaging. Fifty-six consecutive CHM patients attending Oxford Eye Hospital were examined with BCVA, 10-2 microperimetry, optical coherence tomography, and fundus autofluorescence (AF). Microperimetry was repeated in 21 eyes and analyzed with Bland-Altman. Kaplan-Meier survival plots of eyes retaining 20/20 BCVA were created. Intereye symmetry was assessed. Microperimetry coefficient of repeatability was 1.45 dB. Survival analysis showed an indistinguishable pattern between eyes (median survival 39 years). Macular sensitivity showed a similar decline in right and left eyes, with half-lives of 13.6 years. Zonal analysis showed faster decline nasal to the fovea. Intereye symmetry was more consistent for microperimetry sensitivity (r = 0.95, P < 0.001) than BCVA (r = 0.42, P = 0.0006). Near normal foveal sensitivity was maintained when the fovea was at least 2500 μm from the advancing edge of AF. BCVA is a marker of central degeneration and can provide valuable information about the position of the remaining retina as well as a measure of the impact on daily living. Microperimetry represents the global macular region. Both visual functions showed a high degree of intereye symmetry, particularly in early stages, indicating the fellow eye can provide a suitable control for assessing interventions to one eye. The findings may help to tailor visual prognosis and interpret outcomes of trials.


The effects of dehydration, moderate alcohol consumption, and rehydration on cognitive functions.
This study investigated the impact of mild-moderate dehydration on alcohol-induced deteriorations in cognitive functions. Sixteen healthy males participated in a single-blind, placebo-controlled cross-over design study involving 4 experimental trials (separated by ≥7 d). In each trial, participants were dehydrated by 2.5% body mass through exercise. After 1 h recovery in a thermo-neutral environment (22 ± 2 °C, 60-70% relative humidity) 4 tasks from the Cambridge Neuropsychological Test Automated Battery (CANTAB) were administered to the participants (test 1). In two of the trials, participants were provided with water equivalent to either 50% or 150% body mass loss and given salt (NaCl) capsules (50 mmol/L). A set volume of alcohol or placebo was then consumed in each trial, incorporating the conditions: dehydration-placebo (DP), dehydration-alcohol (DA), partial rehydration-alcohol (PA), and full rehydration-alcohol (FA). The same 4 CANTAB tasks were then re-administered (test 2). Subjective ratings of mood and estimates of alcohol intoxication and driving impairment were also recorded in each trial. Alcohol consumption caused deterioration on 3 of the 4 CANTAB measures (viz., choice reaction time, executive function and response inhibition). This reduction in performance was exacerbated when participants were dehydrated compared to trials where full rehydration occurred. Subjective ratings of impairment and intoxication were not significantly different between any of the trials where alcohol was consumed; however ratings for alcohol trials were significantly higher than in the placebo trial. These findings suggest that rehydration after exercise that causes fluid loss can attenuate alcohol-related deterioration of cognitive functions. This may pose implications for post match fluid replacement if a moderate amount of alcohol is also consumed.


Effects of the Gluten-free Diet on Body Mass Indexes in Pediatric Celiac Patients.
The aim of the study was to determine the effects of the gluten-free diet (GFD) on body mass indexes (BMIs) in children with celiac disease at University of Chicago before and after 2011, when processed gluten-free foods became readily available on the market. We conducted a retrospective chart review of children seen at University of Chicago Celiac Center from January 2002 to May 2016. BMI was recorded upon GFD initiation in addition to at least 1 other timepoint: 6 months, 1 year, 2 years, 3 years, and 4+ years. We compared the rate of BMI increase in children who were diagnosed before versus after 2011. A total of 147 children (66% girls) with biopsy-confirmed celiac disease were included in the study. The mean BMI at diagnosis was 17.8 (standard deviation 3.9) for those diagnosed before 2011 and 17.1 (standard deviation 2.7) for those diagnosed after 2011. Based on a mixed-effects random-intercept random-slope regression model, there was no evidence for significant difference in BMI change over time between the 2 groups (P value = 0.36). BMI values overall were noted to increase after starting the GFD, even at the first appointment. Serologies were monitored after patients started the GFD and approached normal values, allowing us to conclude that patients were adherent to the GFD. Although overall we observed no significant changes in BMI before and after 2011, we did notice that in adolescent celiac patients there was a trend toward a higher postdiagnosis BMI in the years after 2011. We speculate that teenagers may be especially vulnerable to choosing quick and easy processed gluten-free options over more healthy, natural alternatives leading to a rise in their BMIs after the 2011 surge in production of processed gluten-free foods on the market. Therefore, special attention must be paid to this population to insure ongoing healthy food choices even after many years on the GFD.


Commensal oral microbiota induces osteoimmunomodulatory effects separate from systemic microbiome in mice.
Commensal microbes critically regulate skeletal homeostasis, yet the impact of specific microbiota communities on osteoimmune response mechanisms is unknown. To discern osteoimmunomodulatory effects imparted by the commensal oral microbiota that are distinct from the systemic microbiota, osteoimmunology studies were performed in both alveolar bone and nonoral skeletal sites of specific pathogen-free (SPF) versus germ-free (GF) mice and SPF mice subjected to saline versus chlorhexidine oral rinses. SPF versus GF mice had reduced cortical/trabecular bone and an enhanced pro-osteoclastic phenotype in alveolar bone. TLR signaling and Th17 cells that have known pro-osteoclastic actions were increased in alveolar BM, but not long BM, of SPF versus GF mice. MHC II antigen presentation genes and activated DCs and CD4+ T cells were elevated in alveolar BM, but not long BM, of SPF versus GF mice. These findings were substantiated by in vitro allostimulation studies demonstrating increased activated DCs derived from alveolar BM, but not long BM, of SPF versus GF mice. Chlorhexidine antiseptic rinse depleted the oral, but not gut, bacteriome in SPF mice. Findings from saline- versus chlorhexidine-treated SPF mice corroborated outcomes from SPF versus GF mice, which reveals that the commensal oral microbiota imparts osteoimmunomodulatory effects separate from the systemic microbiome.


Correlation of oral health status of socially handicapped children with their oral heath knowledge, attitude, and practices from India.
Information on oral health knowledge and practice in orphanage house children is essential for healthcare policy makers to promote oral health resources and address oral health needs of this unprivileged group of society. To assess the source of information, level of knowledge, attitude, and practice toward oral hygiene and oral health among socially handicapped children from city of Mysore, Karnataka state, India. A cross-sectional survey was conducted among 488 children of 12-14 years of age living in five different orphanage houses of Mysore district. Data regarding knowledge and practice were collected through structured questionnaire and oral health status by type III clinical oral examinations by two trained examiners. Decayed, Missing, Filled, Teeth (DMFT) and OH I-S was then correlated with the above information. Spearman's correlation test was used to measure the correlation. The final data analysis included 488 children, of which 216 (44.26%) were boys and 272 (55.74%) were girls. A total of 88.5% children showed one or more decayed teeth in their oral cavity, with an overall mean DMFT of 3.55. Among DMF, component D showed maximum value with mean 3.42, followed by components F and M. Correlation between source of information, knowledge, and attitude for oral health to oral hygiene index (OHI-S; P < 0.05) and gingival index (P < 0.001) showed highly significant negative values. Correlation between oral hygiene practice to OHI-S, DMFT, and gingival index (P < 0.001) showed highly significant (P < 0.001) negative values. Although children of orphanage have positive attitude toward oral health, knowledge and practice among children are still below the satisfactory level. The children did not receive appropriate information or, if informed, were not re-evaluated or reinforced for its practical application by the concerned authority. The findings of this study suggest that awareness on the importance of oral health needs to be enhanced among the orphanage children of Mysore.


Discovery of an RmlC/D fusion protein in the microalga Prymnesium parvum and its implications for NDP-β-l-rhamnose biosynthesis in microalgae.
The 6-deoxy sugar l-rhamnose (l-Rha) is found widely in plant and microbial polysaccharides and natural products. The importance of this and related compounds in host-pathogen interactions often means that l-Rha plays an essential role in many organisms. l-Rha is most commonly biosynthesized as the activated sugar nucleotide uridine 5'-diphospho-β-l-rhamnose (UDP-β-l-Rha) or thymidine 5'-diphospho-β-l-rhamnose (TDP-β-l-Rha). Enzymes involved in the biosynthesis of these sugar nucleotides have been studied in some detail in bacteria and plants, but the activated form of l-Rha and the corresponding biosynthetic enzymes have yet to be explored in algae. Here, using sugar-nucleotide profiling in two representative algae, Euglena gracilis and the toxin-producing microalga Prymnesium parvum, we show that levels of UDP- and TDP-activated l-Rha differ significantly between these two algal species. Using bioinformatics and biochemical methods, we identified and characterized a fusion of the RmlC and RmlD proteins, two bacteria-like enzymes involved in TDP-β-l-Rha biosynthesis, from P. parvum Using this new sequence and also others, we explored l-Rha biosynthesis among algae, finding that although most algae contain sequences orthologous to plant-like l-Rha biosynthesis machineries, instances of the RmlC-RmlD fusion protein identified here exist across the Haptophyta and Gymnodiniaceae families of microalgae. On the basis of these findings, we propose potential routes for the evolution of nucleoside diphosphate β-l-Rha (NDP-β-l-Rha) pathways among algae.


Evolution of resource competition between mutually dependent digital organisms.
We study the emergence and dynamics of competing strains of digital organisms in a world with two depletable resources. Consumption of one resource produces the other resource as a by-product, and vice versa. As a consequence, two types of mutually dependent organisms emerge that each prey on the waste product of the other. In the absence of mutations, that is, in a purely ecological setting, the abundances of the two types of organisms display a wide range of different types of oscillations, from regular oscillations with large amplitude to irregular oscillations with amplitudes ranging from small to large. In this regime, time-averaged abundance levels seem to be controlled by the relative fitness of the organisms in the absence of resources. Under mutational pressure, on the other hand, populations evolve that seem to avoid the oscillations of intermediate to large amplitudes. In this case, the relative fitness of the organisms in the presence of resources plays an important role in the time-averaged abundance levels as well.


A genome-wide survey and functional brain imaging study identify CTNNBL1 as a memory-related gene.
Unbiased genome-wide screens combined with imaging data on brain function may identify novel molecular pathways related to human cognition. Here we performed a dense genome-wide screen to identify episodic memory-related gene variants. A genomic locus encoding the brain-expressed beta-catenin-like protein 1 (CTNNBL1) was significantly (P=7 × 10(-8)) associated with verbal memory performance in a cognitively healthy cohort from Switzerland (n=1073) and was replicated in a second cohort from Serbia (n=524; P=0.003). Gene expression studies showed CTNNBL1 genotype-dependent differences in beta-catenin-like protein 1 mRNA levels in the human cortex. Functional magnetic resonance imaging in 322 subjects detected CTNNBL1 genotype-dependent differences in memory-related brain activations. Converging evidence from independent experiments and different methodological approaches suggests a role for CTNNBL1 in human memory.


Sex counts: An examination of sexual service advertisements in a UK online directory.
Internationally, sex work research, public opinion, policy, laws, and practice are predicated on the assumption that commercial sex is a priori sold by women and bought by men. Scarce attention has been devoted to lesbian, gay, bisexual, transgender, or queer/questioning (LGBTQ) sex working as well as women who pay for sex. This is as much an empirical absence as it is a theoretical one, for the ideological claim that women comprise the "vast majority" of sex workers is rarely, if ever, exposed to empirical scrutiny. Focusing on the UK, we address this major gap in evidence in order to challenge the gendered and heterosexist logics that underpin contemporary debates. We do so by presenting large-scale data gained from the quantitative analysis of 25,511 registered member profiles of an online escort directory. Our findings point to heterogeneity rather than homogeneity in the contemporary sex industry including in terms of gender identity, sexual orientation, and advertised client base. For example, while two-thirds of advertisements self-identify as "Female," one in four are listed as "Male;" less than half list their sexual orientation as "Straight;" and nearly two-thirds advertise to women clients. Our study thus challenges prevailing heteronormative assumptions about commercial sex, which erase LGBTQ sex workers and other non-normative identities and practices, and which we argue have important political, practical, and theoretical consequences.


A theoretical model based upon mast cells and histamine to explain the recently proclaimed sensitivity to electric and/or magnetic fields in humans.
The relationship between exposure to electromagnetic fields (EMFs) and human health is more and more in focus. This is mainly because of the rapid increasing use of such EMFs within our modern society. Exposure to EMFs has been linked to different cancer forms, e.g. leukemia, brain tumors, neurological diseases, such as Alzheimer's disease, asthma and allergy, and recently to the phenomena of 'electrosupersensitivity' and 'screen dermatitis'. There is an increasing number of reports about cutaneous problems as well as symptoms from internal organs, such as the heart, in people exposed to video display terminals (VDTs). These people suffer from subjective and objective skin and mucosa-related symptoms, such as itch, heat sensation, pain, erythema, papules and pustules. In severe cases, people can not, for instance, use VDTs or artificial light at all, or be close to mobile telephones. Mast cells (MCs), when activated, release a spectrum of mediators, among them histamine, which is involved in a variety of biological effects with clinical relevance, e.g. allergic hypersensitivity, itch, edema, local erythema and many types of dermatoses. From the results of recent studies, it is clear that EMFs affect the MC, and also the dendritic cell, population and may degranulate these cells. The release of inflammatory substances, such as histamine, from MCs in the skin results in a local erythema, edema and sensation of itch and pain, and the release of somatostatin from the dendritic cells may give rise to subjective sensations of on-going inflammation and sensitivity to ordinary light. These are, as mentioned, the common symptoms reported from patients suffering from 'electrosupersensitivity'/'screen dermatitis'. MCs are also present in the heart tissue and their localization is of particular relevance to their function. Data from studies made on interactions of EMFs with the cardiac function have demonstrated that highly interesting changes are present in the heart after exposure to EMFs. One could speculate that the cardiac MCs are responsible for these changes due to degranulation after exposure to EMFs. However, it is still not known how, and through which mechanisms, all these different cells are affected by EMFs. In this article, we present a theoretical model, based upon observations on EMFs and their cellular effects, to explain the proclaimed sensitivity to electric and/or magnetic fields in humans.


The evaluation of metabolic function and fat redistribution in clinical trials.
Changes in lipid and carbohydrate metabolism and in body composition associated with the most potent and effective therapies available are being reported with increasing frequency. These changes could potentially lead to increased risks of cardiovascular disease. Clinical trials set up to investigate new antiretroviral therapies need to explore the therapies' impact on these potentially serious adverse outcomes, at least in a subset of patients. The measurements that are feasible to include for all patients clearly differ from those required by a metabolic substudy. We propose the following: firstly, a minimal set of parameters that should be included in all trials; secondly, a desirable set of parameters that should be included whenever possible; and thirdly, a list of exploratory measures that should be considered. These exploratory measures are classified by the different mechanisms for changes in body composition or weight: endocrinal, cardiovascular, sterol and chemokine/cytokine pathways. Standardized instruments for evaluating patients' reports of body changes and potential methods for assessing the risk of cardiovascular disease are described. Minimum and desirable standards for methods of measurement are also proposed. The choice of parameters is based on expert clinical opinion. The experts consulted include investigators from four large ongoing clinical trials with substudies specifically designed to investigate lipid and carbohydrate metabolism and changes in body composition, together with standard parameters for measurement within individual mechanistic pathways. The parameters proposed should be kept under review as the body of knowledge about metabolic function and fat redistribution in HIV infection increases.


Intravenous Cyclophosphamide in Myalgic Encephalomyelitis/Chronic Fatigue Syndrome. An Open-Label Phase II Study.
Introduction: Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) is a disease with high symptom burden, of unknown etiology, with no established treatment. We observed patients with long-standing ME/CFS who got cancer, and who reported improvement of ME/CFS symptoms after chemotherapy including cyclophosphamide, forming the basis for this prospective trial. Materials and methods: This open-label phase II trial included 40 patients with ME/CFS diagnosed by Canadian criteria. Treatment consisted of six intravenous infusions of cyclophosphamide, 600-700 mg/m2, given at four-week intervals with follow-up for 18 months, extended to 4 years. Response was defined by self-reported improvements in symptoms by Fatigue score, supported by Short Form 36 (SF-36) scores, physical activity measures and other instruments. Repeated measures of outcome variables were assessed by General linear models. Responses were correlated with specific Human Leukocyte Antigen (HLA) alleles. Results: The overall response rate by Fatigue score was 55.0% (22 of 40 patients). Fatigue score and other outcome variables showed significant improvements compared to baseline. The SF-36 Physical Function score increased from mean 33.0 at baseline to 51.5 at 18 months (all patients), and from mean 35.0 to 69.5 among responders. Mean steps per 24 h increased from mean 3,199 at baseline to 4,347 at 18 months (all patients), and from 3,622 to 5,589 among responders. At extended follow-up to 4 years 68% (15 of 22 responders) were still in remission. Patients positive for HLA-DQB1*03:03 and/or HLA-C*07:04 (n = 12) had significantly higher response rate compared to patients negative for these alleles (n = 28), 83 vs. 43%, respectively. Nausea and constipation were common grade 1-2 adverse events. There were one suspected unexpected serious adverse reaction (aggravated POTS) and 11 serious adverse events in eight patients. Conclusion: Intravenous cyclophosphamide treatment was feasible for ME/CFS patients and associated with an acceptable toxicity profile. More than half of the patients responded and with prolonged follow-up, a considerable proportion of patients reported ongoing remission. Without a placebo group, clinical response data must be interpreted with caution. We nevertheless believe a future randomized trial is warranted. Clinical Trial Registration: www.ClinicalTrials.gov, identifier: NCT02444091.


Regression of symptomatic multiple cardiac rhabdomyomas associated with tuberous sclerosis complex in a newborn receiving everolimus.
Cardiac rhabdomyoma is the most common primary cardiac tumor, is considered to be a hamartoma of developing cardiac myocytes. Cardiac rhabdomyoma is associated with tuberous sclerosis complex (TSC) in 50-86% of cases. Mutations in TSC-1/TSC-2 genes result in increased mammalian target of rapamycin (mTOR) pathway activation responsible for the hamartomatous lesions of tuberous sclerosis complex. Therapy with mTOR inhibitors is currently under investigation as a treatment option for tumors associated with TSC. In this report we present a case with multiple symptomatic rhabdomyomas associated with tuberous sclerosis complex, deemed to be ineligible for surgical removal, treated with everolimus (mTOR inhibitor). As we observed in our patient, in cases with inoperable symptomatic rhabdomyomas associated with TSC, everolimus, an mTOR inhibitor, may be the treatment of choice, which should be confirmed with additional studies.


The human transferrin receptor gene: genomic organization, and the complete primary structure of the receptor deduced from a cDNA sequence.
Heteroduplex analysis shows that the transferrin receptor gene contains at least 19 distinct coding sequences distributed over 31 kb of genomic DNA. The nucleotide sequence of these coding regions has been determined from a cDNA clone. The sequence contains a single complete open reading frame of 2280 bases which specifies a 760 residue polypeptide with a molecular weight of 85K daltons. The deduced amino acid sequence of the receptor shows that it does not contain an N-terminal hydrophobic signal peptide. We have found a single region of sufficient length and hydrophobicity to span the membrane, located 61 amino acids from the N-terminus. This leads to the prediction that the receptor is oriented in the membrane with a cytoplasmic N-terminus and an extracellular C-terminus. The receptor has no significant homology with transferrin, or with any receptor for which a sequence is available.


Consumer understanding, interpretation and perceived levels of personal responsibility in relation to satiety-related claims.
The aim of this study was to explore (a) whether and how consumers may (over-) interpret satiety claims, and (b) whether and to what extent consumers recognize that personal efforts are required to realize possible satiety-related or weight loss benefits. Following means-end chain theory, we explored for a number of satiety claims the extent of inference-making to higher-level benefits than actually stated in the claim, using internet-based questions and tasks. Respondents (N=1504) in U.K., France, Italy and Germany participated in the study. The majority of these respondents correctly interpret satiety-related claims; i.e. they largely limit their interpretation to what was actually stated. They do not expect a "magic bullet" effect, but understand that personal efforts are required to translate product attributes into potential weight control benefits. Less-restrained eaters were at lower risk for over-interpreting satiety-related claims, whilst respondents with a stronger belief that their weight is something that they can control accept more personal responsibility, and better understand that personal efforts are required to be effective in weight control. Overall, these results indicate there is likely to be a relatively low level of consumer misinterpretation of satiety-related claims on food products.


Redescription of Dexiotricha colpidiopsis (Kahl, 1926) Jankowski, 1964 (Ciliophora, Oligohymenophorea) from a Hot Spring in Iceland with Identification Key for Dexiotricha species.
We isolated an encysted ciliate from a geothermal field in Iceland. The morphological features of this isolate fit the descriptions of Dexiotricha colpidiopsis Kahl, 1926) Jankowski, 1964 very well. These comprise body shape and size in vivo, the number of somatic kineties, and the positions of macronucleus and contractile vacuole. Using state-of-the-art taxonomic methods, the species is redescribed, including phylogenetic analyses of the small subunit ribosomal RNA (SSU rRNA) gene as molecular marker. In the phylogenetic analyses, D. colpidiopsis clusters with the three available SSU rRNA gene sequences of congeners, suggesting a monophyly of the genus Dexiotricha. Its closest relative in phylogenetic analyses is D. elliptica, which also shows a high morphological similarity. This is the first record of a Dexiotricha species from a hot spring, indicating a wide temperature tolerance of this species at least in the encysted state. The new findings on D. colpidiopsis are included in a briefly revision of the scuticociliate genus Dexiotricha and an identification key to the species.


[Prognostic classification of breast cancer and gene expression profiling].
Clinical and pathological heterogeneity of breast cancer, partly responsible of therapeutic failures, reflects complex and combinatory molecular alterations until now poorly documented by classical investigation tools. Thorough molecular typing is crucial. The advent of DNA microarray-based gene expression profiling allowed consistent progresses in this direction. A novel molecular taxonomy of breast cancer has been defined, signatures that predict clinical outcome or therapeutic response have been identified, some of them being tested in ongoing prospective clinical trials. In this review, we present the main results and their potential clinical applications. We also discuss their current limits and future hopes in the therapeutic management of patients.


Consensus Statement- Prehospital Care of Exertional Heat Stroke.
Exertional heat stroke (EHS) is one of the most common causes of sudden death in athletes. It also represents a unique medical challenge to the prehospital healthcare provider due to the time sensitive nature of treatment. In cases of EHS, when cooling is delayed, there is a significant increase in organ damage, morbidity, and mortality after 30 minutes, faster than the average EMS transport and ED evaluation window. The purpose of this document is to present a paradigm for prehospital healthcare systems to minimize the risk of morbidity and mortality for EHS patients. With proper planning, EHS can be managed successfully by the prehospital healthcare provider.


Athermal operation of silicon waveguides: spectral, second order and footprint dependencies.
We report the design criteria and performance of Si ring resonators for passive athermal applications in wavelength division multiplexing (WDM). The waveguide design rules address i) positive-negative thermo-optic (TO) composite structures, ii) resonant wavelength dependent geometry to achieve constant confinement factor (Gamma), and iii) observation of small residual second order effects. We develop exact design requirements for a temperature dependent resonant wavelength shift (TDWS) of 0 pm/K and present prototype TDWS performance of 0.5 pm/K. We evaluate the materials selection tradeoffs between high-index contrast (HIC) and low-index contrast (LIC) systems and show, remarkably, that FSR and footprint become comparable under the constraint of athermal design.


Decay of acquired colostral antibodies to Actinobacillus pleuropneumoniae in pigs.
The main objective of this study was to estimate the decay of acquired colostral antibodies to Actinobacillus pleuropneumoniae serotype 2 in pigs. Data were obtained from pigs in an isolated cohort of 47 pigs born to five sows seropositive to A. pleuropneumoniae serotype 2. The pigs were examined serologically at 18 different times from birth until an age of about 22 weeks, using an A. pleuropneumoniae serotype 2-specific blocking enzyme-linked immunosorbent assay. Antibody concentration was expressed as an OD% derived from the optical density of the sample and the median from eight wells without serum on the same plate. A non-linear mixed model assuming a constant rate of decay (half-life) was specified and fitted to the serological data. To estimate the between-pig variability of different components, between-pig random effects of each component of the model were estimated. The estimated average half-life of acquired colostral antibodies was approximately 2 weeks, but there was a considerable variation between pigs (half-life ranged from 1-3 weeks). The duration until acquired colostral antibodies were no longer detectable ranged from 2 weeks to 2 months postpartum among the pigs in the study, mainly depending on the initial level of acquired colostral antibodies to A. pleuropneumoniae serotype 2.


Unc93B1 biases Toll-like receptor responses to nucleic acid in dendritic cells toward DNA- but against RNA-sensing.
Toll-like receptors (TLRs) 3, 7, and 9 recognize microbial nucleic acids in endolysosomes and initiate innate and adaptive immune responses. TLR7/9 in dendritic cells (DCs) also respond to self-derived RNA/DNA, respectively, and drive autoantibody production. Remarkably, TLR7 and 9 appear to have mutually opposing, pathogenic or protective, impacts on lupus nephritis in MRL/lpr mice. Little is known, however, about the contrasting relationship between TLR7 and 9. We show that TLR7 and 9 are inversely linked by Unc93B1, a multiple membrane-spanning endoplasmic reticulum (ER) protein. Complementation cloning with a TLR7-unresponsive but TLR9-responsive cell line revealed that amino acid D34 in Unc93B1 repressed TLR7-mediated responses. D34A mutation rendered Unc93B1-deficient DCs hyperresponsive to TLR7 ligand but hyporesponsive to TLR9 ligand, with TLR3 responses unaltered. Unc93B1 associates with and delivers TLR7/9 from the ER to endolysosomes for ligand recognition. The D34A mutation up-regulates Unc93B1 association with endogenous TLR7 in DCs, whereas Unc93B1 association with TLR9 was down-regulated by the D34A mutation. Consistently, the D34A mutation up-regulated ligand-induced trafficking of TLR7 but down-regulated that of TLR9. Collectively, TLR response to nucleic acids in DCs is biased toward DNA-sensing by Unc93B1.


Transcriptomic analysis reveals new regulatory roles of Clp signaling in secondary metabolite biosynthesis and surface motility in Lysobacter enzymogenes OH11.
Lysobacter enzymogenes is a bacterial biological control agent emerging as a new source of antibiotic metabolites, such as heat-stable antifungal factor (HSAF) and the antibacterial factor WAP-8294A2. The regulatory mechanism(s) for antibiotic metabolite biosynthesis remains largely unknown in L. enzymogenes. Clp, a cyclic adenosine monophosphate (cAMP)-receptor-like protein, is shown to function as a global regulator in modulating biocontrol-associated traits in L. enzymogenes. However, the genetic basis of Clp signaling remains unclear. Here, we utilized transcriptome/microarray analysis to determine the Clp regulon in L. enzymogenes. We showed that Clp is a global regulator in gene expression, as the transcription of 775 genes belonging to 19 functional groups was differentially controlled by Clp signaling. Analysis of the Clp regulon detected previously characterized Clp-modulated functions as well as novel loci. These include novel loci involved in antibiotic metabolite biosynthesis and surface motility in L. enzymogenes. We further showed experimentally that Clp signaling played a positive role in regulating the biosynthesis of HSAF and WAP-8294A2, as well as surface motility which is a type-IV-pilus-dependent trait. The regulation by Clp signaling of antibiotic (HSAF and WAP-8294A2) biosynthesis and surface motility was found to be independent. Importantly, we identified a factor Lysobacter acetyltransferase (Lat), a homologue of histone acetyltransferase Hpa2, which was regulated by Clp and involved in HSAF biosynthesis, but not associated with WAP-8294A2 production and surface motility. Overall, our study provided new insights into the regulatory role and molecular mechanism of Clp signaling in L. enzymogenes.


User Compliance and Behavioral Adaptation Associated With Supine Avoidance Therapy.
This study investigates behavioral adaptation to vibrotactile position-avoidance therapy during sleep in patients with obstructive sleep apnea (n =135) across 15 to 52 weeks. The overall compliance, based on nights used ≥ 4 hr, was 71%. Overall regular use, that is, ≥ 4 hr/night over 70% of nights, was 88%. Poor early compliance strongly predicted poor long-term treatment adherence, with 92% of those noncompliant across the first 12 weeks of therapy remaining noncompliant. Conversely, 21% of those with compliant utilization in the short term became noncompliant in the long term. It appears that patients do not habituate to the stimulus during sleep, nor was there a training effect associated with long-term use.


Comparability of estimates and trends in adolescent sexual and contraceptive behaviors from two national surveys: National Survey of Family Growth and the Youth Risk Behavior Survey.
To compare adolescents' reports of sexual and contraceptive behaviors between the National Survey of Family Growth (NSFG) and the Youth Risk Behavior Survey (YRBS). For each survey, we estimated the year- and sex-specific prevalence of sexual and contraceptive behaviors among a comparably defined sample of US respondents ages 15-19 currently attending high school. We used logistic regression to test for changes in prevalence from 2007-2019 and conducted sensitivity analyses to investigate between-survey differences. We found differences in both prevalence and trends between the YRBS and NSFG when limited to a comparably defined sample. Compared to the NSFG, adolescents in the YRBS were more likely to report being sexually experienced, less likely to report use of prescription methods for both sexes, and less likely to report condoms among males. Only the YRBS estimated significant declines in sexual experience for both sexes, and significant increases in prescription methods and declines in condom use among males. Differences between surveys in the prevalence of specific contraceptive methods reflected greater combined use of methods in the NSFG. We identified differences in question-wording and other aspects that may influence these differential patterns. The NSFG and YRBS produced inconsistent prevalence estimates and trends for sexual and contraceptive behaviors among in-school adolescents. Further efforts to improve these national surveillance systems are critical to inform policy and research efforts that support adolescent sexual and reproductive health and wellbeing.


Structural and functional outcomes of anaesthetic cornea in children.
To present visual and structural outcomes from the largest series to date of children with absent corneal sensation, and contribute recommendations for management. A two-centre retrospective cohort of 33 eyes of 26 children was studied. Data regarding cause of corneal anaesthesia (CA), visual acuity (VA), complications and management were recorded. The most common underlying causes of CA were posterior fossa tumours (8), CA with somitic abnormalities (5), cerebellar hypoplasia (3), severe head trauma (3) and isolated CA (3). Median follow-up was 36½ months. Coexisting facial palsy was prevalent with 18 patients (69%) being affected. At final follow-up, 4/27 eyes (15%) with VA measures had VA 0.3 logMAR or better; 15/27 (56%) had VA 0.3-1.0 logMAR; and 8/27 (30%) had VA worse than 1.0 logMAR. Seven of eight eyes with final VA worse than 1.0 logMAR had coexisting facial palsy. Only one of these eyes with facial palsy had a permanent tarsorrhaphy before VA fell below 1.0 logMAR. Also, of the nine eyes with facial palsy and a best-recorded VA better than 1.0 logMAR at final follow-up, five had a permanent tarsorrhaphy. Corneal scarring was present in 24/33 (73%) of eyes by final follow-up. 15/33 (45%) had at least one episode of microbial keratitis. The first presentation was with this complication in 9/26 (35%) children. CA in children is a vision-threatening problem, which has a particularly poor prognosis when associated with facial nerve palsy. Earlier tarsorrhaphy should be considered to help preserve vision in eyes with CA and coexisting facial palsy.


Inherited p40phox deficiency differs from classic chronic granulomatous disease.
Biallelic loss-of-function (LOF) mutations of the NCF4 gene, encoding the p40phox subunit of the phagocyte NADPH oxidase, have been described in only 1 patient. We report on 24 p40phox-deficient patients from 12 additional families in 8 countries. These patients display 8 different in-frame or out-of-frame mutations of NCF4 that are homozygous in 11 of the families and compound heterozygous in another. When overexpressed in NB4 neutrophil-like cells and EBV-transformed B cells in vitro, the mutant alleles were found to be LOF, with the exception of the p.R58C and c.120_134del alleles, which were hypomorphic. Particle-induced NADPH oxidase activity was severely impaired in the patients' neutrophils, whereas PMA-induced dihydrorhodamine-1,2,3 (DHR) oxidation, which is widely used as a diagnostic test for chronic granulomatous disease (CGD), was normal or mildly impaired in the patients. Moreover, the NADPH oxidase activity of EBV-transformed B cells was also severely impaired, whereas that of mononuclear phagocytes was normal. Finally, the killing of Candida albicans and Aspergillus fumigatus hyphae by neutrophils was conserved in these patients, unlike in patients with CGD. The patients suffer from hyperinflammation and peripheral infections, but they do not have any of the invasive bacterial or fungal infections seen in CGD. Inherited p40phox deficiency underlies a distinctive condition, resembling a mild, atypical form of CGD.